CN1391473A - VLA-4 inhibitor compounds - Google Patents

VLA-4 inhibitor compounds Download PDF

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Publication number
CN1391473A
CN1391473A CN00809510A CN00809510A CN1391473A CN 1391473 A CN1391473 A CN 1391473A CN 00809510 A CN00809510 A CN 00809510A CN 00809510 A CN00809510 A CN 00809510A CN 1391473 A CN1391473 A CN 1391473A
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China
Prior art keywords
group
mixture
replacement
methoxyl group
pyrrolidinyl
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Pending
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CN00809510A
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Chinese (zh)
Inventor
J·J·巴德温
E·麦当娜德
K·J·莫里尔蒂
C·R·撒尔科
町永信雄
中山敦
千叶淳
饭村信
米田贺行
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Daiichi Pharmaceutical Co Ltd
Pharmacopeia Drug Discovery Inc
Original Assignee
Daiichi Pharmaceutical Co Ltd
Pharmacopeia LLC
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Application filed by Daiichi Pharmaceutical Co Ltd, Pharmacopeia LLC filed Critical Daiichi Pharmaceutical Co Ltd
Publication of CN1391473A publication Critical patent/CN1391473A/en
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    • C07D207/04—Heterocyclic compounds containing five-membered rings not condensed with other rings, with one nitrogen atom as the only ring hetero atom with only hydrogen or carbon atoms directly attached to the ring nitrogen atom having no double bonds between ring members or between ring members and non-ring members
    • C07D207/08—Heterocyclic compounds containing five-membered rings not condensed with other rings, with one nitrogen atom as the only ring hetero atom with only hydrogen or carbon atoms directly attached to the ring nitrogen atom having no double bonds between ring members or between ring members and non-ring members with hydrocarbon radicals, substituted by hetero atoms, attached to ring carbon atoms
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    • C07D401/02—Heterocyclic compounds containing two or more hetero rings, having nitrogen atoms as the only ring hetero atoms, at least one ring being a six-membered ring with only one nitrogen atom containing two hetero rings
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Abstract

Compounds that selectively inhibit the binding of ligands to alpha4beta1 integrin (VLA-4) and methods for their preparation are disclosed. In one embodiment, compounds of the invention are represented by Formula I: As selective inhibitors of VLA-4 mediated cell adhesion, compounds of the present invention are useful in the treatment of conditions associated with such adhesion, including, but not limited to, such conditions as inflammatory and autoimmune responses, diabetes, asthma, psoriasis, inflammatory bowel disease, transplantation rejection, and tumor metastasis. Also disclosed are methods of inhibiting VLA-4 mediated cell adhesion and methods of treating conditions associated with LA-4 mediated cell adhesion.

Description

The VLA-4 inhibitor compound
Invention field
The present invention relates to selectivity and suppress part and adhesion receptor, α 4β 1The bonded chemical compound of integrin (being also referred to as VLA-4).Chemical compound of the present invention is used for the treatment of and cell adhesion relevant pathological changes (for example inflammatory diseases and autoimmune disease) and the neoplasm metastasis of prevention with the VLA-4 mediation.Background of invention
This type of principal character as the pathological changes of inflammatory diseases and autoimmune disease is the gathering of activated leukocyte in affected tissue.The divide a word with a hyphen at the end of a line process of inflammation part of leukocyte self-loopa relates to a series of interaction, and this process is divided into 4 key steps: adhesion and rolling, activation, adhere to securely and divide a word with a hyphen at the end of a line (Springer, T., Ann.Rev.Physiol., 57:827 (1995)).Originally, leukocyte is adhered to lightly on the blood vessel endothelium and along its surface and rolls.It is after the cell-stimulating of the chemotaxis stimulus object mediation of solubility starts the more firm bonded generation between each leukocyte and the endotheliocyte.After this, this firm combination causes that success that leukocyte connects by endotheliocyte adheres to and divides a word with a hyphen at the end of a line.These steps be recur and each step all be essential for dividing a word with a hyphen at the end of a line.This also means divides a word with a hyphen at the end of a line and can be conditioned in each step, thereby provides many potential targets for pharmacology's inhibitory action.
The receptor that relates to leukocytoplania be accredited as to a great extent the adhesion molecule family that belongs to specific (Carlos and Harlan, Blood, 84:2068 (1994)).Initial adhesion and rolling step are called the adjusting of the proteic adhesion receptor of selection family.Firm adhesion is subjected to the interactional adjusting of leukocyte surface integrin and the molecule of the immunoglobulin superfamily of expressing at inner skin surface.Integrin and immunoglobulin-type adhesion molecule also mainly participates in leukocytic dividing a word with a hyphen at the end of a line.After dividing a word with a hyphen at the end of a line, leukocyte relies on integrin and passes extracellular matrix and rest on inflammation part.
Integrin is by two non-covalent bonded subunits, the extended familys of the heterodimer glycoprotein that α and β form (Hynes, R., Cell, 69:11 (1992)).Have 16 kinds of different α subunit (α at least 1-α 9, α L, α M, α D, α X, α E, α IIb, α V) the β subunit (β different with at least 9 kinds 1-β 9).According to the β subunit, integrin is divided into several subfamilies.Leukocyte is expressed different integrins, comprises α 4β 1, α 5β 1, α 6β 1, α 4β 7, α Lβ 2, α Xβ 2And α Vβ 3
α 4β 1Integrin, be also referred to as Vla-4-4 (VLA-4) or CD4 9d/CD29, be expressed on mononuclear cell, lymphocyte, eosinophilic granulocyte and the basophilic granulocyte, all these cells all are important effector lymphocyte (Helmer in various inflammatory diseasess, M. Ann. Rev.Immunol., 8:365 (1990)).α 4β 1Integrin as the receptor to the outer albumen fibronectin (FN) of vascular cell adhesion molecule-1 (VCAM-1) and pair cell work (Elices etc., Cell, 60:577 (1990)).Monoclonal antibody is blocked α in vivo 4β 1Behind/VCAM-1 the approach, confirmed antiinflammatory action and tardy advancing of disease (Lobb etc., J.Clin.Invest., 94:1722-28 (1994)).In pulmonary inflammatory guinea pig model, anti--α 4Inhibition antigen-inductive bronchial hyperreactive and the leukocyte recruitment in bronchoalveolar lavage fluid (Pretolani etc., J.Exp. Med., 180:795 (1994)).To α 4Or the antigen of the antibody of VCAM-1 prevention mice trachea-inductive eosinophilic granulocyte's infiltration (Nakajima etc., J.Exp.Med., 179:1145 (1994)).α 4Or the mab treatment of VCAM-1 also delay or prevented mice and monkey the skin delayed hypersensitivity reaction (Chisholm etc., Eur.J.Immunol., 23:682 (1993); Silber etc., J.Clin.Invest., 93:1554 (1993); Heart allograft rejection in mice, with specific immunosuppressant (Isobe etc., J.Immunol., 153:5810 (1994); Transplanting after the bone marrow transplantation in the mice-right-host disease (Yang etc., Proc.Natl.Acad.Sci. USA, 90:10494 (1993)); And the autoimmunity encephalomyelitis of the experiment in rat and mice (Yednock etc., Nature, 356:63 (1992)); Baron etc., J.Exp.Med., 177:57 (1993)).
Reasonably drug design research has produced the VCAM-Ig fusion rotein of solubility, and this albumen contains the terminal territory of two N-of the people VCAM-1 that merges with human IgG1's constant region.Give in the body autoimmune diabetes of the adoptive transfer that fusion rotein obviously postpones non-non-insulin-dependent diabetes mellitus mice morbidity (Jakubowski etc., J.Immunol., 155:938 (1995)).Another kind method adopts the segmental three-dimensional crystal of VCAM-1 to learn structure synthetic cyclic peptide antagonist, and this antagonist can be simulated α well 4Combining of the ring of integrin and the domain 1 of VCAM-1.Synthetic VCAM-1 peptide CQIDSPC can suppress the VCAM-1 of cell that VLA-4-expresses and purification adhesion (Wang etc., Proc.Natl.Acad.Sci.USA, 92:5714, (1995)).
Another kind method is blocking-up α 4β 1With its other counter receptor, that is, contain the combination (E.A.Wayner, J.Cell.Biol., 116:489 (1992)) in alternative splicing district of the fibronectin of jointing-1 (CS-1) primitive.A kind of synthetic CS-1 tetrapeptide (phenylacetic acid-Leu-Asp-Phe-d-Pro-amide) in the lymphocytic adhesion of vitro inhibition VLA-4-mediation and alleviate the coronary artery disease that the heart allograft of rabbit quickened (Molossi etc., J.Clin.Invest., 95:2601 (1995)).These researchs all provide evidence, promptly optionally suppress α 4β 1The adhesion of/VCAM-1 mediation is the method that obtains proving of treatment autoimmune and allergic inflammation disease.
In addition, though United States Patent (USP) 5821231 and PCT application WO 96/22966, WO97/03094, WO 98/04247 and WO 98/04913 have described in vivo and have shown that VLA-4 suppresses active chemical compound in the binding assay, all do not describe the chemical compound that shows the oral administration effect.
Therefore, although these progress are arranged, still have little, the non-peptide to the dependent cell adhesion of VLA-4, the demand of specific inhibitor, but after the described inhibitor oral administration be biological utilisation and be suitable for long-term treatment to the chronic inflammatory disease pathological changes relevant with leukocytoplania with adhesion with other.The invention summary
Compound selective of the present invention ground suppresses part and α 4β 1Combination, thereby the cell adhesion that is used to suppress, prevent and prevent VLA-4 to mediate reaches the pathological changes relevant with this adhesion, for example, inflammation, asthma, arthritis, diabetes, autoimmunity reaction, multiple sclerosis, psoriasis, graft-rejection and neoplasm metastasis.
In one embodiment, the invention provides compound or its salt by formula I representative, Wherein W is selected from the heteroaryl of aryl, heteroaryl and the replacement of aryl, replacement; W 1Be selected from the inferior heteroaryl of arlydene, inferior heteroaryl and the replacement of arlydene, replacement; A is selected from=O ,=S and=NH; R be selected from a direct key, alkylene group and-(CH 2) n-;
Wherein
N is selected from 1 and 2; X is selected from-C (O)-,-CH 2-and S (O) 2M is selected from Wherein
Figure A0080951000312
Be 4-, 5-, 6-or the 7-unit heterocyclic moiety of bivalence,
Wherein nitrogen-atoms is the junction point with X; R 1, R 2And R 3Independently be selected from-H ,-OH ,-NH 2, halogen atom, alkyl, replacement
The alkoxyl of the aryl of alkyl, aryl, replacement, alkoxyl, replacement, an alkyl
The dialkyl amido of amino, an alkyl amino that replaces, dialkyl amido, replacement,
The cycloalkyl amino of cycloalkyl amino, replacement, alkyl sulfonyl-amino, replacement
The aryl sulfonyl ammonia of alkyl sulfonyl-amino, arlysulfonylamino, replacement
The heteroaryloxy of the aryloxy group of base, aryloxy group, replacement, heteroaryloxy, replacement,
The benzyloxy of benzyloxy and replacement, or R 1, R 2And R 3In two combine and can form by independently being selected from following 1-3
Optional 3-, the 4-that replaces of individual substituent group, 5-, 6-or 7-unit's carbocyclic ring or heterocycle are residual
Base :-OH, halogen atom ,-NH 2, alkyl, alkoxyl, aryl, aryloxy group,
Alkyl amino, benzyloxy and heteroaryl; R 4Be selected from-H and low alkyl group; Y is a direct key or be selected from-C (O)-,-C (O) NH-, alkylene group, inferior alkynyl group
With-(CH 2) kY 2Divalent group,
Wherein
K is selected from 1,2 and 3; With
Y 2Be a direct key or be selected from-O-,-S-,-S (O) ,-S (O) 2-and-NY 3-
Divalent group,
Wherein
Y 3Be selected from-H and low alkyl group;
It is assorted that Z is selected from the Asia of the arlydene of arlydene, replacement, inferior heterocyclic radical, replacement
The cycloalkylidene of cyclic group, cycloalkylidene and replacement; A 1Be a direct key or be selected from alkylene group, inferior alkynyl group ,-(CH 2) t-and-O (CH 2) v
Divalent group,
Wherein
T is selected from 1,2 and 3; With
V is selected from 0,1,2 and 3; And R 5Be selected from-OH, lower alkoxy ,-N (H) OH,
Figure A0080951000322
Wherein Be 4-, 5-, 6-or the 7-unit heterocyclic moiety of bivalence,
Wherein nitrogen-atoms is the junction point with X; R 6And R 7Independently be selected from-H ,-OH, halogen atom, alkyl and alkoxyl; Y 1For be selected from-O-,-S-,-S (O)-,-S (O) 2-and-NY 4-
Divalent group,
Wherein
Y 4Be selected from-H and low alkyl group; Z 1For mixing the Asia of the arlydene that is selected from arlydene, replacement, inferior heterocyclic radical, replacement
The divalent group of the cycloalkylidene of cyclic group, cycloalkylidene and replacement; A 2Be a direct key or be selected from alkylene group, inferior alkynyl group and-(CH 2) eTwo
The valency group,
Wherein
E is selected from 1,2 and 3; And R 8Be selected from-OH, lower alkoxy ,-N (H) OH,
Figure A0080951000332
Wherein L is Wherein Be 4-, 5-, 6-or the 7-unit heterocyclic moiety of bivalence, optional by independently being selected from following 1-3 substituent group replacement: alkyl, alkoxyl, hydroxyalkyl ,-OH, benzyloxy ,-NH 2, halogen atom, aryl and heteroaryl, described part can condense with 1 or 2 other carbocyclic ring or heterocycle residue, and a following 1-3 substituent group is optional to be replaced by independently being selected from for described carbocyclic ring or heterocycle residue: alkyl, aryloxy group, alkoxyl, hydroxyalkyl ,-OH, benzyloxy ,-NH 2, halogen atom, aryl and heteroaryl; M and q independently are selected from 0,1,2 and 3; X 1Be selected from-CH=and-N=; R 9Be selected from-H and low alkyl group; R 10Be selected from-COOH, elementary alkoxy carbonyl, With And Z 2Be selected from-H, COOH and elementary alkoxy carbonyl; With
Figure A0080951000342
R wherein 11Be selected from-O-, With-NR 12-
Wherein
R 12Be selected from-H, the cycloalkanes of the alkyl of alkyl, replacement, cycloalkyl, replacement
The benzyl of the aryl of base, aryl, replacement, benzyl, replacement, rudimentary
The low-grade alkenyl of alkenyl, replacement and alkynyl of low-grade chain;
The left hand key for the junction point right hand key with-X-is and-Z 3Junction point; Z 3Be selected from a direct key, have the divalent aliphatic hydrocarbon part of 1-12 carbon atom,
Wherein
One or more carbon atoms can by-O-or-NR 13-displacement,
Wherein
R 13Be selected from-H and low alkyl group and
The one or more hydrogen atoms that are connected in aliphatic carbon atom can be by lower alkyl
The base displacement; With
Wherein x is selected from 0 and 1; Y is selected from 1,2 and 3; And R 14Be selected from-H ,-OH and halogen atom, With, work as R 11For-NR 12,
Wherein
Figure A0080951000351
Z wherein 4Be selected from
Figure A0080951000352
Wherein
R 14aBe selected from-H ,-OH, low alkyl group and halogen atom;
Figure A0080951000353
With
Figure A0080951000354
Wherein
The left hand key is and R 11Junction point and right hand key is and Q 2
Junction point; Q 2For mixing the Asia of the arlydene that is selected from arlydene, replacement, inferior heterocyclic radical, replacement
The divalent group of the cycloalkylidene of cyclic group, cycloalkylidene, replacement, R wherein 15And R 16Independently be selected from-H, halogen atom
And low alkyl group; With R wherein 17And R 18Independently be selected from-H, the low alkyl group and the low-grade alkenyl of low alkyl group, replacement; And L 1Be selected from-COOH and-COOR 19
Wherein
R 19Be low alkyl group.
In the preferred embodiment of formula I, M is In this embodiment, preferred chemical compound is=O that R is-(CH for A wherein 2) n-and X be-C (O)-chemical compound.Y preferably from alkylene group, inferior alkynyl group ,-(CH 2) kY 2,-CH 2S (O)-and-CH 2O-, more preferably Y is-CH 2O-.
The preferred compound of this embodiment is unsubstituted phenyl for W wherein or has 1 or 2 substituent phenyl that is selected from low alkyl group and halogen atom at its ortho position.W 1Be preferably unsubstituted phenylene or have the substituent phenylene that is selected from methoxyl group, low alkyl group and halogen atom at ortho position with respect to-NH-.
In the preferred compound of this embodiment, A is preferably=O and A 1Be a direct key or-(CH 2) t-.Preferred chemical compound is A wherein 1Be a direct key and R 5Chemical compound for-OH.
The preferred compound of formula I, wherein M is With A is=O to list in the table 1.About-W 1Method for expressing, lower key for the higher key of the junction point of-NH-be junction point with-R-.Be called--R---R 5Clauses and subclauses describe by The part of the particular compound of expression.
Figure A0080951000391
Figure A0080951000411
Figure A0080951000451
Figure A0080951000471
Figure A0080951000481
Figure A0080951000521
Figure A0080951000551
Figure A0080951000591
Figure A0080951000601
Figure A0080951000641
Figure A0080951000661
In another preferred embodiment of formula I, M is
Figure A0080951000662
In this embodiment, preferred chemical compound is=O that R is-(CH for A wherein 2) n-and X be-C (O)-chemical compound.Y preferentially is selected from-O-,-S-,-S (O)-,-S (O) 2-and-NY 4, more preferably-O-.
The preferred compound of this embodiment is unsubstituted phenyl for W wherein or has 1 or 2 substituent phenyl that is selected from low alkyl group and halogen atom at its ortho position.W 1Be preferably unsubstituted phenylene or have the substituent phenylene that is selected from methoxyl group, low alkyl group and halogen atom at ortho position with respect to-NH-.
In this embodiment of formula I, A is preferably=O and A 2Be a direct key or-(CH 2) e-.Preferred chemical compound is A wherein 2Be a direct key and R 8Chemical compound for-OH.
Wherein M is List in the table 2 for the preferred compound of the formula I of=O with A.About-W 1Method for expressing, lower key for the higher key of the junction point of-NH-be junction point with-R-.Be called--R---R 8Clauses and subclauses describe by
Figure A0080951000672
The part of the particular compound of expression.
In another preferred embodiment of formula I, M is
Figure A0080951000674
In this embodiment, preferred chemical compound is=O R for A wherein 10For-CO 2H and q are 0 or 1 chemical compound.More preferably R wherein 10For-CO 2H and q are 0 or 1, most preferably q be 0 and m be 2 chemical compound.
When A be=during O, L preferentially is selected from:
Figure A0080951000681
More preferably, L is selected from Most preferably, L is selected from
The preferred compound of formula I, wherein R is-CH 2-and X be=O, for W wherein is unsubstituted phenyl or has 1 or 2 those chemical compound that are selected from the substituent phenyl of low alkyl group and halogen atom at its ortho position.W 1Be preferably unsubstituted phenylene or have the substituent phenylene that is selected from methoxyl group, low alkyl group and halogen atom at ortho position with respect to-NH-.
The preferred compound of formula I, wherein M is
Figure A0080951000693
, A is=O that R is-CH 2-and X be=O to list in the table 3.About-W 1Method for expressing, lower key for the higher key of the junction point of-NH-be junction point with-R-.Be called--N---Z 2Clauses and subclauses describe by
Figure A0080951000694
The part of the particular compound of expression.
Figure A0080951000711
Figure A0080951000761
Another embodiment preferred of formula I comprises that wherein M is Chemical compound.Preferred A is=O that R is-CH 2-with X be=O.Preferred W is unsubstituted phenyl or has 1 or 2 substituent phenyl that is selected from low alkyl group and halogen atom at its ortho position.Preferred W 1Has the substituent phenylene that is selected from methoxyl group, low alkyl group and halogen atom for unsubstituted phenylene or at ortho position with respect to-NH-.
Q therein 2For And Z 3In the chemical compound for divalent aliphatic hydrocarbon part, preferred chemical compound is R wherein 11For
Figure A0080951000764
Or-NR 12-, more preferably-NR 12Those chemical compounds, R wherein 12Be selected from-H, the low alkyl group of low alkyl group and replacement, most preferably be the dihydroxy low alkyl group.For Z 3Preferred selection be divalent aliphatic hydrocarbon part with 4,5 or 6 carbon atoms.For W 1Preferred a selection be that ortho position with respect to-NH-has the substituent phenylene that is selected from methoxyl group, low alkyl group and halogen atom.
Q therein 2For And Z 3For
Figure A0080951000766
Chemical compound in, R 11Be preferably-NR 12-.In these chemical compounds, x and y are preferably 1.For R 14Preferred selection comprise-H ,-OH and-F.For W 1Preferred a selection be that ortho position with respect to-NH-has the substituent phenylene that is selected from methoxyl group, low alkyl group and halogen atom.
Q therein 2For And Z 3For
Figure A0080951000768
Chemical compound in, R 11Preferentially be selected from-O-and-NR 12-, preferred R wherein 12Be selected from-H and low alkyl group.R 17And R 18Preferably respectively be-H.For W 1Preferred a selection be that ortho position with respect to-NH-has the substituent phenylene that is selected from methoxyl group, low alkyl group and halogen atom.
Q therein 2For And Z 3For Chemical compound in, R 11Be preferably-NR 12-, R wherein 12Be preferably low alkyl group.The preferred compound of this embodiment also comprises wherein R 17And R 18In at least one is those chemical compounds of the low alkyl group of low alkyl group or replacement.
Q therein 2For
Figure A0080951000773
And Z 3For Chemical compound in, R 11Be preferably-NH-and R 17And R 18Respectively be preferably-H.For W 1Preferred a selection be that ortho position with respect to-NH-has the substituent phenylene that is selected from methoxyl group, low alkyl group and halogen atom.
Q therein 2Be selected from aryl, replacement aryl and (more preferably be selected from phenyl and with Z 3The phenyl that replaces of junction point place) chemical compound in, Z 3Be preferably divalent aliphatic hydrocarbon part.
Another embodiment of the invention is the chemical compound of being represented by formula II, Wherein substituent group W, W 1, R 11And Z 3Such as in formula I definition, and L 3Be selected from
Figure A0080951000777
, R wherein 20Preferentially be selected from-H and low alkyl group,
Figure A0080951000778
With
Another embodiment of the invention is the chemical compound of being represented by formula III, Wherein substituent group W, W 1And R 11Such as in formula I definition, and d be selected from 0 and 1 and f be selected from 1 and 2.
The preferred compound of formula I, wherein M is A is=O that R is-CH 2-and X be=O to list in the table 4.About-W 1Method for expressing, lower key for the higher key of the junction point of-NH-be junction point with-R-.Be called--R 11---L 1Clauses and subclauses describe by The part of the particular compound of expression.
Figure A0080951000782
Figure A0080951000791
Figure A0080951000801
Figure A0080951000811
Figure A0080951000821
Figure A0080951000831
Figure A0080951000851
Figure A0080951000881
Figure A0080951000931
Figure A0080951000981
Figure A0080951000991
Main points of the present invention are also included within the prodrug in the formula I scope, and its representational chemical compound comprises wherein R 5Or R 8Be the chemical compound of lower alkoxy, and R wherein 10Or R 19Chemical compound for elementary alkoxy carbonyl.
Main points of the present invention also provide the inhibition cell adhesion, especially at mammal α 4β 1Acceptor site is by the method for the cell adhesion of VLA-4 mediation, and wherein this method comprises the chemical compound of being represented by formula I that gives effective dose.As used herein, suppress the cell adhesion diseases associated that cell adhesion is intended to comprise inhibition, prevention and prevention and VLA-4 mediation, include, but is not limited to inflammation and immunity and the autoimmunity reaction relevant with cell adhesion.
Therefore, main points of the present invention also provide the method for treatment with the cell adhesion diseases associated of VLA-4 mediation, and wherein this method comprises the chemical compound of being represented by formula I that needs the mammal of this kind treatment effective dose.Such disease includes, but is not limited to, for example inflammatory and autoimmunity reaction, diabetes, asthma, arthritis, psoriasis, multiple sclerosis, inflammatory bowel disease, graft-rejection and neoplasm metastasis.As used herein, mammiferous " treatment " also plan to comprise prevention.
Chemical compound of the present invention can be used as monotherapy and gives, or unites with anti-inflammatory agent or immunosuppressant and to give.Such conjoint therapy can comprise the multiple medicine that gives as single dosage form, or as at one time or a plurality of dosage forms of giving of different time.
Can adopt any suitable route of administration, to offer the The compounds of this invention of patient's effective dose.Suitable route of administration can comprise, for example, (as the patch) of oral, rectum, nasal cavity, cheek, parenteral (as in intravenous, the sheath, in subcutaneous, the intramuscular, breastbone, in the liver, in intralesional (intralesional), intracranial, intraarticular and the synovial membrane), percutaneous etc.But preferred oral dosage form such as tablet, dragee, dispersant, suspension, solution, capsule, Perle etc. are because its convenient drug administration.Method and the drug delivery device administration that also can adopt control or slowly discharge.The preparation method of this type of dosage form is well known in the art.
Mix the Pharmaceutical composition of The compounds of this invention, except other therapeutic component, also can comprise excipient, pharmaceutically acceptable carrier.Can suitably select excipient such as starch, sugar, microcrystalline Cellulose, diluent, lubricant, binding agent, coloring agent, flavoring agent, granulating agent, disintegrating agent etc. according to route of administration.Because tablet and capsule are easy to administration, they represent best oral unit dosage form.If desired, can carry out coating to tablet by the water or the non-water technology of standard.
Chemical compound of the present invention can use with the form derived from the pharmaceutically acceptable salt of inorganic base or organic base.Suitable pharmaceutically acceptable base addition salts comprises, but be not limited to, ammonium salt, alkali metal salt, the slaine of making by ammonium, calcium, lithium, magnesium, potassium, sodium and zinc, by chloroprocaine, choline, N, the organic salt that N '-dibenzyl-ethylenediamin, hexanamine, diethanolamine, ethylenediamine, lysine, meglumine (N-methylglucosamine) and procaine are made, and the salt that is become with aminoacid such as arginine, lysine etc.
When The compounds of this invention had basic moiety such as amino, described chemical compound can use with the form of pharmaceutically acceptable, atoxic organic acid or mineral acid.Such acid comprises acetic acid, benzenesulfonic acid, benzoic acid, camphorsulfonic acid, citric acid, ethyl sulfonic acid, methanesulfonic acid, fumaric acid, gluconic acid, glutamic acid, hydrobromic acid, hydrochloric acid, lactic acid, maleic acid, malic acid, mandelic acid, nitric acid, pounces on acid, pantothenic acid, phosphoric acid, succinic acid, sulphuric acid, tartaric acid, right-toluenesulfonic acid etc.Particularly preferred acid is citric acid, hydrochloric acid, maleic acid, fumaric acid, phosphoric acid, sulphuric acid, tartaric acid and right-toluenesulfonic acid.The compounds of this invention also can be the form of hydrate.Detailed Description Of The Invention
Abbreviation and definition
Following term and the abbreviation used in disclosure full text have specified meaning.293E HEK cell=293E HEKC Ac=acetyl group bonded anti--monoclonal antibody of α 4-PE=inhibition of integrins α 4 subunits, in conjunction with
Phycoerythrin bonded anti--monoclonal antibody of β 1-FITC=inhibition of integrins β 1 subunit, in conjunction with
Fluorescein α 5 β 1=beta 2 integrin alpha 5 β 1, fibronectin receptor, VLA-5 α v β 3=beta 2 integrin alpha v β 3, Vitronectic receptor α 4 β 7=integrin alpha-4 β 7Bn=benzyl Boc=tert-butoxycarbonyl BSA=bovine serum albumin c-=ring-cDNA=complementary DNA Chinese hamster ovary celI=Chinese hamster ovary cell p-ClPh=right-chlorophenyl CMV promoter=cytomegalovirus promoter m-CPBA=3-chloro benzylhydroperoxide DAST=diethylamino sulfur trifluoride DCM=dichloromethane=CH 2Cl 2DELFIA=enhanced lanthanide series fluorescence-immunoassay DIAD=diisopropyl azo-2-carboxylic acid DIC=DIC DIEA=N dissociates, N-diisopropylethylamine DMAP=4-N, N-dimethyl aminopyridine DMEM=Dulbecco ' s improvement Eagle ' s culture medium DMF=N, dinethylformamide DTPA=diethylene-triamine pentaacetic acid EDC=1-ethyl-3-(3-dimethylaminopropyl) carbodiimide Et 2O=ether FACS=fluorecyte classification art Fmoc=9-fluorenyl methoxy carbonyl GPIIb/IIIa=beta 2 integrin alpha IIb β 3; fibrinogen deceptor HEPES=N-(2-ethoxy) piperazine-N '-(2-ethanesulfonic acid) HMDS=1; 1; 1; 3; 3; adhesion molecule LDV=Leu-Asp-ValLFA-1 and Mac-1=lymphocyte function-related antigen LiHMDS=1 in 3-HMDS HOAc=acetic acid HOBt=I-hydroxybenzotriazole human IgG1=immunoglobulin G while lICAM=cell; 1; 1; 3; 3,3-HMDS lithium Me=methyl P-MeOPh=right-methoxyphenyl nM=nanomole PBS=phosphate buffered saline (PBS) PEG=Polyethylene Glycol Ph=phenyl PhOH=phenol PyBroP=hexafluorophosphoric acid bromo-three-Bi coughs up the VCAM that Wan Ji Phosphonium RPMI culture medium=Russell Park Memorial Institute culture medium TFA=trifluoroacetic acid TFAA=trifluoroacetic anhydride THF=oxolane TLC=thin layer chromatography TMS=trimethyl silyl Ts=tosyl VCAM-1 (D1D7)=vascular cell adhesion molecule (containing 1-7 immunoglobulin territory) VCAM-IgG merges white 1-7 the immunoglobulin territory (D1D7) of people VCAM-1 that egg=contain is attached on the IgG1 molecule hinge region
The IgG fusion rotein
" alkyl " is intended to comprise the combination of straight or branched alkyl and 1-20 carbon thereof." low alkyl group " refers to about 10 of 1-, preferred 1-about 8 and the alkyl of about 6 carbon atoms of 1-most preferably.Such examples of groups comprises methyl, ethyl, n-pro-pyl, isopropyl, the tert-butyl group, isobutyl group, sec-butyl, the tert-butyl group, amyl group, isopentyl, hexyl, octyl group etc.
" alkylidene " refers to remove a formed divalent group of hydrogen atom from " alkyl ".
" aryl " refers to about 16 carbon atoms by 4-, preferably about 12 carbon atoms of 6-and the more preferably group of the aromatic hydrocarbon ring formation of about 10 carbon atoms of 6-.This ring can be chosen wantonly and is selected from following 1-3 substituent group replacement: alkyl, halogen, hydroxyl, alkoxyl, aryloxy group, haloalkyl, phenyl and heteroaryl.The example of aryl is phenyl, xenyl, 3,4-dichloro-phenyl and naphthyl.
" arlydene " refers to remove a formed divalent group of hydrogen atom from " aryl ".
" aralkyl " refers to comprise the structure of the alkyl that is connected in aromatic ring.Example comprises benzyl, phenethyl, 4-benzyl chloride base etc.
" cycloalkyl " refers to 3-12 carbon atom, the saturated hydrocarbons cyclic group of preferred 3-8 carbon atom.Example comprises cyclopropyl, cyclobutyl, cyclopenta, cyclohexyl, norborny, adamantyl, Fructus Rhodomyrti alkyl (myrtanyl) etc." low-grade cycloalkyl " refers to the cycloalkyl of 3-6 carbon.
" ring alkylidene " means and remove a formed divalent group of hydrogen atom from " cycloalkyl ".
" bivalence C 1-C 20The aliphatic hydrocarbon part " comprise alkylidene, cycloalkylidene, alkylene group, inferior alkynyl group and combination thereof.Example comprises ethylidene, propylidene, inferior propinyl, 2,4-heptadiene subunit etc.
" heterocyclic radical " refers to have 1-6 carbon atom, a preferred 3-6 carbon atom and the 1-4 that is selected from oxygen, a nitrogen and sulfur heteroatomic cyclic group.Example comprises pyrrole radicals, pyridine radicals, pyrazolyl, triazolyl, pyrimidine radicals, pyridazinyl oxazolyl, thiazolyl, pyrazolyl, indyl, thienyl, furyl, azetidinyl, tetrazole radical, the 2-pyrrolinyl, the 3-pyrrolinyl, pyrrolidinyl, 1.3-dioxolanyl, imidazolinyl, imidazolidinyl, pyrazolinyl, pyrazolidinyl isoxazolyl, isothiazolyl, 1,2,3-oxadiazole base, 1,2, the 3-triazolyl, 1,3, the 4-thiadiazolyl group, the 2H-pyranose, the 4H-pyranose, piperidyl, 1,4-dithiane base, morpholinyl, thio-morpholinyl, pyrazinyl, piperazinyl, 1,3, the 5-triazine radical, 1,2,5-trithiane base, benzo (b) thienyl, benzimidazolyl, quinolyl etc.
" inferior heterocyclic radical " means and remove a formed group of hydrogen atom from " heterocyclic radical ".
" heteroaryl " refer to have 1-12 carbon atom, preferred 1-6 carbon atom and be selected from the individual heteroatomic aromatic ring yl of 1-4 of oxygen, nitrogen and sulfur; Or contain 1-4 heteroatomic bicyclo-9-or the hetero-aromatic ring system of 10-unit that is selected from oxygen, nitrogen and sulfur.The structural methine H of heterocyclic radical or heteroaryl atom can be chosen wantonly by alkyl, alkoxy or halogen and replace.Example comprises: imidazole radicals, pyridine radicals, indyl, thienyl, benzopyranyl, thiazolyl, furyl, benzimidazolyl, quinolyl, isoquinolyl, quinoxalinyl, pyrimidine radicals, pyrazinyl, tetrazole radical, pyrazolyl etc.
" inferior heteroaryl " means and remove a formed divalent group of hydrogen atom from " heteroaryl ".
" alkoxyl " refers to the configuration and the combination thereof of straight chain, side chain or cyclic hydrocarbon, comprises 1-20 carbon atom, preferred 1-8 carbon atom, more preferably a 1-4 carbon atom and an oxygen atom on junction point.Suitable alkoxyl comprises methoxyl group, ethyoxyl, positive propoxy, isopropoxy, n-butoxy, isobutoxy, sec-butoxy, tert-butoxy, ring propoxyl group, cyclohexyloxy etc." lower alkoxy " refers to have the alkoxyl of 1-4 carbon atom.
" alkenyl " refers to contain the unsaturated acyclic hydrocarbon base of at least one two key." low-grade alkenyl " refers to contain 2-10 carbon atom, preferred 2-8 carbon atom and more preferably this type of group of 2-6 carbon atom.Suitable non-limiting examples of alkenyls comprises acrylic, butene-1-Ji, isobutenyl, amylene-1-base, 2-methyl butene-1-base, 3-methyl butene-1-base, hexene-1-base, heptene-1-base and octene-1-Ji etc.
" alkylene group " means and remove a formed divalent group of hydrogen atom from " alkenyl ".
" alkynyl group " refers to contain at least one triple-linked unsaturated acyclic hydrocarbon base.Example comprises acetenyl, propinyl etc.
" inferior alkynyl group " refers to remove a formed divalent group of hydrogen atom from " alkynyl group ".
" alkyl of replacement " means wherein the straight or branched alkyl that at least one hydrogen atom that is connected in aliphatic carbons is replaced by following substituent group: for example, and alkyl, amino, alkoxyl, hydroxyl, aryl, cyano group, carboxyl, alkoxy carbonyl, an alkyl amino, alkoxyl, cyano group alkyl, cycloalkyl, alkylthio group, alkyl sulphinyl, alkyl sulphonyl, arylthio, carboxyalkyl, alkoxy carbonyl alkyl, haloalkyl, acyl amino, dialkyl amido, ring amino, halogen atom and nitro.Substituent example like this comprises methyl, isopropyl, methoxyl group, ethyoxyl, propoxyl group, amino, methylamino, phenyl, naphthyl, chlorine, fluorine etc.
" alkylidene of replacement " means wherein the straight or branched alkylidene that at least one hydrogen atom that is connected in aliphatic carbons is replaced by following substituent group: for example, and alkyl, amino, alkoxyl, hydroxyl, aryl, cyano group, carboxyl, alkoxy carbonyl, an alkyl amino, alkoxyl, cyano group alkyl, cycloalkyl, alkylthio group, alkyl sulphinyl, alkyl sulphonyl, arylthio, carboxyalkyl, alkoxy carbonyl alkyl, haloalkyl, acyl amino, dialkyl amido, ring amino, halogen atom and nitro.
" cycloalkyl of replacement " means wherein the cycloalkyl that at least one hydrogen atom that is connected in ring carbon atom is replaced by following substituent group: for example, and alkyl, amino, alkoxyl, hydroxyl, aryl, cyano group, carboxyl, alkoxy carbonyl, an alkyl amino, alkoxyl, cyano group alkyl, cycloalkyl, alkylthio group, alkyl sulphinyl, alkyl sulphonyl, arylthio, carboxyalkyl, alkoxy carbonyl alkyl, haloalkyl, acyl amino, dialkyl amido, ring amino, halogen atom and nitro.
" cycloalkylidene of replacement " means wherein the cycloalkylidene that at least one hydrogen atom that is connected in ring carbon atom is replaced by following substituent group: for example, and alkyl, amino, alkoxyl, hydroxyl, aryl, cyano group, carboxyl, alkoxy carbonyl, an alkyl amino, alkoxyl, cyano group alkyl, cycloalkyl, alkylthio group, alkyl sulphinyl, alkyl sulphonyl, arylthio, carboxyalkyl, alkoxy carbonyl alkyl, haloalkyl, acyl amino, dialkyl amido, ring amino, halogen atom and nitro.
" aryl of replacement " means wherein the aryl that at least one methine hydrogen atom that is connected in aromatic carbon atom is replaced by following substituent group: for example, and alkyl, amino, alkoxyl, hydroxyl, aryl, cyano group, carboxyl, alkoxy carbonyl, an alkyl amino, alkoxyl, cyano group alkyl, cycloalkyl, alkylthio group, alkyl sulphinyl, alkyl sulphonyl, arylthio, carboxyalkyl, alkoxy carbonyl alkyl, haloalkyl, acyl amino, dialkyl amido, ring amino, halogen atom and nitro.
" arlydene of replacement " means wherein the arlydene that at least one hydrogen atom that is connected in aromatic carbon atom is replaced by following substituent group: for example, and alkyl, amino, alkoxyl, hydroxyl, aryl, cyano group, carboxyl, alkoxy carbonyl, an alkyl amino, alkoxyl, cyano group alkyl, cycloalkyl, alkylthio group, alkyl sulphinyl, alkyl sulphonyl, arylthio, carboxyalkyl, alkoxy carbonyl alkyl, haloalkyl, acyl amino, dialkyl amido, ring amino, halogen atom and nitro.
" heteroaryl of replacement " or " heterocyclic radical of replacement " means wherein heteroaryl or the heterocyclic radical that at least one hydrogen atom that is connected in its ring is replaced by following substituent group: for example, and alkyl, amino, alkoxyl, hydroxyl, aryl, cyano group, carboxyl, alkoxy carbonyl, an alkyl amino, alkoxyl, cyano group alkyl, cycloalkyl, alkylthio group, alkyl sulphinyl, alkyl sulphonyl, arylthio, carboxyalkyl, alkoxy carbonyl alkyl, haloalkyl, acyl amino, dialkyl amido, ring amino, halogen atom and nitro.
" inferior heteroaryl of replacement " or " the inferior heterocyclic radical of replacement " means wherein inferior heteroaryl or the inferior heterocyclic radical that at least one hydrogen atom that is connected in its ring is replaced by following substituent group: for example, and alkyl, amino, alkoxyl, hydroxyl, aryl, cyano group, carboxyl, alkoxy carbonyl, an alkyl amino, alkoxyl, cyano group alkyl, cycloalkyl, alkylthio group, alkyl sulphinyl, alkyl sulphonyl, arylthio, carboxyalkyl, alkoxy carbonyl alkyl, haloalkyl, acyl amino, dialkyl amido, ring amino, halogen atom and nitro.
" aralkyl of replacement " means has one or more following substituent aralkyl: for example, and alkyl, amino, alkoxyl, hydroxyl, aryl, cyano group, carboxyl, alkoxy carbonyl, an alkyl amino, alkoxyl, cyano group alkyl, cycloalkyl, alkylthio group, alkyl sulphinyl, alkyl sulphonyl, arylthio, carboxyalkyl, haloalkyl, alkoxy carbonyl alkyl, acyl amino, dialkyl amido, ring amino, halogen atom and nitro.
" halogen " be intended to comprise, for example, and fluorine, chlorine, bromine and iodine.
Term " prodrug " refers to be converted into by metabolic process in vivo the chemical compound of active medicine.[for example seeing N.Boder and J.J.Kaminski, Ann.Rep.Med.Chem.22:303 (1987) and H.Bundgarrd, Adv.Drug Delivery Rev., 3:39 (1989)].The purposes of the prodrug precursor of The compounds of this invention in any method described herein is by well-designed and comprise within the scope of the invention.
Use the term of the functional group that relates to " protection ", " protection " and/or " going protection " in the application's full text.Those skilled in the art should be able to understand these terms well, and they are used for the text description to method, and these methods relate to the successive processing of using a series of reagent.For this reason, blocking group refers to be used to shelter the group of functional group in processing step, otherwise this functional group will react in these steps, and described reaction is undesirable.Blocking group prevents that in this step reaction from taking place, but can remove thereafter, to expose primary functional group.Can remove blocking group in reaction after promptly wherein functional group will interferential reaction finishes and promptly " go protection ".Therefore, when in fact specifying the order of reagent according to method of the present invention, those skilled in the art can easily be contemplated to these groups that will be suitable as " blocking group " of related functional group.
Under situation of the present invention, the functional group that must protect is an amine.Suitable group for this purpose is at the standard textbook of chemical field, as Vitochemical blocking group, T.W.Greene[John Wiley ﹠amp; Sons, New York, 1991] book has description, and this book is attached to herein by reference.Should pay special attention to the chapters and sections of title for " amino protection " (309-405 page or leaf).Preferred blocking group comprises BOC and Fmoc.The protection of these groups and de-protected illustrative methods can be found in Greene and Wuts (318-327 page or leaf).
Carry out synthetic used material described herein and be called solid carrier, beadlet and resin.These terms are intended to comprise: (a) pearl, piller, disk, fiber, gel or granule such as cellulose beadlet, Bio-Glas, silica gel, optional and divinyl benzene crosslinked and optional and the grafted polystyrene bead of Polyethylene Glycol, polyacrylamide beadlet, latex beads, choose wantonly and N, the crosslinked DMAA beadlet of N '-two acryloyl group ethylenediamines, with hydrophobic polymer encapsulated glass granule etc., promptly have the material of rigidity or half-rigid surface; Reach (b) soluble carrier such as Polyethylene Glycol or polystyrene low-molecular-weight, non-crosslinking.Solid carrier can (and usually) have functional group as amino, hydroxyl, carboxyl or halo group, and wherein amino is modal.
Tentagel TMNH 2(Rapp Polymere, Tubingen Germany) are preferred amine-functionalized Polyethylene Glycol-grafted polystyrene resin.Tentagel TM-S-PHB resin has one to the hydroxybenzyl connector, adopts 90% the trifluoroacetic acid in dichloromethane can be with this connector cracking.It is well known in the art making the functionalized technology of solid phase surface.Lysine adhere to (to increase available number of sites) on the beadlet amino acid whose and connector subsequently adhere to and other step in typical combination is synthetic exists, for example, among the PCT application WO95/30642 description is arranged, the content of the disclosure is attached to herein by reference.WO95/30642 describe synthetic in, but described connector is the connector of photodestruciton, but total using priciple of this connector is fully set forth.Optical isomer-diastereomer-geometric isomer
Chemical compounds more described here contain one or more asymmetric centers, thereby can produce enantiomer, diastereomer and other stereoisomer form, can these form aminoacid be defined as (R)-or (S)-aminoacid or be defined as (D)-or (L)-aminoacid according to the absolute stereo chemistry.This invention is intended to comprise diastereomer that all these are possible and their racemic and optically pure form.Optical activity (R)-and (S)-or (D)-and (L)-isomer can be adopted chiral synthon or chiral reagent to prepare, or split with conventional method.When chemical compound described here contains olefinic double bonds or other geometric asymmetry center, except as otherwise noted, then should comprise (E)-and (Z)-two kind of geometric isomer.Equally, plan comprises all tautomeric forms.The configuration that is chosen in any carbon-to-carbon double bond of this appearance does not plan to specify concrete configuration just to convenience; Therefore, arbitrarily be described as the mixture that trans carbon-to-carbon double bond can be any ratio of cis, trans or two kinds of forms at this.
According to top definition, those skilled in the art can easily understand other technical terms of chemistry of using in the application's full text.Term can use separately or use with the form of any combination.Preferred and more preferably the group of chain length be applied to all these type of combinations.Purposes
The compounds of this invention demonstrates the purposes of conduct to the selective depressant of VLA-4 receptor.In direct combination mensuration that adopts other integrin receptor such as β 2 (LFA-1 and Mac-1), β 3 (GPIIb/IIIa and α v β 3) and β 1 (α 4 β 7) and competitive assay, use external test method determination test chemical compound to C 4β 1Inhibition concentration (the IC of receptor 50) and the selectivity of VLA-4.Chemical compound of the present invention has＜K of 1 μ M iValue.Preferred The compounds of this invention is for having K iValue＜300nM, more preferably＜100nM, even more preferably＜50nM, most preferably＜those chemical compounds of 12nM.
Has K iThe embodiment of the preferred compound of value＜50nM shows below.
These embodiment only provide by the mode that illustrates, do not plan to limit the invention.
Figure A0080951001091
Figure A0080951001131
Figure A0080951001151
Figure A0080951001181
Figure A0080951001201
Figure A0080951001221
Figure A0080951001231
Figure A0080951001241
Figure A0080951001251
Figure A0080951001261
Figure A0080951001271
External test
Adopt direct binding assay that the inhibition activity of described chemical compound is carried out quantitatively.In this was measured, the cell inoculation that VLA-4-is expressed was to the microtitration plate of 96-hole.Make cell grow 2 days until converging.The test compound of the various concentration VCAM-IgG fusion rotein with 2nM europium labelling is added.Cell was hatched in micropore 30 minutes at least.After hatching, each micropore of emptying and washing.Determine the bonded amount of VCAM-IgG fusion rotein of europium labelling by the time-resolved fluorescence measurement method.By in the test compound of various concentration each, and the fluorescent quantitation that is incorporated into microtitration plate that does not contain the reference substance of test compound, determine bonded suppression ratio.
The cell that is used for the VLA-4-expression of this algoscopy is personnel selection α 4And β 1The Chinese hamster ovary celI system of the cDNA stable transfection of subunit.The structure of cell line and keeping is described in this assay method.The VCAM IgG fusion rotein that contains 1-7 the immunoglobulin domains (D1D7) of the VCAM-1 that is attached to the people on the IgG1 molecule hinge region carries out labelling with the europium chelate.The preparation of fusion rotein and being marked in this assay method is described.
The adhesion of D1D7-VCAM IgG fusion rotein is come the cell adhesion inhibition activity of determination test chemical compound by blocking-up Jurkat cell.The Jurkat cell is the human lymphocyte system at cell surface expression VLA-4.In this is measured, use 75ng VCAM IgG fusion rotein bag by each hole in the microtitration plate of 96-hole.Remove non-specific adhesion site and seal each hole by adding 1% bovine serum albumin then.Add the test compound of variable concentrations and the Jurkat cell of calcium labelling simultaneously.Under room temperature, make cell adhesion in the dark 1 hour in each hole of VCAM bag quilt.After hatching, be immersed in this plate of washing in the container that fills phosphate buffered saline (PBS) by surface with microtitration plate.Blot each hole with napkin.Determine adherent cell quantity through fluoremetry.The fluorescence that reduces represents that the adhesion of cell is subjected to the inhibition of test compound.
To every kind of test compound at other integrin receptor, promptly among β 2 (LFA-1 and Mac-1), β 3 (GPIIb/IIIa and α v β 3), β 1 (α 5 β 1) and the β 7 (α 4 β 7) to α 4β 1Specificity detect.LFA-1 combines with ICAM-1 and mediated leucocytes is divided a word with a hyphen at the end of a line enters inflammation part.Mac-1 comprises ICAM-1 and Fibrinogen in conjunction with many parts, and is playing an important role aspect the generation of the phagocytosis of neutrophilic granulocyte and oxygen-derived free radicals.The GPIIb/IIIa of platelet surface combines with Fibrinogen in the blood plasma and induced platelet is assembled.α v β 3 comprises vitronectin in conjunction with the albumen of many extracellular matrixs, and mediated cell is divided a word with a hyphen at the end of a line and prevented the death of cell programmatic.α 4 β 7 share identical part such as VLA-4 (VCAM-1, MAdCAM-1 and fibronectin), but have different preferential selections.This receptor is expressed on the lymphoid cell and participates in lymphocyte dividing a word with a hyphen at the end of a line to mucosal tissue.
The mensuration of LFA-1, Mac-1, GPIIb/IIIa and α v β 3 relates to the bag quilt of the receptor of purification to 96-hole microtitration plate.The ligands specific of these receptors europium chelate labelling.In the mensuration of LFA-1 and Mac-1, use the ICAM-1IgG fusion rotein of 1-5 the immunoglobulin domains (D1D5) that contains the H-ICAM-1 who is attached on the IgG1 molecule hinge region.In the mensuration of GPIIb/IIIa and α v β 3, use the Fibrinogen and the vitronectin of europium-labelling respectively.In the presence of the part of europium-labelling, the receptor of purification is hatched together at the test compound of the various concentration of Kong Zhongyu.After hatching, each hole of emptying and washing.Determine the bonded amount of part of europium labelling by the time-resolved fluorescence measurement method.The mensuration of α 4 β 7 is similar to the adhesion inhibition of above-mentioned VLA-4 and measures, and the cell that uses α 4 β 7-to express, RPMI-8886.The MAdCAM-1 IgG fusion rotein that contains 1 and 2 immunoglobulin domains of people MAdCAM-1 and mucin-sample duplicate domain is used as the part corresponding to α 4 β 7.
Be incorporated into the Eu of CHO/VLA-4 cell 3+-VCAM-1 IgG can followingly measure.With 3 * 10 4The concentration in individual/hole is allocated in 4B4 cell (CHO/VLA-4 cell) in each hole of 96-hole microtitration plate.This microtitration plate in 37 ℃, was hatched under 5% carbon dioxide 48 hours, use the lavation buffer solution washed twice then, blot again.To add in each hole with the 50 μ l inhibitor solutions of measuring buffer (2%DMSO final concentration) dilution, then add the 50 μ l Eu that are diluted to 2nM with the mensuration buffer 3+-VCAM-1 IgG.Under room temperature, hatched this plate at least 30 minutes.Every hole reuse lavation buffer solution washing 4 times is also blotted.100 μ l DELFIA are strengthened solution (Enhancement solution) add in each hole, then under room temperature, stirred this plate 5 minutes.Measure then (as DELFIA exometer 1234, Wallace, Inc., USA) fluorescence of each sample.In this was measured, lavation buffer solution contained 25mM HEPES (pH 7.5), 150mM sodium chloride, 1mM calcium chloride, 1mM magnesium chloride and 4mM manganese chloride; Measure buffer and contain 25mM HEPES (pH 7.5), 150mM sodium chloride, 1mM calcium chloride, 1mM magnesium chloride, 4mM manganese chloride, 1%BSA and 20 μ M DTPA.Measure in the body
During can measuring in vivo, further identifies the VLA-4 inhibitor.Such mensuration inspection is soaked into inhibitory action in the bronchoalveolar lavage fluid to the eosinophilic granulocyte in mice (Mus) model.In this is measured, handled animal with cyclophosphamide at the 0th day.At the 2nd day and the 14th day, use ascaris suum (Ascaris suum) extract to animal intraperitoneal immunity inoculation.After 7 days, handle animal with the VLA-4 inhibitor of various dosage.Soon,, the ascaris suum extract attacks animal in the trachea after the administration by being instilled into.After 48 hours, to lung animal is carried out BA lavation by the instillation normal saline.Total cell in the mensuration irrigating solution and eosinophilic granulocyte's counting.
In the mouse model of the inductive bronchitis of ascarid, representational chemical compound (embodiment number 32) suppresses the eosinophilic granulocyte and soaks into and reach 49% when oral dose 30mg/kg.Compare, the representative compounds of a prior art, 4-(N '-2-aminomethyl phenyl urea) phenyl acetyl-LDVP-OH (being described in WO 97/03094), when oral dose 50mg/kg, do not suppress the eosinophilic granulocyte and soak into (% inhibition=-2%).
Other representational chemical compound is also tested in Mus.Dosage, route of administration and the inhibition effect of representative compounds (after this, all test compound the compound number that provides among the synthetic embodiment is provided represent) are shown in Table 5.
Table 5
Dosage/scheme Approach The % that the eosinophilic granulocyte is soaked into suppresses
Chemical compound 79 ?30mg/kg?b.i.d 1X 2 days i.v. ????35.9
Chemical compound 90 ?10mg/kg?t.i.d 2X 2 days p.o. ????18.1
Chemical compound 90 30mg/kg t.i.d x 2 days p.o. ????39.1
Chemical compound 90 50mg/kg t.i.d x 2 days p.o. ????45.9
Chemical compound 90 30mg/kg b.i.d x 2 days i.v. ????47.3
Chemical compound 314 50mg/kg t.i.d x 2 days p.o. ????18.9
Chemical compound 311 50mg/kg t.i.d x 2 days s.c. ????80.2
Chemical compound 311 50mg/kg t.i.d x 2 days p.o. ????42.5
Chemical compound 311 50mg/kg t.i.d x 2 days s.c. ????49.3
1One day twice (0 and 8 hour) 2One day three times (0,8 and 16 hour)
The compounds of this invention also can be measured in other body, in eosinophilic granulocyte's Aggregation Model of testing in rat, carries out further characterized.The compound 48/80 of 50 μ g is expelled in the pleural space of male Sprague Dawley rat.After 24 hours, with twice in each rat thoracic cavity of Hank ' s balanced salt solution washing containing 0.2%EDTA.Measure total cell and eosinophilic granulocyte's counting.Intravenous, oral or subcutaneous giving (b.i.d. was in 0 hour and 8 hours) test compound.The dosage of test compound, route of administration and inhibition effect are shown in Table 6.
Table 6
Dosage/scheme Approach The % that the eosinophilic granulocyte is soaked into suppresses
Chemical compound 90 2 days 30mg/kg of 2 days 10mg/kg of 3mg/kg 2 days ????i.v. ????i.v. ????i.v. ????25.4 ????46.7 ????83.7
Chemical compound 90 50mg/kg 2 days ????p.o. ????50.5
Chemical compound 80 50mg/kg 2 days ????s.c. ????65.3
Chemical compound 92 50mg/kg 2 days ????s.c. ????43.1
Chemical compound 95 50mg/kg 2 days ????s.c. ????40.9
Mice biology-algoscopy
Chemical compound is dissolved in the concentration of 1mg/ml or is suspended in the The suitable solvent.This chemical compound of the female Balb/c mice of orally give (7-9 age in week).After 15 minutes, collect blood sample from the posterior vena cava of anesthetized mice.Preparation serum and-20 ℃ of storages.Through direct binding assay, the cell and the VCAM-IgG fusion rotein that adopt VLA-4 to express, from the serum-concentration of the inhibition determination of activity chemical compound of dilute serum.The serum-concentration that adopts this method to measure is closely related with the concentration of measuring by the LC/MS/MS method.The inhibition effect of the dosage of test compound, route of administration and generation is shown in Table 7.
Table 7
Dosage After the administration minute Serum-concentration (ng/ml)
Chemical compound 58 ?50mg/kg ????30 ????3614
Chemical compound 68 ?10mg/kg ????15 ????261
Chemical compound 78 ?10mg/kg ????15 ????368
Chemical compound 79 ?10mg/kg ????15 ????618
Chemical compound 80 ?10mg/kg ????15 ????693
Chemical compound 90 ?10mg/kg ????15 ????3659
Chemical compound 91 ?10mg/kg ????15 ????2523
Chemical compound 92 ?10mg/kg ????15 ????2162
Chemical compound 96 ?10mg/kg ????15 ????3514
Chemical compound 97 ?10mg/kg ????15 ????1733
Chemical compound 98 ?10mg/kg ????15 ????2796
Chemical compound 102 ?10mg/kg ????15 ????503
Chemical compound 124 ?10mg/kg ????15 ????841
Chemical compound 134 ?10mg/kg ????15 ????224
Chemical compound 146 ?10mg/kg ????15 ????527
Chemical compound 156 ?10mg/kg ????15 ????285
Chemical compound 158 ?10mg/kg ????15 ????301
Chemical compound 166 ?10mg/kg ????15 ????360
Chemical compound 179 ?10mg/kg ????15 ????428
Chemical compound 309 ?10mg/kg ????15 ????669
Chemical compound 311 ?10mg/kg ????15 ????467
Chemical compound 314 ?50mg/kg ????30 ????2309
Chemical compound 318 ?50mg/kg ????30 ????2105
Chemical compound 319 ?50mg/kg ????15 ????603
Chemical compound 323 ?50mg/kg ????15 ????1423
Pharmacokinetics is estimated
The parameter of the pharmacokinetics of the exemplary compounds in mice, rat and monkey model is shown in the table 8,9 and 10.
Table 8
Mice Dosage ????AUC 1(ng·h/mL) ??MRT 2??(hr) ????CL 3?(mL/min/kg)
Chemical compound 68 ?10mg/kg(i.v.) 1595(0-6hr) ????0.2 ????104.5
?1595(0-∞) ????0.2 ????104.5
?20mg/kg(p.o.) 1307(0-6hr) ????1.6 ????637.6
?1751(0-∞) ????4.1 ????476.0
Chemical compound 90 ?10mg/kg(i.v.) 8995(0-6hr) ????0.5 ????18.53
?9219(0-∞) ????0.7 ????18.08
?10mg/kg(p.o.) 2540(0-6hr) ????1.2 ????65.62
?2950(0-∞) ????2.5 ????56.50
Chemical compound 80 ?10mg/kg(i.v.) 5259(0-6hr) ????0.4 ????31.69
?5389(0-∞) ????0.6 ????30.93
?20mg/kg(p.o.) 2190(0-6hr) ????2.3 ????152.24
?3615(0-∞) ????6.5 ????92.21
1In plasma concentration (measuring) through the LC/MS/MS method to the gross area under the time graph 2The average residence time 3Apparent plasma clearance
Table 9
Rat Dosage ????AUC 1??(ng·h/mL) ??MRT 2??(hr) ????CL 3(mL/min/kg)
Chemical compound 90 ?10mg/kg(i.v.) 31915(0-6hr) ????0.8 ????5.23
?33488(0-∞) ????1.1 ????4.98
?10mg/kg(p.o.) 11454(0-6hr) ????1.7 ????17.85
?19968(0-∞) ????8.8 ????9.99
Chemical compound 79 ?10mg/kg(i.v.) 5259(0-6hr) ????0.4 ????31.69
??5389(0-∞) ????0.6 ????30.93
?10mg/kg(p.o.) 22119(0-6hr) ????0.5 ????8.5
?24867(0-∞) ????0.7 ????6.7
Chemical compound 68 ?10mg/kg(i.v.) 6345(0-6hr) ????0.63 ????26.45
??6405(0-∞) ????0.67 ????26.20
?20mg/kg(p.o.) 1867(0-6hr) ????1.1 ????181.1
??2086(0-∞) ????2.0 ????162.7
1In plasma concentration (measuring) through the LC/MS/MS method to the gross area under the time graph 2The average residence time 3Apparent plasma clearance
In monkey model, the representative compounds of single vein dosage (2mg/kg) and ATENOLOL{4-[2 '-hydroxyl-3 '-isopropyl amino) propoxyl group] phenyl-acetamides; SigmaChemical Co., the pharmacokinetic parameter of Code Number A-7655} and the time course of serum-concentration are summarized in table 10 and 11.
Table 10
Monkey AUC 1(ng·h/ml) Cltot 2(ml/min/kg)
Chemical compound 195 ????8405 ????4.0
??ATENOLOL ????5021 ????6.7
1In plasma concentration (measuring) through the LC/MS/MS method to the gross area under the time graph 2Apparent plasma clearance
Table 11
Time 5 minutes 30 minutes 1 hour 2 hours 4 hours 8 hours
The serum-concentration of chemical compound 195 1(ng/ml) 65447 ??3900 ??2225 ?772 ?194 ??28
The serum-concentration of ATENOLOL 1(ng/ml) 4057 ??1189 ??771 ?479 ?348 ?137
1Measure through the LC/MS/MS method VCAM-1 measures the combination of VLA-4 express cell
The preparation of VCAM IgG fusion rotein
In conjunction with in measuring, use the VCAM IgG fusion rotein of 1-7 the immunoglobulin domains (D1D7) of the VCAM-1 that contains the hinge (H), CH2 and the CH3 district that connect the human IgG1.
Express the structure of the stable cell lines of D1D7-VCAM IgG fusion rotein
Transcribing under the control of CMV promoter, with a kind of based on free type plasmid transfection Epstein-Barr virus, that contain D1D7-VCAM IgG fusion gene in the 293E HEKC.Be used in the cell of the 250 μ g/ml hygromycin selection stable transfection among the DMEM that contains 10% hyclone.This cell is secreted into D1D7 VCAM IgG fusion rotein in the culture medium, and its process accumulative total can reach 9 days at most.
The purification of D1D7 VCAM IgG fusion rotein
Cell was cultivated 2 days in the DMEM that contains 10% hyclone, changed into the CCM5 culture medium then and also cultivated again 10 days.This culture is centrifugal, filtration are then with protein A agarose 4 overnight incubation.Thorough washing protein A agarose is with the D1D7 VCAM IgG fusion rotein of 100mM citric acid (pH3) elution of bound.
The europium labelling-preparation of D1D7 VCAM IgG fusion rotein
D1D7-VCAM IgG fusion rotein (1mg/ml) is dialysed to 50mM sodium bicarbonate, 0.9% sodium chloride (pH 8.5).This fusion rotein is added the reagent of an europium labelling, and (test kit of DELFIA labelling derives from Wallac, Gaithersberg, MD; Catalog number (Cat.No.) 1244-302) dark place overnight incubation in the phial and under room temperature.With the albumen of agarose G10 column purification labelling, and measure europium content and protein concentration.Store this albumen until use in-80 ℃.
Express the structure of the cell line of VLA-4 (CHO/VLA-4)
In in conjunction with test, use α 4And β 1The Chinese hamster ovary celI system of cDNA stable transfection.People α 4Gene derive from American Type Culture Collection, and mammiferous expression vector pCI-neo (Promega, Madison, between Xhol WI) and the Xba site again clone.β 1Gene is the pcr amplification by deriving from people's peripheral leukocytes cDNA and through genetic engineering modified, so that start codon places the front and back sequence (context) of total Kozak sequence then.This gene is cloned into again in the pCI-neo downstream of CMV promoter and chimeric intron.
Use coding for alpha 4And β 1The plasmid of gene carries out stable cotransfection to the CHO-K1 cell, selects to express the single cell of high-caliber VLA-4 by fluorecyte classification art (FACS).The antibody that is used for facs analysis has: anti--α 4-PE put together antibody (PharMingen, San Diego, CA) and anti--β 1-FITC put together antibody (Biosource, Camarillo, CA).The α of expression 400,000 and 300,000 sites/cell 4And β 1The cell line 4B4 of subunit is respectively applied in conjunction with in measuring.By facs analysis, adopt Quantum Simply Cellular microballon (Flow CytometryStandards Corporation, Puerto Rico) as standard substance, measure the subunit number.Containing 10% hyclone, 10mM HEPES, in the F12 culture medium of pH7.5,0.5mg/ml G418, adopting goes down to posterity weekly at 1: 48 keeps cell. In conjunction with measuring
The concentration of CHO/VLA-4 cell with 30,000 cells/well is inoculated in the microtitration plate of 96-hole, and, hatches 48 hours under 5% carbon dioxide until fusion in 37 ℃.On the same day of measuring, each hole of emptying is with the 350 μ l lavation buffer solution washed twice that contain 25mM HEPES (pH 7.5), 150mM sodium chloride, 1mM magnesium chloride, 1mM calcium chloride, 2mM manganese chloride.The emptying plate blots to remove buffer with napkin then.
In the presence of the D1D7-VCAM IgG of 2nM europium labelling fusion rotein, with measuring buffer (being added with the lavation buffer solution of 0.1% bovine serum albumin, 20 μ M DTPA and 1% dimethyl sulfoxide) serial dilution test compound.Used final concentration scope is 0.1nM-10 μ M.The equal portions liquid of the mixture of the test compound of 50 μ l is added in the hole of microtitration plate in duplicate.The control wells that is used for total binding is the reception test chemical compound not.Anti--α 4 monoclonal antibodies that the bonded hole of non--specificity is contained (L25.3, Becton Dickinson, Bedford, MA).
Can make cell in the presence of the D1D7-VCAM of europium labelling IgG fusion rotein, at room temperature hatch at least 30 minutes with the mixture of test compound.Use 350 μ l lavation buffer solutions then, employing Skatron plate scrubber washed cell 3 times also blots.The DELFIA enhancing solution of 100 μ l equal portions is added in each hole, then under room temperature, stirred gently 10 minutes.By time-resolved fluorescence measurement method (model: Victor TM, Wallac, Inc., Gaithersberg MD) determines the bonded amount of VCAM-IgG fusion rotein of europium labelling.
As shown in the formula the calculations incorporated percentage rate: [(F T-F NS)-(F I-F NS)]/(F T-F NS) * 100, wherein F TAnd F NSBe respectively and lacking test compound and containing under the situation of anti--α 4 monoclonal antibodies, the D1D7-VCAM IgG fusion rotein of europium labelling is incorporated into the fluorescence signal of cell.F IBe the fluorescence in the hole of containing test compound.By the curve fitting conventional method, (San Diego CA) measures IC to PRIZM for GraphPad Software, Inc. 50(suppress 50%VACM and be incorporated into CHO/VLA-4 cell inhibiting agent concentration). The cell that VLA-4 expresses is to the adhesion of VCAM-1
This subfunction algoscopy is used for the effectiveness of determination test chemical compound in the cell adhesion that suppresses the VLA-4 mediation.
The VCAM bag is by the preparation of plate
(1.5 μ l/ml PBS) add in each hole of 96-hole Costar flat underside (Costar, Franklin, Lakes, NJ, catalog number (Cat.No.) 2580) in phosphate-buffered saline with the D1D7-VCAM IgG fusion rotein of 50 μ l equal portions.Then with this plate in 4 ℃ of overnight incubation.On the same day of measuring, each hole of emptying is with 350 μ l PBS washed twice.1% bovine serum albumin (BSA, Sigma, catalog number (Cat.No.) A9418) with 100 μ l sealed this plate at least 1 hour in PBS under room temperature then.
Cell preparation
Obtain Jurkat cell (clone E6-1) from American Type Cultured Collection, and adopt and go down to posterity at 1: 64 weekly, maintain among RPMI culture medium, 10mM HEPES (pH 7.5), 1mM Sodium Pyruvate, the 10%FCS.Face test before, under room temperature, in the dark use 5 μ M calcein-AM (Molecular Probe, Eugene, OR, catalog number (Cat.No.) C1430) labelling Jurkat cell 30 minutes in the RPMI culture medium.Behind the labelling, with RPMI culture medium twice of washed cell of washing and with 1 * 10 6The concentration resuspending of individual cell/ml.
Cell adhesion is measured
Before facing mensuration, wrap by emptying BSA solution the plate from VCAM.Wash this plate twice with the RPMI culture medium then.The Jurkat cell of the labelling of 100 μ l equal portions is added in each hole, then add the inhibitor solution of 50 μ l.Final inhibitor concentration is 1nM-10 μ M, every kind of concentration retest 3 times.Make inhibitor and cell under room temperature, in the dark hatched 1 hour.After hatching, this plate is immersed in carefully the plate upset is faced down.Draining each Kong Bingyong one deck napkin blots.The 0.1%TritonX-100 of 50 μ l equal portions is added in each hole.In the dark hatched this plate 10 minutes.Go up the adhesion of Jurkat cell is carried out quantitatively at Millipore Cytofluor 2300System plate readout meter (be set in that the 485nM place excites and in the emission of 530nM place).By the curve fitting conventional method, (San Diego CA) measures IC to PRIZM for GraphPadSoftware, Inc. 50(inhibitor concentration that suppresses 50%Jurkat cell adhesion).Synthetic method
The compounds of this invention can pass through the standard chemical synthetic method, and the chemical method of combination, as apply for the method preparation described in the WO 95/30642 at disclosed PCT.Synthetic embodiment
By the general synthetic method of the following example explanation.These specific embodiments only as an illustration mode propose, and are not intended to limit the present invention.To any given raw material or the modification and the variation of processing step will be conspicuous to those skilled in the art.Except as otherwise noted, the solid phase carrier that is used for some embodiment is Tentagel TM-S-PHB resin.This resin has right-hydroxybenzyl connector, it can be cleaved by adopting 90% trifluoroacetic acid in dichloromethane.The load of this resin changes between 0.27-0.30mmol/g, and is not dual load.Embodiment 1
Figure A0080951001411
(1.5g 62.9mmol) packs at the bottom of the 3-neck garden of a 500ml in the flask with 200ml THF and NaH.(6.9g, 62.9mmol) (15.0g, THF 57.3mmol) (100ml) drips of solution was added in this flask with 3-iodobenzoic acid methyl ester with l-vinyl-2-pyrrolidone with 15 minutes.After adding finishes, this reactant mixture is heated to backflow 1 hour.Make reaction vessel be cooled to room temperature, carefully add 6N HCl (100ml) then.This reactant of vacuum concentration adds the 6N HCl (100ml) of another equal portions again to remove THF, and this reactant was refluxed 14 hours.Is 9 to come the quencher reactant by adding sodium bicarbonate until pH, then with this mixture of ethyl acetate extraction 3 times.Through the Organic substance that dried over mgso merges, vacuum concentration obtains a yellow oil.
Then this oil is placed methanol (100ml) to be cooled to-78 ℃.Drip sodium borohydride (3.5g, 96.5) then, made this reactant be warmed to room temperature with 2 hours.Come the quencher reactant by adding 6N HCl until being acidity, make and be alkalescence by adding 40% sodium hydrate aqueous solution then.With this solution of dichloromethane extraction 3 times, through the Organic substance that dried over mgso merges, vacuum concentration obtains the 11.4g yellow oil then.
By top amine is placed 50% dioxane: water (100ml) also adds potassium carbonate and comes this amine is carried out the Boc-protection until being alkalescence.(9.1g 41mmol), stirred 14 hours under room temperature then to add the Boc-acid anhydride in this solution.Come the quencher reactant by adding 1N HCl until being acidity.With this this solution of ethyl acetate extraction 3 times, obtain a sticking yellow oil through dried over mgso and vacuum concentration.This oil reservoir is analysed (25%EtOAc: hexane) obtain 7.0g A.
Hydrogen chloride (gas) bubbling was fed in the 2-bromobenzene guanidine-acetic acid of the 15.8g (73.5mmol) in 100ml methanol 10 minutes.The solution of generation is allocated between 100ml water and the 100ml dichloromethane.Through the dried over mgso organic layer, removal of solvent under reduced pressure, obtain the 2-bromobenzene guanidine-acetic acid methyl ester of 16.8g (73.5mmol), under argon atmospher, it is being mixed in flask at the bottom of the 250ml garden with l-vinyl-2-pyrrolidone and the anhydrous THF of 100ml of 9.0g (80.8mmol).
In this flask, add 3.5g (147mmol) sodium hydride (95%), under room temperature, stirred this solution 10 minutes.Add reflux condenser, with this mixture heated to refluxing 1 hour.This solution is cooled to room temperature, removal of solvent under reduced pressure.Add 30ml aqueous hydrochloric acid solution and 50ml water in the mixture that generates and be heated to backflow under the situation of frozen-free device, reach 96 ℃ until this solution temperature, add condenser this moment, and this solution was refluxed 16 hours.This solution is cooled to room temperature, with 150ml 40% sodium hydrate aqueous solution make be alkalescence, with 3 * 125ml dichloromethane extraction, organic layer with saturated sodium-chloride water solution washing merging, through dried over mgso, removal of solvent under reduced pressure obtains 2-(2-the benzyl bromide)-1-pyrolline of 15.0g (63.0mmol, 86%).
With 15 minutes clockwise 15g (63mmol) 2-(2-benzyl bromide)-1-pyrolline be cooled to 80: 20 methanol of-78 ℃: gradation adds 5.3g (140.0mmol) sodium borohydride in the solution in the acetic acid aqueous solution.This mixture was stirred 1 hour, be warmed to room temperature, this moment, removal of solvent under reduced pressure added 150ml water, made it be alkalescence with sodium hydrate aqueous solution, and it with 10 * 100ml dichloromethane extraction, is obtained emulsion.Organic layer with the saturated sodium bicarbonate aqueous solution washing merges through dried over mgso and solvent removed in vacuo, obtains 14.6g (60.8mmol, 97%) benzyl proline.
In the solution of the 70ml saturated sodium bicarbonate aqueous solution of 14.6g (60.8mmol) benzyl proline and 70ml dioxane, add 15.1g (67.0mmol) Bis(tert-butoxycarbonyl)oxide, under room temperature, stirred this mixture 16 hours.This solution is allocated between 200ml aqueous hydrochloric acid solution and the 200ml ethyl acetate.With 200ml saturated sodium-chloride water solution washing ethyl acetate layer, through dried over mgso and removal of solvent under reduced pressure, obtain residue, through rapid column chromatography (20%-100% ethyl acetate/hexane) purification, obtain the purified A ' of 11.5g (33.8mol, 56%).
Figure A0080951001431
(1.88g 5.0mmol) places at the bottom of the 100ml garden in the flask and is dissolved in DMF (50ml) with A.In this solution, add acid chloride (23mg, 0.3mmol), P (o-Tol) 3(12mg, 0.3mmol), acrylic acid methyl ester. (0.47g, 5.5mmol) and sodium acetate (0.5g, 5.5mmol).Then with this mixture heated to 80 ℃ 14 hours.Make this reactant mixture be cooled to room temperature again, add 1N HCl (100ml).With this solution of ethyl acetate extraction 3 times, through dried over mgso, vacuum concentration obtains a brown oil subsequently then.Chromatography should oil, uses 25% ethyl acetate: the hexane eluting, obtain 1.32g alkene ester, and be the oil of colourless viscosity.
(1.32g 4.1mmol) stands hydrogenization to make this alkene ester then.This alkene is placed Parr hydrogenation bottle, under inert atmosphere, add ethyl acetate (10ml) and 10%Pd/C (100mg).With 45psi hydrogen this bottle is pressurizeed then, jolting is 4 hours under room temperature.By this solution of diatomite filtration, vacuum concentration obtains the 1.29g alkane esters again.
(1.29g 4.0mmol) is dissolved in THF (30ml), methanol (20ml) and the water (10ml) also with Lithium hydrate (200mg, 8.0mmol) saponification to make this chain hydrocarbon ester.Under room temperature, stirred this reactant 3 hours, then among the impouring 1N HCl (50ml).This solution of reuse ethyl acetate extraction 3 times obtains the 1.02g alkanoic acid through dried over mgso and vacuum concentration, is yellow solid.
Made this alkanoic acid in 2 hours (1.02g 3.3mmol) goes protection by adding the 25%TFA/ dichloromethane solution and under room temperature, stirring.The mixture that obtains of vacuum concentration is dissolved in 50% dioxane/water by making de-protected acid then, and (1.08g 4.0mmol) comes to be protected immediately to add potassium carbonate (1.2g) and Fmoc-Cl.Under room temperature, stirred this mixture 14 hours, then among the impouring 1N HCl (100ml).This solution of reuse ethyl acetate extraction 3 times obtains 1 of 495mg through dried over mgso and vacuum concentration, is white crystalline solid.
(500mg 0.14mmol) places a little agitato vase with dried resin.Then with 9ml DMF (2mg, 0.42mmol), DIC (102mg, 0.84mmol) and DMAP (17mg 0.14mmol) adds in this container.Subsequently under room temperature with container jolting 16 hours.Drain content, with DMF, methanol and washed with dichloromethane resin 3 times.By join 10ml 50% piperidines/DMF in the agitato vase and under room temperature jolting removed the Fmoc group in 2 hours.The polyimide resin that obtains with DMF, methanol and washed with dichloromethane 3 times.
Add 9ml DMF, 4-[N '-(o-tolyl urea) in the resin upward]-phenylacetic acid (132mg, 0.42mmol), PyBroP (196mg, 0.42mmol) and DIEA (107mg, 0.84mmol).Under room temperature,, drain then, with DMF, methanol and washed with dichloromethane resin 3 times this content jolting 14 hours.This chemical compound of cracking from the resin is collected filtrate, vacuum concentration then again.By this oil being dissolved in the oil that grinding obtains in the methanol, slowly adding ether and form until precipitation.The collecting precipitation thing, vacuum drying obtains 67mg 1 ', is white crystalline solid.Embodiment 2
(4.2g 11.3mmol) places at the bottom of the 100ml garden in the flask and is dissolved in NEt with A 3(50ml).In this solution, add Pd (PPh 3) 2Cl 2(0.16g, 0.23mmol), CuI (21mg, 0.12mmol) and trimethyl silyl acetylene (1.38g, 13.5mmol).Under room temperature, stirred this mixture 14 hours.By adding 1N HCl (100ml) quencher reactant mixture.With this solution of ethyl acetate extraction 3 times, through dried over mgso, vacuum concentration obtains a yellow oil then then.Chromatography should oil, uses 15% ethyl acetate: the hexane eluting, obtain 3.8g 2, and be the oil of colourless viscosity.
Figure A0080951001451
In 0 ℃, by (12.0ml 12mmol) adds the solution that generates the dicyclohexyl borine in the 6ml anhydrous THF solution of cyclohexene (2.3ml) with borine-THF.Stirred this solution other 1 hour in 0 ℃.(2.0g 5.84mmol), made it be warmed to room temperature with 1 hour then to drip acetylene (2) in 0 ℃ with 15 minutes then.Reuse MeOH (20ml) dilutes this reactant mixture, is cooled to 0 ℃ then.Then drip 2N sodium hydroxide (6ml) and 30%H 2O 2(3.5ml).Again in 0 ℃ of stirred reaction mixture 1 hour, be warmed to then 40 ℃ 2.5 hours.Make this mixture be cooled to room temperature, add the 2N sodium hydroxide of other 6ml.Vacuum is removed Organic substance, with the remaining aqueous solution of ether extraction 3 times, discards Organic substance.With 1N HCl acidify aqueous extract, use ethyl acetate extraction, through dried over mgso, vacuum concentration obtains the 1.7g phenylacetic acid then, is brown crystalline solid.
Made described acid in 2 hours (1.7g 5.6mmol) goes protection by adding the 25%TFA/ dichloromethane solution and under room temperature, stirring.The mixture that obtains of vacuum concentration is dissolved in 50% dioxane/water by making de-protected acid then, and (1.4g 5.5mmol) is protected immediately to add potassium carbonate (15g) and Fmoc-Cl.Under room temperature, stirred this mixture 14 hours, then among the impouring 1NHCl (100ml).This solution of reuse ethyl acetate extraction 3 times, through dried over mgso, vacuum concentration obtains 3 of 1.7g then, is white crystalline solid.
(500mg 0.14mmol) places a little agitato vase with dried resin.Then with 9ml DMF, 3 (180mg, 0.42mmol), DIC (102mg, 0.84mmol) and DMAP (17mg 0.14mmol) adds in this container.Subsequently under room temperature with container jolting 16 hours.Drain content, with DMF, methanol and washed with dichloromethane resin 3 times.By join 10ml 50% piperidines/DMF in the agitato vase and under room temperature jolting removed the Fmoc group in 2 hours.The polyimide resin that obtains with DMF, methanol and washed with dichloromethane 3 times.
Add 9ml DMF, 4-[N '-(o-tolyl urea) in the resin upward]-phenylacetic acid (132mg, 0.42mmol), PyBroP (196mg, 0.42mmol) and DIEA (107mg, 0.84mmol).Under room temperature,, drain then, with DMF, methanol and washed with dichloromethane 3 times this content jolting 14 hours.This chemical compound of cracking from the resin is collected filtrate, vacuum concentration then again.By this oil being dissolved in the oil that grinding obtains in the methanol, slowly adding ether and form until precipitation.Collecting precipitation thing and vacuum drying obtain 52mg 3 ', are white crystalline material.Embodiment 3
Figure A0080951001461
By (0.75g 2.1mmol) places methanol (25ml) and join this potassium hydroxide (1.4g) solution and make acetylene (2) go protection with 2.The solution that stirring generates under room temperature 1 hour.The vacuum concentration reactant mixture is used 1N HCl acidify then.The aqueous solution that generates with ethyl acetate extraction 3 times, through the Organic substance that dried over mgso merges, vacuum concentration obtains the de-protected acetylene of 0.56g again, is brown oil.
(0.56g 2.0mmol) adds among the THF (50ml) and is cooled to-78 ℃ with de-protected acetylene.(1M solution 4.7ml), stirred this reactant 30 minutes to drip LiHMDS then.Again the carbon dioxide bubbling was fed in the reactant mixture 15 minutes, on this reactant impouring carbon dioxide solid.By adding this reactant of 1N HCl (100ml) quencher, with this aqueous solution of ethyl acetate extraction 3 times, through the Organic substance that dried over mgso merges, vacuum concentration obtains propanoic acid (0.71g) then, is white solid.
Made alkanoic acid (0.71g) go protection in 2 hours by adding the 25%TFA/ dichloromethane solution and under room temperature, stirring.The mixture that obtains of vacuum concentration is dissolved in 50% dioxane/water by making de-protected acid then, and (1.29g 4.9mmol) is protected immediately to add potassium carbonate (15g) and Fmoc-Cl.Under room temperature, stirred this mixture 14 hours, then among the impouring 1N HCl (100ml).This solution of reuse ethyl acetate extraction 3 times, through dried over mgso, vacuum concentration obtains 4 then, is brown oil.Chromatography should oil then, and use 5% methanol: the dichloromethane eluting obtains the required chemical compound of 110mg.
Figure A0080951001471
(500mg 0.14mmol) places a little agitato vase with dried resin.Then with 9ml DMF, 4 (184mg, 0.42mmol), DIC (102mg, 0.84mmol) and DMAP (17mg 0.14mmol) adds in this container.Subsequently under room temperature with container jolting 16 hours.Drain content, with DMF, methanol and washed with dichloromethane resin 3 times.By join 10ml 50% piperidines/DMF in the agitato vase and under room temperature jolting removed the Fmoc group in 2 hours.The polyimide resin that obtains with DMF, methanol and washed with dichloromethane 3 times.
Add 9ml DMF, 4-[N '-(o-tolyl urea) in the resin upward]-phenylacetic acid (132mg, 0.42mmol), PyBroP (196mg, 0.42mmol) and DIEA (107mg, 0.84mmol).Under room temperature,, drain then, with DMF, methanol and washed with dichloromethane 3 times this content jolting 14 hours.This chemical compound of cracking from the resin is collected filtrate, vacuum concentration then again.By this oil being dissolved in the oil that grinding obtains in the methanol, slowly adding ether and form until precipitation.Collecting precipitation thing and vacuum drying obtain 27mg 4 ', are white crystalline material.Embodiment 4
(0.5g 1.3mmol) places THF (20ml), is cooled to-78 ℃ with iodide (A).Drip butyl lithium (2.21ml, 1.6M solution), remove cryostat then, bubbling fed gaseous carbon dioxide 10 minutes.In this reactant mixture impouring dry ice, add 1M HCl (100ml).With this mixture of ethyl acetate extraction 3 times, through the Organic substance that dried over mgso merges, vacuum concentration obtains the 0.32g benzoic acid then, is white crystalline solid.
Made this benzoic acid in 2 hours (0.32g 1.68mmol) goes protection by adding the 25%TFA/ dichloromethane solution and under room temperature, stirring.The mixture that obtains of vacuum concentration is dissolved in 50% dioxane/water by making de-protected acid then, and (0.44g 1.67mmol) is protected immediately to add potassium carbonate (15g) and Fmoc-Cl.Under room temperature, stirred this mixture 14 hours, then among the impouring 1N HCl (100ml).This solution of reuse ethyl acetate extraction 3 times, through dried over mgso, vacuum concentration obtains 0.38g 5 then, is white crystalline solid.
Figure A0080951001481
(500mg 0.14mmol) places a little agitato vase with dried resin.Then with 9ml DMF, 5 (173mg, 0.42mmol), DIC (102mg, 0.84mmol) and DMAP (17mg 0.14mmol) adds in this container.Subsequently under room temperature with container jolting 16 hours.Drain content, with DMF, methanol and washed with dichloromethane resin 3 times.By join 10ml 50% piperidines/DMF in the agitato vase and under room temperature jolting removed the Fmoc group in 2 hours.The polyimide resin that obtains with DME, methanol and washed with dichloromethane 3 times.
Add 9ml DMF, 4-[N '-(o-tolyl urea) in the resin upward]-phenylacetic acid (132mg, 0.42mmol), PyBroP (196mg, 0.42mmol) and DIEA (107mg, 0.84mmol).Under room temperature,, drain then, with DMF, methanol and washed with dichloromethane 3 times this content jolting 14 hours.This chemical compound of cracking from the resin is collected filtrate, vacuum concentration then then.By this oil being dissolved in the oil that grinding obtains in the methanol, slowly adding ether and form until precipitation.Collecting precipitation thing and vacuum drying obtain 51mg 5 ', are white crystalline material.Embodiment 5 (E)-4-[2-[1-[4-[N '-(2-aminomethyl phenyl) urea groups] phenyl acetyl]-the 2-pyrrolidinyl] vinyl] benzoic acid
(904mg, (1.0M is in THF, and 3ml 3.00mmol), continues to stir 1 hour under identical temperature to add LiHMDS in THF 3.01mmol) (20ml) solution to 4-(phosphonomethyl) benzoic acid triethyl cold (78 ℃), that stir.Will (500mg 2.51mmol) adds in this mixture, makes this mixture be warmed to room temperature with 1 hour at the N-Boc dried meat ammonium aldehyde (prolinal) among the THF (10ml).Stir after 2 hours, this mixture of water quencher is used ethyl acetate extraction.With saline (200ml) washing extract, through dried over mgso and evaporation.Residue is through silica gel column chromatography, with normal hexane-ethyl acetate (8: 1,, obtain (E)-4-[2-[1-(tert-butoxycarbonyl)-2-pyrrolidinyl of 713mg (82%) v/v) as eluant] vinyl] ethyl benzoate, be the colourless crystallization solid.Mp 68-70 ℃; IR (KBr) 1710,1697,1681cm -1 1H-NMR (CDCl 3) δ 1.39 (12H, a plurality of m), 1.77-1.93 (3H, m), 2.11 (1H, m), 3.47 (2H, m), 4.34-4.54 (be total to 3H, m), 6.22 (1H, m), 6.43 (1H, d, J=14.2Hz), 7.39 (2H, J=8.3Hz), 7.97 (2H, d, J=8.3Hz); MS (FAB) m/z 346 (M ++ 1); C 20H 27NO 4The analytical calculation value: C, 69.54; H, 7.88; N, 4.05.Measured value: C, 69.52; H, 8.08; N, 4.07.
To (E)-4-[2-[1-(the tert-butoxycarbonyl)-2-pyrrolidinyl that stirs] vinyl] (700mg adds TFA (3ml) in dichloromethane 2.03mmol) (3ml) solution to ethyl benzoate, the mixture that stirring obtains 3 hours.Concentrate this mixture, make residue be alkalescence by adding saturated sodium bicarbonate.(2 * 100ml) extract this mixture with chloroform.Through dry extract and the vacuum concentration that merges of sodium carbonate, obtain (E)-4-[2-(2-pyrrolidinyl) vinyl of 434mg (87%)] ethyl benzoate, be brown oil. 1H-NMR (CDCl 3) δ 1.39 (3H, t, J=7.3Hz), 1.52-2.06 (4H, a plurality of m), 2.93-2.99 (1H, m), 3.07-3.13 (1H, m), 3.74 (1H, q, J=7.3Hz), 4.37 (2H, q, J=7.3Hz), 6.34 (1H, dd, J=15.6,7.3Hz), 6.54 (1H, d, J=15.6Hz), 7.41 (2H, d, J=8.3Hz), 7.97 (2H, d, J=8.3Hz).
With (E)-4-[2-(2-pyrrolidinyl) vinyl] ethyl benzoate (434mg, 1.77mmol), 4-[N '-(2-aminomethyl phenyl) urea groups] phenylacetic acid pentafluorophenyl esters (797mg, 1.77mmol), (0.37ml, 2.66mmol) mixture in DMF (15ml) stirred 15 hours triethylamine.(300ml) dilutes this mixture with ethyl acetate.(2 * 200ml) wash this solution, through dried over mgso and vacuum evaporation with saline.Residue as eluant, obtains (E)-4-[2-[1-[4-[N '-(2-aminomethyl phenyl) urea groups of 906mg (q.y.) with chloroform-ethyl acetate (4: 1) through silica gel column chromatography] phenyl acetyl]-the 2-pyrrolidinyl] vinyl] ethyl benzoate, be brown oil. 1H-NMR (CDCl 3) δ 1.39 (3H, t, J=7.3Hz), 1.83-2.20 (4H, a plurality of m), 2.24 (3H, d, J=4.9Hz), 3.63 (4H, m), 4.36 (2H, q, J=7.3Hz), 4.62 and 4.84 (be total to 1H, m), 6.18-6.47 (2H, m), 7.03-8.02 (14H, a plurality of m).
With (E)-4-[2-[1-[4-[N '-(2-aminomethyl phenyl) urea groups] phenyl acetyl]-the 2-pyrrolidinyl] vinyl] (906mg, 1.77mmol) agitate compounds in 0.25N sodium hydroxide (14ml) and THF (14ml) heated 3 days under refluxing ethyl benzoate.In this mixture impouring ice-1N HCl (200ml), the sucking filtration collecting precipitation.With this solid recrystallization from chloroform-methanol-normal hexane, obtain 6 of 453mg (53%), be light yellow crystalline powder.Mp 165-168 ℃; IR (KBr) 3282,2974,2663,2537,1700,1685cm -1 1H-NMR (DMSO-d 6) δ 1.74-2.12 (4H, m), 2.24 (3H, d, J=4.9Hz), 3.35-3.66 (4H, m), 4.67-4.74 (1H, m), 6.25-6.41 (1H, m), 6.53 (1H, s), 6.93 (1H, t, J=7.3Hz), 7.08-7.92 (12H, a plurality of m), 9.00 (1H, m), 12.87 (1H, br s); MS (FAB) m/z 484 (M ++ 1); C 29H 29N 3O 40.5H 2The analytical calculation value of O: C, 70.71; H, 6.14; N, 8.39.Measured value: C, 70.46; H, 6.07; N, 8.39.Embodiment 64-[2-[1-[4-[N '-(2-aminomethyl phenyl) urea groups] phenyl acetyl]-the 2-pyrrolidinyl] ethyl] benzoic acid
Under vigorous stirring; with (E)-4-[2-[1-[4-[N '-(2-aminomethyl phenyl) urea groups] phenyl acetyl]-the 2-pyrrolidinyl] vinyl] benzoate (200mg, 0.414mmol) and the hydrogenation 1 hour under 1 atmospheric pressure of the mixture of 5%Pd/C (200mg) in methanol (20ml).Filter this mixture, concentrated filtrate.Residue as eluant, obtains 7 of 201mg (q.y.) with chloroform-methanol (4: 1) through silica gel column chromatography, is the colourless crystallization powder.Mp 180-190 ℃; IR (KBr) 3345,3124,3060,3027,2960,2927,2875,1706,1672cm -1 1H-NMR (DMSO-d 6) δ 1.04-3.96 (being total to 16H, a plurality of m) 6.91-7.41 (9H, m), 7.79-7.90 (3H, m), 8.23 (1H, br s), 9.31 (1H, br s); MS (FAB) m/z 486 (M ++ 1); C 29H 31N 3O 42.25H 2The analytical calculation value of O: C, 66.21; H, 6.80; N, 7.99.Measured value: C, 65.97; H, 6.20; N, 7.72.Embodiment 7 (S)-4-[2-[1-[4-[N '-(2-aminomethyl phenyl) urea groups] phenyl acetyl] pyrrolidinyl] methoxyl group] benzoic acid
Figure A0080951001511
(S)-4-[2-[1-[3-methoxyl group-4-(N '-the phenyl urea groups) phenyl acetyl] pyrrolidinyl] methoxyl group] benzoic acid
Figure A0080951001512
(S)-and 4-[2-[1-[4-[N '-(2-chlorophenyl) urea groups] phenyl acetyl] pyrrolidinyl] methoxyl group] benzoic acid
8, under 9 and 10 room temperatures to Boc-dried meat ammonia alcohol (prolinol) (3.00g, 14.9mmol), ethyl p-hydroxybenzoate (2.40g, 14.5mmol) and triphenyl phasphine (3.91g, 14.9mol) drip in the stirring the mixture in THF (80ml) diethylazodicarboxylate (2.86g, 16.4mmol).After adding finishes, the mixture that obtains was heated 2 hours under refluxing.After the cooling, this mixture of vacuum concentration.Residue is dissolved in the ethyl acetate, washs in proper order with 1N sodium hydroxide, water, saline.Through dried over mgso ethyl acetate layer and vacuum evaporation.Residue is through purification by silica gel column chromatography, with ethyl acetate-normal hexane (1: 4, v/v) as eluant, obtain (S)-4-(1-tert-butoxycarbonyl-2-pyrrolidinyl) methoxybenzoic acid ethyl ester of 4.88g (93%), be a kind of oil.
Add methanol (100ml) and 1N sodium hydroxide (50ml) in (S)-4-upward (1-tert-butoxycarbonyl-2-pyrrolidinyl) methoxybenzoic acid ethyl ester.Under room temperature, stirred this mixture 15 hours.After methanol is removed in decompression, water (50ml) is added in the residual solution.(x2) washs this aqueous solution with ether, then by adding 1N HCl acidify.With this mixture of ethyl acetate extraction, water, salt water washing, through dried over mgso and vacuum evaporation, obtain (S)-4-(1-tert-butoxycarbonyl-2-pyrrolidinyl) methoxybenzoic acid of 4.26g (95%), be crystalline solid.
Add dichloromethane (10ml) and TFA (10ml) in (S)-4-upward (1-tert-butoxycarbonyl-2-pyrrolidinyl) methoxybenzoic acid.Under room temperature, stir this mixture 1 hour, ether is added in this mixture, collect the solid that obtains.Make in this solid water-soluble (100ml), dioxane (50ml) and the sodium bicarbonate (4.4g).(3.34g 12.9mmol) adds in this solution, the mixture that stirring obtains under room temperature 20 hours with Fmoc-Cl.(x2) washs this mixture with ether, separates the water-bearing layer.By adding 1N HCl acidify water layer.With this mixture of ethyl acetate extraction, water, salt water washing extract through dried over mgso and vacuum evaporation, obtain (S)-4-(1-Fmoc-2-pyrrolidinyl) methoxybenzoic acid of 5.36g (91%), and the oil for viscosity leaves standstill crystallization with it.
Make Wang resin (0.71mmol/g, 400mg) be suspended in (S)-4-(1-Fmoc-2-pyrrolidinyl) methoxybenzoic acid (500mg, 1.13mol), DMAP (35mg, 0.29mmol), HOBt (40mg, 0.30mmol) and DIC (0.45ml is 2.9mmol) in the solution in the mixture of DMF (3ml) and dichloromethane (7ml).This mixture of jolting 20 hours is also drained.With DMF (x3), methanol (x3), dichloromethane (x3) washing resin, drying under reduced pressure obtains the 522mg resin, uses it for preparation 8,9 and 10.
Add piperidines-DMF (50%v/v, 4ml) solution, this mixture of jolting 1 hour in 8 upward the resins (115mg).Wash this resin with DMF (x3), methanol (x3), dichloromethane (x3).In this resin, add DMF (4ml), dichloromethane (2ml), 4-[N '-(2-aminomethyl phenyl) urea groups] phenylacetic acid (70mg, 0.25mmol), PyBrop (115mg, 0.25mmol) and DIEA (0.13ml, 0.75mmol).With this mixture jolting 21 hours and drain.With DMF (x3), methanol (x3), dichloromethane (x3) washing resin.The dichloromethane solution of adding TFA in this resin (50%v/v, 4ml), this mixture of jolting 2 hours.Filter this mixture, vacuum concentrated filtrate.Make residue through the Sep-Pak column purification.Except that after desolvating, ether is added in the residue, collect the solid that produces, obtain 25mg 8, be light yellow crystalline material.MS(FAB)m/z?488(M ++1)
DMF (50%v/v, 3ml) solution, this mixture of jolting 2 hours of adding piperidines in 9 upward the resins (60mg).Wash this resin with DMF (x3), methanol (x3), dichloromethane (x3).In this resin, add DMF (2ml), dichloromethane (1ml), 3-methoxyl group-4-(N '-phenyl urea groups) phenylacetic acid (40mg, 0.13mmol), PyBrop (60mg, 0.13mmol) and DIEA (0.060ml, 0.34mmol).With this mixture jolting 40 hours and drain.With DMF (x3), methanol (x3), dichloromethane (x3) washing resin.The dichloromethane solution of adding TFA in this resin (30%v/v, 3ml), this mixture of jolting 5 hours.Filter this mixture, vacuum concentrated filtrate.Make residue through the Sep-Pak column purification.Except that after desolvating, ether is added in the residue, collect solid, obtain 8mg 9, be crystalline solid.MS(FAB)m/z?504(M ++1)
DMF (50%v/v, 20ml) solution, this mixture of jolting 4 hours of adding piperidines in 10 upward the resins (637mg).Wash this resin with DMF (x3), methanol (x3), dichloromethane (x3).In this resin, add DMF (12ml), dichloromethane (8ml), 4-(Fmoc-amino) phenylacetic acid (530mg, 1.42mmol), PyBrop (660mg, 1.43mmol) and DIEA (0.62ml, 3.56mmol).With this mixture jolting 60 hours and drain.With DMF (x3), methanol (x3), dichloromethane (x3) washing resin, drying under reduced pressure obtains the 617mg resin.DMF (40%v/v, 2ml) solution, this mixture of jolting 1 hour of this resin of 57mg being added piperidines.Wash this resin with DMF (x3), methanol (x3), dichloromethane (x3).(0.050ml 041mmol) adds resin in the suspension of THF (1ml) and dichloromethane (1ml) with Carbimide. 2-chloro phenyl ester.With this mixture jolting 20 hours and drain.Wash this resin with DMF (x3), methanol (x3), dichloromethane (x3).The dichloromethane solution of adding TFA in this resin (25%v/v, 2ml), this mixture of jolting 1.5 hours.Filter this mixture, vacuum concentrated filtrate.Make residue through the Sep-Pak column purification.Except that after desolvating, ether is added in the residue, collect solid and obtain 2mg 10, be crystalline solid.MS (FAB) m/z 508 (M ++ 1) embodiment 8 (S)-3-[2-[1-[3-methoxyl group-4-(N '-phenyl urea groups) phenyl acetyl] pyrrolidinyl] methoxyl group] phenylacetic acid (S)-and 3-[2-[1-[4-[N '-(2-aminomethyl phenyl) urea groups] phenyl acetyl] pyrrolidinyl] methoxyl group] phenylacetic acid
Figure A0080951001542
11 and 12 under room temperature, to Boc-dried meat ammonia alcohol (3.51g, 17.5mmol) ,-the hydroxyphenyl acetic acid methyl ester (2.90g, 17.5mmol), triphenyl phasphine (4.60g, 17.6mol) drip in the stirring the mixture in THF (50ml) diethylazodicarboxylate (3.05g, 17.5mmol).After adding finishes, the mixture that obtains was heated 3 hours under refluxing.After the cooling, this mixture of vacuum concentration.Residue is dissolved in the ethyl acetate, washs in proper order and through dried over mgso with 1N sodium hydroxide, water, saline.Except that after desolvating, make residue through purification by silica gel column chromatography, usefulness ethyl acetate-hexane (1: 4, v/v) as eluant, obtain (S)-3-(1-tert-butoxycarbonyl-2-pyrrolidinyl) methoxyphenyl methyl acetate of 5.49g (90%), be a kind of oil.
The methanol (60ml) of top (S)-3-(1-tert-butoxycarbonyl-2-pyrrolidinyl) methoxyphenyl methyl acetate and the mixture in the 1N sodium hydroxide (20ml) were stirred under room temperature 8 hours.After the removal of solvent under reduced pressure, water (50ml) is added in the residue.(x2) extracts this mixture with ether, by adding 1N HCl acidify water-bearing layer.With this mixture of ethyl acetate extraction.Water, salt water washing extract, through dried over mgso, vacuum concentration then obtains (S)-3-(1-tert-butoxycarbonyl-2-pyrrolidinyl) methoxybenzene guanidine-acetic acid of 4.43g (88%), is a kind of oil of viscosity.
The mixture of top (S)-3-(1-tert-butoxycarbonyl-2-pyrrolidinyl) methoxybenzene guanidine-acetic acid in dichloromethane (10ml) and TFA (10ml) stirred 1 hour under room temperature.Add ether in this mixture and leave standstill.Shift out the upper strata through decant, obtain a kind of oil.Should oil mixture in water (100ml), dioxane (30ml) and sodium bicarbonate (6.0g) add Fmoc-Cl (2.86g, 11.1mmol) in, under room temperature, stirred the mixture 20 hours.(x2) extracts this mixture with ether, by adding 1N HCl acidify water-bearing layer.With this mixture of ethyl acetate extraction.Water, salt water washing extract through dried over mgso and vacuum concentration, obtain (S)-3-(1-Fmoc-2-pyrrolidinyl) methoxybenzene guanidine-acetic acid of 5.08g (81%), are a kind of oil of viscosity.
Make Wang resin (0.71mmol/g, 400mg) be suspended in (S)-3-(1-Fmoc-2-pyrrolidinyl) methoxybenzene guanidine-acetic acid (520mg, 1.14mol), DMAP (35mg, 0.29mmol), HOBt (40mg, 0.30mmol) and DIC (0.45ml is 2.9mmol) in the solution in the mixture of DMF (3ml) and dichloromethane (7ml).This mixture of jolting 20 hours is also drained.With DMF (x3), methanol (x3), dichloromethane (x3) washing resin, drying under reduced pressure obtains the 593mg resin, uses it for preparation 11 and 12.
(40%v/v, 3ml) the mixture jolting in is 1 hour for 11 upward the piperidines-DMF of resin (70mg).Wash this resin with DMF (x3), methanol (x3), dichloromethane (x3).In this resin, add DMF (1.5ml), dichloromethane (1.5ml), 3-methoxyl group-4-(N '-phenyl urea groups) phenylacetic acid (42mg, 0.14mmol), PyBrop (70mg, 0.15mmol) and DIEA (0.065ml, 0.37mmol).With this mixture jolting 15 hours and drain.With DMF (x3), methanol (x3), dichloromethane (x3) washing resin.The dichloromethane solution of adding TFA in this resin (25%v/v, 2ml), this mixture of jolting 3 hours.Filter this mixture, vacuum concentrated filtrate.Make residue through the Sep-Pak column purification.Except that after desolvating, ether is added in the residue, collect solid and obtain 8mg 11, be crystalline solid.MS(FAB)m/z?518(M ++1)
(40%v/v, 3ml) the mixture jolting in is 1 hour for 12 upward the piperidines-DMF of resin (70mg).Wash this resin with DMF (x3), methanol (x3), dichloromethane (x3).In this resin, add DMF (1.5ml), dichloromethane (1.5ml), 4-[N '-(2-aminomethyl phenyl) urea groups] phenylacetic acid (40mg, 0.14mmol), PyBrop (70mg, 0.15mmol) and DIEA (0.065ml, 0.37mmol).With this mixture jolting 15 hours and drain.With DMF (x3), methanol (x3), dichloromethane (x3) washing resin.(25%v/v, 2ml) the mixture jolting in is 3 hours at the TFA-dichloromethane with this resin.Filter this mixture, vacuum concentrated filtrate.Make residue through the Sep-Pak column purification.Except that after desolvating, ether is added in the residue, collect solid and obtain 11mg 12, be crystalline solid.MS (FAB) m/z502 (M ++ 1) embodiment 94-[2-[1-[3-methoxyl group-4-(N '-phenyl urea groups) phenyl acetyl]-the 2-pyrrolidinyl] acetenyl] benzoic acid
To N-boc-dried meat ammonium aldehyde (5.98g, 30mmol) and triphenyl phasphine (62.95g 240mmol) slowly adds CBr in cold (50 ℃) solution of the stirring in dichloromethane (200ml) 4(39.80g, dichloromethane 120mmol) (50ml) solution continue to stir 1 hour in 0 ℃.In this mixture, add saturated sodium bicarbonate, with this mixture of chloroform extraction.Wash extract with water, through dried over mgso and evaporation.Residue is through silica gel column chromatography, with chloroform and normal hexane-ethyl acetate (4: 1, v/v) as eluant, obtain 1-(tert-butoxycarbonyl)-2-(2, the 2-dibromo vinyl) pyrrolidine of 7.84g (74%), be colourless tablet.Mp 61-63; IR (KBr) 1693cm -1 1H-NMR (CDCl 3) δ 1.46 (and 9H, s) 1.72-2.19 (4H, m), 3.35-3.45 (2H, m), 4.35 (1H, br s), 6.36 (1H, br s); MS (FAB) m/z 352,354,356,358; C 11H 17NO 2Br 2The analytical calculation value: C, 37.21; H, 4.83; N, 3.94.Measured value: C, 37.14; H, 4.83; N, 4.00.
1-(the tert-butoxycarbonyl)-2-(2 of cold (78 ℃) that stir with 10 fens clockwise, the 2-dibromo vinyl) pyrrolidine (7.81g, add in THF 22mmol) (200ml) solution n-BuLi (1.59M in hexane, 28ml, 44mmol), continue down to stir 2 hours in uniform temp.By adding this reactant of saturated ammonium chloride quencher, use ethyl acetate extraction.With salt water washing extract, through dried over mgso and evaporation.Residue is through silica gel column chromatography, with normal hexane-ethyl acetate (10: 1, v/v) as eluant, obtain 1-(the tert-butoxycarbonyl)-2-acetenyl pyrrolidine of 4.15g (97%), be light yellow oil. 1H-NMR(CDCl 3)δ1.48(9H，s)，1.82-2.21(4H，m)，3.30-3.45(2H，m)，4.41-4.52(1H，m)。
With 4-iodobenzoic acid ethyl ester (1.7ml, 10mmol), Pd (PPh 3) 4(578mg, 0.5mmol) and CuI (190mmol is 1mmol) at i-Pr 2Suspension among the NH (20ml) stirred 0.5 hour under blanket of nitrogen.With adding 1-(tert-butoxycarbonyl)-2-acetenyl pyrrolidine (1.95g, i-Pr 10mmol) in 10 minutes in this mixture of clockwise 2NH (20ml) solution.After stirring 3 hours under the room temperature, in this mixture impouring water, use ethyl acetate extraction.With salt water washing extract, through dried over mgso and evaporation.Residue is through silica gel column chromatography, with normal hexane-ethyl acetate (10: 1, v/v) as eluant, obtain 1-(tert-butoxycarbonyl)-2-(2-(4-ethoxy carbonyl phenyl) acetenyl) pyrrolidine of 2.77g (81%), be colorless oil. 1H-NMR(CDCl 3)δ1.37(3H，t，J＝6.8Hz)，1.49(9H，s)1.85-2.12(4H，m)，3.37-3.51(2H，m)，4.37(2H，q，J＝6.8Hz)，4.54-4.77(1H，m)，7.44(2H，d，J＝7.8Hz)，7.96(2H，d，J＝7.8Hz)
To 1-(tert-butoxycarbonyl)-2-[2-(4-ethoxy carbonyl phenyl) acetenyl] (2.75g 8mmol) adds TFA (5ml) to pyrrolidine in the agitating solution of dichloromethane (5ml), the mixture that obtains is stirred spend the night.This mixture of vacuum concentration, with saturated sodium bicarbonate make be alkalescence, use chloroform extraction.With salt water washing extract, through dried over mgso, evaporation obtains 2-[2-(the 4-ethoxy carbonyl phenyl) acetenyl of 1.95g (q.y.)] pyrrolidine, be a kind of light yellow oil. 1H-NMR(CDCl 3)δ1.38(3H，t，J＝6.8Hz)，1.82-2.16(4H，m)，3.01-3.48(2H，m)，4.00-4.11(1H，m)，4.37(2H，q，J＝6.8Hz)，4.54-4.77(1H，m)，7.44-7.46(2H，m)，7.95-7.97(2H，m)。
With 3-methoxyl group-4-(N '-phenyl urea groups) phenylacetic acid (180mg, 0.6mmol), 2-(2-(4-ethoxy carbonyl phenyl) acetenyl) pyrrolidine (146mg, 0.6mmol), EDC (173mg, 0.9mmol), DMAP (73mg, 0.6mmol) and the mixture of catalytic HOBt in DMF (10ml) stir and to spend the night.In this mixture impouring 1N HCl, sucking filtration is collected solid.Residue is dissolved in the chloroform, through dried over mgso.Except that after desolvating; residue is through silica gel column chromatography, with chloroform-methanol (10: 1, v/v) as eluant; obtain 4-[2-[1-[3-methoxyl group-4-(N '-phenyl urea groups) phenyl acetyl of 192mg (61%)]-the 2-pyrrolidinyl] acetenyl] ethyl benzoate, be faint yellow amorphous solid. 1H-NMR(CDCl 3)δ1.38(3H，t，J＝6.8Hz)，1.98-2.24(4H，m)，3.48-3.89(2H，m)，3.53(2H，s)，3.62(3H，s)，4.33-4.40(2H，m)，4.78-5.04(1H，m)，6.77-8.00(14H，m)。
To 4-[2-[1-[3-methoxyl group-4-(N '-phenyl urea groups) phenyl acetyl]-the 2-pyrrolidinyl] acetenyl] (184mg adds 0.25N sodium hydroxide (4ml) to ethyl benzoate in the agitating solution of THF 0.35mmol) (5ml).The mixture stirring that obtains is spent the night.In this mixture impouring water, make and be acid by adding 1N HCl (1ml).Sucking filtration is collected solid and is made it to be dissolved in the chloroform.Through this solution of dried over mgso and evaporation.Residue is recrystallization from chloroform-normal hexane, obtains 13 of 65mg (37%), is white crystalline powder.Mp 154-157 ℃; IR (KBr) 3346,2952,2615,1712,1693cm -1 1H-NMR (CDCl 3) δ 1.92-2.29 (4H, m), 3.32-3.82 (2H, m), 3.78 (2H, s), 3.80 (3H, s), 4.87-5.11 (1H, m), 6.77-9.26 (14H, m), 13.10 (1H, br s); MS (FAB) m/z 498 (M ++ 1) embodiment 104-[2-[1-[3-methoxyl group-4-[N '-(2-aminomethyl phenyl) urea groups] phenyl acetyl]-the 2-pyrrolidinyl] acetenyl] benzoic acid
Figure A0080951001581
To 3-methoxyl group-4-[N '-(2-aminomethyl phenyl) urea groups] phenylacetic acid (1.45g, 4.6mmol), 2-(2-(4-ethoxy carbonyl phenyl) acetenyl) pyrrolidine (1.12g, 4.6mmol), (562mg, 4.6mmol) mixture in DMF (20ml) stirs and to spend the night for EDC 1.32g (6.9mmol), DMAP.In this mixture impouring 1N HCl, sucking filtration is collected solid.This solid is dissolved in the chloroform, through dried over mgso.Except that after desolvating; residue is through silica gel column chromatography; with chloroform-methanol (100: 1; v/v) as eluant; obtain 4-[2-[1-[3-methoxyl group-4-[N '-(2-aminomethyl phenyl) urea groups of 2.20g (89%)] phenyl acetyl]-the 2-pyrrolidinyl] acetenyl] ethyl benzoate, be white amorphous solid. 1H-NMR(CDCl 3)δ1.37-1.41(3H，m)，1.94-2.22(4H，m)，2.29(3H，s)，3.41-3.89(2H，m)，3.62(3H，s)，3.69(2H，s)，4.34-4.40(2H，m)，4.72-5.01(1H，m)，6.33(1H，br?s)，6.80-8.06(12H，m)。
To 4-[2-[1-[3-methoxyl group-4-[N '-(2-aminomethyl phenyl) urea groups] phenyl acetyl]-the 2-pyrrolidinyl] acetenyl] ethyl benzoate (2.16g; add 0.25N sodium hydroxide (32ml) in the agitating solution of THF 4mmol) (30ml), and continue to stir and spend the night.In this mixture impouring water, by adding 1N HCl (8ml) acidify.Sucking filtration is collected the precipitation that produces and is made it to be dissolved in the chloroform.Through this solution of dried over mgso and evaporation.Residue is recrystallization from chloroform-normal hexane, obtains 14 of 555mg (27%), is white crystalline powder.Mp 161-164 ℃; IR (KBr) 3338,2954,2875,1707,1691cm -1 1H-NMR (CDCl 3) δ 1.96-2.10 (4H, m), 2.24 (3H, s), 3.32-3.81 (2H, m), 3.62 (2H, s), 3.81 (3H, s), 4.87-5.10 (1H, m), 6.76-8.58 (13H, m); MS (FAB) m/z 512 (M ++ 1) embodiment 113-[2-[1-[3-methoxyl group-4-[N '-(2-aminomethyl phenyl) urea groups] phenyl acetyl]-the 2-pyrrolidinyl] acetenyl] phenylacetic acid
With 3-methoxyl group-4-[N '-(2-aminomethyl phenyl) urea groups] phenylacetic acid (141mg, 0.45mmol), 2-[2-(3-ethoxy carbonyl aminomethyl phenyl) acetenyl] pyrrolidine (116mg, 0.45mmol), EDC (130mg, 0.68mmol), DMAP (55mg, 0.45mmol), the mixture of catalytic HOBt in DMF (10ml) stir and to spend the night.In this mixture impouring 1N HCl, sucking filtration is collected solid.This solid is dissolved in the chloroform, through dried over mgso and evaporation.Residue is through silica gel column chromatography, with chloroform-methanol (100: 1, v/v) as eluant, obtain a kind of oil, it is dissolved among the THF (5ml).Stir down, (4ml) adds in this solution with the 0.25N sodium hydroxide.After stirring is spent the night, in this mixture impouring 1N HCl (20ml).Sucking filtration is collected the precipitation that produces and is made it to be dissolved in the chloroform.Through this solution of dried over mgso and evaporation.Residue is through silica gel column chromatography, with chloroform-methanol (5: 1, v/v) as eluant, obtain 15 of 92mg (39%), be white amorphous solid. 1H-NMR (CDCl 3) δ 1.96-2.18 (7H, m), 3.50-3.88 (9H, m), 4.78-4.98 (2H, m), 6.72-7.99 (14H, m); MS (FAB) m/z526 (M ++ 1) embodiment 124-[1-[4-[N '-(2-aminomethyl phenyl) urea groups] phenyl acetyl]-2-pyrrolidinyl methoxyl group] isophthalic acid
To 4-[N '-(2-aminomethyl phenyl) urea groups] phenylacetic acid (2.32g, 5.15mmol), (S)-4-(2-pyrrolidinyl methoxyl group) dimethyl isophthalate (1.51g, 5.15mmol) the agitating solution of DMF (20ml) in add triethylamine (1.0ml, 6.65mmol), and mixture stirred spend the night.Dilute the mixture that obtains with ethyl acetate.With this solution of salt water washing, through dried over mgso and vacuum evaporation.Residue is through purification by silica gel column chromatography; with chloroform-methanol (10: 1; v/v), obtain 4-[1-[4-[N '-(2-aminomethyl phenyl) urea groups of 1.58g (55%) as eluant] phenyl acetyl]-2-pyrrolidinyl methoxyl group] dimethyl isophthalate, be the yellow crystal solid. 1H-NMR (CDCl 3) δ 1.87-2.25 (m, 7H altogether), 3.50-3.65 (m, 4H), 3.85 (s, 3H), 3.89 (s, 3H), 4.18-4.31 (m, 2H), 4.44 (m, 1H), 6.95-8.45 (m, 13H altogether).
To 4-[1-[4-[N '-(2-aminomethyl phenyl) urea groups] phenyl acetyl]-2-pyrrolidinyl methoxyl group] dimethyl isophthalate (1.56g; 2.79mmol) the agitating solution of THF (30ml) in add 0.25N sodium hydroxide (20ml), with reactant mixture heated overnight under refluxing.With among the mixture impouring 1N HCl that obtains and collect solid.With ether washing crude product solid, obtain 16 of 574mg (39%), be yellow amorphous solid.IR (KBr) 1710cm -1 1H-NMR (DMSO-d 6) δ 1.83-2.18 (m, 4H), 2.24 (s, 3H), 3.36-4.28 (m, 8H), 6.91-9.02 (a plurality of m, 13H altogether), 12.89 (br s, 1H); MS (FAB) m/z 532 (M ++ 1) embodiment 134-[2-[1-[3-methoxyl group-4-[N '-(2-aminomethyl phenyl) urea groups] phenyl acetyl]-the 2-pyrrolidinyl] vinyl] benzoic acid
Figure A0080951001611
In 0 ℃, with 15 fens clockwise 4-[2-[2-(N-tert-butoxycarbonyl) pyrrolidinyl vinyls] benzonitrile (2.26g, and dropping 1.5M hydrogenation diisopropyl aluminum (toluene solution) in the agitating solution of dichloromethane 7.57mmol) (23ml) (6.06ml, 9.09mmol).In 0 ℃ with the solution stirring that generates 3 hours.By adding this solution of saturated ammonium chloride quencher.By the mixture that diatomite filtration obtains, use ethyl acetate extraction filtrate.Use the saline wash filtrate,, obtain 4-[2-[2-(N-tert-butoxycarbonyl) pyrrolidinyl of 1.89g (83%) through dried over sodium sulfate and vacuum evaporation] vinyl] benzaldehyde, be yellow slurry.
In 0 ℃, (1.00g 25.1mmol) added silver nitrate (2.13g, acetonitrile 12.5mmol) (10ml) solution in the agitating solution in water (10ml) to sodium hydroxide with 0.5 hour.In 0 ℃, with dripping 4-[2-[2-(N-tert-butoxycarbonyl) pyrrolidinyl in 20 minutes in the stirred mixture upward] vinyl] benzaldehyde (1.89g, acetonitrile 6.27mmol) (10ml) solution.The mixture that stirring obtains under room temperature is other 3 hours then.This mixture of sucking filtration is used hot wash then.Behind the ethyl acetate wash filtrate, by careful adding 1N HCl acidify water-bearing layer, reuse chloroform extraction.Through dried over sodium sulfate extract and vacuum evaporation, obtain 0.700g 4-[2-[2-(N-tert-butoxycarbonyl) pyrrolidinyl in (two steps 35%)] vinyl] benzoic acid, be faint yellow crystalline material.
Under room temperature, to 4-[2-(2-(N-tert-butoxycarbonyl) pyrrolidinyl) vinyl] benzoic acid (0.700g, and methanol-benzene 2.21mmol) (1: 4, v/v, the TMSCHN of dropping 2M in agitating solution 30ml) 2Hexane solution (1.32ml, 2.65mmol).In stirring this solution under the room temperature after 0.5 hour, this solution of vacuum evaporation.The oily residue that obtains is through silica gel column chromatography, with ethyl acetate-normal hexane (1: 6,, obtain 4-[2-[2-(N-tert-butoxycarbonyl) pyrrolidinyl of 0.64g (88%) v/v) as eluant] vinyl] essence of Niobe, be faint yellow crystalline material.
Under room temperature, to 4-[2-[2-(N-tert-butoxycarbonyl) pyrrolidinyl] vinyl] (0.64g adds TFA (5ml) to essence of Niobe in the agitating solution of dichloromethane 1.93mmol) (5ml).In stirring this mixture under the room temperature after 1 hour, this mixture of vacuum evaporation.Handle this residue with saturated sodium bicarbonate, use dichloromethane extraction.Through dried over sodium sulfate extract and vacuum evaporation, obtain 4-[2-(2-pyrrolidinyl) vinyl of 0.45g (100%)] essence of Niobe, be faint yellow crystalline material.
Under room temperature, to 3-methoxyl group-4-[N '-(2-aminomethyl phenyl) urea groups] phenylacetic acid (285mg, 0.906mmol), 4-[2-(2-pyrrolidinyl) vinyl] essence of Niobe (210mg, 0.906mmol) the stirring the mixture of DMF (4ml) in add 1-ethyl-3-(3-dimethylaminopropyl) carbodiimide (EDC) (209mg, 1.09mmol), I-hydroxybenzotriazole (HOBt) (147mg, 1.09mmol) and 4-dimethylaminopyridine (DMAP) (11mg, 0.0906mmol).In stirring the mixture obtain under the room temperature after 48 hours, in this mixture impouring ice-1N HCl, use ethyl acetate extraction.Through dried over sodium sulfate extract and vacuum evaporation.Residue is through silica gel column chromatography; with acetone-toluene (1: 4-1: 1; v/v), obtain 4-[2-[1-[3-methoxyl group-4-[N '-(2-aminomethyl phenyl) urea groups of 0.47g (98%) as eluant] phenyl acetyl]-the 2-pyrrolidinyl] vinyl] essence of Niobe, be white crystalline material.
To 4-[2-[1-[3-methoxyl group-4-[N '-(2-aminomethyl phenyl) urea groups] phenyl acetyl]-the 2-pyrrolidinyl] vinyl] (0.47g adds 0.25N sodium hydroxide (5.36ml) to essence of Niobe in the agitating solution of THF 0.891mmol) (5ml).The mixture that stirring obtains under room temperature 20 hours.With this mixture of 1N HCl acidify.With this mixture of ethyl acetate extraction.With salt water washing extract, through dried over sodium sulfate and vacuum evaporation, obtain 17 of 430mg (94%), be white crystalline material. 1H-NMR (400MHz, DMSO-d 6) δ 1.78-2.13 (4H, m), 2.50 (3H, s), 3.44-3.68 (4H, m), 3.75 and 3.82 (3H, s), 4.71 (1H, m), 6.26-8.59 (15H, m).Embodiment 144-[2-[1-[3-methoxyl group-4-(N '-the phenyl urea groups) phenyl acetyl]-the 2-pyrrolidinyl] vinyl] benzoic acid
Figure A0080951001631
Under room temperature, to 3-methoxyl group-4-(N '-phenyl urea groups) phenylacetic acid (305mg, 1.01mmol), 4-[2-(2-pyrrolidinyl) vinyl] essence of Niobe (235mg, 1.01mmol) the agitating solution of DMF (4ml) in add 1-ethyl-3-(3-dimethylaminopropyl) carbodiimide (232mg, 1.21mmol), I-hydroxybenzotriazole (HOBt) (164mg, 1.21mmol) and 4-dimethylaminopyridine (DMAP) (12mg, 0.101mmol).Stir this mixture after 48 hours, by adding this mixture of 1N HCl acidify.Use the ethyl acetate extraction mixture.With salt water washing extract,, obtain a kind of oily residue through dried over sodium sulfate and vacuum evaporation.Make residue through silica gel column chromatography; with acetone-toluene (1: 4-1: 1; v/v), obtain 4-[2-[1-[3-methoxyl group-4-(N '-phenyl urea groups) phenyl acetyl of 0.43g (83%) as eluant]-the 2-pyrrolidinyl] vinyl] benzoic acid, be white crystalline material.
Under room temperature, to 4-[2-[1-[3-methoxyl group-4-(N '-phenyl urea groups) phenyl acetyl]-the 2-pyrrolidinyl] vinyl] (0.43g adds 0.25N sodium hydroxide (5.04ml) solution to essence of Niobe in the agitating solution of THF 0.837mmol) (5ml).The mixture that stirring obtains is after 20 hours, by careful this mixture of adding 1N HCl acidify.With this mixture of ethyl acetate extraction.With salt water washing extract, through dried over sodium sulfate and vacuum evaporation, obtain 18 of 397mg (95%), be white crystalline material. 1H-NMR(400MHz，DMSO-d 6)δ?1.78-2.13(4H，m)，3.17-3.68(4H，m)，3.74，3.82(3H，s)，4.71(1H，m)，6.27-9.28(16H，m)。Embodiment 154-[2-[1-[3-methoxyl group-4-[N '-(2-aminomethyl phenyl) urea groups] phenyl acetyl]-the 2-pyrrolidinyl] ethyl] benzoic acid
Figure A0080951001641
Under room temperature; with 4-[2-[1-[3-methoxyl group-4-[N '-(2-aminomethyl phenyl) urea groups] phenyl acetyl]-the 2-pyrrolidinyl] vinyl] benzoic acid (184mg, 0.358mmol) and the hydrogenation under 1 atmospheric pressure of the mixture of 5%Pd-C (368mg) in methanol.In stirring this mixture under the room temperature after 21 hours, remove by filter insoluble catalyst, vacuum evaporation filtrate.Residue is through silica gel column chromatography, with methanol-chloroform (1: 4-1: 3, v/v) as eluant, obtain 19 of 123mg (66%), be white crystalline material. 1H-NMR(DMSO-d 6)δ?1.55-2.03(m，6H)，2.24(s，3H)，2.60(m，2H)，3.17-3.59(m，4H)，3.83(s，3H)，3.97(m，1H)，6.61-8.57(m，13H)；MS(FAB)m/z?516(M ++1)。Embodiment 163-[1-[3-methoxyl group-4-[N '-(2-aminomethyl phenyl) urea groups] phenyl acetyl]-the 2-pyrrolidinyl] methylthio-benzoic acid
To-iodo phenol (20.0g adds 1 in the solution of DMF 90.9mmol) (200ml), 4-diazabicyclo [2,2,2] octane (20.4g, 181.8mmol) and diformazan sulfenyl carbamyl chloride (16.9g, 136.4mmol).In 35 ℃ the turbid solution that generates was stirred 0.5 hour, then in 75 ℃ of heating 0.5 hour.After the cooling, 300ml water is added in this mixture.Collect solid, wash with water and drying under reduced pressure, obtain between the O-of 27.63g (99%)-iodine substituted phenyl dimethyl sulfide aminocarbamic acid ester, be light yellow crystalline powder.IR (KBr) 1540,1463,1278,1193,1166,1124cm -1 1H-NMR (400MHz, CDCl 3) δ 3.33 (s, 3H), 3.45 (s, 3H), 7.05-7.14 (m, 2H), 7.43 (d, J=1.9Hz, 1H), 7.58 (dd, J=1.0,7.8Hz, 1H); MS (FAB) m/z 307 (M ++ 1); C 9H 10The analytical calculation value of INOS: C, 35.19; H, 3.28; N, 4.56.Measured value: C, 35.23; H, 3.40; N, 4.41.
With between O--iodine substituted phenyl dimethyl sulfide aminocarbamic acid ester (10.0g, Ph 32.6mmol) 2O (25ml) solution was in 230 ℃ of heating 10 hours.After the cooling, this reactant mixture is through silica gel column chromatography, with normal hexane-ethyl acetate (5: 1, v/v) as eluant, obtain between the S-of 9.31g (93%)-iodine substituted phenyl dimethyl sulfide aminocarbamic acid ester, be light yellow oil. 1H-NMR(400MHz，CDCl 3)δ3.08(br?s，6H)，7.11(t，J＝7.8Hz，1H)，7.46(d，J＝7.3Hz，1H)，7.71(d，J＝7.3Hz，1H)，7.85(s，1H)；MS(FAB)m/z?307(M ++1)
To between S--iodine substituted phenyl dimethyl sulfide aminocarbamic acid ester (5.01g, and the methanol solution of adding 28%-Feldalat NM in methanol 16.31mmol) (20ml) solution (3.46ml, 17.94mmol).The mixture that stirring obtains under room temperature 3.5 hours is then in 70 ℃ of heated overnight.After the cooling, add 1N HCl.Removal of solvent under reduced pressure, residue dilutes with ethyl acetate.Water, this solution of salt water washing are through dried over sodium sulfate.The concentrating under reduced pressure organic layer.Residue is through silica gel column chromatography, with normal hexane-ethyl acetate (10: 1, v/v) as eluant, obtain between 3.42g (89%)-the iodobenzene thiophenol, be a kind of oil. 1H-NMR(400MHz，CDCl 3)δ3.45(s，1H)，6.95(t，J＝7.8Hz，H)，7.23(d，J＝7.8Hz，1H)，7.48(d，J＝7.3Hz，1H)，7.64(t，J＝1.5Hz，1H)；MS(EI)m/z?236(M +)。
To N-(tert-butoxycarbonyl)-2-pyrrolidinyl methanol (4.30g, add in the agitating solution of pyridine 20.0mmol) (40ml) right-TsCl (5.72g, 30.0mmol).The mixture that stirring obtains under room temperature 3 hours.This reactant mixture of water quencher, revaporization removes and anhydrates.Dilute residue with ethyl acetate.With 1N HCl, salt water washing, through dried over sodium sulfate.Removal of solvent under reduced pressure, residue be through silica gel column chromatography, and (2: 1, v/v) as eluant, N-(the tert-butoxycarbonyl)-2-pyrrolidinyl methyl that obtains 5.76g (81%) was right-tosylate, is a kind of water white oil with normal hexane-ethyl acetate. 1H-NMR (400MHz, CDCl 3) δ 1.36 and 1.41 (respectively be s, be total to 9H), 1.82 (br m, 2H), 1.96 (br m, 2H), 2.44 (s, 3H), 3.30 (br m, 2H), 3.89 (br s, 1H), 3.96 (br s, 1H), 4.09 (br m, 1H), 7.34 (br s, 2H), 7.77 (d, J=8.3Hz, 2H); MS (FAB) m/z 356 (M ++ 1).
To-the iodobenzene thiophenol (2.67g, 11.31mmol) and N-(tert-butoxycarbonyl)-2-pyrrolidinyl methyl right-(3.34g adds 8N potassium hydroxide (1.77ml) to tosylate in the stirring the mixture of pyridine 9.43mmol) (9.4ml).Under room temperature, stir the mixture overnight that obtains.Use the ethyl acetate diluted reaction mixture.Water, saturated ammonium chloride solution, this solution of salt water washing are through dried over sodium sulfate.Concentrating under reduced pressure organic layer, residue be through silica gel column chromatography, with normal hexane-ethyl acetate (5: 1, v/v) as eluant, obtain [N-(tert-butoxycarbonyl)-2-pyrrolidinyl] methyl 3-iodine substituted phenyl sulfide of 1.79g (45%), be a kind of oil. 1H-NMR(400MHz，CDCl 3)δ1.45(s，9H)，1.78-2.01(br?m，4H)，2.71(dt，1H)，3.32-3.49(br?m，3H)，3.90-4.02(br?m，1H)，7.12(d，J＝7.8Hz，1H)，7.18(d，J＝7.8Hz，1H)，7.57(dd，J＝2.0，8.3Hz，2H)；MS(FAB)m/z?420(M ++1)。
To [1-(tert-butoxycarbonyl)-2-pyrrolidinyl] methyl 3-iodine substituted phenyl sulfide (1.76g, add in the agitating solution of DMSO 4.20mmol) (20ml) and methanol (16ml) triethylamine (1.28ml, 9.24mmol), Pd (OAc) 2(47.1mg, 0.21mmol) with 1, (86.6mg, 0.21mmol), bubbling fed CO gas 5 minutes to two (diphenylphosphino) propane of 3-then.In 70 ℃ the mixture stirring that obtains is spent the night.After the cooling, concentrate this mixture.Dilute residue with ethyl acetate.Use the salt water washing, through dried over sodium sulfate.Removal of solvent under reduced pressure, residue be through silica gel column chromatography, with normal hexane-ethyl acetate (5: 1,, obtain 3-[1-(the tert-butoxycarbonyl)-2-pyrrolidinyl of 1.28g (87%) v/v) as eluant] the methylthio-benzoic acid methyl ester, be a kind of oil. 1H-NMR (400MHz, CDCl 3) δ 1.42 and 1.45 (respectively be s, 9H), 1.79-2.05 (br m, 4H), 2.83 (dt, J=10.8,30.3Hz, 1H), 3.34-3.54 (br m, 3H), 3.92 (s, 3H), 3.92 and 4.05 (d, J=7.8Hz, 1H), 7.36 (t, J=7.8Hz, 1H), 7.63 (br d, J=14.7Hz, 1H), 7.83 (br d, J=12.7Hz, 1H), 8.04 (s, 1H); MS (FAB) m/z 352 (M ++ 1).
In 0 ℃ to 3-[1-(tert-butoxycarbonyl)-2-pyrrolidinyl] (1.46g adds TFA (15ml) to the methylthio-benzoic acid methyl ester in the agitating solution of dichloromethane 4.16mmol) (30ml).The mixture that stirring obtains under room temperature 1 hour.Removal of solvent under reduced pressure with 1N naoh treatment residue, is used chloroform extraction.With salt water washing extract, through dried over sodium sulfate and concentrating under reduced pressure, obtain 3-(2-pyrrolidinyl) the methylthio-benzoic acid methyl ester of 947mg (91%), be brown oil. 1H-NMR(400MHz，CDCl 3)δ?1.45-1.54(m，1H)，1.72-2.00(m，4H)，2.88-3.10(m，4H)，3.30(m，1H)，3.92(s，3H)，7.34(t，J＝7.8Hz，1H)，7.52(m，1H)，7.84(m，1H)，8.01(t，J＝2.0Hz，1H)；MS(FAB)m/z?252(M ++1)。
Under room temperature, with 3-methoxyl group-4-[N '-(2-aminomethyl phenyl) urea groups] phenylacetic acid (1.18g, 3.77mmol), EDC (1.08g, 5.65mmol), DMAP (23mg, 0.19mmol) and HOBt (25mg, 0.19mmol) mixture in DMF (5ml) stirred 1 hour, with 3-(2-pyrrolidinyl) methylthio-benzoic acid methyl ester (947mg, 3.77mmol) add in this mixture, the mixture that obtains is stirred spend the night.Add DMAP (460mg, 3.77mmol) and HOBt (835mg, 6.18mmol) after, restir 5 hours.Dilute this reactant mixture with ethyl acetate.With this solution of salt water washing, through dried over sodium sulfate.Removal of solvent under reduced pressure.Residue is through silica gel column chromatography; with normal hexane-ethyl acetate (2: 3; v/v), obtain 3-[1-[3-methoxyl group-4-[N '-(2-aminomethyl phenyl) urea groups of 294.3mg (14%) as eluant] phenyl acetyl]-the 2-pyrrolidinyl] the methylthio-benzoic acid methyl ester, be faint yellow amorphous substance.IR(KBr)2875，1724，1620，1284，1182cm -1； 1H-NMR(400MHz，CDCl 3)δ?1.86-2.05(m，4H)，2.31(s，3H)，2.84(dd，J＝9.3，13.7Hz，1H)，3.43-3.59(m，5H)，3.73(s，3H)，3.92(s，3H)，4.33(m，1H)，6.16(s，1H)，6.77-6.80(m，2H)，7.04(s，1H)，7.1?6(t，J＝8.3Hz，1H)，7.38(t，J＝7.8Hz，1H)，7.49(t，J＝7.8Hz，1H)，7.73(dt，J＝1.0，7.8Hz，1H)，7.79(dd，J＝2.0，6.8Hz，1H)，8.01(d，J＝2.0Hz，1H)，8.05(dd，J＝2.4，7.8Hz，1H)；MS(FAB)m/z?548(M ++1)。
To 3-[1-[3-methoxyl group-4-[N '-(2-aminomethyl phenyl) urea groups] phenyl acetyl]-the 2-pyrrolidinyl] methylthio-benzoic acid methyl ester (148.2mg; 0.271mmol) THF (7.4ml) and the agitating solution of water (1.8ml) in add Lithium hydrate (19.4mg; 0.812mmol), the mixture that stirring obtains under room temperature 9 hours.With 1N HCl treatment mixture, use chloroform extraction.With salt water washing extract, through dried over sodium sulfate.Removal of solvent under reduced pressure, residue be through preparation property TLC purification, with chloroform-methanol (10: 1, v/v) eluting, crystallization obtains 20 of 89.7mg (62%) from normal hexane-ethyl acetate, is white powder.IR(KBr)2960，1708cm -1； 1H-NMR(400MHz，DMSO-d 6)δ?1.82-2.01(m，4H)，2.24(s，3H)，2.93(dd，J＝9.3，12.7Hz，1H)，3.40-3.54(m，5H)，3.86(s，3H)，4.13(br?m，1H)，6.74(d，J＝8.3Hz，1H)，6.87(d，J＝1.5Hz，1H)，6.94(t，J＝7.8Hz，1H)，7.10-7.17(m，2H)，7.42(t，J＝7.8Hz，1H)，7.70(d，J＝7.8Hz，1H)，7.74(d，J＝8.3Hz，1H)，7.79(d，J＝7.8Hz，1H)，7.84(s，1H)，8.00(d，J＝8.3Hz，1H)，8.48(s，1H)，8.57(s，1H)；MS(FAB)m/z?534(M ++1)。Embodiment 173-[[1-[3-methoxyl group-4-[N '-(2-aminomethyl phenyl) urea groups] phenyl acetyl]-the 2-pyrrolidinyl] methyl sulphonyl] benzoic acid
In 0 ℃; to 3-[[1-[3-methoxyl group-4-[N '-(2-aminomethyl phenyl) urea groups] phenyl acetyl]-the 2-pyrrolidinyl] methyl mercapto] essence of Niobe (131.8mg; 0.241mmol) the agitating solution of dichloromethane (3ml) in add m-CPBA (130.5mg, 0.529mmol).Under room temperature, this reactant mixture was stirred 0.5 hour.With chloroform diluted mixture thing, with the quencher of saturated sodium thiosulfate solution.Separate organic layer, with saturated sodium bicarbonate solution, salt water washing, through dried over sodium sulfate.Removal of solvent under reduced pressure obtains 3-[[1-[3-methoxyl group-4-[N '-(2-aminomethyl phenyl) urea groups] phenyl acetyl]-the 2-pyrrolidinyl] methyl sulphonyl] essence of Niobe, be amorphous solid.(17.3mg 0.723mmol), continues to stir under room temperature to spend the night to add Lithium hydrate in the agitating solution of THF of this crude product chemical compound (7.4ml) and water (1.8ml).Use the chloroform diluted reaction mixture, use 1NHCl, use the salt water washing then, through dried over sodium sulfate.Removal of solvent under reduced pressure, residue be through preparation property TLC purification, with chloroform-methanol (10: 1, v/v) as eluant, make crude product solid recrystallization from normal hexane-ethyl acetate obtain 21 of 69.9mg (51%), be white crystalline powder.mp?243-245；IR(KBr)3354，2974，1533cm -1； 1H-NMR(400MHz，DMSO-d 6)δ?1.80-2.00(m，4H)，2.24(s，3H)，3.19-3.62(m，6H)，3.82(s，3H)，4.18(m，1H)，6.67(d，J＝8.8Hz，1H)，6.80(d，J＝1.0Hz，1H)，6.93(t，J＝7.3Hz，1H)，7.10-7.17(m，2H)，7.66(t，J＝7.8Hz，1H)，7.78(d，J＝8.3Hz，1H)，7.91-7.99(m，2H)，8.20(d，J＝7.3Hz，1H)，8.34(s，1H)，8.48(s，1H)，8.56(s，1H)；MS(FAB)m/z?566(M ++1)。C 29H 31N 3O 7S1HCl1H 2The analytical calculation value of O: C, 56.17; H, 5.53; N, 6.78.Measured value: C, 55.92; H, 5.58; N, 6.71.Embodiment 184-[[1-[3-methoxyl group-4-[N '-(2-aminomethyl phenyl) urea groups] phenyl acetyl]-the 2-pyrrolidinyl] methyl sulphonyl] benzoic acid
Figure A0080951001691
In 0 ℃; to 4-[[1-[3-methoxyl group-4-[N '-(2-aminomethyl phenyl) urea groups] phenyl acetyl]-the 2-pyrrolidinyl] methyl mercapto] essence of Niobe (300mg; 0.548mmol) the agitating solution of dichloromethane (6ml) in add m-CPBA (297mg; 1.206mmol), under room temperature, reactant mixture was stirred 1 hour.With chloroform diluted mixture thing, with the quencher of saturated sodium thiosulfate solution.With saturated sodium bicarbonate solution, the isolating organic layer of salt water washing and through dried over sodium sulfate.Removal of solvent under reduced pressure obtains 4-[[1-[3-methoxyl group-4-[N '-(2-aminomethyl phenyl) urea groups] phenyl acetyl]-the 2-pyrrolidinyl] methyl sulphonyl] essence of Niobe, be the crude product yellow oil.(39.4mg 1.643mmol), continues to stir under room temperature to spend the night to add Lithium hydrate in the agitating solution of THF of this crude product chemical compound (4.4ml) and water (1.1ml).Use the chloroform diluted reaction mixture, use 1NHCl, use the salt water washing then, through dried over sodium sulfate.Removal of solvent under reduced pressure, residue be through preparation property TLC purification, with chloroform-methanol (10: 1, v/v) eluting, crystallization obtains 22 of 128.0mg (41%) from normal hexane-ethyl acetate, is white powder.IR(KBr)3388，2974，1537，1298，1155cm -1； 1H-NMR(400MHz，DMSO-d 6)δ?1.80-1.98(m，4H)，2.24(s，3H)，2.54(s，1H)，3.20-3.70(m，5H)，3.82(s，3H)，4.16(br?m，1H)，6.67(dd，J＝1.5，8.3Hz，1H)，6.80(d，J＝1.5Hz，1H)，6.93(d，J＝7.3Hz，1H)，7.10-7.16(m，2H)，7.78(d，J＝7.3Hz，1H)，7.95(d，J＝8.3Hz，2H)，7.98(d，J＝8.3Hz，1H)，8.14(d，J＝8.3Hz，2H)，8.49(s，1H)，8.57(s，1H)；MS(FAB)m/z?566(M ++1)。C 29H 31N 3O 7S3H 2The analytical calculation value of O: C, 56.21; H, 6.02; N, 6.78.Measured value: C, 56.76; H, 5.37; N, 6.70.Embodiment 194-[[1-[3-methoxyl group-4-[N '-(2-aminomethyl phenyl) urea groups] phenyl acetyl]-the 2-pyrrolidinyl] methyl mercapto] benzoic acid
Figure A0080951001701
To right-iodo phenol (20.0g adds 1 in the agitating solution of DMF 90.9mmol) (200ml), 4-diazabicyclo [2,2,2] octane (20.4g, 181.8mmol) and diformazan sulfenyl carbamyl chloride (16.9g, 136.4mmol).In 75 ℃ with the solution stirring that generates 3.5 hours, after the cooling, add 300ml water.Sucking filtration is collected solid, and makes it to be dissolved in the ethyl acetate.Wash ethyl acetate layer with water, through dried over sodium sulfate and reduction vaporization.Crude product solid recrystallization from water, the O-that obtains 26.75g (96%) is right-iodine substituted phenyl dimethyl sulfide aminocarbamic acid ester, be light yellow crystalline powder.IR (KBr) 1479,1207,827cm -1 1H-NMR (400MHz, CDCl 3) δ 3.37 (s, 3H), 3.45 (s, 3H), 6.83 (d, J=8.8Hz, 2H), 7.69 (d, J=8.3Hz, 2H); MS (FAB) m/z 307 (M ++ 1); C 9H 10The analytical calculation value of INOS: C, 35.19; H, 3.28; N, 4.56.Measured value: C, 35.17; H, 3.35; N, 4.44.
With O-right-iodine substituted phenyl dimethyl sulfide aminocarbamic acid ester (10.0g, Ph 32.6mmol) 2O (25ml) agitating solution was in 230 ℃ of heating 5.5 hours.After the cooling, this reactant mixture is through silica gel column chromatography, and (3: 1, v/v) as eluant, the S-that obtains 2.55g (26%) was right-iodine substituted phenyl dimethyl sulfide aminocarbamic acid ester, is white crystalline powder with normal hexane-ethyl acetate.IR (KBr) 3299,1651,1469,1371cm -1 1H-NMR (400MHz, CDCl 3) δ 3.03 (br s, 3H), 3.08 (br s, 3H), 7.21 (d, J=8.3Hz, 2H), 7.70 (d, J=8.3Hz, 2H); MS (FAB) m/z308 (M ++ 1); C 9H 10The analytical calculation value of INOS: C, 35.19; H, 3.28; N, 4.56.Measured value: C, 35.49; H, 3.28; N, 4.43.
To S-right-(2.55g, (495mg 9.14mmol), stirs the mixture that obtains in 70 ℃ and to spend the night iodine substituted phenyl dimethyl sulfide aminocarbamic acid ester to add Feldalat NM in methanol 8.31mmol) (10ml) solution.After the cooling, add 1N HCl.This mixture of concentrating under reduced pressure, residue dilutes with ethyl acetate.Water, salt water washing are through dried over sodium sulfate.Concentrating under reduced pressure organic layer, residue be through silica gel column chromatography, with normal hexane-ethyl acetate (5: 1, v/v) as eluant, obtain 1.75g (89%) right-the iodobenzene thiophenol, be faint yellow crystalline solid.IR(KBr)2559，1097，1002，806cm -1； 1H-NMR(400MHz，CDCl 3)δ?3.43(s，1H)，7.10(d，J＝8.3Hz，2H)，7.53(d，J＝8.3Hz，2H)；MS(FAB)m/z?236(M ++1)。C 6H 5The analytical calculation value of IS: C, 30.53; H, 2.13.Measured value: C, 30.57; H, 2.15.
Under room temperature, to right-iodobenzene thiophenol (1.75g, 7.43mmol) and N-(tert-butoxycarbonyl)-2-pyrrolidinyl methyl p-toluenesulfonic esters (2.39g, 6.75mmol) the stirring the mixture of pyridine (12.7ml) in add 8N potassium hydroxide (1.27ml), under room temperature, stirred the mixture that obtains 4 hours.Use the ethyl acetate diluted reaction mixture.Water, saturated ammonium chloride, this solution of salt water washing are through dried over sodium sulfate.The concentrating under reduced pressure organic layer.Residue is through silica gel column chromatography, with normal hexane-ethyl acetate (5: 1, v/v) as eluant, obtain [N-(tert-butoxycarbonyl)-2-pyrrolidinyl] methyl 4-iodine substituted phenyl sulfide of 1.49g (53%), be a kind of light yellow oil. 1H-NMR(400MHz，CDCl 3)δ(s，9H)，1.78-2.01(br?m，4H)，2.71(dt，1H)，3.32-3.49(br?m，3H)，3.90-4.02(br?m，1H)，7.12(d，J＝7.8Hz，1H)，7.18(d，J＝7.8Hz，1H)，7.57(dd，J＝2.0，8.3Hz，2H)；MS(FAB)m/z?420(M ++1)。
To [1-(tert-butoxycarbonyl)-2-pyrrolidinyl] methyl 4-iodine substituted phenyl sulfide (1.49g, add in the agitating solution of DMSO 3.56mmol) (16ml) and methanol (13ml) triethylamine (1.09ml, 7.84mmol), Pd (OAc) 2(40mg, 0.178mmol) with 1, two (diphenylphosphino) propane of 3-(73.4mg, 0.178mmol).In the mixture that obtains that stirs, fed CO gas 5 minutes.In 70 ℃ the mixture stirring is spent the night.After the cooling, this mixture is concentrated into small size.Dilute residue with ethyl acetate.Use the salt water washing, through dried over sodium sulfate.Removal of solvent under reduced pressure, residue be through silica gel column chromatography, with normal hexane-ethyl acetate (5: 1,, obtain 4-[1-(the tert-butoxycarbonyl)-2-pyrrolidinyl of 1.16g (93%) v/v) as eluant] the methylthio-benzoic acid methyl ester, be a kind of oil. 1H-MR (400MHz, CDCl 3) δ 1.51 and 1.47 (respectively be s, 9H), 1.78-2.05 (br m, 4H), 2.77 (dt, J=10.8,37.1Hz, 1H), 3.34-3.58 (m, 3H), 3.89 (s, 3H), 4.03 (br d, J=27.3Hz, 1H), 7.38 (d, J=7.3Hz, 1H), 7.47 (d, J=7.3Hz, 1H), 7.92 (br s, 2H); MS (FAB) m/z 352 (M ++ 1).
To 4-[1-(tert-butoxycarbonyl)-2-pyrrolidinyl] (1.16g adds TFA (4ml) to the methylthio-benzoic acid methyl ester in the agitating solution of dichloromethane 3.32mmol) (20ml), stirred the mixture under room temperature 1.5 hours.Removal of solvent under reduced pressure is with 1N naoh treatment residue, with this mixture of chloroform extraction.With salt water washing extract, through the dry also concentrating under reduced pressure of potassium hydroxide, obtain 4-(2-pyrrolidinyl) the methylthio-benzoic acid methyl ester of 767mg (92%), be yellow oil. 1H-NMR(400MHz，CDCl 3)δ(dt，J＝3.9，12.7Hz，1H)，1.85-2.09(m，2H)，2.13(m，1H)，3.11-3.27(m，3H)，3.40(dd，J＝6.8，13.2Hz，1H)，3.54(dd，J＝7.3，15.1Hz，1H)，3.89(s，3H)，5.07(br，1H)，7.38(d，J＝8.3Hz，2H)，7.91(d，J＝8.3Hz，2H)；MS(FAB)m/z?252(M ++1)。
In 0 ℃, to 3-methoxyl group-4-[N '-(2-aminomethyl phenyl) urea groups] and phenylacetic acid (1.30g, 4.136mmol), (0.63ml 4.549mmol) adds the trifluoroacetic acid pentafluorophenyl esters to triethylamine in the stirring the mixture of DMF (20ml).Under room temperature, the mixture that obtains was stirred 1 hour.In this mixture impouring water (60ml), the sucking filtration collecting precipitation.With 0.1N HCl, water, normal hexane washing crude product solid,, obtain 3-methoxyl group-4-[N '-(2-aminomethyl phenyl) urea groups of 1.91g (96%) in 40 ℃ of dryings] the phenylacetic acid pentafluorophenyl esters, be light yellow crystalline powder.IR(KBr)1785，1224，1216cm -1； 1H-NMR(400MHz，CDCl 3)δ?2.29(s，3H)，3.76(s，3H)，3.90(s，2H)，6.49(s，1H)，6.81(d，J＝1.5Hz，1H)，6.91(dd，J＝1.5，8.3Hz，1H)，7.15(t，J＝7.3Hz，3H)，7.24(m，1H)，7.50(d，J＝7.8Hz，1H)，8.17(d，J＝7.8Hz，1H)；MS(FAB)m/z?481(M ++1)。C 30H 33N 3O 5S1/4H 2The analytical calculation value of O: C, 57.51; H, 3.57; N, 5.83.Measured value: C, 57.40; H, 3.75; N, 5.68.
Under room temperature, with 3-methoxyl group-4-[N '-(2-aminomethyl phenyl) urea groups] phenylacetic acid pentafluorophenyl esters (1.47g, 3.05mmol), 4-(2-pyrrolidinyl) methylthio-benzoic acid methyl ester (767mg, 3.05mmol), (0.51ml, 3.66mmol) mixture in DMF (15ml) stirs and to spend the night triethylamine.Use the ethyl acetate diluted reaction mixture.Use the salt water washing, through dried over sodium sulfate.Removal of solvent under reduced pressure; residue is through silica gel column chromatography; with normal hexane-ethyl acetate (1: 2; v/v) as eluant; obtain 4-[[1-[3-methoxyl group-4-[N '-(2-aminomethyl phenyl) urea groups of 1.366g (82%)] phenyl acetyl]-the 2-pyrrolidinyl] methyl mercapto] essence of Niobe, be white crystalline powder.IR(KBr)1785，1224，1216cm -1； 1H-NMR(400MHz，CDCl 3)δ?1.88-1.99(m，4H)，2.30(s，3H)，2.75(dd，J＝9.8，13.2Hz，1H)，3.43-3.55(m，3H)，3.56(s，2H)，3.64(dd，J＝1.1，14.2Hz，1H)，3.73(s，3H)，3.88(s，3H)，4.33(m，1H)，6.29(s，1H)，6.78-6.81(m，2H)，7.11-7.26(m，5H)，7.50(d，J＝8.3Hz，3H)，7.93(d，J＝8.8Hz，2H)，8.07(d，J＝7.8Hz，1H)；MS(FAB)m/z?548(M ++1)。C 30H 33N 3O 5S1/4H 2The analytical calculation value of O: C, 65.26; H, 6.12; N, 7.61.Measured value: C, 65.48 H, 6.20; N, 7.47.
To 4-[[1-[3-methoxyl group-4-[N '-(2-aminomethyl phenyl) urea groups] phenyl acetyl]-the 2-pyrrolidinyl] methyl mercapto] essence of Niobe (300mg; 0.548mmol) THF (5.5ml) and the agitating solution of water (1.1ml) in add Lithium hydrate (39.4mg; 1.643mmol); reactant mixture stirred under room temperature spend the night, stirred 9 hours in 50 ℃.Dilute this mixture with chloroform.With 1N HCl, this solution of salt water washing, through dried over sodium sulfate.Removal of solvent under reduced pressure makes the crude product solid recrystallization from normal hexane-ethyl acetate-methanol that obtains obtain 23 of 218.6mg (75%), is white crystalline powder.IR(KBr)3318，2952，1596，1536，1299，1155cm -1； 1H-NMR(400MHz，DMSO-d 6)δ?1.82-2.05(m，4H)，2.25(s，3H)，2.91(dd，J＝9.8，13.2Hz，1H)，3.47-3.52(m，3H)，3.57(s，2H)，3.87(s，3H)，4.14(br?m，1H)，6.76(d，J＝1.5，8.3Hz，1H)，6.89(d，J＝1.5Hz，1H)，6.94(t，J＝7.3Hz，1H)，7.11-7.19(m，2H)，7.57(d，J＝8.3Hz，2H)，7.80(d，J＝8.3Hz，1H)，7.83(d，J＝8.3Hz，2H)，8.02(d，J＝8.3Hz，1H)，8.49(s，1H)，8.58(s，1H)；MS(FAB)m/z?534(M ++1)。C 29H 31N 3O 5S5/4H 2The analytical calculation value of O: C, 62.63; H, 6.07; N, 7.36; S, 5.77.Measured value: C, 62.62; H, 5.74; N, 7.36; S, 5.67.Embodiment 204-[[1-[3-methoxyl group-4-[N '-(2-aminomethyl phenyl) urea groups] phenyl acetyl]-the 2-pyrrolidinyl] methylsulfinyl] benzoic acid
In 0 ℃; to 4-[[1-[3-methoxyl group-4-[N '-(2-aminomethyl phenyl) urea groups] phenyl acetyl]-the 2-pyrrolidinyl] methyl mercapto] essence of Niobe (264mg; 0.482mmol) the agitating solution of dichloromethane (5.2ml) in add m-CPBA (118.8mg; 0.482mmol), under room temperature, mixture was stirred 1 hour.With chloroform diluted mixture thing, with the quencher of saturated sodium thiosulfate solution.With saturated sodium bicarbonate, the isolating organic layer of salt water washing, through dried over sodium sulfate.Removal of solvent under reduced pressure obtains 4-[[1-[3-methoxyl group-4-[N '-(2-aminomethyl phenyl) urea groups] phenyl acetyl]-the 2-pyrrolidinyl] methylsulfinyl] essence of Niobe, be the crude product amorphous solid.(34.6mg 1.45mmol), continues to stir under room temperature to spend the night to add Lithium hydrate in the agitating solution of THF of this crude product chemical compound (4ml) and water (1ml).Dilute this mixture with chloroform, with 1N HCl, salt water washing, through dried over sodium sulfate.Removal of solvent under reduced pressure makes the crude product solid recrystallization from normal hexane-chloroform-methanol that obtains obtain 24 of 193.2mg (73%), is white crystalline powder.IR(KBr)3338，2956，1708，1529，1299，1207，1155cm -1； 1H-NMR(400MHz，DMSO-d 6)δ1.70-2.06(m，4H)，2.24(s，3H)，2.90(dd，J＝8.3，13.2Hz，1H)，3.02-3.08(m，1H)，3.16-3.25(m，1H)，3.41-3.60(m，3H)，3.84(s，3H)，4.40(br?s，1H)，6.74(d，J＝7.8Hz，1H)，6.87(s，1H)，6.94(d，J＝7.3Hz，1H)，7.11-7.17(m，2H)，7.75-7.81(m，3H)，7.98-8.05(m，1H)，8.10(d，J＝8.3Hz，2H)，8.46(s，1H)，8.56(s，1H)；MS(FAB)m/z?550(M ++1)，572(M ++Na)。C 29H 31N 3O 6S3/2H 2The analytical calculation value of O: C, 60.40; H, 5.94; N, 7.29.Measured value: C, 60.15; H, 5.82; N, 6.90.Embodiment 21 (S)-4-[1-[3-methoxyl group-4-[N '-(2-aminomethyl phenyl) urea groups] phenyl acetyl]-2-pyrrolidinyl methoxyl group] benzoic acid
To the 4-methyl hydroxybenzoate (1.96g, 12.88mmol), N-Boc-dried meat ammonia alcohol (2.59g, 12.87mmol) and triphenyl phasphine (4.06g, 15.48mmol) add in the agitating solution in THF (40ml) DIAD (3.10ml, 15.74mmol).The mixture that obtains was heated 14 hours under refluxing.This mixture of vacuum evaporation, residue is through purification by silica gel column chromatography, with normal hexane-ethyl acetate (6: 1, v/v) as eluant, obtain (S)-4-[1-(tert-butoxycarbonyl)-2-pyrrolidinyl methoxyl group of 3.34g (77%)] essence of Niobe, be a kind of oil. 1H-NMR(CDCl 3)δ1.48(s，9H)，1.67(d，J＝9.3Hz，1H)，1.87-2.03(m，3H)，3.36-3.43(m，2H)，3.87-4.09(m，1H)，4.13-4.20(m，2H)，6.94(d，J＝8.3Hz，2H)，7.98(d，J＝8.3Hz，2H)。
Under room temperature, with (S)-4-[1-(tert-butoxycarbonyl)-2-pyrrolidinyl methoxyl group] (3.34g, the mixture in TFA 9.96mmol) (20ml) and the dichloromethane (35ml) stirred 15 hours essence of Niobe.This mixture of vacuum concentration, with saturated sodium bicarbonate make be alkalescence.Use the chloroform extraction mixture, use the salt water washing, through the sodium carbonate drying.The evaporation organic layer obtains (S)-4-(2-pyrrolidinyl methoxyl group) essence of Niobe of 1.70g (73%), is yellow oil. 1H-NMR(CDCl 3)δ1.54-1.61(m，1H)，1.77-1.86(m，2H)，1.87-1.97(m，1H)，2.00(bs，1H)，2.93-3.06(m，2H)，3.52-3.57(m，1H)，3.88(s，3H)，3.90-3.99(m，2H)，6.92(d，J＝9.0Hz，2H)，7.98(d，J＝9.0Hz，2H)
With 3-methoxyl group-4-[N '-(2-aminomethyl phenyl) urea groups] phenylacetic acid (428mg, 1.36mmol), (S)-4-(2-pyrrolidinyl methoxyl group) essence of Niobe (330mg, 1.40mmol), EDC (312mg, 1.63mg), HOBt (220mg, 1.63mmol) and the DMAP of the catalytic amount mixture in DMF (15ml) stirred 6 hours.Dilute the mixture that obtains with ethyl acetate.With 0.5NHCl, saturated sodium bicarbonate, salt water washing, through dried over mgso.Solvent is fallen in vacuum evaporation, obtains the oily residue.With it through the silica gel column chromatography purification; with chloroform-methanol (50: 1; v/v), obtain (S)-4-[1-[3-methoxyl group-4-[N '-(2-aminomethyl phenyl) urea groups of 540mg (75%) as eluant] phenyl acetyl]-2-pyrrolidinyl methoxyl group] essence of Niobe, be grease. 1H-NMR (CDCl 3) δ 1.81-2.12 (m, 4H), 2.88 (bs, 3H), 3.48-3.61 (m, 7H altogether), 3.88 (s, 3H), 4.10-4.21 (m, 2H), 4.42-4.46 (m, 1H), 6.75-8.08 (a plurality of m, 13H altogether).
To (S)-4-[1-[3-methoxyl group-4-[N '-(2-aminomethyl phenyl) urea groups] phenyl acetyl]-2-pyrrolidinyl methoxyl group] (540mg adds 0.25N sodium hydroxide (10ml) to essence of Niobe in the agitating solution of THF 1.02mmol) (10ml).The mixture that obtains was heated 16 hours under refluxing.In this mixture impouring 1N HCl, collect solid.With ether washing crude product solid, obtain 25 of 278mg (53%), be white amorphous solid.IR (KBr) 1708cm -1 1H-NMR (DMSO-d 6) δ 1.83-2.14 (m, 4H), 2.21 (s, 3H), 2.46 (s, 2H), 3.78 (s, 3H), 3.95-4.02 (m, 1H), 4.13-4.16 (m, 1H), 4.24 (bs, 1H), 6.51-7.98 (a plurality of m, 12H), 8.43 (s, 1H), 8.53 (s, 1H), 12.57 (bs, 1H); MS (FAB) m/z 517 (M +).Embodiment 221-[1-[3-methoxyl group-4-[N '-(2-aminomethyl phenyl) urea groups] benzoyl]-the L-prolyl]-4-piperidyl acetic acid
Figure A0080951001761
Under room temperature; with 3-methoxyl group-4-nitrobenzoic acid (229mg; 1.16mmol), 4-(1-prolyl piperidyl) tert-butyl acetate (344mg; 1.16mmol), HOBt (188mg; 1.39mmol), DMAP (14.2mg; 0.116mmol) and EDC (267mg, 1.39mmol) mixture in DMF (7ml) stirred 22 hours.With ethyl acetate (50ml) diluted mixture thing, with 1N HCl, saturated sodium bicarbonate and water washing.Through dried over sodium sulfate organic layer and vacuum evaporation.Residue is used methanol through silica gel column chromatography: and chloroform (1: 30, v/v) as eluant, obtain 1-(3-methoxyl group-4-nitro benzoyl) prolyl-4-(piperidino) tert-butyl acetate of 520mg (94%), be white crystalline material. 1H-NMR (CDCl 3) δ 1.12-1.33 and 1.62-2.23 (respectively be m, 9H), 1.44 (s, 9H), 2.65,3.13,3.47,3.67,4.44 and 4.61 (respectively be m, 8H), 3.99 (s, 3H), 5.05 (m, 1H), 7.21 (d, J=8.3Hz, 1H), 7.31 (s, 1H), 7.86 (d, J=8.3Hz, 1H).
Under room temperature, with 1-(3-methoxyl group-4-nitro benzoyl) prolyl-4-(piperidino) tert-butyl acetate (0.52g, the hydrogenation 94 hours that stirs the mixture under 1 atmospheric pressure 1.09mmol) and in the methanol (10ml) of 5%Pd-C (2.08g).Remove insoluble catalyst, vacuum evaporation filtrate.Residue is through silica gel column chromatography, with methanol-chloroform (1: 40-1: 6, v/v) as eluant, obtain 1-(4-amino-3-anisoyl) prolyl-4-(piperidino) tert-butyl acetate of 279mg (57%), be white crystalline material. 1H-NMR (CDCl 3) δ 1.16-2.17,2.69,3.06,3.67,4.12 and 4.59 (respectively be m, 17H), 3.86 (s, 3H), 5.10 (m, 1H), 6.64 (m, 1H), 7.12 (respectively is m, 2H).
Under room temperature; to 1-(4-amino-3-anisoyl)-L-prolyl-4-(piperidino) tert-butyl acetate (279mg; 0.627mmol) and triethylamine (0.0876ml; 0.627mmol) the agitating solution of THF (4ml) in drip Carbimide. neighbour-tolyl ester (0.0777ml; 0.627mmol), with the mixture that obtains restir 21 hours under room temperature.Frozen water is added in this mixture the sucking filtration collecting precipitation.The crude product solid is through purification by silica gel column chromatography; use methanol: chloroform (1: 40; v/v), obtain 1-[1-[3-methoxyl group-4-[N '-(2-aminomethyl phenyl) urea groups of 254mg (70%) as eluant] benzoyl]-the L-prolyl]-4-(piperidino) tert-butyl acetate, be crystalline solid. 1H-NMR (CDCl 3) δ 1.43 (s, 9H), 1.13-1.25 and 1.76-2.14 (respectively are m, 9H), 2.60 3.18,3.71,4.06 and 4.57 (respectively is m, 8H), 3.67 (s, 3H), 5.06 (m, 1H), 6.63 and 6.90 (s, 2H), 6.98-7.23 and 7.64 (respectively is m, 5H), 7.56 (d, J=7.8Hz, 1H), 8.21 (d, J=8.8Hz, 1H).
Under room temperature, with 1[1-[3-methoxyl group-4-[N '-(2-aminomethyl phenyl) urea groups] benzoyl] prolyl]-4-(piperidino) tert-butyl acetate (254mg, dichloromethane 0.440mmol) (6ml) and TFA (6ml) solution stirring 5 hours.In this mixture impouring frozen water.Sucking filtration is collected solid, washes with water and air-dry, obtains 26 of 179mg (78%), is white crystalline solid. 1H-NMR (DMSO-d 6) δ 0.47,1.05,1.44 and 1.62-1.99 (respectively be m, 9H), 2.49 (s, 3H), 2.15-2.30,2.35,2.56,2.78,3.09,3.04-3.80,4.05,4.15 and 4.32 (respectively is m, 8H), 3.92 (s, 3H), 4.92 (m, 1H), 6.82,6.95,7.11,7.77,8.20,8.57 and 8.75 (m, 9H); MS (FAB) m/z 523 (m ++ 1); C 28H 34N 4O 6The analytical calculation value: C, 64.35; H, 6.56; N, 10.72.Measured value: C, 55.58; H, 5.89; N, 8.75.Embodiment 23 (S)-3-methoxyl group-4-[1-[3-methoxyl group-4-[N '-(2-aminomethyl phenyl) urea groups] phenyl acetyl]-2-pyrrolidinyl methoxyl group] benzoic acid
Figure A0080951001771
In 0 ℃, to the vanillic acid ethyl ester (3.00g, 15.29mmol), (S)-N-Boc-dried meat ammonia alcohol (3.08g, 15.30mmol), triphenyl phasphine (4.81g, 18.34mmol) add in the agitating solution in THF (50ml) DIAD (3.61ml, 18.33mmol).The mixture that obtains was heated 6.5 hours under refluxing.After being cooled to room temperature, evaporate this mixture, through purification by silica gel column chromatography, the usefulness chloroform-methanol (50: 1,, obtain (S)-3-methoxyl group-4-[1-(tert-butoxycarbonyl)-2-pyrrolidinyl methoxyl group v/v) as eluant] ethyl benzoate, be a kind of jelly.Making top (S)-3-methoxyl group-4-[1-(tert-butoxycarbonyl)-2-pyrrolidinyl methoxyl group] ethyl benzoate is dissolved among dichloromethane (50ml) and the TFA (45ml).Under room temperature, this mixture was stirred 2 days.The mixture that vacuum concentration obtains, with saturated sodium bicarbonate make be alkalescence.Use the dichloromethane extraction mixture, use the salt water washing, through dried over mgso.Evaporating solvent, residue be through purification by silica gel column chromatography, with chloroform-methanol (20: 1, v/v) as eluant, obtain 3.27g (S)-3-methoxyl group-4-(2-pyrrolidinyl methoxyl group) ethyl benzoate in (two steps 77%), be yellow oil. 1H-NMR(CDCl 3)δ?1.39(t，3H，J＝7.1Hz)，1.52-1.59(m，1H)，1.76-1.88(m，2H)，1.92-2.01(m，1H)，2.92-3.06(m，2H)，3.56-3.63(m，1H)，3.90(s，3H)，3.91-4.02(m，2H)，4.35(q，2H，J＝7.1Hz)，6.89(d，1H，J＝8.3Hz)，7.54?(d，1H，J＝2.0Hz)，7.65(dd，1H，J＝2.0，8.3Hz)。
To (S)-3-methoxyl group-4-(2-pyrrolidinyl methoxyl group) ethyl benzoate (424mg, 1.52mmol) the agitating solution of DMF (8ml) in add 3-methoxyl group-4-[N '-(2-aminomethyl phenyl) urea groups] pentafluorophenyl esters (728mg of phenylacetic acid, 1.52mmol) and triethylamine (0.26ml, 1.87mmol).Under room temperature, the mixture stirring that obtains is spent the night.Dilute this mixture with ethyl acetate, with 1N HCl, saturated sodium bicarbonate, salt water washing, through dried over mgso.Solvent is fallen in vacuum evaporation; residue is through purification by silica gel column chromatography; with chloroform-methanol (50: 1; v/v) as eluant; obtain (S)-3-methoxyl group-4-[1-[3-methoxyl group-4-[N '-(2-aminomethyl phenyl) urea groups of 830mg (95%)] phenyl acetyl]-2-pyrrolidinyl methoxyl group] ethyl benzoate, be amorphous solid. 1H-NMR(CDCl 3)δ?1.38(t，3H，J＝7.3Hz)，1.88-2.20(m，4H，m)，2.24(m，3H)，3.44-3.50(m，1H)，3.53-3.58(m，7H)，3.82(s，3H)，4.09-4.17(m，1H)，4.22-4.25(m，1H)，4.35(q，2H，J＝7.3Hz)，4.38-4.49(m，1H)，6.71-6.78(m，1H)，6.99(d，1H，J＝8.3Hz)，7.04-7.07(m，1H)，7.16-7.19(m，2H)，7.49-7.66(m，3H)，8.06(d，1H，J＝8.3Hz)。
To (S)-3-methoxyl group-4-[[1-[3-methoxyl group-4-[N '-(2-aminomethyl phenyl) urea groups] phenyl acetyl]-the 2-pyrrolidinyl] methoxyl group] (760mg adds 0.25N sodium hydroxide (10ml) to ethyl benzoate in the agitating solution of THF 1.32mmol) (10ml).With the mixture heated overnight under refluxing that obtains.After being cooled to room temperature, in this mixture impouring 1N HCl (100ml), collect solid.With ether washing crude product solid, obtain 27 of 429mg (59%), be yellow amorphous solid.mp?132-135；IR(KBr)1707?cm -1； 1H-NMR(DMSO-d 6)δ1.84-2.18(m，4H)，2.25(s，3H)，2.49-2.51(m，2H)，3.29-3.59(m，4H)，3.80(s，3H)，3.82(s，3H)，4.00-4.05(m，1H)，6.53-8.01(m，10H)，8.45(s，1H)，8.54(s，1H)，12.63(bs，1H)；MS(FAB)m/z?548(M ++1)。Embodiment 24 (S)-4-[1-[3-methoxyl group-4-[N '-(2-aminomethyl phenyl) urea groups] phenyl acetyl]-2-pyrrolidinyl methoxyl group] phthalic acid
Figure A0080951001791
In 0 ℃, to 4-hydroxyl phthalic dimethyl ester (3.00g, 14.27mmol), N-Boc-dried meat ammonia alcohol (2.87g, 14.26mmol), triphenyl phasphine (4.49g, 17.12mmol) add in the agitating solution in THF (50ml) DIAD (3.40ml, 17.27mmol).With the mixture heated overnight under refluxing that obtains.The mixture that evaporation obtains, residue is through purification by silica gel column chromatography, with normal hexane-ethyl acetate (3: 1, v/v) as eluant, obtain (S)-4-[1-(tert-butoxycarbonyl)-2-pyrrolidinyl methoxyl group of 5.75g (q.y.)] dimethyl phthalate, be a kind of oil. 1H-NMR(CDCl 3)δ?1.47(s，9H)，1.86-2.05(m，4H)，3.36-3.40(m，2H)，3.87(m，3H)，3.91(s，3H)，3.96-4.19(m，3H)，7.03-7.24(m，2H)，7.80(m，1H)。
To (S)-4-[1-(tert-butoxycarbonyl)-2-pyrrolidinyl methoxyl group] (5.75g adds TFA (20ml) to dimethyl phthalate in the solution of dichloromethane 14.62mmol) (25ml), under room temperature, the mixture that obtains was stirred 50 minutes.The mixture that vacuum concentration obtains, with saturated sodium bicarbonate make be alkalescence.Use the dichloromethane extraction mixture, use the salt water washing, through dried over mgso and vacuum evaporation.Residue is through purification by silica gel column chromatography, with chloroform-methanol (50: 1, v/v) as eluant, obtain (S)-4-(2-pyrrolidinyl methoxyl group) dimethyl phthalate of 790mg (18%), be a kind of brown oil. 1H-NMR (CDCl 3) δ 1.48-1.57 (m, 1H), 1.72-1.84 (m, 2H), 1.89-1.98 (m, 2H), 2.91-3.03 (m, 2H), 3.48-3.54 (m, 1H), 3.82-3.97 (m, 8H altogether), 6.98 (dd, 1H, J=2.4,8.8Hz), 7.06 (d, 1H, J=2.4Hz), 7.78 (d, 1H, J=8.8Hz).
To (S)-4-(2-pyrrolidinyl methoxyl group) dimethyl phthalate (212mg, 0.72mmol) the agitating solution of DMF (8ml) in add 3-methoxyl group-4-[N '-(2-aminomethyl phenyl) urea groups] pentafluorophenyl esters (346mg of phenylacetic acid, 0.72mmol) and triethylamine (120ml, 0.86mmol), the mixture stirring is spent the night.With the mixture that the ethyl acetate dilution obtains, use 1N HCl, saturated sodium bicarbonate, salt water washing, through dried over mgso.Solvent is fallen in vacuum evaporation, obtains (S)-4-[1-[3-methoxyl group-4-[N '-(2-aminomethyl phenyl) urea groups of 413mg (97%)] phenyl acetyl]-2-pyrrolidinyl methoxyl group] dimethyl phthalate, be a kind of oil. 1H-NMR (CDCl 3) δ 1.92-2.12 (m, 4H), 2.29 (br s, 3H), 3.51-3.64 (m, 7H), 3.87 (s, 3H), 3.89 (s, 3H), 4.10-4.19 (m, 2H), 4.44 (m, 1H), 6.73-8.02 (a plurality of m, 12H altogether).
Under room temperature; to (S)-4-[1-[3-methoxyl group-4-[N '-(2-aminomethyl phenyl) urea groups] phenyl acetyl]-2-pyrrolidinyl methoxyl group] dimethyl phthalate (413mg; 0.70mmol) the agitating solution of THF (10ml) in add 0.25N sodium hydroxide (10ml), then with the mixture heated overnight under refluxing that obtains.After being cooled to room temperature, in this reactant mixture impouring 1N HCl (100ml).Collect solid, wash with water also air-dry.With ether washing crude product solid, obtain 28 of 310mg (79%), be yellow amorphous solid.IR (KBr) 1701cm -1 1H-NMR (DMSO-d 6) δ 1.87-2.18 (m, 4H), 2.25 (s, 3H), 2.50 (s, 2H), 3.38-3.60 (m, 4H), 3.83 (s, 3H), 4.00-4.14 (m, 1H), 6.74-8.02 (a plurality of m, 10H), 8.46 (s, 1H), 8.54 (s, 1H); MS (FAB) m/z 562 (M ++ 1).Embodiment 253-chloro-4-[[1-[3-methoxyl group-4-[N '-(2-aminomethyl phenyl) urea groups] phenyl acetyl]-the 2-pyrrolidinyl] methoxyl group] benzoic acid Under room temperature, to 3-chloro-4-methyl hydroxybenzoate (600mg, 3.215mmol), N-tert-butoxycarbonyl dried meat ammonia alcohol (647.1mg, 3.215mmol) and triphenyl phasphine (1.01g, 3.858mmol) the agitating solution of THF (10ml) in drip diisopropyl azo-2-carboxylic acid (DIAD) (0.8ml, 3.890mmol), under room temperature, mixture was stirred 3 days, stirred 18 hours in 70 ℃.Vacuum evaporation reactant mixture, residue be through silica gel column chromatography, with normal hexane-ethyl acetate (5: 1, v/v) as eluant, obtain 3-chloro-[1-(the tert-butoxycarbonyl)-2-pyrrolidinyl] methoxyl methyl benzoate of 1.147g (97%), be a kind of oil. 1H-NMR (400MHz, CDCl 3) δ 1.46,1.48 (respectively be s, 9H), 1.59-1.63 (br, 1H), 1.88 (br s, 1H), 2.05 (s, 1H), 2.05-2.21 (m, 2H), 3.34-3.45 (br m, 1.5H), 3.89 (s, 3H), 3.97 (br m, 0.5H), 4.21 (br s, 2H), 7.05 (d, J=8.8Hz, 1H), 7.90 (dd, J=2.0,8.8Hz, 1H), 8.04 (d, J=2.0Hz, 1H); MS (FAB) m/z 370 (M ++ 1).
In 0 ℃, (1.14g adds TFA (5ml) in the agitating solution of dichloromethane 3.10mmol) (20ml), stirred reaction mixture is 2 hours under room temperature to 3-chloro-[1-(tert-butoxycarbonyl)-2-pyrrolidinyl] methoxyl methyl benzoate.Removal of solvent under reduced pressure is with 1N naoh treatment residue, with this mixture of chloroform extraction.With salt water washing extract, through the dry also concentrating under reduced pressure of potassium hydroxide, obtain 3-chloro-4-(2-pyrrolidinyl) methoxyl methyl benzoate of 741mg (89%), be yellow oil. 1H-NMR(400MHz，CDCl 3)δ?1.60-1.67(m，1H)，1.78-2.02(m，3H)，2.93-2.98(m，1H)，3.03-3.09(m，1H)，3.59(dt，J＝2.0，9.3Hz，1H)，3.89(s，3H)，3.98(dd，J＝6.3，8.8Hz，1H)，4.05(dd，J＝4.9，9.3Hz，1H)，6.93(d，J＝8.8Hz，1H)，7.90(dd，J＝2.0，8.8Hz，1H)，8.04(d，J＝2.0Hz，1H)；MS(FAB)m/z?270(M ++1)。
Under room temperature, with 3-methoxyl group-4-[N '-(2-aminomethyl phenyl) urea groups] phenylacetic acid pentafluorophenyl esters (500mg, 1.04mmol), 3-chloro-4-(2-pyrrolidinyl) methoxyl methyl benzoate (281mg, 1.04mmol), (0.17ml, 1.25mmol) mixture in DMF (5ml) stirred 1 hour triethylamine.With ethyl acetate diluted mixture thing, use the salt water washing, through dried over sodium sulfate.Removal of solvent under reduced pressure; residue is through silica gel column chromatography; with normal hexane-ethyl acetate (1: 3; v/v) as eluant; obtain 3-chloro-4-[[1-[3-methoxyl group-4-[N '-(2-aminomethyl phenyl) urea groups] phenyl acetyl]-the 2-pyrrolidinyl] methoxyl group] essence of Niobe (620mg; 1.04mmol), be white crystalline solid.(74.9mg 3.126mmol), stirs this mixture 2 days under room temperature to add Lithium hydrate in the agitating solution of THF of this chemical compound (8ml) and water (2ml).Dilute this mixture with chloroform, handle with 1N HCl.With this solution of salt water washing, through dried over sodium sulfate and vacuum evaporation.Make crude product solid recrystallization from normal hexane-ethyl acetate-chloroform obtain 29 of 561.2mg (98%), be white crystalline material.IR(KBr)1676，1599，1487，1267，758，754cm -1； 1H-NMR(400MHz，DMSO-d 6)δ?1.82-2.24(m，4H)，2.25(s，3H)，3.48-3.60(m，4H)，3.78(s，3H)，4.18(m，2H)，4.31(m，1H)，6.74(dd，J＝1.5，8.3Hz，1H)，6.84(d，J＝2.0Hz，1H)，6.91-6.95(m，1H)，7.11-7.17(m，3H)，7.79(dd，J＝2.0，8.3Hz，2H)，7.85(d，J＝2.0Hz，1H)，7.98(d，J＝8.3Hz，1H)，8.53(s，1H)，8.58(s，1H)；MS(FAB)m/z?552(M ++1)。Embodiment 263,5-two chloro-4-[[1-[3-methoxyl group-4-[N '-(2-aminomethyl phenyl) urea groups] phenyl acetyl]-the 2-pyrrolidinyl] methoxyl group] benzoic acid
Under room temperature, to 3,5-two chloro-4-methyl hydroxybenzoate (600mg, 2.714mmol), N-tert-butoxycarbonyl dried meat ammonia alcohol (546mg, 2.714mmol) and triphenyl phasphine (854mg drips DIAD (0.68ml in the agitating solution of THF 3.257mmol) (10ml), 3.283mmol), under room temperature, mixture was stirred 3 days, stirred 18 hours in 70 ℃.Concentrated reaction mixture, residue are used normal hexane-ethyl acetate (6: 1 through silica gel column chromatography, v/v) as eluant, obtain 4-[1-(the tert-butoxycarbonyl)-2-pyrrolidinyl of 988.8mg (90%)] methoxyl group-3, the 5-methyl p-dichlorobenzene is a kind of light yellow oil. 1H-NMR(400MHz，CDCl 3)δ?1.44(s，9H)，1.88-2.15(br?m，3H)，2.34(br?s，1H)，3.40-3.44(m，2H)，3.92(s，3H)，3.92，4.14(m，1H)，4.18(br?s，2H)，7.98(s，2H)；MS(FAB)m/z?404(M ++1)。
In 0 ℃, to 4-[1-(tert-butoxycarbonyl)-2-pyrrolidinyl] methoxyl group-3, (988mg adds TFA (5ml) to the 5-methyl p-dichlorobenzene in the agitating solution of dichloromethane 3.248mmol) (20ml), stirred reaction mixture is 2 hours under room temperature.Removal of solvent under reduced pressure is with 1N naoh treatment residue, with this solution of chloroform extraction.With salt water washing extract, through dried over sodium sulfate and concentrating under reduced pressure, obtain 3 of 672mg (68%), 5-two chloro-4-(2-pyrrolidinyl) methoxyl methyl benzoate is light yellow oil. 1H-NMR(400MHz，CDCl 3)δ?1.62-1.69(m，1H)，1.78-1.86(m，2H)，1.89-1.99(m，1H)，2.92-2.98(m，1H)，3.04-3.09(m，1H)，3.55-3.60(m，1H)，3.91(s，3H)，4.01(dd，J＝6.8，8.8Hz，1H)，4.08(dd，J＝4.9，8.8Hz，1H)，7.97(s，2H)；MS(FAB)m/z?304(M ++1)。
Under room temperature, with 3-methoxyl group-4-[N '-(2-aminomethyl phenyl) urea groups] phenylacetic acid pentafluorophenyl esters (385.8mg, 0.803mmol), 3,5-two chloro-4-(2-pyrrolidinyl) methoxyl methyl benzoate (244.3mg, 0.803mmol), (0.13ml, 0.964mmol) mixture in DMF (4ml) stirred 1 hour triethylamine.With ethyl acetate diluted mixture thing, use the salt water washing, through dried over sodium sulfate.Removal of solvent under reduced pressure, residue are used normal hexane-ethyl acetate (1: 2 through silica gel column chromatography; v/v) as eluant; obtain 3,5-two chloro-4-[[1-[3-methoxyl group-4-[N '-(2-aminomethyl phenyl) urea groups] phenyl acetyl]-the 2-pyrrolidinyl] methoxyl group] essence of Niobe, be grease.(57.7mg 2.409mmol), spends the night this mixture stirring under room temperature to add Lithium hydrate in the agitating solution of THF of this chemical compound (8ml) and water (2ml).This mixture of vacuum concentration dilutes residue with chloroform, with 1N HCl, this solution of salt water washing, through dried over sodium sulfate.Removal of solvent under reduced pressure makes the crude product solid recrystallization from normal hexane-methanol-chloroform that obtains obtain 30 of 428.2mg (91%), is white crystalline powder.IR(KBr)1618，1535，1454，1257，754cm -1； 1H-NMR(400MHz，DMSO-d 6)δ?1.83-2.24(m，4H)，2.24(s，3H)，3.50-3.58(m，4H)，3.84(s，3H)，3.98-4.05(m，1H)，4.15(dd，J＝2.9，8.7Hz，1H)，4.29(br?m，1H)，6.74(d，J＝8.3Hz，1H)，6.87(s，1H)，6.93(t，J＝7.3Hz，1H)，7.11(d，J＝7.8Hz，1H)，7.16(d，J＝8.3Hz，1H)，7.79(d，J＝8.3Hz，1H)，7.86(s，1H)，7.87(d，J＝9.8Hz，1H)，7.99(d，J＝8.3Hz，1H)，8.49(s，1H)，8.58(s，1H)；MS(FAB)m/z?586(M ++1)。Embodiment 274-[1-[3-methoxyl group-4-[N '-(2-aminomethyl phenyl) urea groups] phenyl acetyl]-2-pyrrolidinyl methoxyl group]-the 3-nitrobenzoic acid
Figure A0080951001841
Under room temperature, to 4-hydroxyl-3-nitrobenzoic acid (3.00g, methanol-benzene 0.0164mol) (1: 4, drip the TMSCHN of 2.0M in agitating solution v/v) 2Hexane solution (8.2ml, 0.0164mol).In the solution that stir to generate under the room temperature after 4 hours, this mixture of vacuum evaporation.The oily residue as eluant, obtains 4-hydroxyl-3-nitrobenzoic acid methyl ester of 4.23g (79%) with chloroform through silica gel column chromatography, is faint yellow crystalline material.
In 0 ℃, to N-tert-butoxycarbonyl dried meat ammonia alcohol (1.02g, 5.07mmol), 4-hydroxyl-3-nitrobenzoic acid methyl ester (1.00g, 5.07mmol) and triphenyl phasphine (1.46g, 5.58mmol) the stirring the mixture of THF (10ml) in drip diisopropyl azo-2-carboxylic acid (DIAD) (95%) (1.16ml, 5.58mmol).The mixture that obtains was heated 46 hours under refluxing.After the cooling, the vacuum evaporation mixture.Residue is dissolved in the dichloromethane (10ml), adds TFA (10ml).In stirring this solution under the room temperature after 0.5 hour, this solution of vacuum evaporation.Water added in the residue and with ethyl acetate wash.By adding in the saturated sodium bicarbonate and the water-bearing layer, use ethyl acetate extraction.Through the dried over sodium sulfate extract, vacuum evaporation obtains 3-nitro-4-(2-pyrrolidinyl methoxyl group) essence of Niobe of 0.698g (49%), is a kind of jelly.
Under room temperature, with 3-nitro-4-(2-pyrrolidinyl methoxyl group) essence of Niobe (0.668g, 2.38mmol), 3-methoxyl group-4-[N '-(2-aminomethyl phenyl) urea groups] phenylacetic acid (1.12g, 3.57mmol), I-hydroxybenzotriazole (HOBt) (0.482g, 3.57mmol), 4-dimethylaminopyridine (DMAP) (43.6mg, 0.357mmol) and 1-ethyl-3-(3-dimethylaminopropyl) carbodiimide (EDC) (0.684g, 3.57mmol) mixture in DMF (10ml) stirred 15 hours.Ethyl acetate is added in this mixture, wash in proper order with 1N HCl, saturated sodium bicarbonate and saline.Through dried over sodium sulfate organic layer and vacuum evaporation.Residue is through silica gel column chromatography; with ethanol-chloroform (1: 20; v/v), obtain 4-[1-[3-methoxyl group-4-[N '-(2-aminomethyl phenyl) urea groups of 0.927g (68%) as eluant] phenyl acetyl]-2-pyrrolidinyl methoxyl group]-3-nitrobenzoic acid methyl ester, be the yellow crystal material.
With 4-[1-[3-methoxyl group-4-[N '-(2-aminomethyl phenyl) urea groups] phenyl acetyl]-2-pyrrolidinyl methoxyl group]-3-nitrobenzoic acid methyl ester (0.917g; 1.59mmol) in THF (10ml) and 1N sodium hydroxide (2.38ml, the 2.38mmol) mixture in heating 2 hours under refluxing.After the cooling, in this mixture impouring frozen water, use ethyl acetate extraction.With salt water washing extract, through dried over sodium sulfate and vacuum evaporation, obtain 31 of 0.826g (92%), be the yellow crystal solid. 1H-NMR (400MHz, CDCl 3) δ 1.91,2.09 (1H, 3H respectively are m), 2.28 (3H, s), 3.54-3.62 (4H, m), 3.64 (3H, s), 4.15,4.59 (respectively being 1H, respectively is d, J=7.8Hz), 4.46 (1H, m), 6.66,7.22 (respectively being 1H, they respectively is s), 6.72 (1H, d, J=8.3Hz), 7.11-7.28 (4H, m), 7.46 (1H, d, J=7.8Hz), 7.74 (1H, d, J=7.8Hz), 7.85 (1H, s), 8.17 (1H, dd, J=2.0,8.8Hz), 8.48 (1H, d, J=2.4Hz); MS (FAB) m/z 563 (M ++ 1).Embodiment 283-amino-4-[1-[3-methoxyl group-4-[N '-(2-aminomethyl phenyl) urea groups] phenyl acetyl]-2-pyrrolidinyl methoxyl group] benzoic acid
Figure A0080951001851
With 4-[1-[3-methoxyl group-4-[N '-(2-aminomethyl phenyl) urea groups] phenyl acetyl]-2-pyrrolidinyl methoxyl group]-3-nitrobenzoic acid (101mg, the hydrogenation 48 hours that stirs the mixture under 1 atmospheric pressure 0.190mmol) and in the methanol of 5%Pd-C (0.247g).Remove insoluble catalyst, vacuum evaporation filtrate.Residue is through silica gel column chromatography, with ethanol-chloroform (1: 1, v/v) as eluant, obtain 32 of 61.0mg (60%), be crystalline material. 1H-NMR (400MHz, DMSO-d 6) δ 1.95 (4H, m), 2.23 (3H, s), 3.60,3.91,4.10,4.34 (5H respectively is m), 3.81 (3H, s), 4.88 (2H, m), 6.74 (1H, d, J=8.3Hz), and 6.86-7.28 (5H, m), 7.78 (1H, d, J=7.8Hz), 7.99 (1H, d, J=8.3Hz), 8.30 (1H, s), 8.45,8.55 (respectively being 1H, respectively is s); MS (FAB) m/z 533 (M ++ 1).Embodiment 294-[2-[1-[4-[N '-(2-fluoro phenyl) urea groups]-3-methoxyphenyl acetyl group]-the 2-pyrrolidinyl] acetenyl] benzoic acid
Figure A0080951001861
To 4-amino-3-methoxyphenyl benzyl acetate (1.36g, add in the agitating solution of THF 5mmol) (20ml) Carbimide. 2-fluoro phenyl ester (561 μ l, 5mmol) and the triethylamine of catalytic amount.The mixture that stirring obtains 3 hours.By adding this mixture of entry (10ml) quencher, use ethyl acetate extraction.With salt water washing extract, through dried over mgso and evaporation, residue as eluant, obtains 4-[N '-(2-fluoro phenyl) urea groups of 2.06g (q.y.) with chloroform through silica gel column chromatography]-3-methoxyphenyl benzyl acetate, be green grease. 1H-NMR(CDCl 3)δ?3.63(2H，s)，3.82(3H，s)，5.14(2H，s)，6.79-7.37(12H，m)，8.01(1H，d，J＝7.8Hz)，8.09-8.14(1H，m)。
To 4-[N '-(2-fluoro phenyl) urea groups]-(2.04g adds 0.25N sodium hydroxide (40ml) to 3-methoxyphenyl benzyl acetate in the agitating solution of THF 5mmol) (40ml).The mixture stirring that obtains is spent the night.In this mixture impouring 1N HCl (10ml), sucking filtration is collected the precipitation that produces.Make residue recrystallization from chloroform-ethanol, obtain 4-[N '-(2-fluoro phenyl) urea groups of 1.04g (66%)]-3-methoxybenzene guanidine-acetic acid, be white crystalline powder.Mp 185-188 (d); 1H-NMR (DMSO-d 6) δ 3.50 (2H, s), 3.82 (3H, s), 6.78 (1H, dd, J=1.4 and 8.3Hz), 6.92 (1H, d, J=1.4Hz), and 6.95-7.01 (1H, m), 7.10-7.14 (1H, m), and 7.19-7.24 (1H, m), 8.01 (1H, d, J=8.3Hz), 8.14-8.18 (1H, m), 8.72 (1H, s), 9.17 (1H, s); MS (FAB) m/z 319 (M ++ 1).C 16H 15N 2O 4The analytical calculation value of F: C, 60.37; H, 4.75; N, 8.80.Measured value: C, 60.20; H, 4.82; N, 8.67.
With 4-[N '-(2-fluoro phenyl) urea groups]-3-methoxybenzene guanidine-acetic acid (255mg, 0.8mmol), 2-[2-(4-ethoxy carbonyl phenyl) acetenyl) pyrrolidine (195mg, 0.8mmol), EDC (230mg, 1.2mmol), (98mg, 0.8mmol) mixture in DMF (20ml) stirs and to spend the night DMAP.In this reactant mixture impouring 1N HCl, sucking filtration is collected the precipitation that produces, and it is dissolved in the chloroform.Through this solution of dried over mgso and evaporation.Residue is through silica gel column chromatography, with chloroform-methanol (100: 1, v/v) as eluant, obtain required compound, it is dissolved among the THF (8ml).0.25N sodium hydroxide (8ml) is added in this solution, stir the mixture overnight that obtains.In this mixture impouring 1N HCl, use chloroform extraction.With salt water washing extract, through dried over mgso and evaporation.Make residue recrystallization from chloroform-normal hexane, obtain 33 of 144mg (37%), be light yellow crystalline powder.Mp 152-155 (d); 1H-NMR (DMSO-d 6) δ 1.92-2.27 (4H, m), 2.50 (2H, s), 3.33-3.78 (2H, m), 3.80 and 3.82 (3H respectively is s altogether), 4.88-5.12 (1H, m), and 6.77-7.24 and 7.99-8.20 (be total to 7H, m), 7.48 and 7.52 (2H, d, J=8.3Hz, separately), 7.91 (2H, d, J=8.3Hz), 8.72 (1H, s), 9.18 (1H, s), 13.11 (1H, br-s); MS (FAB) m/z 516 (M ++ 1).C 29H 26N 3O 5F2H 2The analytical calculation value of O: C, 63.15; H, 5.48; N, 7.62.Measured value: C, 63.58; H, 5.15; N, 7.22.Embodiment 304-[[1-[3-methoxyl group-4-[N '-(2-aminomethyl phenyl) urea groups] phenyl acetyl]-the 2-pyrrolidinyl] methoxyl group]-the 3-ar-Toluic acid
Under room temperature, to 4-iodo-2-methylphenol (465mg, 1.987mmol), N-tert-butoxycarbonyl dried meat ammonia alcohol (400mg, 1.987mmol) and triphenyl phasphine (625mg, 2.384mmol) the agitating solution of THF (7ml) in drip DIAD (0.5ml, 2.404mmol), in 70 ℃, mixture was stirred 13 hours.The vacuum concentration reactant mixture, residue is through silica gel column chromatography, with normal hexane-ethyl acetate (9: 1, v/v) as eluant, obtain 4-[1-(the tert-butoxycarbonyl)-2-pyrrolidinyl of 645.3mg (78%)] methoxyl group-1-iodo-3-methylbenzene, be a kind of light yellow oil. 1H-NMR (400MHz, CDCl 3) δ 1.47 (s, 9H), 1.83-1.89 (m, 1H), 1.96-2.04 (m, 3H), 2.16 (s, 3H), 3.37-3.43 (br m, 2H), 3.81,3.94 (respectively be br m, 1H), 4.08-4.18 (m, 2H), 6.62 (br s, 1H), 7.42 (s, 2H); MS (FAB) m/z 418 (M ++ 1).
To 4-[1-(tert-butoxycarbonyl)-2-pyrrolidinyl] methoxyl group-1-iodo-3-methylbenzene (645.3mg, add in the agitating solution of DMSO 1.546mmol) (7ml) and methanol (6ml) triethylamine (0.47ml, 3.401mmol), Pd (OAc) 2(17.4mg, 0.077mmol) with 1, two (diphenylphosphino) propane of 3-(31.46mg, 0.077mmol).In stirred mixture, fed CO gas 10 minutes.In 70 ℃ mixture was stirred 2 days and concentrated.Dilute residue with ethyl acetate, use the salt water washing, through dried over sodium sulfate.Removal of solvent under reduced pressure, residue be through silica gel column chromatography, with normal hexane-ethyl acetate (5: 1,, obtain 4-[1-(the tert-butoxycarbonyl)-2-pyrrolidinyl of 301.6mg (56%) v/v) as eluant] methoxyl group-3-methyl toluate, be a kind of oil. 1H-NMR (400MHz, CDCl 3) δ 1.47 (s, 9H), 1.86-2.10 (br m, 4H), 2.33 (s, 3H), 3.32-3.50 (br m, 2H), 3.88 (s, 3H), 3.88,4.04 (respectively be br m, 1H), 4.13-4.20 (m, 2H), 6.89 (br m, 1H), 7.82 (s, 1H), 7.85 (dd, J=2.0,8.8Hz, 1H); MS (FAB) m/z350 (M ++ 1).
In 0 ℃, to 4-[1-(tert-butoxycarbonyl)-2-pyrrolidinyl] (301.6mg adds TFA (1.2ml) to methoxyl group-3-methyl toluate in the agitating solution of dichloromethane 0.863mmol) (6ml), stirred the mixture under room temperature 1 hour.Removal of solvent under reduced pressure makes residue be alkalescence by adding the 1N sodium hydroxide.With this mixture of chloroform extraction.With salt water washing extract, through dried over sodium sulfate and concentrating under reduced pressure, obtain 3-methyl-4-(2-pyrrolidinyl) methoxyl methyl benzoate of 192.5mg (90%), be a kind of oil. 1H-NMR(400MHz，CDCl 3)δ?1.58-1.65(m，1H)，1.78-2.00(m，3H)，2.24(s，3H)，2.97(dt，J＝6.8，10.2Hz，1H)，3.05(dt，J＝5.9，6.8Hz，1H)，3.54-3.58(m，1H)，3.87(s，3H)，3.92(dd，J＝6.3，9.3Hz，1H)，3.99(dd，J＝4.9，9.3Hz，1H)，6.81(d，J＝8.3Hz，1H)，7.83(s，1H)，7.85(dd，J＝2.0，8.3Hz，1H)；MS(FAB)m/z?250(M ++1)。
Under room temperature, with 3-methoxyl group-4-[N '-(2-aminomethyl phenyl) urea groups] phenylacetic acid pentafluorophenyl esters (211.3mg, 0.44mmol), 3-methyl-4-(2-pyrrolidinyl) methoxyl methyl benzoate (109.7mg, 0.44mmol), (73.6ul, 0.528mmol) mixture in DMF (2ml) stirred 1.5 hours triethylamine.Use the ethyl acetate diluted reaction mixture, with this solution of salt water washing, through dried over sodium sulfate.Removal of solvent under reduced pressure; residue is through silica gel column chromatography; with normal hexane-ethyl acetate (1: 3; v/v) as eluant; obtain 4-[[1-[3-methoxyl group-4-[N '-(2-aminomethyl phenyl) urea groups] phenyl acetyl]-the 2-pyrrolidinyl] methoxyl group]-3-methyl toluate (241.6mg; q.y.), be a kind of oil.(32mg 1.32mmol), spends the night this reactant mixture stirring under room temperature to add Lithium hydrate in the agitating solution of THF of this chemical compound (4.4ml) and water (1.1ml).With chloroform diluted mixture thing, by adding 1N HCl acidify.With this solution of salt water washing, through dried over sodium sulfate.Removal of solvent under reduced pressure makes the crude product solid recrystallization from normal hexane-ethyl acetate-chloroform-methanol that obtains obtain 34 of 126.3mg (54%), is white crystalline powder.IR(KBr)1685，1606，1454，1257，752cm -1； 1H-NMR(400MHz，DMSO-d 6)δ?1.87-2.10(m，4H)，2.12(s，3H)，2.25(s，3H)，3.51-3.71(m，4H)，3.76(s，3H)，4.08-4.18(m，2H)，4.34(m，1H)，6.74(dd，J＝1.5，9.8Hz，1H)，6.84(d，J＝1.5Hz，1H)，6.94(t，J＝6.8Hz，1H)，7.06(d，J＝8.8Hz，1H)，7.12(d，J＝7.8Hz，1H)，7.16(d，J＝7.8Hz，1H)，7.72(s，1H)，7.76(dd，J＝2.0，8.3Hz，1H)，7.79(d，J＝7.8Hz，1H)，7.99(d，J＝8.3Hz，1H)，8.46(s，1H)，8.54(s，1H)；MS(FAB)m/z?532(M ++1)。C 30H 33N 3O 61/2H 2The analytical calculation value of O: C, 66.65; H, 6.34; N, 7.77.Measured value: C, 66.16; H, 6.37; N, 7.50.Embodiment 31 (S)-4-[1-[3-methoxyl group-4-[N '-(2-aminomethyl phenyl) urea groups] phenyl acetyl]-2-pyrrolidinyl methoxyl group] isophthalic acid
(1.52g adds saturated sodium bicarbonate in the solution of methanol 6.03mmol) (70ml), under room temperature, the mixture that obtains is stirred 3 hours to 4-acetoxyl group dimethyl isophthalate.In the mixture impouring 1N HCl that obtains, use ethyl acetate extraction.With saturated sodium bicarbonate, salt water washing extract, through dried over sodium sulfate.Evaporating solvent obtains the 4-hydroxyisophthalic acid dimethyl ester of 1.27g (q.y.), is white crystalline powder. 1H-NMR(CDCl 3)δ?3.91(s，3H)，3.99(s，3H)，7.01(d，1H，J＝8.8Hz)，8.11(dd，1H，J＝2.4，8.8Hz)，8.55(d，1H，J＝2.4Hz)
Under room temperature, to 4-hydroxyisophthalic acid dimethyl ester (1.27g, 6.04mmol), (S)-N-Boc-dried meat ammonia alcohol (1.22g, 6.06mmol), triphenyl phasphine (1.90g, 7.24 add in the agitating solution of THF mmol) (30ml) DIAD (1.43ml, 7.26mmol).The stirred mixture that obtains was heated 15 hours under refluxing.After being cooled to room temperature, the mixture that evaporation obtains, residue is through purification by silica gel column chromatography, with normal hexane-ethyl acetate (3: 1, v/v) as eluant, obtain (S)-4-[1-(tert-butoxycarbonyl)-2-pyrrolidinyl methoxyl group of 2.10g (88%)] dimethyl isophthalate, be a kind of yellow oil. 1H-NMR(CDCl 3)δ?1.26(s，9H)，1.85-2.16(m，3H)，3.36-3.46(m，2H)，3.90(s，6H)，4.11-4.31(m，2H)，4.95-5.02(m，2H)，7.09(dd，1H，J＝9.3，24.9Hz)，8.11-8.14(m，1H)，8.46(d，1H，J＝9.3Hz)
Under room temperature, with (S)-4-[1-(tert-butoxycarbonyl)-2-pyrrolidinyl methoxyl group] and dimethyl isophthalate (2.01g, 5.11mmol), the mixture of TFA (20ml) and dichloromethane (25ml) stirred 1.5 hours.The mixture that vacuum concentration obtains, with saturated sodium bicarbonate make be alkalescence.Use the dichloromethane extraction mixture, use the salt water washing, through dried over sodium sulfate and evaporation.Residue is through purification by silica gel column chromatography, with chloroform-methanol (9: 1, v/v) as eluant, obtain (S)-4-(2-pyrrolidinyl methoxyl group) dimethyl isophthalate of 0.80g (53%), be a kind of yellow oil. 1H-NMR(CDCl 3)δ?1.71(m，1H)，1.89(m，2H)，2.00(m，1H)，3.05-3.13(m，2H)，3.67(m，1H)，3.90(s，3H)，3.91(s，3H)，4.05-4.18(m，2H)，7.00(d，1H，J＝8.8Hz)，8.14(dd，1H，J＝2.4，8.8Hz)，8.50(d，1H，J＝2.4Hz)。
To (S)-4-(2-pyrrolidinyl methoxyl group) dimethyl isophthalate (616mg, 2.10mmol) the agitating solution of DMF (13ml) in add 3-methoxyl group-4-[N '-(2-aminomethyl phenyl) urea groups] five fluoroester (1.00g of phenylacetic acid, 2.08mmol) and triethylamine (425 μ l, 3.12mmol), the mixture that obtains was stirred under room temperature 3.5 hours.With the mixture that the ethyl acetate dilution obtains, use 1N HCl, saturated sodium bicarbonate, salt water washing, through dried over sodium sulfate.Except that after desolvating; residue is through purification by silica gel column chromatography; with chloroform-methanol (50: 1; v/v) as eluant; obtain (S)-4-[1-[3-methoxyl group-4-[N '-(2-aminomethyl phenyl) urea groups of 1.41g (q.y.)] phenyl acetyl]-2-pyrrolidinyl methoxyl group] dimethyl isophthalate, be a kind of yellow oil. 1H-NMR (CDCl 3) δ 1.86-2.29 (m, 4H), 2.30 (s, 3H), 3.47-3.57 (m, 2H), 3.58 (s, 3H), 3.59 (s, 2H), 3.83 (s, 3H), 3.91 (s, 3H), 4.22-4.37 (m, 2H), 4.42-4.47 (m, 1H), 6.44-8.46 (a plurality of m, 12H).
To (S)-4-[1-[3-methoxyl group-4-[N '-(2-aminomethyl phenyl) urea groups] phenyl acetyl]-2-pyrrolidinyl methoxyl group] dimethyl isophthalate (1.41g; 2.39mmol) the agitating solution of THF (20ml) in add 0.25N sodium hydroxide (20ml), with the mixture heated overnight under refluxing that obtains.After being cooled to room temperature, in this mixture impouring 1N HCl (150ml), collect solid.Make crude product solid recrystallization from chloroform-methanol, obtain 35 of 140mg (10%), be white crystalline powder. 1H-NMR (CDCl 3) δ 1.83-2.18 (m, 4H), 2.24 (s, 3H), 3.44-3.55 (m, 4H), 3.59 (s, 2H), 3.80 (s, 3H), 4.05-4.24 (m, 2H), 4.28-4.32 (m, 1H), 6.73-8.55 (a plurality of m, 12H altogether).MS(FAB)m/z?562(M ++1)。C 30H 31N 3O 84H 2The analytical calculation value of O: C, 56.87; H, 6.20; N, 6.63.Measured value: C, 56.73; H, 5.56; N, 6.52.Embodiment 323-methoxyl group-4-[2-[1-[3-methoxyl group-4-[N '-(2-aminomethyl phenyl) urea groups] phenyl acetyl]-the 2-pyrrolidinyl] acetenyl] benzoic acid
(1.20g, the hydrogenation that stirs the mixture under 1 atmospheric pressure 5.7mmol) and among the ethanol (30ml) of 5%Pd-C (1.0g) and the THF (20ml) is spent the night with 3-methoxyl group-4-nitrobenzoic acid methyl ester.Filter this mixture, evaporated filtrate.Residue is through silica gel column chromatography, and as eluant, the solid that obtains is further purified by recrystallization from chloroform-normal hexane, obtains 4-amino-3-methoxyl methyl benzoate of 805mg (78%), is white plates with chloroform.Mp 126-128; IR (KBr) 3475,1700cm -1 1H-NMR (CDCl 3) δ 3.86 (3H, s), 3.89 (3H, s), 4.21 (2H, br s), 6.66 (1H, d, J=8.3Hz), 7.45 (1H, d, J=1.9Hz), 7.54 (1H, dd, J=1.9 and 8.3Hz); MS (FAB) m/z 182 (M ++ 1); C 9H 11NO 3The analytical calculation value: C, 59.66; H, 6.12; N, 7.73.Measured value: C, 59.65; H, 6.15; N, 7.65.
In 0 ℃, (725mg, the agitating solution of ethanol 4mmol) (10ml) add in the dilute sulfuric acid (by 0.5ml sulphuric acid and the preparation of 10ml water) with 4-amino-3-methoxyl methyl benzoate.(331mg, water 4.8mmol) (10ml) solution adds in this mixture with sodium nitrite.After uniform temp down stirs 0.5 hour, with KI this mixture impouring cold (0 ℃), that stir (1.83g, 11mmol) and Catalysts Cu in the aaerosol solution of water (100ml).This mixture of vigorous stirring is 1 hour under room temperature, uses chloroform extraction.With salt water washing extract, through dried over mgso and evaporation.Residue is through silica gel column chromatography, with normal hexane-ethyl acetate (10: 1, v/v) as eluant, obtain the mixture (748mg) of 4-iodo-3-methoxyl methyl benzoate and 3-methoxyl methyl benzoate, be a kind of water white oil.
In this oil, add Pd (PPh 3) 4(150mg, 0.13mmol), CuI (57mg, 0.3mmol) and i-Pr 2NH (10ml).Under blanket of nitrogen, stirred this mixture 1 hour, with 1-(tert-butoxycarbonyl)-2-acetenyl pyrrolidine (488mg, i-Pr 2.5mmol) 2Solution among the NH (10ml) adds in this mixture.Stir after 2 hours, in this mixture impouring water, use ethyl acetate extraction.With salt water washing extract, through dried over mgso and evaporation.Residue is through silica gel column chromatography, with normal hexane-ethyl acetate (5: 1,, obtain 1-(tert-butoxycarbonyl)-2-[2-(2-methoxyl group-4-methoxycarbonyl phenyl) acetenyl of 431mg (48%) v/v) as eluant] pyrrolidine, be a kind of yellow oil. 1H-NMR(CDCl 3)δ?1.49(9H，s)，1.77-2.14(4H，m)，3.36-3.51(2H，m)，3.90(3H，s)，3.91(3H，s)，4.60-4.81(1H，m)，7.36-7.39(1H，m)，7.51(1H，s)，7.55-7.57(1H，m)。
To 1-(tert-butoxycarbonyl)-2-[2-(2-methoxyl group-4-methoxycarbonyl phenyl) acetenyl] (395mg adds TFA (3ml) to pyrrolidine in the agitating solution of dichloromethane 1.1mmol) (3ml).The mixture that stirring obtains 1 hour.This mixture of vacuum concentration makes and is alkalescence by adding saturated sodium bicarbonate, uses chloroform extraction.Wash extract with water,, obtain 2-[2-(2-methoxyl group-4-methoxycarbonyl phenyl) acetenyl of 238mg (84%) through dried over mgso and evaporation] pyrrolidine, be a kind of yellow oil. 1H-NMR(CDCl 3)δ1.81-2.16(4H，m)，2.97-3.17(2H，m)，3.91(6H，s)，4.13-4.15(1H，m)，7.41(1H，d，J＝8.3Hz)，7.51(1H，s)，7.56(1H，d，J＝8.3Hz)。
With 2-[2-(2-methoxyl group-4-methoxycarbonyl phenyl) acetenyl] pyrrolidine (233mg, 0.9mmol), 3-methoxyl group-4-[N '-(2-aminomethyl phenyl) urea groups] phenylacetic acid (314mg, 1mmol), EDC (268mg, 1.4mmol), (110mg, 0.9mmol) mixture in DMF (10ml) stirs and to spend the night DMAP.In this mixture impouring 1N HCl, sucking filtration is collected the solid that produces.The solid that obtains is dissolved in the chloroform, through this solution of dried over mgso and evaporation.Residue as eluant, obtains a kind of oil with ethyl acetate through silica gel short column chromatography.This oil is dissolved among the THF (5ml), under stirring the 0.25N sodium hydroxide is added in this solution.To obtain solid among this solution impouring ice-1N HCl.Collect this solid, wash with water also air-dry.Make crude product solid recrystallization from chloroform-normal hexane obtain 36 of 215mg (44%), be white crystalline powder.mp?141-145；IR(KBr)3338，2956，2935，2875，2593，1711cm -1； 1H-NMR(DMSO-d 6)δ1.91-2.14(4H，m)，2.24(3H，s)，3.38-3.68(4H，m)，4.88-5.08(1H，m)，6.76-8.56(12H，m)；MS(FAB)m/z?542(M ++1)。Embodiment 333-N, N-dimethylamino-4-[1-[3-methoxyl group-4-[N '-(2-aminomethyl phenyl) urea groups] phenyl acetyl]-2-pyrrolidinyl methoxyl group] benzoic acid
Under room temperature, with 4-hydroxyl-3-nitrobenzoic acid methyl ester (3.22g, the hydrogenation 70 hours that stirs the mixture under 1 atmospheric pressure 0.0163mol) and in the methanol (30ml) of 5%Pd-C (12.9g).Remove insoluble catalyst, vacuum evaporation filtrate.Residue is through silica gel column chromatography, with ethanol-chloroform (1: 20, v/v) as eluant, obtain the 3-amino-4-hydroxy essence of Niobe of 1.89g (69%), be the light brown slurry.
Under room temperature, with 3-amino-4-hydroxy essence of Niobe (1.07g, 6.40mmol) and 5%Pd-C (2.14g) in methanol (20ml), 37% formalin (1.08ml, the hydrogenation 26 hours that stirs the mixture under 1 atmospheric pressure 0.0122mol) and among the 1NHCl (6.1ml).Remove insoluble catalyst, vacuum evaporation filtrate.Residue is through silica gel column chromatography, with ethyl acetate-normal hexane (1: 10, v/v) as eluant, obtain 3-(N, N-the dimethylamino)-4-methyl hydroxybenzoate of 0.817g (70%), be a kind of slurry.
In 0 ℃, to 3-(N, the N-dimethylamino)-4-methyl hydroxybenzoate (0.817g, 4.18mmol), N-tert-butoxycarbonyl dried meat ammonia alcohol (0.926g, 4.60mmol), triphenyl phasphine (1.21g, 4.60mmol) the stirring the mixture of THF (20ml) in drip DIAD (95%) (0.953ml, 4.60mmol).The mixture that obtains was heated 41 hours under refluxing.After the cooling, the vacuum evaporation mixture.Residue is through silica gel column chromatography, with ethyl acetate-normal hexane (1: 10-1: 6, v/v) as eluant, obtain a kind of slurry, this slurry can need not to be further purified and be used for subsequent reaction.This slurry is dissolved in the dichloromethane (10ml), adds TFA (10ml).In stirring this solution under the room temperature after 5 hours, this solution of vacuum evaporation.Water is added in the residue, wash with chloroform.By adding in the saturated sodium bicarbonate and the water-bearing layer, use chloroform extraction.Through the dried over sodium sulfate extract, vacuum evaporation obtains 3-(N, N-dimethylamino)-4-(the 2-pyrrolidinyl methoxyl group) essence of Niobe of 1.03g (89%), is a kind of jelly.
Under room temperature, with 3-(N, the N-dimethylamino)-4-(2-pyrrolidinyl methoxyl group) essence of Niobe (0.529g, 1.90mmol), 3-methoxyl group-4-[N '-(2-aminomethyl phenyl) urea groups] phenylacetic acid methyl ester (0.597g, 1.90mmol), HOBt (0.308g, 2.28mmol), 4-dimethylaminopyridine (DMAP) (23.2mg, 0.190mmol) and 1-ethyl-3-(3-dimethylaminopropyl) carbodiimide (EDC) (0.437g, 2.28mmol) mixture in DMF (10ml) stirred 15 hours, by careful adding 1N HCl this mixture that neutralizes, use ethyl acetate extraction.Through dried over sodium sulfate extract and vacuum evaporation; obtain the 3-N of 0.607g (56%); N-dimethylamino-4-[1-[3-methoxyl group-4-[N '-(2-aminomethyl phenyl) urea groups] phenyl acetyl]-2-pyrrolidinyl methoxyl group] essence of Niobe, be white crystalline material.
Under room temperature; with 3-N; N-dimethylamino-4-[1-[3-methoxyl group-4-[N '-(2-aminomethyl phenyl) urea groups] phenyl acetyl]-2-pyrrolidinyl methoxyl group] essence of Niobe (0.600g; 1.04mmol) (5ml, 1.25mmol) mixture in stirred 21 hours at THF (10ml) and 0.25N sodium hydroxide.Chloroform is added in this mixture, and the mixture of water (100ml)-1N sodium hydroxide (4ml) extracts.With in the saturated ammonium chloride and extract, use chloroform extraction.Through dried over sodium sulfate extract and vacuum evaporation, obtain 37 of 428mg (70%), be white crystalline solid. 1H-NMR (400MHz, DMSO-d 6) δ 1.88,1.99 and 2.11 (4H respectively is m), 2.24 (3H, s), 2.67 (6H, s), 3.33 (2H, m), 3.58 (2H, m), 4.05-4.32 (3H, m), 6.75 (1H, d, J=7.3Hz), 6.92-6.95 (1H, m), 7.05 (1H, d, J=8.3Hz), 7.11-7.17 (2H, m), 7.42 (1H, s), 7.52 (1H, d, J=7.8Hz), 7.79 (1H, d, J=7.8Hz), 8.00 (1H, d, J=7.8Hz), 8.31 (1H, s), 8.46,8.55 (respectively being 1H, they respectively is s); MS (FAB) m/z 533 (M ++ 1).Embodiment 343-fluoro-4-[[1-[3-methoxyl group-4-[N '-(2-aminomethyl phenyl) urea groups] phenyl acetyl]-the 2-pyrrolidinyl] methoxyl group] benzoic acid
Under room temperature, to 4-bromo-2-fluorophenol (217ul, 2.002mmol), N-tert-butoxycarbonyl dried meat ammonia alcohol (403mg, 2.002mmol) and triphenyl phasphine (630mg, 2.403mmol) the agitating solution of THF (7ml) in add DIAD (477ul, 2.423mmol).The mixture that obtains was stirred under room temperature 6 hours, spend the night in 70 ℃ of stirrings then.Vacuum concentrated mixture, residue be through silica gel column chromatography, with normal hexane-ethyl acetate (5: 1,, obtain 1-bromo-4-[1-(tert-butoxycarbonyl)-2-pyrrolidinyl of 549.4mg (73%) v/v) as eluant] methoxyl group-3-fluorobenzene, be a kind of oil. 1H-NMR (400MHz, CDCl 3) δ 1.46 (s, 9H), 1.85 (br m, 1H), 1.90-2.10 (br s, 3H), 3.30-3.47 (m, 2H), 3.85,4.04 (respectively be br s, 1H), 4.11-4.20 (m, 2H), 6.82-6.98 (m, 1H), 7.13-7.26 (m, 2H); MS (FAB) m/z 374 (M ++ 1).
To 1-bromo-4-[1-(tert-butoxycarbonyl)-2-pyrrolidinyl] methoxyl group-3-fluorobenzene (549.4mg, add in the agitating solution of DMSO 1.468mmol) (6ml) and methanol (5ml) triethylamine (448ul, 3.229mmol), Pd (OAc) 2(36.2mg, 0.161mmol) with 1, two (diphenylphosphino) propane of 3-(66.4mg, 0.161mmol).In this mixture, fed CO gas 10 minutes.Under the CO air-flow, the mixture that obtains was stirred 2 days in 70 ℃.After concentrating this mixture, dilute residue with ethyl acetate.With this solution of salt water washing, through dried over sodium sulfate.Removal of solvent under reduced pressure, residue be through silica gel column chromatography, with normal hexane-ethyl acetate (5: 1,, obtain 4-[1-(the tert-butoxycarbonyl)-2-pyrrolidinyl of 323.0mg (62%) v/v) as eluant] methoxyl group-3-fluorophenyl carbamate, be light yellow oil. 1H-NMR (400MHz, CDCl 3) δ 1.47 (and s, 9H), 1.87 (br s, 1H), 1.95-2.10 (m, 3H), 3.34-3.44 (br m, 2H), 3.89 (s, 3H), 3.94 and 4.11-4.26 (respectively be br m, 3H), 7.03-7.11 (m, 1H), 7.75-7.80 (m, 2H); MS (FAB) m/z 354 (M ++ 1).
In 0 ℃, to 4-[1-(tert-butoxycarbonyl)-2-pyrrolidinyl] (323.0mg adds TFA (1.3ml) to methoxyl group-3-fluorophenyl carbamate in the agitating solution of dichloromethane 0.914mmol) (6.5ml), stirred the mixture under room temperature 1.5 hours.Removal of solvent under reduced pressure makes residue be alkalescence by adding the 1N sodium hydroxide.With this mixture of chloroform extraction.With salt water washing extract, through dried over sodium sulfate and concentrating under reduced pressure, obtain 3-fluoro-4-(2-pyrrolidinyl) methoxyl methyl benzoate of 174.8mg (76%), be a kind of brown oil. 1H-NMR(400MHz，CDCl 3)δ1.54-1.63(m，1H)，1.76-2.02(m，3H)，2.93-3.07(m，2H)，3.57(ddd，J＝4.9，6.9，14.3Hz，1H)，3.89(s，3H)，3.97(dd，J＝6.8，9.3Hz，1H)，4.04(dd，J＝5.0，8.8Hz，1H)，6.98(t，J＝17.6Hz，1H)，7.73(dd，J＝2.0，11.7Hz，1H)，7.78(dt，J＝2.0，8.8Hz，1H)；MS(FAB)m/z?253(M ++1)。
Under room temperature, with 3-methoxyl group-4-[N '-(2-aminomethyl phenyl) urea groups] phenylacetic acid pentafluorophenyl esters (324.5mg, 0.676mmol), 3-fluoro-4-(2-pyrrolidinyl) methoxyl methyl benzoate (171.1mg, 0.676mmol), (113ul, 0.811mmol) mixture in DMF (5ml) stirred 2 hours triethylamine.With ethyl acetate diluted mixture thing, use the salt water washing, through dried over sodium sulfate.Removal of solvent under reduced pressure; residue is through silica gel column chromatography; with normal hexane-ethyl acetate (1: 2; v/v) as eluant; obtain 3-fluoro-4-[[1-[3-methoxyl group-4-[N '-(2-aminomethyl phenyl) urea groups] phenyl acetyl]-the 2-pyrrolidinyl] methoxyl group]-essence of Niobe (365.1mg; 0.664mmol), be a kind of oil.(46.3mg 1.932mmol), spends the night this reactant mixture stirring under room temperature to add Lithium hydrate in the agitating solution of THF of this chemical compound (4.4ml) and water (1.1ml).With chloroform diluted mixture thing, by adding 1N HCl acidify.With the isolating organic layer of salt water washing, through dried over sodium sulfate.Removal of solvent under reduced pressure makes the crude product solid recrystallization from normal hexane-ethyl acetate-chloroform that obtains obtain 38 of 102mg (30%), is white crystalline powder.mp?123-126；IR(KBr)1616，1537，1282，756cm -1； 1H-MR(400MHz，DMSO-d 6)δ1.87-2.09(m，4H)，2.25(s，3H)，3.48-3.57(m，2H)，3.60(s，2H)，3.83(s，3H)，4.11-4.16(m，1H)，4.24(dd，J＝2.9，9.8Hz，1H)，4.28-4.34(br?s，1H)，6.74(dd，J＝1.5，8.3Hz，1H)，6.87(s，1H)，6.94(t，J＝7.3Hz，1H)，7.12(d，J＝7.8Hz，1H)，7.1?5(t，J＝8.3Hz，1H)，7.34(t，J＝8.8Hz，1H)，7.66(dd，J＝2.0，12.2Hz，1H)，7.73(d，J＝9.3Hz，1H)，7.79(d，J＝8.3Hz，1H)，7.99(d，J＝7.8Hz，1H)，8.46(s，1H)，8.55(s，1H)；MS(FAB)m/z?536(M ++1)。C 29H 30FN 3O 60.5H 2The analytical calculation value of O: C, 63.96; H, 5.74; N, 7.72; F, 3.49.Measured value: C, 64.11; H, 5.80; N, 7.39; F, 3.54.Embodiment 354-[1-[4-[N '-(2-fluoro phenyl) urea groups]-3-methoxyphenyl acetyl group]-the 2-pyrrolidinyl] methoxyl group-3-methoxybenzoic acid
With 4-[N '-(2-fluoro phenyl) urea groups]-3-methoxybenzene guanidine-acetic acid (318mg, 1mmol), 2-(2-methoxyl group-4-ethoxy carbonyl) phenoxymethyl pyrrolidine (279mg, 1mmol), EDC (288mg, 1.5mmol) and DMAP (122mg, 1mmol) mixture in DMF (20ml) stirs and to spend the night.In this mixture impouring 1N HCl, sucking filtration is collected the solid that produces.Solid is dissolved in the chloroform, through dried over mgso.Remove desolvate after, residue is through silica gel column chromatography, with chloroform-methanol (100: 1, v/v) as eluant, obtain a kind of oil, make this oil be dissolved in THF: methanol (4: 1, v/v, 10ml) in.(8ml) adds in this solution with the 0.25N sodium hydroxide, and the stirred mixture that obtains was heated 3 hours under refluxing.In this mixture impouring 1N HCl.Sucking filtration is collected the solid that produces.Make to be dissolved in the chloroform, through dried over mgso and evaporation.Residue recrystallization from chloroform-normal hexane-ether obtains 39 of 329mg (60%), is white crystalline powder.mp140-144；IR(KBr)3338，2956，2875，2607，1709cm -1； 1H-NMR(CDCl 3)δ1.95-2.25(4H，m)，3.45-4.50(12H，m)，6.66-8.15(12H，m)；MS(FAB)m/z552(M ++1)。Embodiment 362-[[1-[3-methoxyl group-4-[N '-(2-aminomethyl phenyl) urea groups] phenyl acetyl]-the 2-pyrrolidinyl] methoxyl group] pyridine-5-formic acid
Figure A0080951001981
In 0 ℃, (500mg drips TMSCHN in the agitating solution of benzene 3.594mmol) (8ml) and methanol (2ml) to the 6-hydroxy niacin 2(1.97ml 3.953mmol), stirs mixture and to spend the night under room temperature.By adding acetic acid quencher reactant mixture, the mixture that vacuum concentration obtains.Residue is through silica gel column chromatography, with normal hexane-ethyl acetate (1: 3, v/v) as eluant, obtain 2 hydroxy pyrimidine-5-methyl formate of 269.8mg (49%), be white crystalline powder.IR(KBr)3062，1657，1654，1612，1435，1300，1113，775，642cm -1； 1H-NMR(400MHz，CDCl 3)δ?3.87(s，3H)，6.58(d，J＝9.8Hz，1H)，7.99(dd，J＝2.4，9.8Hz，1H)，8.19(d，J＝2.4Hz，1H)；MS(FAB)m/z?154(M ++1)。C 7H 7NO 31/4H 2The analytical calculation value of O: C, 53.33; H, 4.80; N, 8.89.Measured value: C, 53.58; H, 4.65; N, 8.87.
Under room temperature, to 2 hydroxy pyrimidine-5-methyl formate (269.8mg, 1.762mmol), N-tert-butoxycarbonyl dried meat ammonia alcohol (354.6mg, 1.762mmol) and triphenyl phasphine (554.6mg, 2.114mmol) the agitating solution of THF (10ml) in slowly add DIAD (0.42ml, 2.114mmol), the mixture that obtains was stirred 6 hours in 70 ℃.Concentrated reaction mixture, residue be through silica gel column chromatography, with normal hexane-ethyl acetate (5: 1,, obtain 2-[[1-(the tert-butoxycarbonyl)-2-pyrrolidinyl of 262.5mg (44%) v/v) as eluant] methoxyl group] pyridine-5-methyl formate, be a kind of oil. 1H-NMR(400MHz，CDCl 3)δ1.47(s，9H)，1.85-1.98(m，4H)，3.37(br?s，2H)，3.92(s，3H)，4.12-4.33(br?m，2H)，4.4(br?s，1H)，6.75(m，1H)，8.15(m，1H)，8.79(m，1H)；MS(FAB)m/z?337(M ++1)。
In 0 ℃, to 2-[[1-(tert-butoxycarbonyl)-2-pyrrolidinyl] methoxyl group] pyridine-5-methyl formate (262.5mg, 0.870mmol) the agitating solution of dichloromethane (5.3ml) in add TFA (1.1ml), under room temperature, stirred the mixture that obtains 1 hour.Removal of solvent under reduced pressure makes residue be alkalescence by adding the 1N sodium hydroxide, uses chloroform extraction.With salt water washing extract,, obtain the 2-[(2-pyrrolidinyl of 173.1mg (94%) through dried over sodium sulfate and concentrating under reduced pressure) methoxyl group] pyridine-5-methyl formate, be light yellow oil. 1H-NMR(400MHz，CDCl 3)δ1.49-1.58(ddt，J＝6.8，8.8Hz，1H)，1.72-1.87(m，2H)，1.90-1.99(m，1H)，2.92-3.05(m，2H)，3.50-3.57(ddd，J＝4.4，7.3，15.1Hz，1H)，3.91(s，3H)，4.23(dd，J＝7.8，10.7Hz，1H)，4.38(dd，J＝4.4，10.7Hz，1H)，6.78(d，J＝8.8Hz，1H)，8.15(dd，J＝2.4，8.8Hz，1H)，8.80(d，J＝2.4Hz，1H)；MS(FAB)m/z237(M ++1)。
Under room temperature, with 3-methoxyl group-4-[N '-(2-aminomethyl phenyl) urea groups] phenylacetic acid pentafluorophenyl esters (351.7mg, 0.732mmol), the 2-[(2-pyrrolidinyl) methoxyl group] pyridine-5-methyl formate (173.0mg, 0.732mmol), (122.4 μ l, 0.878mmol) mixture in DMF (5.2ml) stirred 1 hour triethylamine.With ethyl acetate diluted mixture thing, use the salt water washing, through dried over sodium sulfate.Removal of solvent under reduced pressure; residue is through silica gel column chromatography; with normal hexane-ethyl acetate (1: 5; v/v) as eluant; obtain 2-[[1-[3-methoxyl group-4-[N '-(2-aminomethyl phenyl) urea groups] phenyl acetyl]-the 2-pyrrolidinyl] methoxyl group] pyridine-5-methyl formate (338.4mg; 87%), is a kind of oil.(45.7mg 1.91mmol), spends the night this reactant mixture stirring under room temperature to add Lithium hydrate in the agitating solution of THF of this chemical compound (5.6ml) and water (1.4ml).With chloroform diluted mixture thing, handle with saturated ammonium chloride, use the salt water washing, through dried over sodium sulfate.Removal of solvent under reduced pressure makes the crude product solid recrystallization from normal hexane-E-C-methanol that obtains obtain 40 of 193.8mg (59%), is white crystalline powder.mp?125-128；IR(KBr)1716，1600，1533，1255cm -1； 1H-NMR(400MHz，DMSO-d 6)δ?1.67-2.03(m，4H)，2.50(s，3H)，3.33-3.42(m，1H)，3.52(m，2H)，3.58(d，J＝4.4Hz，1H)，3.83(s，3H)，4.27-4.31(m，2H)，4.42-4.47(m，1H)，6.73(d，J＝7.8Hz，1H)，6.87-6.95(m，3H)，7.11-7.17(m，2H)，7.79(d，J＝8.3Hz，1H)，7.99(d，J＝8.3Hz，1H)，8.14(dd，J＝2.0，8.8Hz，1H)，8.46(s，1H)，8.56(s，1H)，8.69(d，J＝2.0Hz，1H)，13.06(br?s，1H)；MS(FAB)m/z?519(M ++1)。C 28H 30N 4O 61/2H 2The analytical calculation value of O: C, 63.75; H, 5.92; N, 10.62.Measured value: C, 63.61; H, 5.94; N, 10.27.Embodiment 373-methoxyl group-4-[2-[4-[N '-(2-aminomethyl phenyl) urea groups] benzyl]-the 4-thiazolyl] methoxybenzoic acid
Under room temperature, to phosphorus pentasulfide (27.4g, 123.34mmol) and freshly prepared anhydrous sodium sulfide (4.8g, add in the agitating solution of THF 61.67mmol) (200ml) right-nitrobenzyl cyanide (2.0g, 12.33mmol).Under room temperature, the mixture that obtains was stirred 17 hours.Dilute this mixture with ethyl acetate, wash with 10% potassium phosphate.Use the dichloromethane extraction water layer.Through dried over sodium sulfate extract and vacuum concentration.Residue is through silica gel column chromatography, with normal hexane-ethyl acetate (5: 1-2: 1, v/v) as eluant, obtain the 4-nitrobenzyl thioformamide (carbothioamide) of 1.53g (64%), be faint yellow crystalline material.IR (KBr) 1529,1446,1326,1315,858cm -1 1H-NMR (400MHz, CDCl 3) δ 4.15 (s, 2H), 7.51 (d, J=8.3Hz, 2H), 8.24 (d, J=8.8Hz, 2H); MS (FAB) m/z197 (M ++ 1); C 8H 8N 2O 2The analytical calculation value of S: C, 48.97; H, 4.11; N, 14.28; S, 16.34.Measured value: C, 48.69; H, 4.06; N, 14.07; S, 16.10.
(502.0mg adds 1 in the agitating solution of ethanol 2.558mmol) (5ml), and (649.6mg 5.16mmol), heats this mixture 1 hour under refluxing 3-two chloro-2-acetone to 4-nitrobenzyl thioformamide.Concentrate this mixture, dilute residue with chloroform.With 1N sodium hydroxide, this solution of salt water washing, through dried over sodium sulfate.Removal of solvent under reduced pressure, residue be through silica gel column chromatography, with normal hexane-ethyl acetate (4: 1,, obtain 495.2mg (72%) 4-[2-(4-nitrobenzyl) thiazolyl v/v) as eluant] methyl chloride, be a kind of light yellow oil. 1H-NMR(400MHz，CDCl 3)δ?4.43(s，2H)，4.68(s，2H)，7.23(s，1H)，7.49(d，J＝8.8Hz，2H)，8.20(d，J＝8.8Hz，2H)；MS(FAB)m/z?269(M ++1)。
In 0 ℃, to ethyl vanillate (308.0mg, 1.570mmol) and Feldalat NM (89mg is added in 4-[2-(4-nitrobenzyl) thiazolyl in the methanol (1.4ml) in the agitating solution in methanol 1.570mmol) (6.5ml)] and methyl chloride (211.0mg, 0.785mmol).The mixture that stirring obtains under room temperature 16 hours, heating is 1 day under refluxing.Removal of solvent under reduced pressure is used the chloroform extraction residue.Water, salt water washing extract are through dried over sodium sulfate.Removal of solvent under reduced pressure, residue be through silica gel column chromatography, with normal hexane-ethyl acetate (2: 1,, obtain 201.7mg (60%) 3-methoxyl group-4-[2-(4-nitrobenzyl)-4-thiazolyl v/v) as eluant] the methoxybenzoic acid ethyl ester, be a kind of light yellow oil. 1H-NMR(400MHz，CDCl 3)δ1.39(t，J＝7.3Hz，3H)，3.93(s，3H)，4.36(q，J＝7.3Hz，2H)，4.44(s，2H)，5.31(s，2H)，6.97(d，J＝8.3Hz，1H)，7.28(s，1H)，7.48(d，J＝8.8Hz，2H)，7.57(d，J＝2.0Hz，1H)，7.64(dd，J＝2.0，8.3Hz，1H)，8.20(d，J＝8.8Hz，2H)；MS(FAB)m/z?429(M ++1)。
With 3-methoxyl group-4-[2-(4-aminobenzyl)-4-thiazolyl] and the methoxybenzoic acid ethyl ester (201.7mg, 0.471mmol) and the hydrogenation 24 hours under 1 atmospheric pressure of the agitating solution of 5%Pd/C (40mg) in ethanol (8ml).Filter this mixture, concentrated filtrate.Residue is through silica gel column chromatography, with normal hexane-ethyl acetate (1: 1,, obtain 3-methoxyl group-4-[2-(4-aminobenzyl)-4-thiazolyl of 87.8mg (47%) v/v) as eluant] the methoxybenzoic acid ethyl ester, be the yellow crystal powder. 1H-NMR(400MHz，CDCl 3)δ?1.38(t，J＝7.3Hz，3H)，3.93(s，3H)，4.21(s，2H)，4.35(q，J＝7.3Hz，2H)，5.30(s，2H)，6.66(dd，J＝2.0，6.4Hz，2H)，6.97(d，J＝8.3Hz，1H)，7.09(d，J＝8.3Hz，2H)，7.18(s，1H)，7.56(d，J＝2.0Hz，1H)，7.64(dd，J＝2.0，8.3Hz，1H)；MS(FAB)m/z?399(M ++1)。
To 3-methoxyl group-4-[2-(4-aminobenzyl)-4-thiazolyl] methoxybenzoic acid ethyl ester (87.8mg, 0.220mmol) oxolane (2.0ml) solution in add triethylamine (30.5ul, 0.220mmol) and Carbimide. neighbour-tolyl ester (30 μ l), reactant mixture was stirred under room temperature 21 hours.Reactant mixture is added in the frozen water precipitation that elimination produces.Use chloroform extraction filtrate.Water, salt water washing.Through the dried over sodium sulfate extract.Removal of solvent under reduced pressure obtains 110.4mg (94%) 3-methoxyl group-4-[2-[4-[N '-(2-aminomethyl phenyl) urea groups] benzyl]-the 4-thiazolyl] the methoxybenzoic acid ethyl ester, be light yellow crystalline powder. 1H-NMR(400MHz，CDCl 3)δ1.38(t，J＝7.3Hz，3H)，2.28(s，3H)，3.92(s，3H)，4.28(s，2H)，4.35(q，J＝7.3Hz，2H)，5.29(s，2H)，6.20(s，1H)，6.47(s，1H)，6.97(d，J＝8.8Hz，1H)，7.20(s，1H)，7.24(s，2H)，7.27(s，2H)，7.33(d，J＝8.3Hz，2H)，7.49(d，J＝7.3Hz，1H)，7.56(d，J＝2.0Hz，1H)，7.63(dd，J＝2.0，8.3Hz，1H)；MS(FAB)m/z532(M ++1)。
To 3-methoxyl group-4-[2-[4-[N '-(2-aminomethyl phenyl) urea groups] benzyl]-the 4-thiazolyl] add Lithium hydrate (6.0mg in the THF (1.6ml) of methoxybenzoic acid ethyl ester and the agitating solution of water (0.4ml), 0.249mmol), under room temperature, this mixture was stirred 1 hour, under refluxing, heated 8 hours again.Concentrate this mixture, dilute with chloroform.Make this solution be alkalescence by adding the 1N sodium hydroxide.By adding 1N HCl acidify aqueous extract, use chloroform extraction.With salt water washing extract, through dried over sodium sulfate.Removal of solvent under reduced pressure makes the crude product solid recrystallization from normal hexane-ethyl acetate-ethanol that obtains obtain 41 of 59.6mg (57%), is white crystalline powder.mp?243-245；IR(KBr)3282，1685，1637，1600，1554，1516，1278，1234，763，748cm -1； 1H-NMR(400MHz，DMSO-d 6)δ2.27(s，3H)，3.83(s，3H)，4.30(s，2H)，5.21(s，2H)，6.97(t，J＝7.3Hz，1H)，7.15-7.24(m，3H)，7.29(d，J＝8.8Hz，2H)，7.47(d，J＝8.3Hz，2H)，7.50(s，1H)，7.58(d，J＝8.3Hz，1H)，7.62(s，1H)，7.86(d，J＝7.8Hz，1H)，7.97(s，1H)，9.09(s，1H)，12.70(br?s，1H)；MS(FAB)m/z504(M ++1)。C 27H 25N 3O 5S1/4H 2The analytical calculation value of O: C, 63.83; H, 5.06; N, 8.27.Measured value: C, 63.74; H, 4.99; N, 8.10.Embodiment 384-[[1-[3-methoxyl group-4-[N '-(2-aminomethyl phenyl) urea groups] phenyl acetyl]-2-methyl-2-pyrrolidinyl] methoxyl group]-the 3-nitrobenzoic acid
Under room temperature, to N-tert-butoxycarbonyl proline (6.00g, methanol 0.0279mol): benzene (1: 4, drip TMSCHN in agitating solution v/v) 2The 2.0M hexane solution (16.7ml, 0.0334mol).After stirring 1 hour under the room temperature, this mixture of vacuum evaporation obtains the N-tert-butoxycarbonyl proline methyl ester of 6.39g (100%), is yellow slurry with the solution that obtains. 1H-NMR(CDCl 3)δ1.41(s，9H)，1.85-1.98(m，4H)，2.21-2.28(m，2H)，3.72(s，3H)，4.29(m，1H)。
In-78 ℃, with 5 fens clockwise diisopropylamine (2.02ml, drip in the agitating solution of THF 0.0144mol) (30ml) n-BuLi the 1.59M hexane solution (9.06ml, 0.0144mol).The solution that obtains in-78 ℃ of stirrings 20 minutes.In-78 ℃, with dripping N-tert-butoxycarbonyl proline methyl ester (3.00g, THF 0.0131mmol) (30ml) solution in 5 fens this solution of clockwise.The solution that obtains in-78 ℃ of stirrings 10 minutes.In-78 ℃, in this solution, be added dropwise to MeI (0.900ml, 0.0144mol).The solution that obtains in-78 ℃ of stirrings 30 minutes.By adding this solution of saturated ammonium chloride quencher.The mixture that obtains with chloroform extraction.Wash extract with water,, obtain 3.20g (q.y.) N-tert-butoxycarbonyl-2-methylproline methyl ester, be yellow slurry through dried over sodium sulfate and vacuum evaporation. 1H-NMR(CDCl 3)δ1.33(s，9H)，1.38(s，3H)，1.72-2.20(m，4H)，3.27-3.59(m，2H)，3.63(d，J＝6.3Hz，3H)。
Under room temperature, (3.20g adds 1N sodium hydroxide (15.7ml) in the agitating solution of THF 0.013lmol) (20ml) to N-tert-butoxycarbonyl-2-methylproline methyl ester.The mixture that stirring obtains is after 24 hours, and this mixture of dilute with water washs with ethyl acetate.By adding the isolating water-bearing layer of 1N HCl acidify, use ethyl acetate extraction.Through dried over sodium sulfate and vacuum evaporation extract, obtain N-tert-butoxycarbonyl-2-methylproline of 1.71g (57%), be yellow slurry. 1H-NMR(CDCl 3)δ1.42(s，9H)，1.48(s，3H)，1.88-2.31(m，4H)，3.34-3.57(m，2H)，9.35(br?s，1H)。
Under room temperature, (1.10g drips BH in the agitating solution of THF 4.80mmol) (20ml) to N-tert-butoxycarbonyl-2-methylproline 3-SMe 2(0.546ml, 5.76mmol).In mixture that 80 ℃ of stirrings obtain after 6 hours, this mixture of vacuum evaporation.Dilute residue with methanol,, obtain N-tert-butoxycarbonyl-2-methylol-2-crassitude of 0.648g (60%), be yellow slurry with normal hexane (3 *) washing, vacuum evaporation. 1H-NMR(CDCl 3)δ1.47(s，9H)，1.76-2.05(m，4H)，3.28-3.48(m，2H)，3.66(m，2H，d)。
In 0 ℃, to N-tert-butoxycarbonyl-2-methylol-2-crassitude (0.648g, 3.01mmol), 4-hydroxyl-3-nitrobenzoic acid methyl ester (0.593g, 3.01mmol) and triphenyl phasphine (0.868g, 3.31mmol) the agitating solution of THF (10ml) in drip DIAD (95%) (0.686ml, 3.31mmol).In mixture that 80 ℃ of stirrings obtain after 24 hours, this mixture of vacuum evaporation.With dichloromethane (5ml) dilution residue, add TFA (5ml).In stirring the mixture obtain under the room temperature after 2 hours, this mixture of vacuum evaporation.With 0.5N HCl dilution residue, use chloroform extraction.With in the saturated sodium bicarbonate and water-bearing layer and use chloroform extraction.Through the dried over sodium sulfate extract, and vacuum evaporation, obtain 3-nitro-4-(2-methyl-2-pyrrolidinyl methoxyl group) essence of Niobe of 0.188g (21%), be yellow slurry.
Under room temperature, with 3-nitro-4-(2-methyl-2-pyrrolidinyl methoxyl group) essence of Niobe (0.188g, 0.920mmol), 3-methoxyl group-4-[N '-(2-aminomethyl phenyl) urea groups] phenylacetic acid (0.289g, 0.920mmol), HOBt (0.149g, 1.10mmol), DMAP (11.2mg, 0.0920mmol) and EDC (0.211g, 1.10mmol) mixture in DMF (5ml) stirred 14 hours.Ethyl acetate is added in this mixture, wash this solution in proper order with 0.5N HCl, saturated sodium bicarbonate and saline.Through dried over sodium sulfate organic layer and vacuum evaporation; obtain 4-[[1-[3-methoxyl group-4-[N '-(2-aminomethyl phenyl) urea groups of 0.489g (q.y.)] phenyl acetyl]-2-methyl-2-pyrrolidinyl] methoxyl group]-3-nitrobenzoic acid methyl ester, be the yellow crystal material. 1H-NMR(CDCl 3)δ1.26(d，J＝5.9Hz)，1.85-4.50(m，10H)，2.30(s，3H)，3.67(s，2H)，3.92(s，3H)，6.36(s，2H)，6.75-7.52(m，7H)，8.02(d，J＝7.8Hz，1H)，8.15(d，J＝8.8Hz，1H)，8.47(s，1H)。
With 4-[1-[3-methoxyl group-4-[N '-(2-aminomethyl phenyl) urea groups] phenyl acetyl]-2-methyl-2-pyrrolidinyl] methoxyl group]-3-nitrobenzoyl acid esters (0.489g, 0.828mmol) heated 2 hours under refluxing by stirring the mixture in methanol (5ml) and 1N sodium hydroxide (1.24ml).After the cooling, the dilute with water mixture is used chloroform extraction.With 1N HCl acidify water-bearing layer, use chloroform extraction.Through dried over sodium sulfate extract and vacuum evaporation, obtain 42 of 0.366g (99%), be the yellow crystal material.Embodiment 394-[4-hydroxyl-1-[3-methoxyl group-4-[N '-(2-aminomethyl phenyl) urea groups] phenyl acetyl]-2-pyrrolidinyl methoxyl group]-the 3-methoxybenzoic acid
Figure A0080951002041
With 4-[4-benzyloxy-1-[3-methoxyl group-4-[N '-(2-aminomethyl phenyl) urea groups] phenyl acetyl]-2-pyrrolidinyl methoxyl group]-3-methoxybenzoic acid ester (440mg; 0.645mmol) and 5%Pd/C (400mg) at acetic acid: ethanol (1: 1; v/v, the hydrogenation 5 hours that stirs the mixture under 1 atmospheric pressure in 100ml).Filter this mixture to remove catalyst, vacuum concentrated filtrate.Residue is through silica gel column chromatography; use chloroform: ethanol (10: 1; v/v) as eluant; obtain 4-[4-hydroxyl-1-[3-methoxyl group-4-[N '-(2-aminomethyl phenyl) urea groups of 90mg (24%)] phenyl acetyl]-2-pyrrolidinyl methoxyl group]-3-methoxybenzoic acid ethyl ester, be faint yellow oily thing. 1H-NMR (CDCl 3) (J=7.3Hz), 2.04-2.37 (is total to 5H, m) to δ 1.39 for 3H, t, (3.44-4.70 16H, a plurality of m), 6.63 (1H, s), 6.70-6.80 (2H, m), 6.84 (1H, d, J=8.3Hz), 7.11 (1H, t, J=7.8Hz), 7.20-7.24 (3H, m), 7.45 (1H, d, J=2.0Hz), 7.59 (2H, dd, J=8.3,2.0Hz), 8.01 (1H, d, J=7.8Hz).
With 4-[4-hydroxyl-1-[3-methoxyl group-4-[N '-(2-aminomethyl phenyl) urea groups] phenyl acetyl]-2-pyrrolidinyl methoxyl group]-3-methoxybenzoic acid ethyl ester (90mg; 0.152mmol) the 0.25N sodium hydroxide (5ml, 1.25mmol) and the heated overnight under refluxing of the stirred mixture among the THF (5ml).In this mixed liquor impouring ice-1N HCl (200ml).The sucking filtration collecting precipitation, recrystallization obtains 43 of 40mg (47%) from chloroform-methanol-normal hexane, is colourless amorphous solid. 1H-NMR (DMSO-d 6) δ 1.92-2.11 (2H, m), 2.24 (3H, s), 3.31-5.07 (14H, a plurality of m), 6.73 (1H, d, J=8.3Hz), 6.84 (1H, s), 6.93 (1H, t, J=7.8Hz), 7.01-7.17 (3H, m), 7.44 (1H, s), 7.52 (1H, d, J=8.8Hz), 7.79 (1H, d, J=8.3Hz), 7.99 (1H, d, J=7.8Hz), 8.46 (1H, s), 8.55 (1H, s), 12.67 (1H, br s); MS (FAB) m/z 564 (M ++ 1).Embodiment 40 (2S, 4R)-3-amino-4-[4-hydroxyl-1-[3-methoxyl group-4-[N '-(2-aminomethyl phenyl) urea groups] phenyl acetyl]-the 2-pyrrolidinyl] methoxybenzoic acid
To (2S, 4R)-4-benzyloxy-1-(tert-butoxycarbonyl)-2-dried meat ammonia alcohol (891mg, 2.9mmol), 4-hydroxyl-3-nitrobenzoic acid methyl ester (572mg, 2.9mmol) and triphenyl phasphine (839mg, 3.2mmol) the agitating solution of THF (6ml) in add DIAD (630ml, 3.2mmol), with this mixture heated overnight under refluxing.Except that after desolvating, residue is through silica gel column chromatography, use normal hexane: ethyl acetate (1: 1) and toluene: ethyl acetate (10: 1, v/v) as eluant, obtain (the 2S of 700mg (50%), 4R)-and 4-[4-benzyloxy-1-(tert-butoxycarbonyl)-2-pyrrolidinyl] methoxyl group-3-nitrobenzoic acid methyl ester, be faint yellow oily thing.
To (2S, 4R)-4-[4-benzyloxy-1-(tert-butoxycarbonyl)-2-pyrrolidinyl] (681mg adds TFA (2ml) to methoxyl group-3-nitrobenzoic acid methyl ester in the agitating solution of dichloromethane 1.4mmol) (2ml), the mixture that obtains was stirred 2 hours.Behind the concentrated reaction mixture, make residue be alkalescence, use chloroform extraction by adding saturated sodium bicarbonate.Wash extract with water, through dried over mgso and the evaporation, obtain 511mg (95%) (2S, 4R)-4-[4-benzyloxy-2-pyrrolidinyl] methoxyl group-3-nitrobenzoic acid methyl ester, be yellow oil.
With 3-methoxyl group-4-[N '-(2-aminomethyl phenyl) urea groups] phenylacetic acid (409mg, 1.3mmol), (2S, 4R)-4-(benzyloxy-2-pyrrolidinyl) methoxyl group-3-nitrobenzoic acid methyl ester (502mg, 1.3mmol), EDC (383mg, 2mmol) and DMAP (159mg, 1.3mmol) mixture in DMF (20ml) stirred 3 days, in this mixture impouring 1N HCl, sucking filtration is collected the precipitation that produces.Residue is dissolved in the chloroform, through dried over mgso.Except that after desolvating; residue is through silica gel column chromatography; with chloroform-methanol (200: 1; v/v) as eluant; obtain (the 2S of 680mg (91%); 4R)-4-amino-4-[4-benzyl Oxy-1-[3-methoxyl group-4-[N '-(2-aminomethyl phenyl) urea groups] phenyl acetyl]-the 2-pyrrolidinyl] methoxyl group-3-nitrobenzoic acid methyl ester, be white amorphous solid.
With (2S; 4R)-4-[4-benzyl Oxy-1-[3-methoxyl group-4-[N '-(2-aminomethyl phenyl) urea groups] phenyl acetyl]-the 2-pyrrolidinyl] methoxyl group-3-nitrobenzoic acid methyl ester (676mg; 0.99mmol) and the ethanol of 5%Pd-C (1g): acetic acid (1: 1; v/v, 30ml) solution hydrogenation 6 hours under 1 atmospheric pressure.Filter this mixture, evaporated filtrate obtains a kind of grease, makes it be alkalescence by adding saturated sodium bicarbonate.With this mixture of ethyl acetate extraction.With salt water washing extract, through dried over mgso and evaporation.Make residue recrystallization from the eluent of chloroform-ethanol-normal hexane, obtain 44 of 120mg (22%), be light yellow crystalline powder; MS (FAB) m/z 549 (M ++ 1).Embodiment 414-[[4-fluoro-1-[3-methoxyl group-4-[N '-(2-aminomethyl phenyl) urea groups] phenyl acetyl]-the 2-pyrrolidinyl] methoxyl group]-the 3-methoxybenzoic acid
Figure A0080951002071
With 4-[4-benzyloxy-1-(tert-butoxycarbonyl)-2-pyrrolidinyl] methoxyl group-3-methoxybenzoic acid ethyl ester (1.189g, 2.449mmol) and stir the mixture in room temperature under the hydrogenation of 5% Pd-C (240mg) in ethanol (10ml) spend the night.Filter this mixture to remove catalyst, vacuum concentrated filtrate obtains 4-[1-(tert-butoxycarbonyl)-4-hydroxyl-2-pyrrolidinyl] methoxyl group-3-methoxybenzoic acid ethyl ester (735.3mg, 76%), be faint yellow oily thing.(0.491ml, dichloromethane (2ml) solution of this chemical compound of dropping in dichloromethane 3.718mmol) (7.4ml) solution spend the night the mixture stirring that obtains to cold (78 ℃) DAST that stirs.This mixture of water quencher is used chloroform extraction.With salt water washing extract, through dried over sodium sulfate.Removal of solvent under reduced pressure, residue is used normal hexane through silica gel column chromatography: and ethyl acetate (3: 1, v/v) as eluant, obtain 4-[1-(the tert-butoxycarbonyl)-4-fluoro-2-pyrrolidinyl of 418.7mg (57%)] methoxyl group-3-methoxybenzoic acid ethyl ester, be a kind of grease. 1H-NMR(400MHz，CDCl 3)δ1.39(t，J＝7.3Hz，3H)，1.49(s，9H)，2.16(br?m，1H)，2.58(dd，J＝15.6，19.0Hz，1H)，3.60-3.75(m，2H)，3.91(s，3H)，3.97(t，J＝9.3Hz，1H)，4.35(q，J＝7.3Hz，2H)，4.33-4.53(m，2H)，5.25(d，J＝52.7Hz，1H)，7.04(dd，J＝7.8，56.2Hz，1H)，7.55(s，1H)，7.65(br?s，1H)；MS(FAB)m/z?398(M ++1)。
In 0 ℃, to 4-[1-(tert-butoxycarbonyl)-4-fluoro-2-pyrrolidinyl] methoxyl group-3-methoxybenzoic acid ethyl ester (482.2mg, 1.213mmol) the agitating solution of dichloromethane (10.0ml) in add TFA (1.9ml), under room temperature, mixture was stirred 2 hours.Removal of solvent under reduced pressure makes residue be alkalescence by adding the 1N sodium hydroxide, uses chloroform extraction.With salt water washing extract, through dried over sodium sulfate and concentrating under reduced pressure, obtain 4-(the 4-fluoro-2-pyrrolidinyl) methoxyl group-3-methoxybenzoic acid ethyl ester of 348.7mg (97%), be brown oil. 1H-NMR (400MHz, CDCl 3) δ 1.39 (t, J=6.8Hz, 3H), 1.97 (ddt J=1.5 5.4,14.7Hz, 1H), 2.27 (dddd, J=5.9,8.8,14.7,32.7Hz, 1H), 3.02 (ddd, J=3.9,13.1,35.2Hz, 1H), 3.36 (dd, J=12.7,21.5Hz, 1H), 3.65 (m, 1H), 3.90 (s, 3H), 4.09 (m, 1H), 4.35 (q, J=6.8Hz, 2H), 5.17,5.31 (respectively be br m, 1H), 6.90 (d, J=8.3Hz, 1H), 7.75 (d, J=2.0Hz, 1H), 7.65 (dd, J=2.0,8.3Hz, 1H); MS (FAB) m/z 298 (M ++ 1).
Under room temperature, with 3-methoxyl group-4-[N '-(2-aminomethyl phenyl) urea groups] phenylacetic acid phenyl-pentafluoride ester (404.0mg, 0.840mmol), 4-(4-fluoro-2-pyrrolidinyl) methoxyl group-3-methoxybenzoic acid ethyl ester (250.0mg, 0.840mmol), (141 μ l, 1.009mmol) mixture in DMF (4.0ml) stirred 1 hour triethylamine.With ethyl acetate diluted mixture thing, water, salt water washing are through dried over sodium sulfate.Removal of solvent under reduced pressure; residue is through silica gel column chromatography; with normal hexane-ethyl acetate (1: 3; v/v) eluting; obtain 4-[[4-fluoro-1-[3-methoxyl group-4-[N '-(2-aminomethyl phenyl) urea groups of 502mg (q.y.)] phenyl acetyl]-the 2-pyrrolidinyl] methoxyl group]-3-methoxybenzoic acid ethyl ester, be a kind of yellow oil.(60.4mg 2.520mmol), spends the night this mixture stirring under room temperature, stirs 1 day in 50 ℃ to add Lithium hydrate in the agitating solution of THF of this chemical compound (8.0ml) and water (2.0ml).With chloroform diluted mixture thing, use the 1N sodium hydroxide extraction.By adding 1N HCl acidify water-bearing layer, use chloroform extraction.With salt water washing extract, through dried over sodium sulfate.Removal of solvent under reduced pressure makes the crude product solid recrystallization from ethyl acetate-chloroform-ethanol-normal hexane that obtains obtain 45 of 294.8mg (62%), is white crystalline powder.IR(KBr)2958，2937，1687，1601，1531，1454，1419，1267，1214，1029cm -1； 1H-NMR(400MHz，DMSO-d 6)δ1.86-2.09(m，5H)，2.06(s，3H)，2.25(s，3H)，3.47-3.67(m，6H)，3.76(s，3H)，4.05-4.12(m，2H)，4.30-4.31(m，1H)，6.51(s，1H)，6.55(s，1H)，6.73-6.95(m，2H)，7.11-7.17(m，2H)，7.64(s，1H)，7.79(d，J＝7.8Hz，1H)，7.99(d，J＝7.8Hz，1H)，847(s，1H)，8.55(s，1H)；MS(FAB)m/z?566(M ++1)。C 30H 32FN 3O 71/2H 2The analytical calculation value of O: C, 62.71; H, 5.79; F, 3.31; N, 7.31.Measured value: 63.13; H, 6.17; F, 3.12; N, 7.04.Embodiment 423-acetylaminohydroxyphenylarsonic acid 4-[1-[3-methoxyl group-4-[N '-(2-aminomethyl phenyl) urea groups] phenyl acetyl]-the 2-pyrrolidinyl] methoxyl group] benzoic acid
Under room temperature; with 3-amino-4-[1-[3-methoxyl group-4-[N '-(2-aminomethyl phenyl) urea groups] phenyl acetyl]-2-pyrrolidinyl methoxyl group] benzoic acid (130mg; 0.244mmol) and DMAP (2.9mg, pyridine 0.0244mmol) (5ml) and acetic anhydride (5ml) solution stirring 2 hours.This mixture of vacuum evaporation (excessive acetic anhydride via and methylbenzene azeotropic are removed).Water is added in the residue, use chloroform extraction.Through dried over sodium sulfate extract and vacuum evaporation.Residue is used methanol through silica gel column chromatography: and chloroform (1: 15-1: 1, v/v) as eluant, obtain 46 of 29mg (21%), be white crystalline material. 1H-NMR (DMSO-d 6) δ 1.80-2.30 (m, 4H), 2.04 (s, 3H), 2.26 (s, 3H), 3.33 (s, 3H), 3.40-4.80 (m, 7H), 6.59 (s, 1H), 6.74 (d, J=8.3Hz, 1H), 6.79 (d, J=8.8Hz, 1H), 7.07-7.57 (m, 6H), 7.75 (d, J=8.8Hz, 1H), 8.07 (d, J=8.3Hz, 1H), 8.41 and 8.96 (they respectively are s, are 1H); MS (FAB) m/z575 (M ++ 1).Embodiment 433-chloro-2-[[1-[3-methoxyl group-4-[N '-(2-aminomethyl phenyl) urea groups] phenyl acetyl]-the 2-pyrrolidinyl] methoxyl group] pyridine-5-formic acid
Figure A0080951002092
In 0 ℃, (1g drips TMSCHN in the agitating solution of benzene 5.762mmol) (16ml) and methanol (4ml) to 3-chloro-2 hydroxy pyrimidine-5-formic acid 2(3.17ml 6.338mmol), stirs the mixture that obtains and to spend the night under room temperature.By adding acetic acid quencher reactant mixture, evaporate this mixture.Residue is suspended in water and collecting precipitation.With ether washing crude product solid, drying under reduced pressure, obtain 3-chloro-2 hydroxy pyrimidine-5-methyl formate of 728.1mg (67%), be white crystalline powder.IR(KBr)1655，1282，1245，769cm -1； 1H-NMR(400MHz，DMSO-d 6)δ3.79(s，3H)，8.01(s，1H)，8.06(s，1H)；MS(FAB)m/z?188(M ++1)。C 7H 6ClNO 3The analytical calculation value: C, 44.82; H, 3.22; Cl, 18.90; N, 7.47.Measured value: C, 44.74; H, 3.22; Cl, 19.00; N, 7.34.
Under room temperature, to 3-chloro-2 hydroxy pyrimidine-5-methyl formate (300mg, 1.599mmol), N-tert-butoxycarbonyl dried meat ammonia alcohol (321.9mg, 1.599mmol) and triphenyl phasphine (503mg, 1.919mmol) the agitating solution of THF (3ml) in slowly add DIAD (378 μ l, 1.919mmol).Mixture was stirred 13 hours in 70 ℃.Enriched mixture, residue is through silica gel column chromatography, with normal hexane-ethyl acetate (3: 1, v/v) as eluant, obtain 3-chloro-2-[[1-(tert-butoxycarbonyl)-2-pyrrolidinyl of 235.6mg (40%)] methoxyl group] pyridine-5-methyl formate, be a kind of light yellow oil. 1H-NMR (400MHz, CDCl 3) δ 1.46 (s, 9H), 1.87 (m, 1H), 2.05 (br s, 3H), 3.43 (br s, 2H), 3.92 (s, 3H), 4.17,4.26 (respectively be br s, 1H), 4.45-4.5 1 (m, 1H), 4.50 (s, 1H), 8.21 (s, 1H), 8.67 (d, J=2.0Hz, 1H); MS (FAB) m/z 371 (M ++ 1).
In 0 ℃, to 3-chloro-2-[[1-(tert-butoxycarbonyl)-2-pyrrolidinyl] methoxyl group] pyridine-5-methyl formate (235.6mg, 0.635mmol) the agitating solution of dichloromethane (5.0ml) in add TFA (1.0ml), under room temperature, stirred the reactant mixture that obtains 2 hours.Removal of solvent under reduced pressure makes residue be alkalescence by adding the 1N sodium hydroxide, uses chloroform extraction.Through the dried over sodium sulfate extract, concentrating under reduced pressure obtains the 3-chloro-2-[(2-pyrrolidinyl of 172.3mg (q.y.)) methoxyl group] pyridine-5-methyl formate, be light yellow oil. 1H-NMR(400MHz，CDCl 3)δ1.55-1.63(m，1H)，1.76-1.99(m，3H)，2.93-2.99(m，1H)，3.02-3.08(m，1H)，3.57-3.62(m，1H)，3.92(s，3H)，4.33(dd，J＝7.3，10.7Hz，1H)，4.44(dd，J＝4.4，10.7Hz，1H)，8.21(d，J＝2.0Hz，1H)，8.67(d，J＝2.0Hz，1H)；MS(FAB)m/z?271(M ++1)。
Under room temperature, with 3-methoxyl group-4-[N '-(2-aminomethyl phenyl) urea groups] phenylacetic acid pentafluorophenyl esters (317.0mg, 0.660mmol), 3-chloro-2-[(2-pyrrolidinyl) methoxyl group] pyridine-5-methyl formate (172.0mg, 0.635mmol), (105 μ l, 0.756mmol) mixture in DMF (2.0ml) stirred 1 hour triethylamine.With ethyl acetate diluted mixture thing, use the salt water washing, through dried over sodium sulfate.Removal of solvent under reduced pressure obtains 3-chloro-2-[[1-[3-methoxyl group-4-[N '-(2-aminomethyl phenyl) urea groups] phenyl acetyl]-the 2-pyrrolidinyl] methoxyl group] pyridine-5-methyl formate, be a kind of brown oil.(45.3mg 1.89mmol), stirs this reactant mixture 5 hours under room temperature to add Lithium hydrate in the agitating solution of THF of this chemical compound (6.0ml) and water (2.0ml).With normal hexane diluted mixture thing, use the 1N sodium hydroxide extraction.By adding 1N HCl acidify water-bearing layer, use chloroform extraction.With salt water washing extract, through dried over sodium sulfate.Removal of solvent under reduced pressure makes the crude product solid recrystallization from normal hexane-ethyl acetate-ethanol that obtains obtain 47 of 242.2mg (70%), is orange crystalline powder.mp?122-125；IR(KBr)3354，1709，1593，1535，1454，1257cm -1； 1H-NMR(400MHz，DMSO-d 6)δ1.67-2.03(m，4H)，2.50(s，3H)，3.33-3.42(m，1H)，3.52(m，2H)，3.58(d，J＝4.4Hz，1H)，3.83(s，3H)，4.27-4.31(m，2H)，4.42-4.47(m，1H)，6.73(d，J＝7.8Hz，1H)，6.87-6.95(m，3H)，7.11-7.17(m，2H)，7.79(d，J＝8.3Hz，1H)，7.99(d，J＝8.3Hz，1H)，8.14(dd，J＝2.0，8.8Hz，1H)，8.46(s，1H)，8.56(s，1H)，8.69(d，J＝2.0Hz，1H)，13.06(br?s，1H)；MS(FAB)?m/z?553(M ++1)。C 28H 29ClN 4O 6The analytical calculation value: C, 60.81; H, 5.29; N, 10.31.Measured value: 60.98; H, 5.50; N, 9.46.Embodiment 442-[[1-[4-[N '-(2-fluoro phenyl) urea groups]-3-methoxyphenyl acetyl group]-the 2-pyrrolidinyl] methoxyl group] pyridine-5-formic acid
In 0 ℃, (2g drips TMSCHN in the agitating solution of benzene 14.38mmol) (32ml) and methanol (8ml) to the 6-hydroxy niacin 2(1.97ml 3.953mmol), stirred the mixture that obtains 2 hours under room temperature.By adding acetic acid quencher mixture, concentrating under reduced pressure.Residue is suspended in water and collect solid.With ether washing crude product solid, drying under reduced pressure, obtain 2 hydroxy pyrimidine-5-methyl formate of 1.566g (71%), be the light brown crystalline powder.IR(KBr)1655，1645，1610，1433，1300，1113，777，642cm -1； 1H-NMR(400MHz，DMSO-d 6)δ3.77(s，3H)，6.37(d，J＝9.8Hz，1H)，7.99(dd，J＝2.4，9.8Hz，1H)，8.03(d，J＝2.4Hz，1H)；MS(FAB)m/z?154(M ++1)。C 7H 7NO 3The analytical calculation value: C, 54.90; H, 4.61; N, 9.15.Measured value: C, 54.89; H, 4.60; N, 9.13.
Under room temperature, to 2 hydroxy pyrimidine-5-methyl formate (1.00g, 6.529mmol), N-tert-butoxycarbonyl dried meat ammonia alcohol (1.31g, 6.529mmol) and triphenyl phasphine (2.06g, 7.836mmol) the agitating solution of THF (10ml) in add DIAD (1.54ml, 7.836mmol), the mixture that obtains was stirred 13 hours in 70 ℃.Enriched mixture, residue be through silica gel column chromatography, with normal hexane-ethyl acetate (3: 1,, obtain 2-[[1-(the tert-butoxycarbonyl)-2-pyrrolidinyl of 712.3mg (32%) v/v) as eluant] methoxyl group] pyridine-5-methyl formate, be a kind of light yellow oil. 1H-NMR(400MHz，CDCl 3)δ1.47(s，9H)，1.85-1.98(m，4H)，3.37(br?s，2H)，3.92(s，3H)，4.12-4.33(br?m，2H)，4.48(br?s，1H)，6.75(m，1H)，8.15(m，1H)，8.79(m，1H)；MS(FAB)m/z?337(M ++1)。
In 0 ℃, to 2-[[1-(tert-butoxycarbonyl)-2-pyrrolidinyl] methoxyl group] (232.3mg adds TFA (0.9ml) to pyridine-5-methyl formate in the agitating solution of dichloromethane 0.691mmol) (4.6ml), stirred reaction mixture is 2 hours under room temperature.Removal of solvent under reduced pressure makes residue be alkalescence by adding the 1N sodium hydroxide, uses the chloroform extraction aqueous solution, uses the salt water washing, through dried over sodium sulfate.Solvent evaporated under reduced pressure obtains 2-(2-pyrrolidinyl) methoxypyridine-5-methyl formate of 146.2mg (90%), is a kind of oil. 1H-NMR(400MHz，CDCl 3)δ1.49-1.58(m，1H)，1.72-2.18(m，3H)，2.92-3.05(m，2H)，3.50-3.57(m，1H)，3.91(s，3H)，4.23(dd，J＝8.0，10.7Hz，1H)，4.38(dd，J＝4.4，10.3Hz，1H)，6.78(d，J＝8.8Hz，1H)，8.15(dd，J＝2.4，8.8Hz，1H)8.80(d，J＝2.4Hz，1H)；MS(FAB)m/z237(M ++1)。
Under room temperature, with 4-[N '-(2-fluoro phenyl) urea groups]-3-methoxyl group-phenylacetic acid pentafluorophenyl esters (314.8mg, 0.650mmol), the 2-[(2-pyrrolidinyl) methoxyl group] pyridine-5-methyl formate (146.2mg, 0.619mmol), (103 μ l, 0.743mmol) mixture in DMF (1.5ml) stirred 1 hour triethylamine.With ether diluted mixture thing, use the salt water washing, through dried over sodium sulfate.Removal of solvent under reduced pressure obtains 2-[[1-[4-[N '-(2-fluoro phenyl) urea groups]-3-methoxyphenyl acetyl group]-the 2-pyrrolidinyl] methoxyl group] pyridine-5-methyl formate, be faint yellow crude product oil.
(44.5mg 1.857mmol), stirs this reactant mixture 17 hours under room temperature to add Lithium hydrate in the agitating solution of THF of this chemical compound (6.0ml) and water (2.0ml).With normal hexane diluted mixture thing, make and be alkalescence by adding the 1N sodium hydroxide.With 1N HCl acidify water-bearing layer, use chloroform extraction.With salt water washing extract, through dried over sodium sulfate.Removal of solvent under reduced pressure makes the crude product solid recrystallization from normal hexane-ethyl acetate-ethanol that obtains obtain 48 of 202.5mg (63%), is white crystalline powder.IR(KBr)1602，1537，1456，1265，752cm -1； 1H-NMR(400MHz，DMSO-d 6)δ1.67-2.03(m，4H)，2.50(s，3H)，3.33-3.42(m，1H)，3.52(m，2H)，3.58(d，J＝4.4Hz，1H)，3.83(s，3H)，4.27-4.31(m，2H)，4.42-4.47(m，1H)，6.73(d，J＝7.8Hz，1H)，6.87-6.95(m，3H)，7.11-7.17(m，2H)，7.79(d，J＝8.3Hz，1H)，7.99(d，J＝8.3Hz，1H)，8.14(dd，J＝2.0，8.8Hz，1H)，8.46(s，1H)，8.56(s，1H)，8.69(d，J＝2.0Hz，1H)，13.06(br?s，1H)；MS(FAB)m/z?523(M ++1)。C 27H 27FN 4O 61/2H 2The analytical calculation value of O: C, 61.01; H, 5.31; N, 10.54.Measured value: C, 61.52; H, 5.39; N, 10.01.Embodiment 454-[1-[3-methoxyl group-4-[N '-(2-aminomethyl phenyl) urea groups] phenyl acetyl]-2-pyrrolidinyl methyl]-]-piperazinyl acetic acid
Figure A0080951002131
Under room temperature, to the 1-benzyl diethylenediamine (5g, 28.4mmol) and potassium carbonate (5.89g, add in the stirring suspension solution of DMF 42.6mmol) (30ml) bromoethyl acetate (4.74g, 28.4mmol).Other 3 hours of the mixture that stirring obtains.(300ml) dilutes this mixture with ethyl acetate, and (2 * 100ml) washings are through dried over mgso and evaporation with saline.Residue is through silica gel column chromatography, with chloroform-ethanol (10: 1, v/v) as eluant, obtain 4-benzyl-1-piperazinyl ethyl acetate of 7.45g (q.y.), be a kind of yellow oil. 1H-NMR(CDCl 3)δ1.27(3H，t，J＝7.3Hz)，2.88-2.96(8H，m)，3.20(2H，s)，3.52(2H，s)，4.18(2H，q，J＝7.3Hz)，7.22-7.32(5H?m)。
With 4-benzyl-1-piperazinyl ethyl acetate (2.00g, 7.62mmol) and the acetic acid of 5%Pd/C (2g): ethanol (1: 1, the 40ml) agitating solution in hydrogenation 8 hours under 1 atmospheric pressure.Filter this mixture, vacuum concentrated filtrate.Make residue be alkalescence by adding saturated sodium bicarbonate.(2 * 200ml) extract with chloroform.Through dry extract that merges of potassium carbonate and evaporation, obtain the 1-piperazinyl ethyl acetate of 1.16g (88%), be yellow oil. 1H-NMR(CDCl 3)δ1.26-1.30(3H，m)，1.67(1H，br?s)，2.55(4H，m)，2.92-2.96(4H，m)，3.19-3.20(2H，m)，4.16-4.22(2H?m)。
Under room temperature, (1.00g, 5.02mmol) (864mg, methanol 5.02mmol): (10: 1, v/v added NaBH in agitating solution 11ml) to acetic acid with 1-piperazinyl ethyl acetate to N-Boc-L-dried meat ammonia alcohol 3CN (664mg, 10.0mmol).After stirring is spent the night, in this mixture impouring frozen water (100ml), make and be alkalescence by adding sodium bicarbonate.(2 * 200ml) extract this mixture with chloroform.Through dry extract that merges of sodium carbonate and evaporation.Residue is through silica gel column chromatography, with chloroform-ethanol (10: 1,, obtain 4-[1-(the tert-butoxycarbonyl)-2-pyrrolidinyl methyl of 1.20g (67%) v/v) as eluant]-1-piperazinyl ethyl acetate, be a kind of water white oil. 1H-NMR (CDCl 3) δ 1.27 (3H, t, J=7.3Hz), 1.46-1.47 (9H, m), 1.79-3.96 (total 19H, a plurality of m), 4.19 (2H, q, J=7.3Hz).
With 4-[1-(tert-butoxycarbonyl)-2-pyrrolidinyl methyl]-(1.20g, 3.38mmol) mixture in TFA (5ml) and dichloromethane (5ml) stirs and spends the night 1-piperazinyl ethyl acetate.Except that after desolvating, make residue be alkalescence by adding saturated sodium bicarbonate.(2 * 200ml) extract this mixture with chloroform.Through dry extract that merges of sodium carbonate and evaporation, obtain 4-(2-pyrrolidinyl the methyl)-1-piperazinyl ethyl acetate of 386mg (45%), be yellow oil.MS(FAB)256(M ++1)。
To 4-(2-pyrrolidinyl methyl)-1-piperazinyl ethyl acetate (380mg, 1.49mmol) and 3-methoxyl group-4-[N '-(2-aminomethyl phenyl) urea groups] phenylacetic acid (468mg, 1.49mmol) the agitating solution of DMF (10ml) in add EDC.HCl (428m, 2.24mmol), HOBt and DMAP (cat.).After stirring was spent the night, (300ml) diluted this mixture with ethyl acetate, and (2 * 200ml) washings are through the magnesium carbonate drying with saline.Except that after desolvating; residue is through silica gel column chromatography, with chloroform-ethanol (9: 1, v/v) as eluant; obtain 4-[1-[3-methoxyl group-4-[N '-(2-aminomethyl phenyl) urea groups of 257mg (31%)] phenyl acetyl]-2-pyrrolidinyl methyl]-1-piperazinyl ethyl acetate, be yellow foam thing. 1H-NMR (CDCl 3) δ 1.24-1.29 (3H, m), 1.69-4.24 (being total to 29H, a plurality of m), 6.41 (1H, m), 6.81 (2H, m), 7.13-7.26 (4H, m), 7.52 (1H, d, J=7.3Hz), 8.04 (1H, d, J=8.3Hz).
To 4-[1-[3-methoxyl group-4-[N '-(2-aminomethyl phenyl) urea groups] phenyl acetyl]-2-pyrrolidinyl methyl]-(250mg adds 0.25N sodium hydroxide (3.6ml) to 1-piperazinyl ethyl acetate in the agitating solution of THF 0.453mmol) (4ml).The mixture stirring that obtains is spent the night.By adding 1N HCl this mixture is adjusted to pH 7.5, (4: 1,3 * 100ml) extracted with chloroform-methanol.The extract that merges through dried over mgso also evaporates.Make crude product solid recrystallization from chloroform-methanol-normal hexane, obtain 49 of 40mg (17%), be the colourless crystallization powder.Mp 160-170 ℃; 1H-NMR (DMSO-d 6) δ 1.74-4.08 (being total to 27H, a plurality of m), 6.73 (1H, d, J=7.8Hz), 6.87 (1H, s), 6.93 (1H, t, J=7.8Hz), 7.11-7.17 (2H, m), 7.79 (1H, d, J=7.8Hz), 8.00 (1H, dd, J=7.8,2.4Hz), 8.47 (1H, s), 8.56 (1H, s); MS (FAB) 524 (M ++ 1).C 28H 37N 5O 5HClH 2The analytical calculation value of O: C, 58.17; H, 6.97; N, 12.11.Measured value: C, 58.26; H, 7.26; N, 11.53.Embodiment 46
To 4-aminophenyl acetic acid (10g, 66mmol) at 1: 1 dichloromethane: add in the suspension in the acetone (100ml) Carbimide. neighbour-tolyl ester (8.8g, 66mmol).This mixture heated to refluxing 4 hours, is formed white precipitate this moment.Filtering precipitate is with 1: 1 a large amount of dichloromethane: this solid of washing with acetone.Make this solid recrystallization with hot methanol, vacuum drying obtains the required 4-of 14.1g (yield 75%) (neighbour-tolyl urea groups) phenylacetic acid 50.Embodiment 47
To 2-amino-4-thiazolyl acetic acid (4g, 25mmol) at 1: 1 dichloromethane: add in the suspension in the acetone (100ml) Carbimide. neighbour-tolyl ester (3.5g, 26mmol).This mixture heated to refluxing 8 hours, is formed yellow mercury oxide this moment.Filtering precipitate is with 1: 1 a large amount of dichloromethane: this solid of washing with acetone.Make this solid recrystallization with hot methanol, vacuum drying obtains the required 2-of 4.8g (yield 66%) (neighbour-tolyl urea groups)-4-thiazolyl acetic acid 51.Embodiment 484-[[1-[3-methoxyl group-4-[N '-(2-aminomethyl phenyl) urea groups] phenyl acetyl]-2-pyrrolidinyl methylamino] benzoic acid
(1.52g, 10.04mmol) (3.00g, 15.06mmol) stirring the mixture in toluene (30ml) heated 3 hours under refluxing with 1-tert-butoxycarbonyl dried meat ammonium aldehyde with the 4-Methyl anthranilate.After being cooled to room temperature, vacuum evaporating solvent.This solid is dissolved in methanol (27ml) and the acetic acid (3ml), then with NaBH 3(1.33g 20.08mmol) adds in this mixture CN, under room temperature the mixture stirring that obtains is spent the night.Water quencher reactant mixture, removal of solvent under reduced pressure.Water is added in the residue, use ethyl acetate extraction.Water, salt water washing extract are through dried over sodium sulfate.Removal of solvent under reduced pressure, residue be through silica gel column chromatography, with normal hexane-ethyl acetate (3: 1,, obtain 4-[1-(the tert-butoxycarbonyl)-2-pyrrolidinyl methylamino of 2.17g (65%) v/v) as eluant] benzoate, be a kind of light yellow oil. 1H-NMR (400MHz, CDCl 3) δ 1.48 (s, 9H), 1.51-2.09 (m, 4H), 3.05-3.07 and 3.43-3.48 (br m, 1H), 3.18 (brs, 1H), 3.36 (br s, 2H), 3.84 (s, 1H), 4.06-4.08,4.20-4.24 (respectively is br m, 1H), 6.49-6.65 (m, 2H), 7.84 (d, J=8.3Hz, 2H); MS (FAB) m/z335 (M ++ 1).
In 0 ℃, to 4-[1-(tert-butoxycarbonyl)-2-pyrrolidinyl methylamino] (2.17g adds TFA (8.7ml) to essence of Niobe in the agitating solution in dichloromethane 6.490mmol) (44ml), under room temperature the mixture stirring that obtains is spent the night.Removal of solvent under reduced pressure is with 1N naoh treatment residue.With this mixture of dichloromethane extraction.With salt water washing extract, through dried over sodium sulfate, removal of solvent under reduced pressure obtains 4-(2-pyrrolidinyl the methylamino)-essence of Niobe of 1.34g (88%), is brown oil, and it can need not to be further purified and be used for subsequent reaction.
With top 4-(2-pyrrolidinyl methylamino)-essence of Niobe (397.8mg, 1.69mmol), 3-methoxyl group-4-[N '-(2-aminomethyl phenyl) urea groups] phenylacetic acid (587.1mg, 1.87mmol), EDC (HCl) (486mg, 2.54mmol), HOBt (23mg, 0.17mmol) and DMAP (21mg, 0.17mmol) mixture in DMF (4ml) stirs under room temperature and spends the night.Dilute this mixture with ethyl acetate, use the salt water washing, through dried over sodium sulfate.Removal of solvent under reduced pressure.Residue is through silica gel column chromatography; with chloroform-methanol (50: 1; v/v) as eluant; obtain 4-[1-[3-methoxyl group-4-[N '-(2-aminomethyl phenyl) urea groups of 882mg (98%)] phenyl acetyl]-2-pyrrolidinyl methylamino] essence of Niobe; be brown amorphous solid, it can need not to be further purified and be used for subsequent reaction.
To 4-[1-[3-methoxyl group-4-[N '-(2-aminomethyl phenyl) urea groups] phenyl acetyl]-2-pyrrolidinyl methylamino] essence of Niobe (882mg; 1.662mmol) THF (18ml) and the agitating solution of methanol (5.0ml) in add the 1N sodium hydroxide (5.0ml, 5.000mmol).Mixture was heated 3 days under refluxing.Concentrate this mixture.Dichloromethane extraction is handled and used to residue with 1N HCl.With salt water washing extract, through dried over sodium sulfate and vacuum evaporation.Make solid recrystallization from normal hexane-diisopropyl ether-chloroform-methanol, obtain 52 of 180.5mg (21%), be faint yellow amorphous solid.IR (KBr) 1604,1535,1511,1454,1255,1224,1174cm -1 1H-NMR (400MHz, DMSO-d 6) δ 1.79-1.99 (br m, 4H), 2.25 (s, 3H), 2.90-2.94 (m, 1H), 3.35-3.62 (m, 6H), 3.87 (s, 3H), 4.12-4.15 (br s, 1H), and 6.63-6.78 (m, 4H), 6.89-6.95 and 7.11-7.17 (respectively are m, 3H), 7.65 (d, J=8.3Hz, 2H), 7.80 (d, J=8.3Hz, 1H), 8.02 (d, J=8.3Hz, 1H), 8.47 (s, 1H), 8.57 (s, 1H), 12.0 (br s, 1H); MS (FAB) m/z 517 (M ++ 1).C 29H 32N 4O 51H 2The analytical calculation value of O: C, 65.15; H, 6.41; N, 10.48.Measured value: C, 65.45; H, 6.33; N, 10.02.Embodiment 494-[N-[1-[3-methoxyl group-4-[N '-(2-aminomethyl phenyl) urea groups] phenyl acetyl]-2-pyrrolidinyl methyl]-the N-methylamino] benzoic acid
To 4-[N-(2-pyrrolidinyl) methylamino] essence of Niobe (600mg, 1.794mmol), 37%-formaldehyde (1.79ml, 23.32mmol) and NaBH 3(368mg, (0.205ml 3.588mmol), stirs the mixture that obtains 2 hours under room temperature CN to drip acetic acid in the mixture of acetonitrile 5.561mmol) (6.0ml).By adding saturated sodium bicarbonate quencher reactant mixture, use ethyl acetate extraction.With salt water washing organic layer, through dried over sodium sulfate.Removal of solvent under reduced pressure, residue is through silica gel column chromatography, with normal hexane-ethyl acetate (3: 1, v/v) as eluant, obtain 4-[N-[1-(the tert-butoxycarbonyl)-2-pyrrolidinyl methyl of 645mg (100%)]-the N-methylamino] essence of Niobe, be a kind of water white oil. 1H-NMR(400MHz，CDCl 3)δ1.50(s，9H)，1.76-1.91(m，4H)，3.07(s，3H)，3.15-3.43(m，3H)，3.67-3.71(m，1H)，3.85(s，3H)，4.11-4.17(m，1H)，4.37(s，1H)，6.75(d，J＝8.3Hz，2H)，7.89(d，J＝8.8Hz，2H)；MS(FAB)m/z349(M ++1)。
In 0 ℃, to 4-[N-[1-(tert-butoxycarbonyl)-2-pyrrolidinyl methyl]-the N-methylamino] essence of Niobe (645mg, 1.794mmol) dichloromethane (6.5ml) in agitating solution in add TFA (1.3ml), under room temperature, mixture stirred and spends the night.Removal of solvent under reduced pressure is handled residue with the 1N sodium hydroxide solution.With this mixture of dichloromethane extraction.With salt water washing extract, through dried over sodium sulfate, the concentrating under reduced pressure solvent obtains 4-[N-(2-pyrrolidinyl) methyl-N-methyl of 363.2mg (82%)] Methyl anthranilate, be yellow oil, it can need not to be further purified and be used for subsequent reaction.
With 4-[N-(2-pyrrolidinyl) methyl-N-methyl) Methyl anthranilate (191.8mg, 0.772mmol), 3-methoxyl group-4-(N '-2-aminomethyl phenyl urea groups) phenylacetic acid (258.1mg, 0.811mmol), EDC (hydrochlorate) (221.9mg, 1.158mmol), HOBt (10.0mg, 0.077mmol) and DMAP (9.4mg, 0.077mmol) mixture in DMF (2.0ml) stirred 3 hours under room temperature.Dilute this reactant mixture with ether, use the salt water washing, through dried over sodium sulfate.Removal of solvent under reduced pressure; obtain 4-[N-[1-[3-methoxyl group-4-[N '-(2-aminomethyl phenyl) urea groups of 482.5mg] phenyl acetyl]-2-pyrrolidinyl methyl]-the N-methylamino] essence of Niobe; be white amorphous powder, it can need not to be further purified and be used for subsequent reaction.
To 4-[N-[1-[3-methoxyl group-4-[N '-(2-aminomethyl phenyl) urea groups] phenyl acetyl]-2-pyrrolidinyl methyl]-the N-methylamino] add in the agitating solution of THF (5.0ml) of essence of Niobe the 1N sodium hydroxide (6.2ml, 6.2mmol).Mixture was heated 3 days under refluxing.This reactant mixture of vacuum concentration.Residue is used dichloromethane extraction with 1N HCl neutralization.With saturated ammonium chloride, salt water washing extract, through dried over sodium sulfate and vacuum evaporation.Make crude product solid recrystallization from normal hexane-chloroform-methanol-isopropyl ether, obtain 53 of 102.8mg (25%, two step), be faint yellow amorphous solid.mp?144-146；IR(KBr)3325，1600，1529，1454，1284，1257，1184cm -1； 1H-NMR(400MHz，DMSO-d 6)δ1.73-1.91(m，3H)，2.03-2.11(m，1H)，3.03(s，3H)，3.16(dd，J＝9.3，14.2Hz，1H)，3.37-3.60(m，4H)，3.76-3.80(m，1H)，3.86(s，3H)，4.25(br?s，1H)，6.75(dd，J＝1.5，8.3Hz，1H)，6.86(d，J＝1.5Hz，1H)，6.90(d，J＝8.8Hz，2H)，6.95-7.01(m，1H)，7.12(t，J＝7.8Hz，1H)，7.20-7.25(m，1H)，7.73(d，J＝8.8Hz，2H)，8.01(d，J＝7.8Hz，1H)，8.16-8.20(m，1H)，8.73(s，1H)，9.19(d，J＝2.0Hz，1H)，12.0(br?s，1H)；MS(FAB)m/z?535(M ++1)。C 29H 31FN 4O 51/2H 2The analytical calculation value of O: C, 64.08; H, 5.93; N, 10.31, F, 3.49.Measured value: C, 64.17; H, 5.84; N, 10.06; F, 3.26.Embodiment 504-[N-[1-[4-[N '-(2-fluoro phenyl) urea groups]-3-methoxyphenyl acetyl group]-2-pyrrolidinyl methyl]-the N-methylamino]-the 3-nitrobenzoic acid To 4-fluoro-3-nitrobenzoic acid methyl ester (1.58g, 4.666mmol) and [I-(tert-butoxycarbonyl)-2-pyrrolidinyl] methylamine (500mg, 2.333mmol) add potassium carbonate (967mg in the mixture in DMF (8.0ml), 6.999mmol), the mixture that stirring obtains under room temperature 3 hours.Use the ethyl acetate diluted reaction mixture, wash with water, through dried over sodium sulfate.Removal of solvent under reduced pressure, residue is through silica gel column chromatography, with normal hexane-ethyl acetate (3: 1, v/v) as eluant, obtain 4-[N-[1-(the tert-butoxycarbonyl)-2-pyrrolidinyl methyl of 834.9mg (91%)]-the N-methyl] amino-3-nitrobenzoic acid methyl ester, be a kind of light yellow oil, it can need not to be further purified and be used for subsequent reaction.
Under room temperature, add TFA (1.7ml) in the ice-cooling solution in the dichloromethane of grease upward (8.3ml), under room temperature, the reactant mixture stirring is spent the night.Removal of solvent under reduced pressure.With 1N naoh treatment residue, use chloroform extraction.With salt water washing extract,, obtain 4-[N-(2-pyrrolidinyl the methyl)-N-methyl of 553.6mg (90%) through dried over sodium sulfate and reduction vaporization] amino-3-nitrobenzoic acid methyl ester, be faint yellow oily thing. 1H-NMR(400MHz，CDCl 3)δ1.31-1.40(m，1H)，1.74-2.05(m，4H)，2.73-2.79(m，1H)，2.81-2.99(m，1H)，2.94(s，3H)，3.29-3.55(m，2H)，3.89(s，3H)，7.14(d，J＝9.3Hz，1H)，7.98(dd，J＝2.0，8.8Hz，1H)，8.42(d，J＝2.0Hz，1H)；MS(FAB)m/z294(M ++1)。
With 3-methoxyl group-4-[N '-(2-fluoro phenyl) urea groups] phenylacetic acid (630.0mg, 1.979mmol), 3-nitro-4-[N-(2-pyrrolidinyl) methyl-N-methylamino] essence of Niobe (553.0mg, 1.885mmol), EDC (hydrochlorate) (542.0mg, 2.827mmol), HOBt (25.5mg, 0.189mmol) and DMAP (23.1mg, 0.189mmol) mixture in DMF (5.0ml) stirred 2 hours under room temperature.Dilute this mixture with ether, use the salt water washing, through dried over sodium sulfate.Removal of solvent under reduced pressure; residue is through silica gel column chromatography; with chloroform-methanol (30: 1; v/v) as eluant; obtain 1.18g (100%) 4-[N-[1-(3-methoxyl group-4-[N '-(2-fluoro phenyl) urea groups] phenyl acetyl]-2-pyrrolidinyl methyl]-the N-methylamino]-3-nitrobenzoic acid methyl ester; be yellow foam thing, it can need not to be further purified and be used for subsequent reaction.
To 4-[N-[1-[3-methoxyl group-4-[N '-(2-fluoro phenyl) urea groups] phenyl acetyl]-2-pyrrolidinyl methyl]-the N-methylamino]-3-nitrobenzoic acid methyl ester (2.50mg; 0.421mmol) the agitating solution of THF (3.0ml) in add the 1N sodium hydroxide (1.5ml, 1.500mmol).With mixture heated overnight under refluxing.After the cooling, this mixture is concentrated into small size.Residue is handled with 1N HCl, uses chloroform extraction.With salt water washing extract, through dried over sodium sulfate and vacuum evaporation.Make crude product solid recrystallization from normal hexane-E-C-methanol, obtain 54 of 194.9mg (80%), be yellow amorphous solid.IR(KBr)1685，1610，1529，1454，1284，1259，1228cm -1； 1H-NMR(400MHz，DMSO-d 6)δ1.63-1.91(m，3H)，2.04-2.07(br?s，1H)，2.60(br?s，1H)，2.80(s，1H)，2.99(s，2H)，3.05-3.10(m，1H)，3.32-3.58(m，3H)，3.76-3.81(m，1H)，3.81(s，3H)，4.25(br?s，1H)，6.68(t，J＝3.9Hz，1H)，6.71(d，J＝8.8Hz，1H)，6.81-6.96(m，1H)，7.07(t，J＝7.3Hz，1H)，7.17(dd，J＝7.8，9.8Hz，1H)，7.48(t，J＝7.8Hz，1H)，7.85(d，J＝8.8Hz，1H)，7.97(t，J＝8.8Hz，1H)，8.11-8.20(m，2H)，8.68(s，1H)，9.14(s，1H)，12.8(br?s，1H)；MS(FAB)m/z580(M ++1)。C 29H 30FN 5O 71/4H 2The analytical calculation value of O: C, 59.63; H, 5.26; N, 11.99, F, 3.25.Measured value: C, 59.68; H, 5.34; N, 11.80, F, 3.21.Embodiment 513-amino-4-[N-methyl-[1-[4-[N '-(2-fluoro phenyl) urea groups]-3-methoxyl group-phenyl acetyl]-2-pyrrolidinyl methyl]-the N-methylamino] benzoic acid
Figure A0080951002211
With 4-[N-[1-[4-[N '-(2-fluoro phenyl) urea groups]-3-methoxyl group-phenyl acetyl]-2-pyrrolidinyl methyl]-the N-methylamino]-(901.0mg, the solution of the stirring in methanol 1.518mmol) (18.0ml) spends the night through 5%Pd-C (1.35g) hydrogenation under 45psi 3-nitrobenzoic acid methyl ester.Filter this mixture, vacuum concentrated filtrate.Make residue be alkalescence with the 1N sodium hydroxide solution, use chloroform extraction.With salt water washing extract, through dried over sodium sulfate, removal of solvent under reduced pressure.Residue is through silica gel column chromatography; with chloroform-methanol (24: 1; v/v) as eluant; obtain 3-amino-4-[N-[1-[4-[N '-(2-fluoro phenyl) urea groups of 283.7mg (48%)]-3-methoxyl group-phenyl acetyl]-2-pyrrolidinyl methyl]-the N-methylamino] essence of Niobe; be brown amorphous solid, it can need not to be further purified and be used for subsequent reaction.
(1.5ml 1.500mmol), spends the night this mixture backflow to add the 1N sodium hydroxide solution upward in the agitating solution of the THF of chemical compound (3.0m1).Concentrate this mixture, handle, use chloroform extraction with 1N HCl.With salt water washing extract, through dried over sodium sulfate and reduction vaporization.Make solid recrystallization from normal hexane-E-C-methanol, obtain 55 of 179.8mg (65%), be white amorphous solid.IR(KBr)1614，1601，1537，1454，1228，1219，1184cm -1； 1H-NMR(400MHz，DMSO-d 6)δ1.60-2.20(m，4H)，2.61-2.68(m，1H)，2.89(s，3H)，3.13-3.18(m，1H)，3.40-3.61(m，4H)，3.85(s，3H)，4.01(br?m，1H)，4.93(br?s，2H)，6.50-7.31(m，8H)，8.01(dd，J＝2.9，8.3Hz，1H)，8.18(t，J＝8.3Hz，1H)，8.71(s，1H)，9.17(d，J＝1.5Hz，1H)，12.3(br?s，1H)；MS(FAB)m/z550(M ++1)。C 29H 32FN 5O 51/4H 2The analytical calculation value of O: C, 62.86; H, 5.91; N, 12.64, F, 3.43.Measured value: C, 62.71; H, 6.00; N, 12.39, F, 3.16.Embodiment 524-[1-[4-[N '-(2-aminomethyl phenyl) urea groups] phenyl acetyl]-the 2-pyrrolidinyl] methylamino] benzoic acid
Figure A0080951002221
Under room temperature, to the 4-[(2-pyrrolidinyl) methylamino] essence of Niobe (220mg, 0.94mmol), 4-[N '-(2-aminomethyl phenyl) urea groups] phenylacetic acid (285mg, 0.94mmol), 4-DMAP (140mg, 1.13mmol) and the stirring the mixture of the DMF (7ml) of the HOBT of catalytic amount in add EDCHCl (220mg, 1.13mmol).Under room temperature, the mixture that obtains was stirred 20 hours.This mixture is poured in the frozen water.Collect solid, wash with water also air-dry.The crude product solid is through silica gel (20ml) column chromatography purification; with chloroform-ethyl acetate (3: 1; v/v) to chloroform-ethanol (9: 1; v/v) as eluant; obtain 4-[1-[4-[N '-(2-aminomethyl phenyl) urea groups] phenyl acetyl]-the 2-pyrrolidinyl] methylamino] essence of Niobe (400mg; 85%), is a kind of slurry. 1H-NMR (CDCl 3) δ 1.75-2.05 (a plurality of m, 4H), 2.24 (s, 3H), 3.18 and 3.27 (respectively is m, respectively be 1H), 3.51 (m, 2H), 3.60 (s, 2H), 3.83 (s, 3H), 4.52 (m, 1H), 6.52 (m, 3H), 6.81 (s, 1H), and 7.11-7.25 (a plurality of m, 7H), 7.53 (m, 1H), 7.81 (d, J=8.8Hz, 2H).
In 60-70 ℃; with 4-[1-[4-[N '-(2-aminomethyl phenyl) urea groups] phenyl acetyl]-the 2-pyrrolidinyl] methylamino] essence of Niobe (280mg; 0.56mmol) THF (3ml) and the 0.25N sodium hydroxide (6.8ml, the mixture in 1.75mmol) stirred 3 hours.After the cooling, in this mixture impouring ice-1N HCl (3ml).Collect solid, wash with water also air-dry.Make crude product crystalline material recrystallization from chloroform-ethanol-IPE, obtain 56 (180mg, 66%), be thin needle.MW 486.56 IR (KBr) n 3367,3294,1712,1606,1539cm -1 1H-NMR (CDCl 3-DMSO-d 6) δ 1.80-2.05 (a plurality of m, 4H), 2.26 (s, 3H), 2.94 (m, 1H), 3.38 and 3.56 (a plurality of m, 3H), 3.57 (s, 2H), 4.23 (m, 2H), 6.48 (br s, 1H), 6.69 (d, J=8.8Hz, 2H), 6.91 (t, J=7Hz, 1H), 6.91 (m, 4H), 7.39 (d, J=8.3Hz, 2H), 7.66 (d, J=8.8Hz, 2H), 7.80 (m, 2H), 8.88 (s, 1H), 11.76 (s, 1H); MS (FAB) m/z 487 (M ++ 1).C 28H 30N 4O 40.75 * H 2The analytical calculation value of O: C, 67.24; H, 6.45; N, 11.20.Measured value: C, 67.13; H, 6.32; N, 11.01.Embodiment 534-[1-[4-[N '-(2-chlorophenyl) urea groups]-3-methoxyphenyl acetyl group]-(2S)-and pyrrolidinyl] methoxyl methyl benzoate
Figure A0080951002231
4-[1-[4-[N '-(2-chlorophenyl) urea groups]-3-methoxyphenyl acetyl group]-(2S)-and pyrrolidinyl] methoxybenzoic acid
(15.1g adds in the agitating solution of dioxane 149.5mmol) (100ml) (Boc) to the 2S-pyrrolidine carbinol 2O (32.6g, dioxane 164.4mmol) (100ml) solution.Under room temperature, reactant mixture was stirred 18 hours and concentrating under reduced pressure.Residue is through purification by silica gel column chromatography, and (1: 5, (31.6g quantitatively), was a kind of colorless oil to methanol, to obtain (1-tert-butoxycarbonyl-(2S)-pyrrolidinyl) v/v) as eluant with ethyl acetate-hexane. 1H-NMR(CDCl 3)δ1.47(s，9H)，1.60-2.00(m，3H)，3.25-3.70(4H，m)，3.92-4.00(m，1H)。
In 0 ℃, to (1-tert-butoxycarbonyl-(2S)-pyrrolidinyl) methanol (4.02g, 20.0mmol), the 4-methyl hydroxybenzoate (3.04g, 20.0mmol) and triphenyl phasphine (6.28g, 24.0mmol) the agitating solution of THF (50ml) in add DIAD (4.85g, 24.0mmol).Under room temperature, this reactant mixture was stirred 18 hours.This mixture of vacuum concentration.Residue is through purification by silica gel column chromatography, with normal hexane-ethyl acetate (5: 1, v/v) as eluant, obtain 4-(1-tert-butoxycarbonyl-(2S)-pyrrolidinyl) methoxyl methyl benzoate (5.4g, 81%), be a kind of faint yellow oily thing. 1H-NMR(CDCl 3)δ1.47(s，9H)，1.88-2.04(m，4H)，3.41(m，2H)，3.91(s，3H)，3.90-3.92(m，1H)，4.11-4.16(m，2H)，6.94(d，J＝8.6Hz，2H)，7.94(d，J＝8.3Hz，2H)。
In 0 ℃, (2.1g adds TFA (6.0ml) in the agitating solution of dichloromethane 6.27mmol) (9.0ml) to 4-(1-tert-butoxycarbonyl-2S-pyrrolidinyl) methoxyl methyl benzoate.Under room temperature, this reactant mixture was stirred 0.5 hour.This mixture of vacuum concentration.Saturated sodium bicarbonate is added in the residue, use dichloromethane extraction.With salt water washing extract, through dried over sodium sulfate and vacuum concentration.The crude product product can need not to be further purified and be used for subsequent reaction.Under 0 ℃, to crude product (470mg, 2.0mmol), 4-[N '-(2-chlorophenyl) urea groups]-3-methoxybenzene guanidine-acetic acid (669mg, 2.0mmol), HOBt (405mg, 3.0mmol) and triethylamine (554ml, 4.0mmol) THF (10.0ml) and the agitating solution of MeCN (10.0ml) in add EDCHCl (576mg, 3.0mmol).Under room temperature, reactant mixture was stirred 16 hours vacuum concentration.Water is added in the residue, use ethyl acetate extraction.Wash extract with saturated sodium bicarbonate.Through dried over sodium sulfate and vacuum concentration.Residue is through purification by silica gel column chromatography, with normal hexane-ethyl acetate (1: 4, v/v) as eluant, obtain 57 (900mg, 82%), be colorless oil.MW?552.02 1H-NMR(CDCl 3)δ2.04-2.10(m，4H)，3.51-3.70(m，6H)，3.87(s，3H)，4.11-4.18(m，2H)，6.77-6.88(m，4H)，7.23-7.34(m，4H)，7.91-7.96(m，2H)，8.17-8.19(m，1H)。
This mixture was stirred 24 hours in 70 ℃.This mixture of vacuum concentration adds water wherein, with 1N HCl neutralization.The solid that collection obtains washes with water, and vacuum drying obtains 58 (640mg, 94%), is white crystalline solid.MW?537.99?mp?126-130℃；IR(KBr)3324，2938，2877，1604，1533，1249，1166，750cm -1； 1H-NMR(DMSO-d 6)δ1.93-2.05(m，4H)，3.52-3.61(m，5H)，3.82(s，3H)，3.99-4.01(m，2H)，4.18-4.20(m，1H)，4.29(m，1H)，6.74-6.76(d，1H，J＝8.3Hz)，6.87(s，1H)，6.99-7.04(m，3H)，7.25-7.29(m，1H)，7.41-7.43(d，1H，J＝8.1Hz)，7.86-7.91(m，2H)，7.95-7.97(m，1H)，8.09-8.11(d，1H，J＝8.3Hz)，8.87-8.92(m，1H)；MS(FAB)?m/z?538(M ++1)。C 28H 28N 3O 60.5H 2The analytical calculation value of O: C, 61.48; H, 5.34; N, 7.68; Cl, 6.48.Measured value: C, 61.46; H, 5.36; N, 7.62; Cl, 6.50.Sodium salt for 58: C 28H 27N 3O 6Na1.5H 2The analytical calculation value of O: C, 57.29; H, 5.15; N, 7.16.Measured value: C, 57.48; H, 5.04; N, 6.99.Embodiment 544-[1-[4-[N '-(2-bromo phenyl) urea groups]-3-methoxyphenyl acetyl group]-(2S)-and pyrrolidinyl] methoxybenzoic acid
In 0 ℃, to 4-(2S-pyrrolidinyl) methoxyl methyl benzoate (470mg, 2.0mmol), 4-[N '-(2-bromo phenyl) urea groups]-3-methoxybenzene guanidine-acetic acid (758mg, 2.0mmol), HOBt (405mg, 3.0mmol) and triethylamine (554ml, 3.0mmol) THF (10.0ml) and the agitating solution of MeCN (10.0ml) in add EDCHCl (576mg, 3.0mmol).Under room temperature, reactant mixture was stirred 16 hours vacuum concentration.Water is added in the residue, use ethyl acetate extraction.Wash extract with saturated sodium bicarbonate, then through dried over sodium sulfate and vacuum concentration.Residue is through purification by silica gel column chromatography; with normal hexane-ethyl acetate (1: 4,, obtain 4-[1-[4-[N '-(2-bromo phenyl) urea groups v/v) as eluant]-3-methoxyphenyl acetyl group]-(2S)-pyrrolidinyl] methoxyl methyl benzoate (1.0g; 84%), is colorless oil. 1H-NMR(CDCl 3)δ2.04-2.10(m，4H)，3.52-3.54(m，1H)，3.62(s，2H)，3.70(s，3H)，3.88(s，3H)，4.13-4.19(m，2H)，6.79-6.94(m，4H)，7.20-7.31(m，1H)，7.91-8.12(m，2H)，8.13-8.15(m，1H)。
To 4-[1-[4-[N '-(2-bromo phenyl) urea groups]-3-methoxyphenyl acetyl group]-(2S)-and pyrrolidinyl] methoxyl methyl benzoate (780mg; 1.31mmol) THF (10.0ml) and the agitating solution of MeOH (5.0ml) in add the 1N sodium hydroxide (2.0ml, 2.0mmol).This mixture was stirred 24 hours in 70 ℃.This mixture of vacuum concentration adds water wherein, with 1N HCl neutralization.The solid that collection obtains washes with water, and vacuum drying obtains 59 (730mg, 96%), is white crystalline solid.MW?582.44，mp?120-125℃；IR(KBr)3318，2938，1604，1529，1166，1025cm -1； 1H-NMR(DMSO-d 6)δ1.92-1.96(m，4H)，3.52-3.60(m，5H)，3.82(s，3H)，3.98-4.02(m，1H)，4.16-4.19(m，1H)，4.29(m，1H)，6.75(d，J＝8.3Hz，1H)，6.87(m，1H)，6.94-7.04(m，3H)，7.29-7.33(m，1H)，7.57-7.59(m，1H)，7.85-7.96(m，4H)，8.72(s，1H)，8.91(s，1H)；MS(FAB)m/z582(M ++1)。C 28H 28N 3O 6Br1.0H 2The analytical calculation value of O: C, 56.01; H, 5.04; N, 7.00; Br, 13.31.Measured value: C, 56.12; H, 4.98; N, 6.96; Br, 13.57.Embodiment 553-amino-4-[1-[4-[N '-(2-hydroxy phenyl) urea groups]-3-methoxyphenyl acetylamino]-2-pyrrolidinyl methoxyl group] benzoic acid
Figure A0080951002261
In 0 ℃, to the 2-nitrophenol (10.0g, 72.0mmol) and potassium carbonate (9.96g drips benzyl bromide a-bromotoluene (9.40ml, 79.2 mmol) in the agitating solution of DMF 72.0mmol) (150ml).After stirring 3 hours under the room temperature, the dilute with water reactant mixture is used ether extraction with it.With salt water washing extract,, be concentrated into dried through dried over sodium sulfate.Residue is through chromatography, with normal hexane-ethyl acetate (2: 1, v/v) as eluant, obtain 2-benzyloxy Nitrobenzol (14.7g, 89%), be yellow oil. 1H-NMR(CDCl 3)δ5.24(s，2H)，7.04(t，J＝7.8Hz，1H)，7.12(d，J＝7.8Hz，1H)，7.31-7.50(m，5H)，7.51(d，J＝1.5Hz，1H)，7.86(dd，J＝7.8，1.5Hz，1H)。
In 0 ℃, to 2-benzyloxy Nitrobenzol (9.92g, 43.3mmol) and NiCl 2(20.3g, drip in the agitating solution of methanol 157mmol) (350ml) sodium borohydride (8.09g, 214mmol).(through the TLC monitoring) evaporated this mixture after raw material disappeared.The precipitation of black is dissolved among the 1N HCl, by adding 1N sodium hydroxide this acid solution that alkalizes, uses ethyl acetate extraction then.With salt water washing extract, through dried over sodium sulfate and be concentrated into dried.Residue as eluant, obtains 2-benzyloxy-aniline (8.60g, 100%) with chloroform through chromatography, is reddish oil. 1H-NMR (CDCl 3) δ 3.71 (wide s, 2H), 5.06 (s, 2H), 6.68-6.86 (m, 4H), 7.32-7.44 (m, 5H).FAB-MS?m/z?200(M ++1)。
In 0 ℃, to the 2-benzyloxy-aniline (1.15g, add in benzene 5.77mmol) (60ml) solution triphosgene (1.27g, 6.35mmol) and triethylamine (2.60ml, 17.3mmol).This reactant mixture was heated 20 hours under refluxing.The mixture that filtration obtains is used hexane wash, and concentrated filtrate obtains reddish oil, with it through chromatography, with hexane-ethyl acetate (4: 1, v/v) as eluant, obtain 4-[N '-(2-benzyloxy phenyl) urea groups]-3-methoxyphenyl tert-butyl acetate (2.38g, 89%), be yellow oil. 1H-NMR(CDCl 3)δ1.44(s，9H)，3.44(s，2H)，3.78(s，3H)，5.07(s，2H)，6.73(dd，J＝8.0，1.7Hz，1H)，6.78(d，J＝1.7Hz，1H)，6.90-6.98(m，3H)，7.07(s，1H)，7.29(s，1H)，7.33-7.38(m，5H)，7.91(d，J＝8.0Hz，1H)，8.14(m，1H)。
In 0 ℃, to 4-[N '-(2-benzyloxy phenyl) urea groups]-(2.35g adds TFA (25ml) in dichloromethane 5.08mmol) (25ml) solution to 3-methoxyphenyl tert-butyl acetate.After stirring 3 hours under the room temperature, concentrate this mixture.Residue is dissolved in the 1N sodium hydroxide, washs with ether.In alkaline water layer impouring ice-1N HCl, and the usefulness chloroform-methanol (4: 1, v/v) extract the mixture that obtains.With this extract of salt water washing, through dried over sodium sulfate and be evaporated to dried.Residue is dissolved in the isopropyl ether, hexane is added in this solution until finishing crystallization.Collect solid, obtain 4-[N '-(2-benzyloxy phenyl) urea groups]-3-methoxybenzene guanidine-acetic acid (1.59g, 77%), be brown solid. 1H-NMR (DMSO-d 6) δ 3.50 (s, 2H), 3.85 (s, 3H), 5.26 (s, 2H), 6.76 (d, J=8.3Hz, 1H), 6.83-6.89 (m, 2H), 6.91 (s, 1H), 7.01 (dd, J=8.3,2.3Hz, 1H), 7.31 (t, J=7.3Hz, 1H), 7.39 (t, J=7.3Hz, 12H), 7.49 (d, J=7.3Hz, 2H), 7.97 (dd, J=8.3Hz, 1H), 8.04 (d, J=8.3Hz, 1H), 8.80 (s, 1H), 8.86 (s, 1H), 12.24 (wide s, 1H); FAB-MS m/z 407 (M ++ 1).
To 4-[N '-(2-benzyloxy phenyl) urea groups]-3-methoxybenzene guanidine-acetic acid (1.12g, 2.76mmol), 4-(2-pyrrolidinyl methoxyl group)-3-nitrobenzoic acid methyl ester (890mg, 2.76mmol), HOBt (74.0mg, 0.55mmol), DMAP (67.0mg, 0.55mmol) and triethylamine (0.58ml, 4.13mmol) the solution of THF (15ml) in add EDCHCl (792mg, 4.13mmol).After stirring 12 hours under the room temperature, the dilute with water reactant mixture is used ethyl acetate extraction.With salt water washing extract, through dried over sodium sulfate and be evaporated to dried.Residue is through chromatography, with ethyl acetate as eluant, obtain 4-[1-[4-[N '-(2-benzyloxy phenyl) urea groups]-3-methoxyphenyl acetylamino]-2-pyrrolidinyl methoxyl group]-3-nitrobenzoic acid methyl ester (1.52g, 82%), be yellow amorphous solid. 1H-NMR(CDCl 3)δ1.91(m，1H)，1.95-2.17(m，3H)，3.47-3.53(m，2H)，3.56(s，2H)，3.60(m，1H)，3.68(s，3H)，3.90(s，3H)，4.11(d，J＝7.3Hz，1H)，4.45(m，1H)，5.08(s，2H)，6.70(dd，J＝8.3，1.9Hz，1H)，6.75(d，J＝1.9Hz，1H)，6.91-6.99(m，3H)，7.16(s，1H)，7.18(d，J＝8.3Hz，1H)，7.33-7.40(m，6H)，7.91(d，J＝8.3Hz，1H)，8.11-8.16(m，2H)，8.46(s，1H)。
Under room temperature, with 4-[1-[4-[N '-(2-benzyloxy phenyl) urea groups]-3-methoxyphenyl acetylamino]-2-pyrrolidinyl methoxyl group]-3-nitrobenzoic acid methyl ester (1.52g, 2.27mmol) methanol (20ml) and THF (5ml) solution (wet through 5%Pd-C, 52.2%, 1.21g) in nitrogen atmosphere (4kg/cm 2) hydrogenation down.Stir after 17 hours, the elimination catalyst, concentrated filtrate is to doing.Residue as eluant, obtains 3-amino-4-[1-[4-[N '-(2-hydroxy phenyl) urea groups with ethyl acetate through chromatography]-3-methoxyphenyl acetylamino]-2-pyrrolidinyl methoxyl group] essence of Niobe (1.12g, 90%), be brown amorphous solid. 1H-NMR (CDCl 3) δ 1.97-2.10 (m, 4H), 3.44 (s, 3H), 3.52-3.63 (m, 2H), 3.85 (s, 3H), 4.10-4.18 (m, 2H), 4.53 (m, 1H), 6.65-6.67 (m, 4H), 6.93-7.02 (m, 3H), 7.33 (d, J=2.2Hz, 1H), 7.36 (dd, J=8.3,2.2Hz, 1H), 7.60 (s, 1H), 7.69 (d, J=8.3Hz, 1H), 8.08 (s, 1H), 9.47 wide s, 1H); FAB-MS m/z 549 (M ++ 1).
To 3-amino-4-[1-[4-[N '-(2-hydroxy phenyl) urea groups]-3-methoxyphenyl acetylamino]-2-pyrrolidinyl methoxyl group] essence of Niobe (1.12g, 2.04mmol) THF-methanol (4: 1, v/v, add 1N sodium hydroxide (4.20ml in solution 20ml), 4.20mmol), after stirring 24 hours under the room temperature, concentrated reaction mixture.The dilute with water residue neutralizes with 1N HCl in 0 ℃.With chloroform-methanol (4: 1, v/v) extract this mixture, use the salt water washing, through dried over sodium sulfate and be evaporated to dried.Residue is through chromatography, with chloroform-methanol (5: 1, v/v) as eluant, obtain 60 (352mg, 32%), be faint yellow amorphous solid.MW 534.56 IR (KBr) 3282,3062,3025,2952,2865,1629,1546,1509,1454,1419cm -1 1H-NMR (DMSO-d 6) δ 1.87-2.04 (m, 4H), 3.48-3.57 (m, 2H), 3.60 (s, 2H), 3.79 (s, 3H), 3.94 (dd, J=9.5,7.6Hz, 1H), 4.12 (dd, J=9.5,3.9Hz), 4.35 (m, 1H), 4.87 (wide s, 1H), 6.70-6.91 (m, 6H), 7.16 (dd, 1H, J=8.3,2.0Hz, 1H), 7.26 (d, J=2.0Hz, 1H), 7.96 (d, J=8.3Hz, 1H), 7.97 (d, J=8.3Hz, 1H), 8.80 (s, 1H), 8.82 (s, 1H); FAB-MS m/z 535 (M ++ 1).C 28H 30N 4O 74.5H 2The analytical calculation value of O: C, 55.63; H, 6.39; N, 9.10.Measured value: C, 55.08; H, 5.06; N, 8.69.Embodiment 565-[[1-[4-[N '-(2-chlorophenyl) urea groups]-3-methoxyphenyl acetyl group]-the 2-pyrrolidinyl] methylamino] pyridine-2-formic acid 5-[[1-[3-methoxyl group-4-[N '-(2-aminomethyl phenyl) urea groups] phenyl acetyl]-the 2-pyrrolidinyl] methylamino] pyridine-2-formic acid
Under room temperature, to 5-(methoxycarbonyl) pyridine-2-formic acid (2.5g, 13.8mmol) and 4-DMAP (340mg adds Boc in the agitating solution of tert-butyl alcohol 2.8mmol) (15ml) 2O (6g, 27.6mmol).After stirring 2 hours under the room temperature, will ice and add in this mixture with 0.2N HCl (20ml) and use dichloromethane extraction.Wash extract with saturated sodium bicarbonate, through dried over sodium sulfate and evaporation.Residue as eluant, obtains 6-tert-butoxycarbonyl methyl nicotinate (2.92g, 89%) with dichloromethane through silica gel (50ml) chromatography, is spicule.IR(KBr)2729，1736，1720，1590，1570cm -1；MS(FAB)m/z?238(M ++1)。C 12H 15NO 4The analytical calculation value: C, 60.75; H, 6.37; N, 5.90.Measured value: C, 60.72; H, 6.46; N, 5.78.
Under room temperature, (1.2g, 5.06mmol) (40ml, 10mol) mixture in stirred 0.5 hour at THF (15ml) and 0.25N sodium hydroxide with 6-tert-butoxycarbonyl methyl nicotinate.In this mixture impouring ice-1N HCl (10ml).Collect solid, wash with water also air-dry.Make crude product solid recrystallization from chloroform-ethanol-IPE, obtain 6-tert-butoxycarbonyl nicotinic acid (850mg, 76%), be spicule.IR (KBr) n3095,1728,1705cm -1 1H-NMR (DMAO-d 6) δ 1.63 (s, 9H), 8.09 (m, 1H), 8.17 (m, 1H), 8.42 (dt, J=2.4 and 8.3Hz, 1H), 9.21 (t, J=2.4 and 8.8Hz, 1H); MS (FAB) m/z 224 (M ++ 1).C 29H 33N 3O 6The analytical calculation value: C, 36.18; H, 3.18; N, 3.84.Measured value: C, 36.85; H, 3.35; N, 3.79.
Under room temperature; to 6-tert-butoxycarbonyl nicotinic acid (1.9g; 8.51mmol) and triethylamine (1.17g 11.49mmol) adds diphenyl phosphoryl azide (2.93g, toluene 10.64mmol) (3ml) solution in the stirred mixture in the tert-butyl alcohol (30ml) and toluene (30ml).Then this mixture was heated 5 hours under refluxing.After the cooling, will ice and 1N HCl (5ml) adds in this mixture and uses methylbenzene extraction.With salt water washing extract, through dried over sodium sulfate and evaporation.Residue is used toluene through silica gel (50ml) chromatography: and ethyl acetate (5: 1, v/v) as eluant, obtain 5-tert-butoxycarbonyl amino-2-pyridine carboxylic acid tert-butyl ester (1.9g, 76%), be slurry. 1H-NMR(CDCl 3)δ1.53(s，9H)，1.63(s，9H)，6.82(br?s，1H)，8.01(d，J＝8.8Hz，1H)，8.17(m，1H)，8.46(d，J＝2.4Hz，1H)。
(1.9g, dichloromethane 6.45mmol) (20ml) add TFA (5ml) in stirring the mixture to 5-tert-butoxycarbonyl amino-2-pyridine carboxylic acid tert-butyl ester.Evaporate this mixture, make residue be dissolved in ethanol (30ml).HCl-gas is fed in this solution, stirred 10 minutes in 0-10 ℃ simultaneously.Then the stirred mixture that obtains was heated 10 hours under refluxing.After the cooling, feed nitrogen, evaporate this mixture to remove a large amount of excessive HCl-gases 15 minutes.Make the residue alkalization by adding saturated sodium bicarbonate, use dichloromethane extraction.With salt water washing extract, through dried over sodium sulfate and evaporation, residue is through silica gel (30ml) chromatography, with chloroform-ethanol (98: 2, v/v) as eluant, obtain 5-amino-2-pyridine carboxylic acid ethyl ester (700mg, 65%), be crystalline material.IR (KBr) n 3423,3190,1708,1657,1587,3338,3296,1691,1641cm -1 1H-NMR (CDCl 3) δ 1.42 (t, J=7.0Hz, 3H), 4.11 (br s, 1H), 4.43 (q, J=7.0Hz, 2H), 6.99 (dd, J=2.7 and 8.5Hz, 1H), 7.95 (d, J=8.5Hz, 1H), 8.16 (d, J=2.7Hz, 1H); MS (FAB) m/z 167 (M +); C 27H 29ClN 4O 6The analytical calculation value: C, 57.47; H, 6.63; N, 16.76.Measured value: C, 57.27; H, 5.99; N, 16.72.
With 5-amino-2-pyridine carboxylic acid ethyl ester (660mg, 3.95mmol) and 1-tert-butoxycarbonyl dried meat ammonium aldehyde (1.1g, 5.33mmol) stirring the mixture under refluxing heating 1 hour in toluene (10ml), use Dean-Stark water knockout drum azeotropic removal of water during this period.After the cooling, this mixture of vacuum evaporation.Make residue be dissolved in methanol-acetic acid (9: 1, v/v, 30ml) in.In 0-5 ℃, in the solution of this stirring, add NaBH 3CN (500mg, 7.90mmol).The mixture that restir obtains under room temperature 12 hours.In this mixture impouring ice-saturated sodium bicarbonate (50ml), use dichloromethane extraction.With salt water washing extract, through dried over sodium sulfate and evaporation.Residue is through silica gel (50ml) chromatography, with chloroform-ethyl acetate (98: 2,, obtain 5-[N-[2-(1-tert-butoxycarbonyl) pyrrolidinyl v/v) as eluant] methylamino] pyridine-2-Ethyl formate (1.1g, 70%), be jelly. 1H-NMR (CDCl 3) δ 1.38 (s, 9H), 1.42 (s, 6H), 3.93 (s, 3H), 4.29 (s, 2H), 4.67 (br s, 1H), 7.15 (d, J=8.8Hz, 1H), 8.18 (dd, J=1.7 and 8.8Hz, 1H), 8.52 (d, J=1.7Hz, 1H).
Under room temperature, with 5-[[2-(1-tert-butoxycarbonyl) pyrrolidinyl] methylamino] (800mg, 2.29mmol) mixture in dichloromethane (17ml) and TFA (3ml) stirred 3 hours pyridine-2-Ethyl formate.Evaporate this mixture, make residue be alkalescence with saturated sodium bicarbonate.Use the dichloromethane extraction mixture.With salt water washing extract, dry and evaporation obtains the 5-[(2-pyrrolidinyl through sodium sulfate-sodium carbonate) methylamino] pyridine-2-Ethyl formate (460mg, 81%), be jelly. 1H-NMR (CDCl 3) δ 1.32 (t, J=7Hz, 3H), 1.58-2.10 (a plurality of m, 4H), and 3.12-3.28 (a plurality of m, 3H), 3.65 (m, 1H), 4.30 (be q, J=7Hz, 2H), 6.27 (br, 1H), 6.59 (dd, J=2.4 and 8.5Hz, 1H), 7.65 (d, J=8.5Hz, 1H), 7.94 (d, J=2.4Hz, 1H).
Under room temperature, to the 5-[(2-pyrrolidinyl) methylamino] pyridine-2-Ethyl formate (220mg, 0.88mmol), 4-[N '-(2-chlorophenyl) urea groups]-3-methoxybenzene guanidine-acetic acid (300mg, 0.88mmol), 4-DMAP (135mg, 1.10mmol) add in the stirring the mixture in DMF (7ml) EDCHCl (215mg, 1.10mmol).The mixture that stirring obtains under room temperature 20 hours.In this mixture impouring ice-water.Collect solid, wash with water also air-dry.The crude product solid is through silica gel (30ml) column chromatography purification; with chloroform-ethanol (98: 2; v/v) as eluant; obtain 5-[1-[4-[N '-(2-chlorophenyl) urea groups with the ether crystallization]-3-methoxyphenyl acetyl group]-the 2-pyrrolidinyl] methylamino] pyridine-2-formic acid esters (420mg; 84%), is thin spicule.IR (KBr) 3319,1703,1628,1585,1529cm -1 1H-NMR (CDCl 3) δ 1.38 (t, J=7Hz, 3H), 1.73-2.17 (a plurality of m, 4H), 3.19 and 3.54 (respectively being m, respectively is 1H), 3.63 (s, 2H), 3.70 (s, 3H), 4.39 (be q, J=7Hz, 2H), 4.55 (m, 1H), 6.02 (br s, 1H), 6.78-6.84 (a plurality of s and m, 3H), 6.98 (dt, J=2.4 and 8.0Hz, 1H), 7.16 (s, 1H), and 7.21-7.26 (a plurality of m, 3H), 7.34 (dd, J=2.4 and 8.0Hz, 1H), 7.90 (d, J=8.3Hz, 1H), 7.98 (m, 2H), 8.16 (dd, J=1.2 and 8.8Hz, 1H); MS (FAB) m/z 566 (M ++ 1).C 29H 32ClN 5O 5H 2The analytical calculation value of O: C, 59.63; H, 5.87; N, 12.37.Measured value: C, 60.06; H, 5.76; N, 11.95.
Under room temperature; with 5-[[1-[4-[N '-(2-chlorophenyl) urea groups]-3-methoxyphenyl acetyl group]-the 2-pyrrolidinyl] methylamino] pyridine-2-Ethyl formate (300mg; 0.53mmol) THF: methanol (1: 1; v/v; 16ml) (11ml, 2.75mmol) mixture in stirred 3 hours with the 0.25N sodium hydroxide.In this mixture impouring ice-1N HCl (3ml), collect solid, wash with water also air-dry.Collect rough crystalline material with dichloromethane-ether, obtain 61 (180mg, 63%), be amorphous solid.MW 537.99 IR (KBr) 3319,1701,1620,1585,1533cm -1 1H-NMR (CDCl 3) δ 1H-NMR (DMSO-d 6) δ 1.80-2.05 (a plurality of m, 4H), 2.99 (m, 1H), 3.50-3.59 (a plurality of m, 3H), 3.60 (s, 2H), 3.86 (s, 3H), 4.11 (m, 1H), 6.78 (d, J=8.5Hz, 1H), 6.91 (s, 1H), 6.94 (m, 1H), 7.02 (m, 1H), 7.14 (dd, J=2.5 and 8.5Hz, 1H), 7.28 (t, J=7.0Hz, 1H), 7.45 (d, J=8.0Hz, 1H), 7.78 (d, J=8.8Hz, 1H), 7.97 (d, J=8.3Hz, 1H), 8.08 (br s, 1H), 8.11 (m, 1H), 8.89 (s, 1H), 8.94 (s, 1H); MS (FAB) m/z538 (M ++ 1).C 27H 28ClN 5O 51.5 * H 2The analytical calculation value of O: C, 57.39; H, 5.53; N, 12.39.Measured value: C, 57.37; H, 5.54; N, 11.74.
Under room temperature, to the 5-[(2-pyrrolidinyl) methylamino] pyridine-2-Ethyl formate (230mg, 0.923mmol), 3-methoxyl group-4-[N '-(2-aminomethyl phenyl) urea groups] phenylacetic acid ethyl ester (290mg, 0.923mmol), 4-DMAP (145mg, 1.15mmol) add in the stirring the mixture in DMF (7ml) EDCHCl (225mg, 1.15mmol).The mixture that stirring obtains under room temperature 20 hours.In this mixture impouring ice-water.Collect solid, wash with water also air-dry.The crude product solid is through silica gel (30ml) column chromatography purification; with chloroform-ethanol (98: 2; v/v) as eluant; obtain 5-[[1-[3-methoxyl group-4-[N '-(2-aminomethyl phenyl) urea groups] phenyl acetyl]-the 2-pyrrolidinyl] methylamino] pyridine-2-Ethyl formate (400mg; 80%), is thin spicule.IR (KBr) n 3325,1709,1618,1585,1531cm -1 1H-NMR (CDCl 3) δ 1.39 (t, J=7Hz, 3H), 1.73-2.07 (a plurality of m, 4H), 2.28 (s, 3H), 3.12 and 3.49 (respectively is m, respectively be 1H), 3.60 (s, 2H), 4.39 (br q, J=7Hz, 2H), 4.53 (m, 1H), 6.07 (br s, 1H), 6.23 (br s, 1H), 6.75-6.77 (a plurality of s and m, 2H), 6.82 (dd, J=3.0 and 8.5Hz, 1H), and 7.09-7.22 (a plurality of m, 3H), 7.49 (d, J=8.0Hz, 1H), 7.90 (d, J=8.5Hz, 1H), 7.98 (d, J=2.6Hz, 1H), 8.06 (d, J=8.8Hz, 1H); MS (FAB) m/z546 (M ++ 1).C 30H 35N 5O 51.5 * H 2The analytical calculation value of O: C, 52.92; H, 6.69; N, 12.23.Measured value: C, 63.11; H, 6.48; N, 11.96.
Under room temperature; with 5-[[1-[3-methoxyl group-4-[N '-(2-aminomethyl phenyl) urea groups] phenyl acetyl]-the 2-pyrrolidinyl] methylamino] pyridine-2-Ethyl formate (D91-4596) (290mg; 0.53mmol) THF: methanol (1: 1; v/v; 16ml) (11ml, 2.75mmol) mixture in stirred 3 hours with the 0.25N sodium hydroxide.In this mixture impouring ice-1N HCl (3ml), collect solid, wash with water also air-dry.Collect rough crystalline material with dichloromethane-ether, obtain 62 (170mg, 62%), be amorphous solid.MW 517.58 IR (KBr) 3283,1701,1618,1529 cm -1 1H-NMR (CDCl 3) δ 1H-NMR (DMSO-d 6) δ 1.78-2.04 (a plurality of m, 4H), 2.25 (s, 3H), 2.95-3.55 (a plurality of m, 4H), 3.59 (s, 2H), 3.87 (s, 3H), 4.11 (m, 1H), 6.75-7.24 (a plurality of m, 7H), 7.83-7.97 (a plurality of m, 3H), 8.01 (d, J=8.3Hz, 1H), 8.13 (d, J=2.6Hz, 1H), 8.11 (m, 1H), 8.47 (s, 1H), 8.57 (s, 1H); MS (FAB) m/z 518 (M ++ 1).C 28H 31N 5O 52.5 * H 2The analytical calculation value of O: C, 60.50; H, 6.39; N, 12.60.Measured value: C, 60.31; H, 6.28; N, 12.10.Embodiment 572-[1-[[4-[N '-(2-chlorophenyl) urea groups]-3-methoxyphenyl acetyl group]-the 2-pyrrolidinyl] methoxyl group] pyridine-5-formic acid
Figure A0080951002341
In 0-10 ℃, to 2-hydroxyl-5-pyridine carboxylic acid methyl ester (2.0g, 13.06mmol), triphenyl phasphine (4.2g, 15.93mmol) and the 1-tert-butoxycarbonyl-(L)-dried meat ammonia alcohol (2.63g, 13.06mmol) the agitating solution of THF (25ml) in add DIAD (3.3g, THF 15.67mmol) (5ml) solution.Stirring the mixture of will obtaining then heated 3 hours under refluxing.After the cooling, this mixture of vacuum evaporation.Residue is through silica gel (120ml) chromatography, with normal hexane-ethyl acetate (4: 1,, obtain 2-[2-(1-tert-butoxycarbonyl) pyrrolidinyl v/v) as eluant] methoxypyridine-5-methyl formate (3.0g, 68%), be jelly. 1H-NMR (CDCl 3) δ 1.46 (s, 9H), 1.82-2.04 (a plurality of m, 4H), 3.45 (m, 2H), 3.91 (s, 3H), 4.10-4.32 (a plurality of m, 2H), 4.48 (m, 1H), 6.32 (br, 1H), 6.75 (d, J=8.8Hz, 1H), 8.15 (dd, J=2 and 8.8Hz, 1H), 8.79 (d, J=2Hz, 1H).
Under room temperature, with 2-[2-(1-tert-butoxycarbonyl) pyrrolidinyl] (2.9g, 8.62mmol) mixture in dichloromethane (80ml) and TFA (20ml) stirred 3 hours methoxypyridine-5-methyl formate.Evaporate this mixture, make the residue alkalization with saturated sodium bicarbonate.With this mixture of dichloromethane extraction, with salt water washing extract, dry and evaporation obtains 2-(2-pyrrolidinyl) methoxypyridine-5-methyl formate (1.2g, 59%) through sodium sulfate-sodium carbonate, is a kind of jelly. 1H-NMR (CDCl 3) δ 1.58-2.050 (a plurality of m, 4H), 2.90-3.02 (a plurality of m, 2H), 3.87 and 3.90 (respectively be s, 3H), 4.23 (m, 1H), 4.37 (m, 1H), 6.33 (br, 1H), 6.78 (d, J=8.5Hz, 1H), 8.15 (dd, J=2.2 and 8.8Hz, 1H), 8.79 (d, J=2.2Hz, 1H).
Under room temperature, to 2-(2-pyrrolidinyl) methoxypyridine-5-methyl formate (370mg, 1.57mmol), 4-[N '-(2-chlorophenyl) urea groups]-3-methoxybenzene guanidine-acetic acid (525mg, 1.57mmol), 4-DMAP (230mg, 1.88mmol) add in the stirring the mixture in DMF (10ml) EDCHCl (360mg, 1.88mmol).The mixture that stirring obtains under room temperature 20 hours.In this mixture impouring ice-water.Collect solid, wash with water also air-dry.The crude product solid is through silica gel (30ml) column chromatography purification; with chloroform-ethanol (98: 2; v/v) as eluant; obtain 2-[1-[4-[N '-(2-chlorophenyl) urea groups with the ether crystallization]-3-methoxyphenyl acetyl group]-2-pyrrolidinyl methoxyl group] pyridine-5-methyl formate (600mg; 69%), is amorphous solid. 1H-NMR (CDCl 3) δ .21 and 2.01 (respectively be m, 4H), 3.45-4.50 (a plurality of s and m, 13H, amide containing isomer), 6.58-8.83 (a plurality of s and m, 12H, amide containing isomer).
Under room temperature; with 2-[1-[4-[N '-(2-chlorophenyl) urea groups]-3-methoxyphenyl acetyl group]-2-pyrrolidinyl methoxyl group] pyridine-5-methyl formate (230mg; 0.415mmol) THF (1ml) and 0.25N sodium hydroxide (4ml; mixture 1mmol) stirred 14 hours, stirred 3 hours in 60 ℃.After the cooling, in this mixture impouring ice-1N HCl (2ml), collect solid, wash with water also air-dry.By preparation property TLC plate purification crude product crystalline material, use chloroform: ethanol (9: 1, v/v) as eluant, obtain 63 (150mg, 67%) with the ether crystallization, be amorphous solid.MW 538.98; IR (KBr) 3329,1709,1601,1533 cm -1 1H-NMR (CDCl 3) δ 1.85-2.05 (a plurality of m, 4H), 3.50-3.60 (a plurality of m, 2H), 3.82 (s, 3H), 3.86 (s, 2H), 4.29 (m, 1H), 4.42 (m, 1H), 6.72-7.05 (a plurality of m, 4H), 7.28 (m, 1H), 7.43 (d, J=8Hz, 2H), 7.95 (d, J=8.3Hz, 1H), 8.09 (d, J=8.3Hz, 2H), 8.64 (m, 1H), 8.89 (s, 1H), 8.93 (s, 1H); MS (FAB) m/z 539; (M +).C 27H 28ClN 4O 61.3 * H 2The analytical calculation value of O: C, 57.55; H, 5.47; N, 9.94.Measured value: C, 57.94; H, 5.00; N, 9.45.Embodiment 585-[1-[4-[N '-(2-bromo phenyl) urea groups]-3-methoxyphenyl acetyl group]-2-pyrrolidinyl methoxyl group] pyridine-2-formic acid
Figure A0080951002351
Under room temperature, to 5-(2-pyrrolidinyl) methoxypyridine-2-methyl formate (370mg, 1.57mmol), 4-[N '-(2-bromo phenyl) urea groups]-3-methoxybenzene guanidine-acetic acid (595mg, 1.57mmol), 4-DMAP (230mg, 1.88mmol) add in the stirring the mixture in DMF (10ml) EDCHCl (360mg, 1.88mmol).The mixture that stirring obtains under room temperature 20 hours.In this mixture impouring ice-water.Collect solid, wash with water also air-dry.The crude product solid is through silica gel (30ml) column chromatography purification; with chloroform-ethanol (98: 2; v/v) as eluant; obtain 5-[1-[4-[N '-(2-bromo phenyl) urea groups with the ether crystallization]-3-methoxyphenyl acetyl group]-2-pyrrolidinyl methoxyl group] pyridine-2-methyl formate (650mg; 69%), is amorphous solid.IR (KBr) n 3323,1720,1624,1601,1527 cm -1 1H-NMR (CDCl 3) δ 1.22 and 2.00 (respectively be m, 4H), 3.48-4.55 (a plurality of s and m, 13H, amide containing isomer), 6.93-8.82 (a plurality of s and m, 12H, amide containing isomer).MS (FAB) m/z 597 (M +-1) and 599 (M ++ 1).C 28H 30BrN 4O 61.0 * H 2The analytical calculation value of O: C, 54.55; H, 5.23; N, 9.09.Measured value: C, 54.13; H, 5.03; N, 9.33.
Under room temperature; with 5-[1-[4-[N '-(2-bromo phenyl) urea groups]-3-methoxyphenyl acetyl group]-2-pyrrolidinyl methoxyl group] pyridine-2-methyl formate (300mg; 0.5mmol) THF: methanol (1: 1; v/v; 2ml) with 0.25N sodium hydroxide (4ml; mixture 1mmol) stirred 3 hours, stirred 5 hours in 60 ℃.After the cooling, in this mixture impouring ice-1N HCl (2ml).Collect solid, wash with water also air-dry.By preparation property TLC plate purification crude product crystalline material, use chloroform: ethanol (9: 1, v/v) as eluant, obtain 64 (180mg, 62%) with the ether crystallization, be amorphous solid.MW 583.43 IR (KBr) n 3319,1705,1685,1601,1529 cm -1 1H-NMR (DMSO-d 6) δ 1.82-2.05 (a plurality of m, 4H), 3.48-3.58 (a plurality of m, 2H), 3.82 (s, 3H), 3.86 (s, 2H), 4.42-4.55 (a plurality of m, 3H), 6.72-6.98 (a plurality of m, 4H), 7.32 (t, J=8Hz, 1H), 7.60 (d, J=8Hz, 1H), 7.95 (m, 2H), 8.08 (m, 1H), 8.63 (m, 1H), 8.64 (m, 1H), 8.89 (s, 1H), 8.93 (s, 1H); MS (FAB) m/z583 (M +); C 27H 28BrN 4O 62.0 * H 2The analytical calculation value of O: C, 52.26; H, 5.20; N, 9.03.Measured value: C, 52.72; H, 4.63; N, 8.50.Embodiment 594-[1-[3-[N '-(2-bromo phenyl) urea groups]-2-methoxyl group-6-pyridine radicals acetyl group]-(4S)-and fluoro-(2S)-pyrrolidinyl methoxyl group] benzoic acid
To 3-amino-2-methoxyl group-6-pyridine radicals ethyl acetate (1.61g, add in the agitating solution of THF 7.66mmol) (10ml) Carbimide. 2-bromo phenyl ester (948ml, 7.66mmol) and triethylamine (107ml, 0.776mmo1).After stirring was spent the night, in this mixture impouring water (100ml), (4: 1,2 * 200ml) extracted with chloroform-methanol.The extract that merges through dried over mgso also evaporates.Make residue recrystallization from chloroform-methanol-hexane, obtain 3-[N '-(2-bromo phenyl) urea groups]-2-methoxyl group-6-pyridine radicals ethyl acetate (2.91g, 93%), be the colourless crystallization powder.mp?160-163℃； 1H-NMR(DMSO-d 6)δ1.19(dt，J＝7.1，0.7Hz，3H)，3.69(s，2H)，3.95(s，3H)，4.07-4.13(m，2H)，6.90(d，J＝7.8Hz，1H)，6.99(t，J＝7.8Hz，1H)，7.33(t，J＝7.8Hz，1H)，7.61(d，J＝7.8Hz，1H)，7.96(dd，J＝7.8，1.5Hz，1H)，8.3?1(d，J＝7.8Hz，1H)，8.82(s，1H)，9.12(s，1H)。MS(FAB)m/z408(M +)，410(M ++2)。C 17H 18BrN 3O 40.25H 2The analytical calculation value of O: C, 49.47; H, 4.52; N, 9.96.Measured value: C, 49.34; H, 4.48; N, 9.96.
With 3-[N '-(2-bromo phenyl) urea groups]-2-methoxyl group-6-pyridine radicals ethyl acetate (2.90g, 7.10mmol), the 0.25N sodium hydroxide (56.8ml, 14.2mmol) and the mixture of THF (50ml) stirred 5 hours.With 1N HCl this mixture that neutralizes, filter and collect the precipitation that produces.Make residue recrystallization from chloroform-methanol-hexane, obtain 3-[N '-(2-bromo phenyl) urea groups]-2-methoxyl group-6-pyridine radicals acetic acid (2.40g, 89%), be the colourless crystallization powder.Mp 195-197 ℃; 1H-NMR (DMSO-d 6) δ 3.59 (s, 2H), 3.95 (s, 3H), 6.88 (d, J=8.1Hz, 1H), 6.97-7.01 (m, 1H), 7.33 (t, J=7.3Hz, 1H), 7.61 (d, J=8.1Hz, 1H), and 7.95-7.97 (m, 1H), 8.29 (d, J=8.1Hz, 1H), 8.81 (s, 1H), 9.10 (s, 1H), 12.35 (br s, 1H); C 15H 14BrN 3O 4The analytical calculation value: C, 47.39; H, 3.71; N, 11.05.Measured value: C, 47.27; H, 3.59; N, 10.86.
To 3-[N '-(2-bromo phenyl) urea groups]-2-methoxyl group-6-pyridine radicals acetic acid (751mg, 1.97mmol) and (4S)-fluoro-(2S)-pyrrolidinyl methoxyl methyl benzoate (500mg, 1.97mmol) the agitating solution of DMF (10ml) in add EDCHCl (566mg, 2.96mmol), DMAP (cat.) and HOBt (cat.).After stirring is spent the night, this mixture is allocated between ethyl acetate (200ml) and the saline (200ml).Separate each phase.With saline (100ml) washing organic facies, through dried over mgso and evaporation.Make the residue that obtains through silica gel column chromatography; with chloroform-methanol (20: 1) as eluant; obtain 4-[1-[3-[N '-(2-bromo phenyl) urea groups]-2-methoxyl group-6-pyridine radicals acetyl group]-(4S)-and fluoro-(2S)-pyrrolidinyl methoxyl group] essence of Niobe (1.16g; 96%), is yellow viscous solid.
With 4-[1-[3-[N '-(2-bromo phenyl) urea groups]-2-methoxyl group-6-pyridine radicals acetyl group]-(4S)-and fluoro-(2S)-pyrrolidinyl methoxyl group] essence of Niobe (1.16g; 1.88mmol), the 0.25N sodium hydroxide (15ml, 3.75mmol) and the mixture of THF (15ml) stir and to spend the night.With 1N HCl this mixture that neutralizes, (4: 1,2 * 200ml) extracted with chloroform-methanol.The extract that merges through dried over mgso also evaporates.The residue that obtains is through silica gel column chromatography, with chloroform-methanol (40: 1-10: 1) as eluant, obtain 65, be faint yellow amorphous solid.MW 601.42, 1H-NMR (DMSO-d 6) δ 2.27-2.39 (m, 2H), 3.33-4.84 (a plurality of m, 10H), 5.33-5.53 (m, 1H), 6.87-6.90 (m, 1H), 6.99 (t, 1H, J=7.6Hz), 7.08 (d, 2H, J=9.0Hz), 7.34 (t, 1H, J=7.6Hz), 7.61 (d, 1H, J=7.8Hz), 7.88 (d, 2H, J=9.0Hz), 7.97 (d, 1H, J=8.3Hz), 8.28-8.32 (m, 1H), 8.81-8.82 (m, 1H), 9.10-9.12 (m, 1H), 12.66 (br s, 1H); MS (FAB) m/z 601 (M +), 603 (M ++ 2).C 27H 26BrFN 4O 6The analytical calculation value: C, 53.92; H, 4.36; N, 9.32.Measured value: C, 52.37; H, 4.62; N, 8.38.Embodiment 604-[(4S)-and fluoro-1-[4-[N '-(2-aminomethyl phenyl) urea groups] phenyl acetyl]-(2S)-and the pyrrolidinyl methoxyl group] benzoic acid
Figure A0080951002381
To 4-[N '-(2-aminomethyl phenyl) urea groups] phenylacetic acid (337mg, 1.18mmol) and 4-[(4S)-fluoro-(2S)-pyrrolidinyl methoxyl group] essence of Niobe (300mg, 1.18mmol) the agitating solution of DMF (10ml) in add EDCHCl (339mg, 1.77mmol), HOBt (cat.) and DMAP (cat.).This reactant mixture stirring is spent the night.This mixture is allocated between ethyl acetate (200ml) and the water (200ml), separates organic facies.With saline (200ml) washing organic facies, through dried over mgso and evaporation.Make residue through silica gel column chromatography; with chloroform-methanol (50: 1) as eluant; obtain 4-[(4S)-fluoro-1-[4-[N '-(2-aminomethyl phenyl) urea groups] phenyl acetyl]-(2S)-and the pyrrolidinyl methoxyl group] (613mg quantitatively), is the grease of yellow viscosity to essence of Niobe. 1H-NMR (CDCl 3) δ 2.03-2.55 (a plurality of m, 5H altogether), 3.47-4.21 (a plurality of m, 7H altogether), 4.44-4.60 (m, 3H), 5.21 with 5.34 (they respectively are m, are total to 1H), and 6.87-7.16 (m, 8H), 7.52-7.55 (m, 3H), 7.93 (d, J=8.8Hz, 2H), 7.99 (d, J=8.8Hz, 1H).
To 4-[(4S)-fluoro-1-[4-[N '-(2-aminomethyl phenyl) urea groups] phenyl acetyl]-(2S)-and the pyrrolidinyl methoxyl group] essence of Niobe (613mg; 1.18mmol) the agitating solution of THF (10ml) in add the 0.25N sodium hydroxide (9.4ml, 2.36mmol).This mixture was refluxed 1 day.After being cooled to room temperature, in this mixture impouring 1N HCl (50ml), (5: 1,2 * 200ml) extracted with chloroform-methanol.The extract that merges through dried over mgso also evaporates.The residue that obtains as eluant, obtains 66 (378mg, 63%) with chloroform-methanol (10: 1) through silica gel column chromatography, is colourless amorphous solid.MW 505.54, 1H-NMR (DMSO-d 6) δ .08-2.30 (m, 5H altogether), 3.47-4.63 (a plurality of m, 7H), 5.30-5.50 (m, 1H), 6.94 (t, J=7.3Hz, 1H), 7.02-7.17 (m, 6H), 7.37-7.41 (m, 2H), 7.82-7.96 (m, 4H), 9.05 (s, 1H); MS (FAB) m/z506 (M ++ 1).C 28H 28FN 3O 51.75H 2The analytical calculation value of O: C, 62.62; H, 5.91; N, 7.82.Measured value: C, 62.23; H, 5.63; N, 7.18.Embodiment 614-[(4S)-and fluoro-1-[3-methoxyl group-4-[N '-(2-aminomethyl phenyl) urea groups] phenyl acetyl]-(2S)-and the pyrrolidinyl methoxyl group] benzoic acid
Under room temperature, (1.85g adds BH in the agitating solution of THF 7.93mmol) (15ml) to 1-(tert-butoxycarbonyl)-(4S)-fluoro-(2S)-proline 3DMS (0.75ml, 7.93mmol).Stirring down, reflux made this mixture be cooled to room temperature, vacuum concentration after 5 hours.By adding the residue that entry (100ml) quencher obtains, (2 * 200ml) extract with chloroform.Extract with saline (100ml) washing merges also evaporates through dried over mgso.Residue is through silica gel column chromatography, and as eluant, (1.76g quantitatively), is colorless oil to obtain 1-(tert-butoxycarbonyl)-(4S)-fluoro-(2S)-pyrrolidinyl methanol with chloroform-ethyl acetate (4: 1). 1H-NMR (CDCl 3) δ 1.48 (s, 9H), 1.64 (m, 1H), 1.97-2.28 (m, 2H), 3.53-3.87 (a plurality of m, 4H), 4.09-4.25 (m, 1H), 5.09 and 5.22 (they respectively are m, are total to 1H).
Under room temperature, to 1-(tert-butoxycarbonyl)-(4S)-fluoro-(2S)-pyrrolidinyl methanol (500mg, 2.28mmol), 4-methyl hydroxybenzoate (416mg, 2.74mmol), triphenyl phasphine (719mg, 2.74mmol) the stirring the mixture of THF (10ml) in add DIAD (0.54ml, 2.74mmol).Stir down, with mixture heated to refluxing 5 hours.After being cooled to room temperature, this mixture of vacuum concentration.Residue is through silica gel column chromatography, with chloroform-ethyl acetate (10: 1-4:, obtain 4-[1-(tert-butoxycarbonyl)-(4S)-fluoro-(2S)-pyrrolidinyl methoxyl group 1) as eluant] essence of Niobe (597mg, 74%), be colorless oil. 1H-NMR (CDCl 3) δ 1.49-1.59 (m, 9H), 2.05-2.21 (m, 1H), 3.56-4.43 (a plurality of m, 8H), 5.19 and 5.32 (they respectively are m, are total to 1H), 6.97 (m, 2H), 7.98 (d, J=8.5Hz, 2H).
With 4-[1-(tert-butoxycarbonyl)-(4S)-fluoro-(2S)-pyrrolidinyl methoxyl group] essence of Niobe (590mg, 1.67mmol) and the mixture of TFA (5ml) in dichloromethane (5ml) stirred 3 hours.Behind this mixture of vacuum concentration, make residue be alkalescence with saturated sodium bicarbonate.(2 * 200ml) extract with chloroform.Through dry extract that merges of potassium carbonate and evaporation, obtain 4-[(4S)-fluoro-(2S)-pyrrolidinyl methoxyl group] essence of Niobe (414mg, 98%), be yellow solid. 1H-NMR (CDCl 3) δ 1.89-2.02 (m, 1H), 2.16-2.31 (m, 1H), 2.98 (m, 1H), 3.35 (m, 1H), 3.46-3.68 (m, 1H), 3.86 (s, 3H), 4.00-4.08 (m, 2H), 5.16 and 5.29 (respectively be t, J=4.7Hz, be total to 1H), 6.91 (d, J=8.8Hz, 2H), 7.96 (d, J=8.8Hz, 2H).
With 4-[(4S)-fluoro-(2S)-pyrrolidinyl methoxyl group] essence of Niobe (205mg, 0.810mmol), 3-methoxyl group-4-[N '-(2-aminomethyl phenyl) urea groups] phenylacetic acid (254mg, 0.810mmol), EDCHCl (233mg, 1.22mmol), HOBt (cat.) and DMAP (cat.) mixture in DMF (10ml) stirs and spend the night.(200ml) dilutes this mixture with ethyl acetate, and (2 * 100ml) washings are through dried over mgso and evaporation with saline.Residue is through silica gel column chromatography; with chloroform-methanol (20: 1) as eluant; obtain 4-[(4S)-fluoro-1-[3-methoxyl group-4-[N '-(2-aminomethyl phenyl) urea groups] phenyl acetyl]-(2S)-and the pyrrolidinyl methoxyl group] (445mg quantitatively), is the light brown goo to essence of Niobe. 1H-NMR (CDCl 3) δ 2.05-2.55 (m, 6H altogether), and 3.55-4.13 (m, 11H), 4.48-4.60 (m, 2H), 5.20 and 5.33 (they respectively are m, are total to 1H), 6.29 (s, 1H), 6.79 (m, 2H), 6.96 (d, J=8.8Hz, 2H), 7.11-7.25 (m, 3H), 7.48 (d, J=7.6Hz, 1H), and 7.93-8.09 (m, 4H).
Under room temperature; with 4-[(4S)-fluoro-1-[3-methoxyl group-4-[N '-(2-aminomethyl phenyl) urea groups] phenyl acetyl]-(2S)-and the pyrrolidinyl methoxyl group] essence of Niobe (445mg; 1.62mmol) and 0.25N sodium hydroxide (15ml; 3.75mmol) mixture in THF (15ml) stirs and spend the night, reflux then and stirred 2 hours down.With this mixture of 1H HCl acidify, (4: 1,2 * 200ml) extracted with chloroform-methanol.The extract that merges through dried over mgso also evaporates.Residue as eluant, obtains 67 (260mg, 30%) with chloroform-methanol (10: 1) through silica gel column chromatography, is faint yellow amorphous solid.MW 535.56, 1H-NMR (DMSO-d 6) δ 2.25-2.51 (m, 5H), 3.33-4.41 (a plurality of m, 10H), 5.30-5.50 (m, 1H), 6.75-7.17 (m, 7H), 7.79 (d, J=8.1Hz, 1H), 7.87-8.04 (m, 3H), 8.48 (m, 1H), 8.58 (m, 1H); MS (FAB) m/z536 (M ++ 1).C 29H 30FN 3O 6H 2The analytical calculation value of O: C, 62.92; H, 5.83; N, 7.59.Measured value: C, 62.40; H, 5.82; N, 6.93.Embodiment 624-[1-[4-[N '-(2-bromo phenyl) urea groups]-3-methoxyphenyl acetyl group]-(4S)-and fluoro-(2S)-pyrrolidinyl methoxyl group] benzoic acid
Figure A0080951002411
With 4-[(4S)-fluoro-(2S)-pyrrolidinyl methoxyl group] essence of Niobe (501mg, 1.98mmol), 4-[N '-(2-bromo phenyl) urea groups]-3-methoxybenzene guanidine-acetic acid (750mg, 1.98mmol), EDCHCl (569mg, 2.97mmol), HOBt (cat.) and DMAP (cat.) mixture in DMF (10ml) stirs and spend the night.(300ml) dilutes this mixture with ethyl acetate, with saline (100ml) washing, through dried over mgso and evaporation.Residue is through silica gel column chromatography; with chloroform-ethyl acetate (4: 1) to chloroform-methanol (10: 1) as eluant; obtain 4-[1-[4-[N '-(2-bromo phenyl) urea groups]-3-methoxyphenyl acetyl group]-(4S)-and fluoro-(2S)-pyrrolidinyl methoxyl group] essence of Niobe (1.29g; quantitatively), be the grease of brown viscosity. 1H-NMR (CDCl 3) δ 2.05-2.58 (m, 2H), 3.49-4.17 (a plurality of m, 12H), 4.52-4.65 (m, 2H), 6.82-7.33 (a plurality of m, 8H), 7.53 (dd, J=8.1,1.5Hz, 1H), 7.95-8.02 (m, 4H), 8.14 (dd, J=8.3,1.7Hz, 1H).
With 4-[1-[4-[N '-(2-bromo phenyl) urea groups]-3-methoxyphenyl acetyl group]-(4S)-and fluoro-(2S)-pyrrolidinyl methoxyl group] essence of Niobe (1.29g; 2.10mmol) and the 0.25N sodium hydroxide (17ml, 4.20mmol) mixture in THF (20ml) under agitation refluxed 5 hours.In this mixture impouring ice-refrigerative 1N HCl (100ml), (4: 1,2 * 200ml) extracted with chloroform-methanol.The extract that merges through dried over mgso also evaporates.Residue as eluant, obtains 68 (860mg, 68%) with chloroform-methanol (10: 1) through silica gel column chromatography, is colourless amorphous solid.MW 600.43 1H-NMR (DMSO-d 6) δ 2.24-2.31 (m, 2H), 3.21-4.63 (a plurality of m, 10H), 5.31-5.51 (m, 1H), 6.74-7.10 (m, 5H), 7.32 (t, J=7.8Hz, 1H), 7.60 (d, J=7.8Hz, 2H), 7.87-7.99 (m, 4H), 8.74-8.75 (m, 1H), 8.92-8.94 (m, 1H); MS (FAB) m/z 601 (M ++ 1).C 28H 27BrFN 3O 62H 2The analytical calculation value of O: C, 52.84; H, 4.91; N, 6.60.Measured value: C, 52.38; H, 4.62; N, 5.99.Sodium salt for 68: mp 180-182 ℃; C 28H 27BrFN 3NaO 60.75H 2The analytical calculation value of O: C, 52.88; H, 4.36; N, 6.61.Measured value: C, 52.97; H, 4.36; N, 6.61.Embodiment 634-[1-[4-[N '-(2-chlorophenyl) urea groups]-3-methoxyphenyl acetyl group]-(4S)-and fluoro-(2S)-pyrrolidinyl methoxyl group] benzoic acid
With 4-[(4S)-fluoro-(2S)-pyrrolidinyl methoxyl group] essence of Niobe (205mg, 0.810mmol), 3-methoxyl group-4-[N '-(2-aminomethyl phenyl) urea groups] phenylacetic acid (254mg, 0.810mmol), EDCHCl (233mg, 1.22mmol), HOBt (cat.) and DMAP (cat.) mixture in DMF (10ml) stirs and spend the night.(200ml) dilutes this mixture with ethyl acetate, and (2 * 100ml) washings are through dried over mgso and evaporation with saline.Residue is through silica gel column chromatography; with chloroform-methanol (20: 1) as eluant; obtain 4-[1-[4-[N '-(2-chlorophenyl) urea groups]-3-methoxyphenyl acetyl group]-(4S)-and fluoro-(2S)-pyrrolidinyl methoxyl group] essence of Niobe (376mg, 81%), be yellow foam thing. 1H-NMR (CDCl 3) δ 2.07-2.56 (m, 2H), 3.57-4.14 (a plurality of m, 11H), 4.50-4.61 (m, 2H), 5.22 and 5.35 (a plurality of m, 1H altogether), 6.80-7.33 (a plurality of m, 9H), 7.93-8.00 (m, 3H), 8.16 (d, J=8.1Hz, 1H).
Under room temperature; with 4-[1-[4-N '-(2-chlorophenyl) urea groups]-3-methoxyphenyl acetyl group]-(4S)-and fluoro-(2S)-pyrrolidinyl methoxyl group] essence of Niobe (376mg; 0.660mmol) and 0.25N sodium hydroxide (15ml; 3.75mmol) mixture in THF (15ml) stirs and spend the night, reflux then and stirred 2 hours down.With this mixture of 1H HCl acidify, (4: 1,2 * 200ml) extracted with chloroform-methanol.The extract that merges through dried over mgso also evaporates.Residue as eluant, obtains 69 (260mg, 30%) with chloroform-methanol (20: 1) through silica gel column chromatography, is faint yellow amorphous solid.MW 555.98, 1H-NMR (DMSO-d 6) δ 2.24-2.501 (m, 2H), 3.48-4.65 (a plurality of m, 10H), 5.30-5.50 (m, 1H), 6.75-7.08 (m, 5H), 7.29 (t, J=7.3Hz, 1H), 7.43-7.45 (m, 1H), and 7.89-7.98 (m, 2H), 7.99 (d, J=8.3Hz, 1H), 8.09 (d, J=7.1Hz, 1H), and 8.90-8.96 (m, 2H); MS (FAB) m/z 556 (M ++ 1).C 28H 27ClFN 3O 61/4H 2The analytical calculation value of O: C, 60.00; H, 4.95; N, 7.50.Measured value: C, 59.67; H, 5.08; N, 7.10.Embodiment 644-[1-[4-[N '-(2-bromo phenyl) urea groups] phenyl acetyl]-(4S)-and fluoro-(2S)-pyrrolidinyl methoxyl group] benzoic acid
Figure A0080951002441
To 4-[1-(4-benzyloxycarbonyl aminophenyl acetyl group)-(4S)-fluoro-(2S)-pyrrolidinyl methoxyl group] essence of Niobe (300mg; 0.576mmol) in added ethanol-THF (5: 1; 30ml), stir down, with this solution through 5%Pd/C (300ml) hydrogenation 12 hours.Filter this mixture to remove catalyst.Concentrating under reduced pressure filtrate.Residue as eluant, obtains 4-[1-(4-aminophenyl acetyl group)-(4S)-fluoro-(2S)-pyrrolidinyl methoxyl group with chloroform-methanol (20: 1) through silica gel column chromatography] essence of Niobe (200mg, 90%), be yellow oil. 1H-NMR (CDCl 3) δ 2.01-2.56 (a plurality of m, 2H), 3.50-4.14 (a plurality of m, 5H), 4.45-4.62 (m, 2H), 5.21 with 5.34 (they respectively are m, are total to 1H), and 6.60-6.65 (m, 2H), 6.88 (d, J=8.8Hz, 0.5H), 6.99-7.05 (m, 3.5H), 7.95-8.00 (m, 2H).
With 4-[1-(4-aminophenyl acetyl group)-(4S)-fluoro-(2S)-pyrrolidinyl methoxyl group] (200mg 0.518mmol) is dissolved among the THF (10ml) essence of Niobe.(108 μ l, 0.776mmol) (96 μ l 0.776mmol) add in this solution with Carbimide. 2-bromo phenyl ester with triethylamine.The mixture stirring is spent the night, with ethyl acetate (200ml) dilution.(100ml) washs this solution with saline, through dried over mgso and removal of solvent under reduced pressure.Residue is through silica gel column chromatography; with chloroform-ethyl acetate (4: 1) to chloroform-methanol (10: 1) as eluant; obtain 4-[1-[4-[N '-(2-bromo phenyl) urea groups] phenyl acetyl]-(4S)-and fluoro-(2S)-pyrrolidinyl methoxyl group] essence of Niobe (303mg; quantitatively), be yellow oil. 1H-NMR (CDCl 3) δ 2.08-2.60 (a plurality of m, 2H), 3.56-4.69 (a plurality of m, 10H), 5.28 and 5.40 (respectively is m, be total to 1H), and 6.84-6.92 (m, 3H), 7.03-7.10 (m, 3H), 7.14 (d, J=8.1Hz, 1H), 7.23 (t, J=8.1Hz, 1H), 7.39-7.44 (m, 2H), 7.89 (d, J=8.1Hz, 1H), 7.98-8.03 (m, 2H), 8.09 (d, J=8.1Hz, 1H).
With 4-[1-[4-[N '-(2-bromo phenyl) urea groups] phenyl acetyl]-(4S)-and fluoro-(2S)-pyrrolidinyl-methoxyl group] essence of Niobe (300mg; 0.513mmol) be dissolved among the THF (5ml); (4.0ml 1.00mmol) adds in this solution with the 0.25N sodium hydroxide.Stir after 3 days, in this mixture impouring 1N HCl (100ml), (5: 1,2 * 200ml) extracted with chloroform-methanol.Through the extract that dried over mgso merges, removal of solvent under reduced pressure.Residue as eluant, obtains 70 (209mg, 71%) with chloroform-methanol (10: 1) through silica gel column chromatography, is colourless amorphous solid.MW 570.41, 1H-NMR (DMSO-d 6) δ 2.24-2.51 (m, 2H), 3.36-4.64 (a plurality of m, 7H), 5.31-5.50 (m, 1H), 6.97 (t, J=7.8Hz, 1H), 7.04 (d, J=8.5Hz, 1H), 7.09 (d, J=8.8Hz, 1H), 7.14-7.20 (m, 2H), 7.34 (t, J=7.8Hz, 1H), 7.38-7.43 (m, 2H), 7.61 (d, J=8.1Hz, 1H), 7.87-7.92 (m, 2H), 8.08 (d, J=8.1Hz, 1H), 8.1 5 (s, 1H), 9.45-9.47 (m, 1H), 12.66 (br s, 1H); MS (FAB) m/z572 (M ++ 2), 570 (M +).C 27H 25BrFN 3O 51.5H 2The analytical calculation value of O: C, 54.28; H, 4.72; N, 7.03.Measured value: C, 54.67; H, 4.51; N, 6.61.Embodiment 654-[1-[4-[N '-(2-iodine substituted phenyl) urea groups]-3-methoxyphenyl acetyl group]-(4S)-and fluoro-(2S)-pyrrolidinyl methoxyl group] benzoic acid
To 4-amino-3-methoxyphenyl tert-butyl acetate (1.94g, add in the agitating solution of THF 8.16mmol) (20ml) Carbimide. 2-iodo phenyl ester (2.0g, 8.16mmol) and triethylamine (114 μ l, 0.816mmol).After stirring is spent the night, with this mixture impouring 1N HCl (200ml).Filter and collect the precipitation that produces, and make it to be dissolved in the chloroform (200ml).Through this solution of dried over mgso and evaporation, obtain 4-[N '-(2-iodine substituted phenyl) urea groups]-(3.93g quantitatively), is faint yellow amorphous solid to 3-methoxyphenyl tert-butyl acetate. 1H-NMR(CDCl 3)δ1.44(s，9H)，3.49(s，2H)，3.85(s，3H)，6.78-6.88(m，4H)，7.07(s，1H)，7.31-7.35(m，1H)，7.76(dd，J＝7.8，1.5Hz，1H)，7.95(d，J＝8.3Hz，1H)，7.99(dd，J＝8.3，1.5Hz，1H)；MS(ESI)m/z483(M ++1)。
With 4-[N '-(2-iodine substituted phenyl) urea groups]-3-methoxyphenyl tert-butyl acetate (3.93g, 8.16mmol) and TFA (5ml) stirring the mixture in dichloromethane (5ml) refluxed 3 hours.After being cooled to room temperature, this mixture of vacuum concentration, (50ml) adds in this residue with water.Filter and collect the precipitation that produces,, as eluant, obtain 4-[N '-(2-iodine substituted phenyl) urea groups with chloroform-methanol (9: 1) through purification by silica gel column chromatography]-3-methoxybenzene guanidine-acetic acid (2.89g, 83%), be light yellow crystalline powder. 1H-NMR(DMSO-d 6)δ3.62(s，2H)，3.88(s，3H)，6.78(d，J＝8.3Hz，1H)，6.83-6.87(m，1H)，6.94(s，1H)，7.32-7.36(m，1H)，7.69(dd，J＝8.3，1.5Hz，1H)，7.84(dd，J＝8.3，1.5Hz，1H)，7.97-8.00(m，1H)，8.55(m，1H)，8.82(m，1H)，12.26(br?s，1H)。
With 4-[N '-(2-iodine substituted phenyl) urea groups]-3-methoxybenzene guanidine-acetic acid (505mg, 1.18mmol), 4-[(4S)-fluoro-(2S)-pyrrolidinyl methoxyl group] essence of Niobe (300mg, 1.18mmol), EDCHCl (339mg, 1.77mmol), DMAP (catalytic amount) and HOBt (catalytic amount) mixture in DMF (10ml) stirs and spend the night.(300ml) dilutes this mixture with ethyl acetate, with saline (2 * 200ml) washings.Through this solution of dried over mgso and evaporation.The residue that obtains is through silica gel column chromatography; with chloroform-ethyl acetate (4: 1) as eluant; obtain 4-[1-[4-[N '-(2-iodine substituted phenyl) urea groups]-3-methoxyphenyl acetyl group]-(4S)-and fluoro-(2S)-pyrrolidinyl methoxyl group] essence of Niobe (500mg; 64%), is the oil of colourless viscosity. 1H-NMR (CDCl 3) δ 07-2.58 (m, 2H), 3.59-4.20 (m, 11H), 4.51-4.64 (m, 2H), 5.24 and 5.37 (they respectively are m, are total to 1H), 6.80-6.91 (m, 5H), 6.98 (d, J=8.8Hz, 2H), 7.34 (t, J=7.8Hz, 1H), 7.78 (dd, J=7.8,1.2Hz, 1H), and 7.95-8.02 (m, 4H).
To 4-[1-[4-[N '-(2-iodine substituted phenyl) urea groups]-3-methoxyphenyl acetyl group]-(4S)-and fluoro-(2S)-pyrrolidinyl methoxyl group] (500mg adds 0.25N sodium hydroxide (6ml) to essence of Niobe in the agitating solution of THF 0.756mmol) (6ml).Under room temperature, continue to stir to spend the night, reflux then and stirred 5 hours down.After being cooled to room temperature, in this solution impouring 1N HCl (100ml), (4: 1,2 * 200ml) extracted with chloroform-methanol.The extract that merges through dried over mgso also evaporates.Residue as eluant, obtains 71 (295mg, 60%) with chloroform-methanol (10: 1) through silica gel column chromatography, is colourless amorphous solid.MW 647.43 1H-NMR (DMSO-d 6) δ 2.2.09-2.31 (m, 2H), 3.33-4.41 (a plurality of m, 10H), 5.30-5.50 (m, 1H), 6.77-6.92 (m, 3H), 7.03-7.09 (m, 2H), 7.34 (t, J=8.1Hz, 1H), 7.69 (dd, J=8.3,1.5Hz, 1H), and 7.83-7.99 (m, 4H), 8.54 (m, 1H), 8.82 (m, 1H); MS (FAB) m/z 648 (M ++ 1).C 28H 27FIN 3O 4The analytical calculation value: C, 51.94; H, 4.20; N, 6.49.Measured value: C, 51.17; H, 4.53; N, 5.76.Embodiment 664-[(4S)-fluoro-1-[4-(N '-the phenyl urea groups) phenyl acetyl]-(2S)-and the pyrrolidinyl methoxyl group] benzoic acid
To 4-aminophenyl ethyl acetate (6.43g, 35.9mmol) and triethylamine (5.50ml, (3.90ml, 35.9mmol), stirred reaction mixture is 4 days under room temperature to add carbanil in the agitating solution of THF 39.5mmol) (70ml).The precipitation that decompression collect to produce is used the normal hexane wash filtrate, obtains 4-(N '-phenyl urea groups) phenylacetic acid ethyl ester (9.64g, 90%), is white crystalline powder.mp?153-155℃； 1H-NMR(CDCl 3)δ1.26(t，J＝7.1Hz，3H)，3.52(s，2H)，4.15(q，J＝7.1Hz，2H)，6.98-7.04(m，1H)，7.07-7.11(m，4H)，7.18-7.25(m，5H)，7.42(s，1H)；MS(FAB)m/z?299(M ++1)。C 17H 18N 2O 3The analytical calculation value: C, 68.44; H, 6.08; N, 9.39.Measured value: C, 68.22; H, 6.10; N, 9.36.
(9.64g adds 0.5N sodium hydroxide (80ml) in the agitating solution of THF 32.3mmol) (80ml), down this reactant mixture was heated 5 hours in refluxing to 4-(N '-phenyl urea groups) phenylacetic acid ethyl ester.After being cooled to room temperature, in this mixture impouring ice-1N HCl.The precipitation that decompression collect to produce, crude product solid recrystallization from methanol-chloroform obtains 4-(N '-phenyl urea groups) phenylacetic acid (8.14g, 93%), is white crystalline powder.MS (FAB) m/z 271 (M ++ 1); C 15H 14N 2O 3The analytical calculation value: C, 66.66; H, 5.22; N, 10.36.Measured value: C, 66.45; H, 5.22; N, 10.30.
Under room temperature, with 4-(N '-phenyl urea groups) phenylacetic acid (310mg, 1.15mmol), 4-[(4S)-fluoro-(2S)-pyrrolidinyl methoxyl group] essence of Niobe (287mg, 1.13mmol), EDCHCl (260mg, 1.36mmol), HOBt (185mg, 1.37mmol) and triethylamine (190ml, 1.36mmol) mixture in DMF (5ml) stirs and to spend the night.This mixture of dilute with water is used ethyl acetate extraction.With salt water washing extract, through dried over sodium sulfate and evaporation.Residue is through purification by silica gel column chromatography; with chloroform-methanol (100: 1-50: 1,, obtain 4-[(4S)-fluoro-1-[4-(N '-phenyl urea groups) phenyl acetyl v/v) as eluant]-(2S)-and the pyrrolidinyl methoxyl group] essence of Niobe (570mg; 99%), is the weak yellow foam thing. 1H-NMR (CDCl 3) δ 2.07-2.58 (m, 2H), 3.55-3.56 (m, 1H), (3.69-3.98 a plurality of s and m, 6H altogether), 4.01-4.08 and 4.21-4.25 (respectively are m, 1H), 4.46-4.65 (m, 2H), 5.23-5.25 and 5.38 (respectively is m, 1H), 6.88-7.07 (m, 7H), 7.15-7.20 (m, 2H), 7.28-7.30 (m, 2H), 7.34 and 7.40 (respectively is s, 1H), 7.71 and 7.81 (respectively be s, 1H), 7.91-7.95 and 7.99-8.01 (respectively are m, 2H); MS (ESI) m/z 506 (M ++ 1).
To 4-[(4S)-fluoro-1-[4-(N '-the phenyl urea groups) phenyl acetyl]-(2S)-and the pyrrolidinyl methoxyl group] essence of Niobe (570mg; 1.13mmol) the agitating solution of THF (5ml) in add 0.5N sodium hydroxide (5ml), with the heating 5 hours under refluxing of this reactant mixture.After being cooled to room temperature, in this mixture impouring ice-1N HCl, the precipitation that produces is collected in decompression.The crude product solid is recrystallization from methanol-chloroform-IPE, obtains 72 (348mg, 63%), is white crystalline powder.MW491.51, mp 169-171 ℃; 1H-NMR (DMSO-d 6) δ 2.24-2.36 (m, 2H), 3.47-4.08 (m, 5H), 4.20-4.64 (m, 2H), 5.31-5.50 (m, 1H), 6.94-7.46 (a plurality of m, 11H altogether), 7.87-7.92 (m, 2H), 8.64-8.67 (m, 2H), 12.63 (br s, 1H); MS (FAB) m/z 492 (M ++ 1).C 27H 26FN 3O 61/4H 2The analytical calculation value of O: C, 65.38; H, 5.38; N, 8.47; F, 3.83.Measured value: C, 65.13; H, 5.38; N, 8.25; F, 3.78.Embodiment 674-[(4S)-fluoro-1-[3-methyl-4-(N '-the phenyl urea groups) phenyl acetyl]-(2S)-and the pyrrolidinyl methoxyl group] benzoic acid
Figure A0080951002481
To 4-amino-3-aminomethyl phenyl tert-butyl acetate (1.20g, 5.42mmol) and triethylamine (830ml, (650ml, 5.98mmol), stirred reaction mixture spends the night under room temperature to add carbanil in the agitating solution of THF 5.95mmol) (10ml).This reactant mixture is concentrated into small size, dilutes with normal hexane.The precipitation that decompression collect to produce is used the normal hexane wash filtrate, obtains 3-methyl-4-(N '-phenyl urea groups) phenylacetic acid tert-butyl ester (1.12g, 61%), is white crystalline powder.mp?143-145℃； 1H-NMR(CDCl 3)δ1.47(s，9H)，2.09(s，3H)，3.47(s，2H)，6.44(s，1H)，7.01-7.07(m，4H)，7.16-7.27(m，2H)，7.30-7.33(m，2H)，7.45-7.47(m，1H)。
(1.12g adds TFA (10ml) in the agitating solution of dichloromethane 3.29mmol) (10ml), under room temperature, this reactant mixture was stirred 4 hours to 3-methyl-4-(N '-phenyl urea groups) phenylacetic acid tert-butyl ester.This reactant mixture is concentrated in small size and the impouring ice-water.The precipitation that decompression collect to produce, crude product solid recrystallization from methanol-chloroform obtains 3-methyl-4-(N '-phenyl urea groups) phenylacetic acid (680mg, 73%), is the white needles thing. 1H-NMR(DMSO-d 6)δ2.22(s，3H)，3.46(s，2H)，6.93-7.05(m，3H)，7.25-7.29(m，2H)，7.43-7.46(m，2H)，7.72-7.74(m，1H)，7.90(s，1H)，8.98(s，1H)，12.26(brs，1H)。C 16H 16N 2O 3The analytical calculation value: C, 67.59; H, 5.67; N, 9.85.Measured value: C, 67.47; H, 5.68; N, 9.73.
Under room temperature, with 3-methyl-4-(N '-phenyl urea groups) phenylacetic acid (301mg, 1.06mmol), 4-[(4S)-fluoro-(2S)-pyrrolidinyl methoxyl group] essence of Niobe (268mg, 1.06mmol), EDCHCl (243mg, 1.27mmol), HOBt (172mg, 1.27mmol) and triethylamine (180ml, 1.29mmol) mixture in DMF (5ml) stirs and to spend the night.This mixture of dilute with water is used ethyl acetate extraction.With salt water washing extract, through dried over sodium sulfate and evaporation.Residue is through purification by silica gel column chromatography; with chloroform-methanol (100: 1-60: 1,, obtain 4-[(4S)-fluoro-1-[3-methyl-4-(N '-phenyl urea groups) phenyl acetyl v/v) as eluant]-(2S)-and the pyrrolidinyl methoxyl group] essence of Niobe (550mg; q.y.), be the white foam thing. 1H-NMR (CDCl 3) δ 1.79 and 1.87 (respectively be s, 3H), 2.04-2.61 (m, 2H), 3.52-3.54 (m, 1H), 3.73-4.27 (a plurality of s and m, 7H altogether), 4.47-4.67 (m, 2H), 5.26-5.27 and 5.40 (respectively is m, 1H), 6.79-6.99 (m, 6H), 7.14-7.18 (m, 2H), 7.27-7.31 (m, 2H), 7.40-7.44 (m, 1H), and 7.89-8.01 (m, 3H); MS (ESI) m/z 520 (M ++ 1).
To 4-[(4S)-fluoro-1-[3-methyl-4-(N '-the phenyl urea groups) phenyl acetyl]-(2S)-and the pyrrolidinyl methoxyl group] essence of Niobe (550mg; 1.06mmol) the agitating solution of THF (5ml) in add 0.5N sodium hydroxide (5ml), with the heating 2 hours under refluxing of this reactant mixture.After being cooled to room temperature, in this mixture impouring ice-1N HCl, the precipitation that produces is collected in decompression.The crude product solid is recrystallization from methanol-chloroform-IPE, obtains 73 (226mg, 42%), is white crystalline powder.MW 505.54, mp 130-135 ℃; 1H-NMR (DMSO-d 6) δ 2.18-2.30 (a plurality of s and m, 5H altogether), 3.47-3.92 (a plurality of m, 5H altogether), 4.03-4.63 (m, 2H), 5.31-5.50 (m, 1H), 6.94-7.10 (m, 5H), 7.26-7.30 (m, 2H), 7.45-7.47 (m, 2H), 7.70-7.75 (m, 1H), 7.87-7.92 (m, 3H), 8.96-8.98 (m, 1H), 12.63 (br s, 1H); MS (ESI) m/z506 (M ++ 1).C 28H 28FN 3O 51/2H 2The analytical calculation value of O: C, 65.36; H, 5.68; N, 8.17; F, 3.69.Measured value: C, 65.61; H, 5.71; N, 7.84; F, 3.60.Embodiment 684-[(4S)-fluoro-1-[4-[N '-(2-fluoro phenyl) urea groups]-3-aminomethyl phenyl acetyl group]-(2S)-and the pyrrolidinyl methoxyl group] benzoic acid
To 4-amino-3-aminomethyl phenyl tert-butyl acetate (1.09g, 4.93mmol) and triethylamine (755ml, 5.42mmol) the agitating solution of THF (10ml) in add Carbimide. 2-fluoro phenyl ester (610 μ l, 5.44mmol), stirred reaction mixture spend the night under room temperature.This reactant mixture is concentrated into small size, dilutes with normal hexane.The precipitation that produces is collected in decompression, uses the normal hexane wash filtrate, obtains 4-[N '-(2-fluoro phenyl) urea groups]-3-aminomethyl phenyl tert-butyl acetate (1.31g, 74%), be white crystalline powder.mp?89-91℃； 1H-NMR(CDCl 3)δ1.47(s，9H)，2.06(s，3H)，3.49(s，2H)，6.62(s，1H)，6.92-7.09(m，5H)，7.21(br?s，1H)，7.49-7.51(m，1H)，8.10-8.15(m，1H)。C 20H 23FN 2O 3The analytical calculation value: C, 67.02; H, 6.47; N, 7.82; F, 5.30.Measured value: C, 66.74; H, 6.35; N, 7.85; F, 5.69.
To 4-[N '-(2-fluoro phenyl) urea groups]-(1.25g adds TFA (10ml) to 3-aminomethyl phenyl tert-butyl acetate in the agitating solution of dichloromethane 3.49mmol) (10ml), under room temperature, this reactant mixture stirring is spent the night.This reactant mixture is concentrated in small size and the impouring ice-water.The precipitation that produces is collected in decompression, and the crude product solid is recrystallization from methanol-chloroform-IPE, obtains 4-[N '-(2-fluoro phenyl) urea groups]-3-aminomethyl phenyl acetic acid (830mg, 79%), be the white needles thing. 1H-NMR(DMSO-d 6)δ2.23(s，3H)，3.47(s，2H)，6.96-7.30(m，5H)，7.74-7.76(m，1H)，8.17-8.20(m，1H)，8.33(s，1H)，8.94(s，1H)，12.27(brs，1H)。C 16H 15FN 2O 3The analytical calculation value: C, 63.57; H, 5.00; N, 9.27, F, 6.28.Measured value: C, 63.28; H, 5.00; N, 9.14, F, 6.43.
Under room temperature, with 4-[N '-(2-fluoro phenyl) urea groups]-3-aminomethyl phenyl acetic acid (321mg, 1.06mmol), 4-[(4S)-fluoro-(2S)-pyrrolidinyl methoxyl group] essence of Niobe (269mg, 1.06mmol), EDCHCl (244mg, 1.27mmol), HOBt (172mg, 1.27mmol) and triethylamine (177ml, 1.27mmol) mixture in DMF (5ml) stirs and to spend the night.This mixture of dilute with water is used ethyl acetate extraction.With salt water washing extract, through dried over sodium sulfate and evaporation.Residue is through purification by silica gel column chromatography; with chloroform-methanol (100: 1; v/v) as eluant; obtain 4-[(4S)-fluoro-1-[4-[N '-(2-fluoro phenyl) urea groups]-3-aminomethyl phenyl acetyl group]-(2S)-and the pyrrolidinyl methoxyl group] essence of Niobe (560mg; 98%), is the white foam thing. 1H-NMR (CDCl 3) δ 1.78 and 1.86 (respectively be s, 3H), 2.16-2.65 (m, 2H), 3.58-3.61 (m, 1H), 3.74-4.15 (a plurality of s and m, 7H altogether), 4.29-4.34 and 4.46-4.49 (respectively are m, 1H), 4.64-4.73 (m, 1H), 5.29-5.34 and 5.43-5.47 (respectively are m, 1H), 6.84-6.97 (m, 6H), 7.04-7.07 (m, 1H), 7.21 (br s, 1H), 7.55-7.59 (m, 1H), 7.85-8.02 (m, 3H), 8.18-8.22 (m, 1H); MS (ESI) m/z538 (M ++ 1).
To 4-[(4S)-fluoro-1-[4-[N '-(2-fluoro phenyl) urea groups]-3-aminomethyl phenyl acetyl group]-(2S)-and the pyrrolidinyl methoxyl group] essence of Niobe (560mg; 1.04mmol) the agitating solution of THF (5ml) in add 0.5N sodium hydroxide (5ml), with the heating 5 hours under refluxing of this reactant mixture.After being cooled to room temperature, in this mixture impouring ice-1N HCl, the precipitation that produces is collected in decompression.The crude product solid is recrystallization from methanol-chloroform-IPE, obtains 74 (297mg, 42%), is white crystalline powder.MW 523.53, mp 137-143 ℃; 1H-NMR (DMSO-d 6) δ 2.20-2.31 (a plurality of s and m, 5H altogether), 3.56-3.92 (a plurality of m, 5H altogether), 4.03-4.63 (m, 2H), 5.31-5.50 (m, 1H), (6.96-7.26 a plurality of m, 7H altogether), and 7.72-7.77 (m, 1H), 7.87-7.92 (m, 2H), 8.17-8.22 (m, 1H), 8.32-8.36 (m, 1H), 8.94-8.95 (m, 1H), 12.66 (br s, 1H); MS (ESI) m/z 524 (M ++ 1).C 28H 27F 2N 3O 5The analytical calculation value: C, 64.24; H, 5.20; N, 8.03; F, 7.26.Measured value: 64.44; H, 5.75; N, 7.40; F, 6.73.Embodiment 694-[(4S)-and fluoro-1-[4-[N '-(2-trifluoromethyl) urea groups] phenyl acetyl]-(2S)-and the pyrrolidinyl methoxyl group] benzoic acid
Figure A0080951002521
To 4-aminophenyl ethyl acetate (1.13g, 6.31mmol) and triethylamine (965ml, (953ml, 6.31mmol), stirred reaction mixture is 2 days under room temperature to add Carbimide. 2-trifluoromethyl phenyl ester in the agitating solution of THF 6.92mmol) (10ml).The precipitation that produces is collected in decompression, uses the normal hexane wash filtrate, obtains 4-[N '-(2-trifluoromethyl) urea groups] phenylacetic acid ethyl ester (1.93g, 84%), be the white needles thing.mp?137-139℃； 1H-NMR(CDCl 3)δ1.25-1.29(m，3H)，3.59(s，2H)，4.15-4.20(m，2H)，7.05(br?s，1H)，7.13-7.23(m，6H)，7.47-7.51(m，1H)，7.54-7.56(m，1H)，8.01-8.03(m，1H)。
To 4-[N '-(2-trifluoromethyl) urea groups] (1.93g adds 1N sodium hydroxide (10ml) to the phenylacetic acid ethyl ester in the agitating solution of THF 5.27mmol) (10ml), this reactant mixture was heated 5 hours under refluxing.After being cooled to room temperature, in this mixture impouring ice-1N HCl.The precipitation that produces is collected in decompression, and the crude product solid is recrystallization from methanol-chloroform-IPE, obtains 4-[N '-(2-trifluoromethyl) urea groups] phenylacetic acid (910mg, 51%), be white crystalline powder.mp?224-225℃； 1H-NMR(DMSO-d 6)δ3.50(s，2H)，7.18(d，J＝8.3Hz，2H)，7.25-7.29(m，1H)，7.40(d，J＝8.3Hz，2H)，7.62-7.69(m，2H)，7.95-7.97(m，1H)，8.06(s，1H)，9.37(s，1H)，12.27(br?s，1H)。C 16H 13F 3N 2O 3The analytical calculation value: C, 56.81; H, 3.87; N, 8.28, F, 16.85.Measured value: C, 56.68; H, 3.87; N, 8.16, F, 16.89.
Under room temperature, with 4-[N '-(2-trifluoromethyl) urea groups] phenylacetic acid (302mg, 0.89mmol), 4-[(4S)-fluoro-(2S)-pyrrolidinyl methoxyl group] essence of Niobe (226mg, 0.89mmol), EDCHCl (205mg, 1.07mmol), HOBt (145mg, 1.07mmol) and triethylamine (150ml, 1.08mmol) mixture in DMF (5ml) stirred 3 days.This mixture of dilute with water is used ethyl acetate extraction.With salt water washing extract, through dried over sodium sulfate and evaporation.Residue is through purification by silica gel column chromatography; with chloroform-methanol (100: 1-60: 1; v/v) as eluant; obtain 4-[(4S)-fluoro-1-[4-[N '-(2-trifluoromethyl) urea groups] phenyl acetyl]-(2S)-and the pyrrolidinyl methoxyl group] essence of Niobe (463mg; 90%), is the weak yellow foam thing. 1H-NMR (CDCl 3) δ 2.09-2.60 (m, 2H), 3.56-4.12 (a plurality of s and m, 8H altogether), 4.26-4.65 (m, 2H), 5.26-5.29 and 5.39-5.42 (respectively be m, be total to 1H), 6.87-6.93 (m, 2H), 6.99-7.13 (m, 5H), 7.30-7.33 (m, 1H), 7.44-7.53 (m, 2H), 7.88-7.92 (m, 1H), 7.99-8.04 (m, 2H), and 8.09-8.15 (m, 1H); MS (ESI) m/z574 (M ++ 1).
To 4-[(4S)-fluoro-1-[4-[N '-(2-trifluoromethyl) urea groups] phenyl acetyl]-(2S)-and the pyrrolidinyl methoxyl group] essence of Niobe (460mg; 0.80mmol) the agitating solution of THF (5ml) in add 0.5N sodium hydroxide (5ml), with the heating 5 hours under refluxing of this reactant mixture.After being cooled to room temperature, in this mixture impouring ice-1N HCl, the precipitation that produces is collected in decompression.The crude product solid is recrystallization from methanol-chloroform-IPE, obtains 75 (169mg, 38%), is white crystalline powder.MW 559.51 mp 130-135 ℃; 1H-NMR (DMSO-d 6) δ 2.24-2.30 (m, 2H), 3.51-4.24 (a plurality of m, be total to 5H), 4.38-4.40 and 4.61 (respectively be m, be total to 2H), 5.31-5.50 (m, 1H), 7.03-7.42 (a plurality of m, be total to 7H), and 7.62-7.69 (m, 2H), 7.87-8.07 (m, 4H), 9.36-9.37 (m, 1H), 12.64 (br s, 1H); MS (ESI) m/z560 (M ++ 1).C 28H 25F 4N 3O 5The analytical calculation value: C, 60.11; H, 4.50; N, 7.51; F, 13.58.Measured value: C, 60.10; H, 4.85; N, 7.01; F, 12.90.Embodiment 704-[(4S)-and fluoro-1-[3-methoxyl group-4-[N '-(2-trifluoromethyl) urea groups] phenyl acetyl]-(2S)-and the pyrrolidinyl methoxyl group] benzoic acid
To 4-amino-3-methoxyphenyl tert-butyl acetate (1.11g, 4.68mmol) and triethylamine (720ml, 5.17mmol) the agitating solution of THF (10ml) in add Carbimide. 2-trifluoromethyl phenyl ester (707ml, 4.68mmol), stirred reaction mixture be 2 days under room temperature.This reactant mixture is concentrated into small size, dilutes with normal hexane.The precipitation that produces is collected in decompression, with the normal hexane washing, obtains 3-methoxyl group-4-[N '-(2-trifluoromethyl) urea groups] the phenylacetic acid tert-butyl ester (1.11g, 56%), be white crystalline powder.mp?131-133℃； 1H-NMR(CDCl 3)δ1.44(s，9H)，3.49(s，2H)，3.85(s，3H)，6.83-6.88(m，3H)，6.98(br?s，1H)，7.17-7.21(m，1H)，7.52-7.59(m，2H)，7.89-7.91(m，1H)，8.04-8.06(m，1H)。
To 3-methoxyl group-4-[N '-(2-trifluoromethyl) urea groups] (1.11g adds TFA (10ml) to the phenylacetic acid tert-butyl ester in the agitating solution of dichloromethane 2.62mmol) (10ml), under room temperature, this reactant mixture was stirred 4 hours.This reactant mixture is concentrated in small size and the impouring ice-water.The precipitation that produces is collected in decompression, and the crude product solid is recrystallization from methanol-chloroform-IPE, obtains 3-methoxyl group-4-[N '-(2-trifluoromethyl) urea groups] phenylacetic acid (839mg, 87%), be white crystalline powder.Mp 218-220 ℃; 1H-NMR (DMSO-d 6) δ 3.51 (s, 2H), 3.87 (s, 3H), 6.76-6.79 (m, 1H), 6.93-6.94 (m, 1H), 7.27-7.30 (m, 1H), 7.61-7.69 (m, 2H), 7.82-7.84 (m, 1H), 7.97-7.99 (m, 1H), 8.71 (s, 1H), 8.89 (s, 1H), 12.30 (br s, 1H); C 17H 15F 3N 2O 4The analytical calculation value: C, 55.44; H, 4.11; N, 7.61; F, 15.47.Measured value: C, 55.30; H, 4.08; N, 7.63; F, 15.13.
Under room temperature, with 3-methoxyl group-4-[N '-(2-trifluoromethyl) urea groups] phenylacetic acid (353mg, 0.96mmol), 4-[(4S)-fluoro-(2S)-pyrrolidinyl methoxyl group] essence of Niobe (243mg, 0.96mmol), EDCHCl (221mg, 1.15mmol), HOBt (156mg, 1.15mmol) and triethylamine (160ml, 1.15mmol) mixture in DMF (5ml) stirs and to spend the night.This mixture of dilute with water is used ethyl acetate extraction.With salt water washing extract, through dried over sodium sulfate and evaporation.Residue is through purification by silica gel column chromatography; with chloroform-methanol (100: 1-60: 1; v/v) as eluant; obtain 4-[(4S)-fluoro-1-[3-methoxyl group-4-[N '-(2-trifluoromethyl) urea groups] phenyl acetyl]-(2S)-and the pyrrolidinyl methoxyl group] essence of Niobe (570mg; 98%), is the white foam thing. 1H-NMR (CDCl 3) δ 2.05-2.58 (m, 2H), 3.56-4.21 (a plurality of m, 11H altogether), 4.05-4.64 (m, 2H), 5.23-5.25 and 5.36-5.37 (respectively be m, be total to 1H), 6.79-6.82 (m, 2H), 6.89-7.00 (m, 2H), and 7.16-7.20 (m, 2H), 7.39-7.43 (m, 1H), 7.51-7.59 (m, 2H), and 7.93-8.02 (m, 4H); MS (ESI) m/z640 (M ++ 1).
To 4-[(4S)-fluoro-1-[3-methoxyl group-4-[N '-(2-trifluoromethyl) urea groups] phenyl acetyl]-(2S)-and the pyrrolidinyl methoxyl group] essence of Niobe (570mg; 0.94mmol) the agitating solution of THF (5ml) in add 0.5N sodium hydroxide (5ml), with the heating 2 hours under refluxing of this reactant mixture.After being cooled to room temperature, in this mixture impouring ice-1N HCl, the precipitation that produces is collected in decompression.The crude product solid is recrystallization from methanol-chloroform-IPE, obtains 76 (234mg, 42%), is white crystalline powder.MW?589.54?mp?129-132℃。 1H-NMR (DMSO-d 6) δ 2.23-2.29 (m, 2H), 3.54-4.38 (a plurality of s and m, 8H altogether), 4.40-4.61 (m, 2H), 5.30-5.36 and 5.43-5.49 (respectively be m, be total to 1H), 6.72-6.91 (m, 2H), 7.02-7.08 (m, 2H), 7.25-7.29 (m, 1H), 7.59-7.67 (m, 2H), 7.81-7.99 (m, 4H), 8.69-8.70 (m, 1H), 8.87-8.90 (m, 1H), 12.67 (br s, 1H); MS (ESI) m/z 589 (M ++ 1).C 29H 27F 4N 3O 6The analytical calculation value: C, 59.08; H, 4.62; N, 7.13.Measured value: C, 59.22; H, 5.10; N, 6.58.Embodiment 714-[(4S)-and fluoro-1-[3-methyl-4-[N '-(2-trifluoromethyl) urea groups] phenyl acetyl]-(2S)-and the pyrrolidinyl methoxyl group] benzoic acid
Figure A0080951002551
To 4-amino-3-aminomethyl phenyl tert-butyl acetate (927mg, 4.19mmol) and triethylamine (645 μ l, 4.63mmol) the agitating solution of THF (10ml) in add Carbimide. 2-trifluoromethyl phenyl ester (633 μ l, 4.19mmol), stirred reaction mixture be 2 days under room temperature.This reactant mixture is concentrated into small size, dilutes with normal hexane.The precipitation that produces is collected in decompression, uses the normal hexane wash filtrate, obtains 3-methyl-4-[N '-(2-trifluoromethyl) urea groups] the phenylacetic acid tert-butyl ester (1.06g, 62%), be white crystalline powder.mp?178-180℃； 1H-NMR(CDCl 3)δ1.44(s，9H)，2.25(s，3H)，3.51(s，2H)，6.38(br?s，1H)，7.12-7.18(m，3H)，7.36-7.37(m，1H)，7.49-7.53(m，2H)，8.13-8.16(m，1H)。
To 3-methyl-4-[N '-(2-trifluoromethyl) urea groups] (1.06g adds TFA (10ml) to the phenylacetic acid tert-butyl ester in the agitating solution of dichloromethane 2.60mmol) (10ml), under room temperature, this reactant mixture was stirred 4 hours.This reactant mixture is concentrated in small size and the impouring ice-water.The precipitation that produces is collected in decompression, and the crude product solid is recrystallization from methanol-chloroform-IPE, obtains 3-methyl-4-[N '-(2-trifluoromethyl) urea groups] phenylacetic acid (702mg, 77%), be white crystalline powder.mp?262-263℃； 1H-NMR(DMSO-d 6)δ2.24(s，3H)，3.48(s，2H)，7.03(d，J＝8.3Hz，1H)，7.08(s，1H)，7.26-7.30(m，1H)，7.61-7.69(m，3H)，7.88(d，J＝8.3Hz，1H)，8.39(s，1H)，8.55(s，1H)，12.28(br?s，1H)。C 17H 15F 3N 2O 3The analytical calculation value: C, 57.96; H, 4.29; N, 7.95; F, 16.18.Measured value: C, 57.73; H, 4.23; N, 7.92; F, 16.05.
Under room temperature, with 3-methyl-4-[N '-(2-trifluoromethyl) urea groups] phenylacetic acid (359mg, 1.02mmol), 4-[(4S)-fluoro-(2S)-pyrrolidinyl methoxyl group] essence of Niobe (258mg, 1.02mmol), EDCHCl (234mg, 1.22mmol), HOBt (165mg, 1.22mmol) and triethylamine (170 μ l, 1.22mmol) mixture in DMF (5ml) stirs and to spend the night.This mixture of dilute with water is used ethyl acetate extraction.With salt water washing extract, through dried over sodium sulfate and evaporation.Residue is through purification by silica gel column chromatography; with chloroform-methanol (100: 1-60: 1; v/v) as eluant; obtain 4-[(4S)-fluoro-1-[3-methyl-4-[N '-(2-trifluoromethyl) urea groups] phenyl acetyl]-(2S)-and the pyrrolidinyl methoxyl group] essence of Niobe (612mg; q.y.), be the white foam thing. 1H-NMR (CDCl 3) δ 1.92 and 2.00 (respectively be s, be total to 3H), 2.09-2.61 (m, 2H), 3.56-4.29 (a plurality of m, 8H altogether), 4.45-4.48 and 4.59-4.64 (respectively be m, altogether 2H), 5.24-5.30 and 5.38-5.44 (respectively are m, be total to 1H), and 6.90-7.14 (m, 5H), 7.22-7.53 (m, 5H), 7.90-7.92 (m, 1H), and 8.00-8.06 (m, 2H); MS (ESI) m/z588 (M ++ 1).
To 4-[(4S)-fluoro-1-[3-methyl-4-[N '-(2-trifluoromethyl) urea groups] phenyl acetyl]-(2S)-and the pyrrolidinyl methoxyl group] essence of Niobe (610mg; 1.04mmol) the agitating solution of THF (5Hl) in add 0.5N sodium hydroxide (5ml), with the heating 2 hours under refluxing of this reactant mixture.After being cooled to room temperature, in this mixture impouring ice-1N HCl, the precipitation that produces is collected in decompression.The crude product solid is recrystallization from methanol-chloroform-IPE, obtains 77 (186mg, 31%), is white crystalline powder.MW?573.54?mp?123-126℃。 1H-NMR (DMSO-d 6) δ 2.19-2.29 (a plurality of s and m, 5H altogether), 3.64-4.21 (a plurality of m, 5H altogether), 4.36-4.60 (m, 2H), 5.30-5.36 and 5.43-5.49 (respectively be m, be total to 1H), 6.97-7.08 (m, 4H), 7.24-7.28 (m, 1H), 7.59-7.68 (m, 3H), 7.85-7.90 (m, 3H), 8.37-8.39 (m, 1H), 8.54-8.55 (m, 1H), 12.67 (br s, 1H); MS (ESI) m/z573 (M +).Embodiment 724-[(4S)-and fluoro-1-[3-methyl-4-[N '-(2-aminomethyl phenyl) urea groups] phenyl acetyl]-(2S)-and pyrrolidinyl] methoxybenzoic acid
Under room temperature, with 3-methyl-4-[N '-(2-aminomethyl phenyl) urea groups] phenylacetic acid (250mg, 0.84mmol), 4-[(4S)-fluoro-(2S)-pyrrolidinyl] methoxyl methyl benzoate (400mg, 1.06mmol), EDCHCl (242mg, 1.26mmol) and DMAP (154mg, 1.26mmol) mixture in DMF (5ml) stirred 21 hours.In this mixture impouring frozen water, use ethyl acetate extraction.With the extract of frozen water and salt water washing merging, after dried over sodium sulfate, the vacuum concentration extract.Residue is through silica gel column chromatography [50g; chloroform/methanol (50/1)]; then through TLC[chloroform/acetone (10/1)] purification; obtain 4-[(4S)-fluoro-1-[3-methyl-4-[N '-(2-aminomethyl phenyl) urea groups] phenyl acetyl]-(2S)-and pyrrolidinyl] methoxyl methyl benzoate (342mg; 76%), is colourless amorphous solid.IR(KBr)3356，2951，1716，1651，1604，1537，1252cm -1； 1H-NMR(CDCl 3)δ2.07(d，J＝6.6Hz，2H)，2.12(s，3H)，2.27(m，1H)，2.24(s，3H)，2.30-2.59(m，1H)，3.60(d，J＝5.3Hz，1H)，3.65-4.23(m，3H)，3.87(s，3H)，4.50-4.62(m，1H)，5.31(d，J＝52.4Hz，1H)，6.23(d，J＝11.2Hz，1H)，6.26(d，J＝11.9Hz，1H)，6.87-7.27(m，8H)，7.54-7.65(m，3H)，7.94-8.01(m，2H)；MS(FAB)m/z534(M ++1)。C 30H 32FN 3O 50.7H 2The analytical calculation value of O: C, 65.97; H, 6.16; F, 3.48; N, 7.69.Measured value: C, 66.04; H, 6.07; F, 3.55; N, 7.64.
To 4-[(4S)-fluoro-1-[3-methyl-4-[N '-(2-aminomethyl phenyl) urea groups] phenyl acetyl]-(2S)-and pyrrolidinyl] (227mg adds 0.25N sodium hydroxide (3.4ml) to methoxyl methyl benzoate in the agitating solution of THF 0.425mmol) (3.4ml).After stirring 4 days under the room temperature,, extract with chloroform-methanol (10/1) with this mixture of 1N HCl acidify.Extract and vacuum concentration through the dried over sodium sulfate merging.Residue is through preparation property-TLC[chloroform/methanol (10/1)] purification, obtain 78 (190mg, 86%), be colourless amorphous solid.MW?519.56?IR(KBr)3356，2974，1604，1537，1454，1252?cm -1； 1H-NMR(DMSO-d 6)δ2.24(s，3H)，2.26(s，3H)，3.60(d，J＝3.7Hz，2H)，3.65-4.65(m，8H)，5.31-5.50(m，1H)，6.92-7.18(m，7H)，7.67-7.92(m，4H)，8.22-8.32(m，2H)；MS(FAB)m/z?520(M ++1)。C 29H 30FN 3O 71.1H 2The analytical calculation value of O: C, 64.58; H, 6.02; F, 3.52; N, 7.79.Measured value: C, 64.71; H, 5.90; F, 3.24; N, 7.51.Embodiment 734-[1-[4-[N '-(2-chlorophenyl) urea groups]-3-aminomethyl phenyl acetyl group]-(4S)-and fluoro-(2S)-pyrrolidinyl] methoxybenzoic acid
Under room temperature, to 4-amino-3-aminomethyl phenyl tert-butyl acetate (1.00g, 4.52mmol), Carbimide. 2-chloro phenyl ester (0.55ml, 4.52mmol) add in the stirring the mixture in THF (10ml) triethylamine (0.13ml, 0.90mmol).Stir after 6 hours the vacuum concentration reactant mixture.Grind residue by adding normal hexane, obtain 4-[N '-(2-chlorophenyl) urea groups]-3-aminomethyl phenyl tert-butyl acetate (1.57g, 93%), be pale yellow powder.Mp 104-106 ℃ (decomposition); 1H-NMR (CDCl 3) δ 1.45 (s, 9H), 2.28 (s, 3H), 3.51 (s, 2H), 6.33 (br, 1H), 6.96 (t, J=7.6Hz, 1H), 7.08 (br, 1H), 7.16-7.30 (m, 4H), 7.42 (m, 1H), 8.2 (d, J=8.1Hz, 1H).
Under room temperature, to 4-[N '-(2-chlorophenyl) urea groups]-(1.57g adds TFA (6ml) to 3-aminomethyl phenyl tert-butyl acetate in the agitating solution of dichloromethane 4.19mmol) (10ml).Stir after 4 hours vacuum concentrated mixture.Grind residue by adding entry, obtain 4-[N '-(2-chlorophenyl) urea groups]-3-aminomethyl phenyl acetic acid (1.33g, 100%), be pale yellow powder.Mp243-245 ℃ (decomposition); 1H-NMR (CDCl 3) δ 2.24 (s, 3H), 3.47 (s, 2H), 6.99-7.08 (m, 3H), 7.28 (t, J=7.6Hz, 1H), 7.44 (dt, J=8.0,2.4Hz, 1H), 7.66 (dd, J=8.3,1.9Hz, 1H), 8.13 (dd, J=6.1,1.7Hz, 1H), 8.61 (d, J=6.3Hz, 2H); MS (ESI), m/z 319 (M ++ 1), 321 (M ++ 3); C 16H 15ClN 2O 30.7TFA the analytical calculation value: C, 59.33; H, 4.65; Cl, 10.85; N, 8.57.Measured value: C, 59.23; H, 4.64; Cl, 10.90; N, 8.40.
With 4-[N '-(2-chlorophenyl) urea groups]-3-aminomethyl phenyl acetic acid (252mg, 0.79mmol), 4-[(4S)-fluoro-(2S)-pyrrolidinyl] methoxyl methyl benzoate (200mg, 0.79mmol), EDCHCl (227mg, 1.20mmol) and DMAP (147mg, 1.20mmol) mixture in DMF (5ml) stirred 17 hours.In this mixture impouring frozen water, use ethyl acetate extraction.Extract with frozen water and salt water washing merging.After dried over sodium sulfate, the vacuum concentration extract.Residue is through TLC[chloroform/acetone (10/1)] purification; obtain 4-[1-[4-[N '-(2-chlorophenyl) urea groups]-3-aminomethyl phenyl acetyl group]-(4S)-and fluoro-(2S)-pyrrolidinyl] methoxyl methyl benzoate (390mg; 89%), is colourless amorphous solid.IR(KBr)3340，2951，1712，1624，1604，1533，1438?cm -1； 1H-NMR(CDCl 3)δ1.92-2.05(m，3H)，2.07-2.63(m，2H)，3.61(d，2H，J＝8.8Hz)，3.70-4.15(m，5H)，4.25-4.67(m，2H)，5.26-5.44(m，1H)，6.84-8.19(m，13H)；MS(FAB)m/z?554(M ++1)，556(M ++3)。
To 4-[1-[4-[N '-(2-chlorophenyl) urea groups]-3-aminomethyl phenyl acetyl group]-(4S)-and fluoro-(2S)-pyrrolidinyl] (268mg adds 0.25N sodium hydroxide (3.8ml) to methoxyl methyl benzoate in the agitating solution of THF 0.484mmol) (3.8ml).After stirring 1 day under the room temperature,, extract with chloroform-methanol (10/1) with this mixture of 1N HCl acidify.Extract and vacuum concentration through the dried over sodium sulfate merging.Residue is through TLC[chloroform/methanol (10/1)] purification, obtain 79 (124mg, 47%), be colourless amorphous solid.MW 539.98 IR (KBr) 3346,2976,1709,1685,1604,1533,1439cm -1 1H-NMR (DMSO-d 6) δ 2.20 (s, 3H, one of isomer), 2.24 (s, 3H, one of isomers), 2.30 (m, 1H), 3.60 (s, 2H), 3.71-4.62 (m, 6H), 5.30-5.50 (m, 1H), 7.01-7.09 (m, 5H), 7.28 (t, J=7.8Hz, 1H), 7.44 (d, J=8.1Hz, 1H), 7.66 (t, J=8.1Hz, 1H), 7.87 (d, J=7.1Hz, 2H), 8.13 (d, J=7.9Hz, 1H), 8.62 (d, J=6.1Hz, 2H); MS (FAB) m/z540 (M ++ 1), 542 (M ++ 3).For sodium salt, C 28H 27ClFN 3O 7Na0.5EtOH1.5H 2The analytical calculation value of O: C, 56.91; H, 5.27; Cl, 5.79; F, 3.10; N, 6.87.Measured value: C, 56.60; H, 4.98; Cl, 5.88; F, 3.08; N, 6.52.Embodiment 744-[1-[4-[N '-(2-bromo phenyl) urea]-3-aminomethyl phenyl acetyl group]-(4S)-and fluoro-(2S)-pyrrolidinyl] methoxybenzoic acid
Figure A0080951002601
Under room temperature, to 4-amino-3-aminomethyl phenyl tert-butyl acetate (780mg, 3.30mmol), Carbimide. 2-bromo phenyl ester (0.41ml, 3.30mmol) add in the stirring the mixture in THF (7ml) triethylamine (0.092ml, 0.66mmol).Stir after 3 hours the vacuum concentration reactant mixture.Grind residue by adding normal hexane, obtain 4-[N '-(2-bromo phenyl) urea groups]-3-aminomethyl phenyl tert-butyl acetate (1.57g, 93%), be pale yellow powder.Mp 138-145 ℃ (decomposition); 1H-NMR (CDCl 3) δ 1.44 (s, 9H), 2.33 (s, 3H), 3.51 (s, 2H), 6.90 (dt, J=9.0,1.4Hz, 1H), 6.98 (br, 1H), 7.18-7.31 (m, 4H), 7.39 (dd, J=8.1,2.9Hz, 1H), 7.44 (d, J=8.0Hz, 1H), 8.22 (d, J=8.3Hz, 2H); C 20H 22BrN 2O 30.2H 2The analytical calculation value of O: C, 56.80; H, 5.58; N, 6.62.Measured value: C, 56.85; H, 5.42; N, 6.62.
Under room temperature, to 4-[N '-(2-bromo phenyl) urea groups]-(1.27g adds TFA (5ml) to 3-aminomethyl phenyl tert-butyl acetate in the agitating solution of dichloromethane 3.03mmol) (10ml).Stir after 1 hour vacuum concentrated mixture.Grind residue by adding entry, obtain 4-[N '-(2-bromo phenyl) urea groups]-3-aminomethyl phenyl acetic acid (1.05g, 95%), be pale yellow powder.Mp245-248 ℃ (decomposition); 1H-NMR (CDCl 3) δ 2.24 (s, 3H), 3.48 (s, 2H), 6.96 (dt, J=7.3,1.5Hz, 1H), 7.02 (d, J=8.3Hz, 1H), 7.07 (s, 1H), 7.32 (t, J=8.1Hz, 1H), 7.59-7.66 (m, 2H), 8.44 (s, 1H), 8.62 (s, 1H); MS (ESI), m/z 363 (M ++ 1), 365 (M ++ 3); C 16H 15BrN 2O 30.7H 2The analytical calculation value of O: C, 51.13; H, 4.40; Br, 21.26; N, 7.45.Measured value: C, 50.84; H, 4.62; Br, 21.72; N, 7.18.
With 4-[N '-(2-bromo phenyl) urea groups]-3-aminomethyl phenyl acetic acid (287mg, 0.79mmol), 4-[(4S)-fluoro-(2S)-pyrrolidinyl] methoxyl methyl benzoate (200mg, 0.79mmol), EDCHCl (228mg, 1.20mmol), HOBT (160mg, 1.19mmol) and triethylamine (0.55ml, 3.95mmol) mixture in DMF (5ml) stirred 4 days under room temperature.In this mixture impouring frozen water, use ethyl acetate extraction.With the extract of frozen water and salt water washing merging, after dried over sodium sulfate, the vacuum concentration extract.Residue is through TLC[chloroform/acetone (10/1)] purification; obtain 4-[(2S; 4S)-1-[4-[N '-(2-chlorophenyl) urea groups]-3-aminomethyl phenyl acetyl group]-4-fluoro-2-pyrrolidinyl] methoxyl methyl benzoate (440mg, 93%), be colourless amorphous solid. 1H-NMR (CDCl 3) δ 1.90 and 1.97 (respectively being s, 3H, amide isomers), 2.05-2.62 (m, 2H), 3.58 (d, J=8.1Hz, 1H), 3.77 (m, 1H), 3.86 and 3.89 (respectively be s, 3H, amide isomers), and 3.92-4.64 (m, 5H), 5.24-5.42 (m, 1H), and 6.83-7.23 (m, 6H), 7.41-7.62 (m, 4H), and 7.86-8.09 (m, 3H); MS (ESI) m/z598 (M ++ 1), 600 (M ++ 3).
To 4-[1-[4-[N '-(2-bromo phenyl) urea groups]-3-aminomethyl phenyl acetyl group]-(4S)-and fluoro-(2S)-pyrrolidinyl] (440mg adds 0.25N sodium hydroxide (6.0ml) to methoxyl methyl benzoate in the agitating solution of THF 0.74mmol) (6.0ml).After stirring 1 day under the room temperature,, extract with chloroform-methanol (10/1) with this mixture of 1N HCl acidify.Extract and vacuum concentration through the dried over sodium sulfate merging.Residue is through TLC[chloroform/methanol (10/1)] purification, obtain 80 (229mg, 53%), be colourless amorphous solid.MW?584.44?IR(KBr)3325，2972，1709，1604，1529，1252cm -1； 1H-NMR(DMSO-d 6)δ2.25(s，3H)，2.31(m，1H)，3.17(s，1H)，3.60(d，J＝4.7Hz，2H)，3.83-4.67(m，5H)，5.31-5.51(m，1H)，6.97(t，J＝7.3Hz，1H)，7.02-7.09(m，5H)，7.33(t，J＝8.0Hz，1H)，7.61(d，J＝7.8Hz，1H)，7.64(d，J＝8.3Hz，1H)，7.87(d，J＝8.3Hz，2H)，7.90(d，J＝8.8Hz，1H)，8.44-8.65(m，2H)；MS(ESI)，m/z?584(M ++1)，586(M ++3)。C 28H 27BrFN 3O 70.4H 2The analytical calculation value of O: C, 56.84; H, 4.74; Br, 13.51; F, 3.21; N, 7.10.Measured value: C, 56.91; H, 4.93; Br, 13.23; F, 3.15; N, 6.88.Sodium salt for 80: C 28H 27BrFN 3O 7Na1.8H 2The analytical calculation value of O: C, 52.64; H, 4.67; Br, 12.51; F, 2.97; N, 6.58.Measured value: C, 53.04; H, 4.67; Br, 12.95; F, 3.28; N, 6.11.Embodiment 754-[1-[4-[N '-(2-chlorophenyl) urea groups]-3-aminomethyl phenyl acetyl group]-(2S)-and pyrrolidinyl] methoxybenzoic acid
Figure A0080951002621
In 0 ℃, (2.0g 5.9mmol) adds concentrated hydrochloric acid (3.0ml) in the agitating solution in ethanol (10.0ml) to methoxyl methyl benzoate to 4-(1-tert-butoxycarbonyl-(2S)-pyrrolidinyl).Stirred reaction mixture is 4.0 hours under room temperature.Vacuum concentrated mixture.Collect the solid that produces,, obtain 4-(2S-pyrrolidinyl) methoxyl methyl benzoate hydrochlorate (1.4g, 87%), be white crystalline solid with the alcohol-ether washing. 1H-NMR(CDCl 3)δ1.90-2.25(m，4H)，3.25-3.45(m，2H)，3.88(s，3H)，3.90-4.00(m，1H)，4.25-4.45(m，2H)，6.96(d，J＝8.5Hz，2H)，7.95(d，J＝8.5Hz，2H)。
In 0 ℃, to 4-[(2S)-pyrrolidinyl] methoxyl methyl benzoate hydrochlorate (135mg, 0.5mmol), 4-[N '-(2-chlorophenyl) urea groups]-3-aminomethyl phenyl acetic acid (159mg, 0.5mmol), HOBt (68mg, 0.5mmol) and triethylamine (278ml, 2.0mmol) add in the agitating solution in THF (5.0ml) and MeCN (5.0ml) EDCHCl (144mg, 0.75mmol).Under room temperature, stirred this reactant mixture 16 hours, vacuum concentration.Water is added in the residue, use ethyl acetate extraction.Wash extract with saturated sodium bicarbonate, then through dried over sodium sulfate and vacuum concentration.Residue is through purification by silica gel column chromatography; with normal hexane-ethyl acetate (1: 2,, obtain 4-[1-[4-[N '-(2-chlorophenyl) urea groups v/v) as eluant]-3-aminomethyl phenyl acetyl group]-the 2-pyrrolidinyl] methoxyl methyl benzoate (220mg; 82%), is colorless oil. 1H-NMR (CDCl 3) δ 1.91 and 1.97 (respectively be s, be total to 3H), and 2.00-2.20 (m, 4H), 3.55-3.65 (m, 4H), 3.87 and 3.89 (they respectively are s, are total to 3H), 4.10-4.20 (m, 2H), 4.51 (m, 1H), 6.86-7.04 (m, 6H), 7.20-7.53 (m, 4H), and 7.89-8.01 (m, 2H), 8.22 (d, J=8.3Hz, 1H).
To 4-[1-[4-[N '-(2-chlorophenyl) urea groups]-3-aminomethyl phenyl acetyl group]-the 2S-pyrrolidinyl] methoxyl methyl benzoate (220mg; 0.41mmol) THF (8.0ml) and the agitating solution of methanol (4.0ml) in add the 1N sodium hydroxide (0.8ml, 0.8mmol).Stirred this mixture 24 hours in 70 ℃.This mixture of vacuum concentration adds water wherein, with 1N HCl neutralization.Collect the solid that produces, wash with water, vacuum drying obtains 81, and (220mg quantitatively), is white crystalline solid.MW 521.99, mp 122-124 ℃; IR (KBr) 3340,1710,1685,1604,1533,1511,1438 cm -1 1H-NMR (DMSO-d 6) δ 1.81-2.11 (m, 4H), 2.18 and 2.20 (they respectively are s, are total to 3H), 3.45-3.80 (m, 4H), and 3.95-4.05 (m, 1H), 4.12-4.20 (m, 1H), 4.21-4.31 (m, 1H), 6.99-7.06 (m, 5H), 7.26-7.30 (m, 1H), 7.44 (d, J=7.8Hz, 1H), 7.62-7.64 (m, 1H), and 7.85-7.90 (m, 2H), 8.13 (d, J=6.8Hz, 1H), 8.60-8.62 (m, 2H); MS (FAB) m/z 522 (M ++ 1); C 28H 28N 3O 5Cl0.2H 2The analytical calculation value of O: C, 63.99; H, 5.45; N, 7.99.Measured value: C, 63.90; H, 5.40; N, 7.72.Embodiment 764-[1-[4-[N '-(2-bromo phenyl) urea groups]-3-aminomethyl phenyl acetyl group]-(2S)-and pyrrolidinyl] methoxybenzoic acid
Figure A0080951002631
In 0 ℃, to [(2S)-and pyrrolidinyl] methoxyl methyl benzoate hydrochlorate (135mg, 0.5mmol), 4-[N '-(2-bromo phenyl) urea groups]-3-aminomethyl phenyl acetic acid (181mg, 0.5mmol), HOBt (68mg, 0.5mmol) and triethylamine (278ml, 2.0mmol) add in the agitating solution in THF (5.0ml) and MeCN (5.0ml) EDCHCl (144mg, 0.75mmol).Under room temperature, stirred this reactant mixture 16 hours, vacuum concentration.Water is added in the residue, use ethyl acetate extraction.Wash extract with saturated sodium bicarbonate, then through dried over sodium sulfate and vacuum concentration.Residue is through purification by silica gel column chromatography; with normal hexane-ethyl acetate (1/2,, obtain 4-[1-[4-[N '-(2-bromo phenyl) urea groups v/v) as eluant]-3-aminomethyl phenyl acetyl group]-(2S)-pyrrolidinyl] methoxyl methyl benzoate (290mg; quantitatively), be colorless oil. 1H-NMR (CDCl 3) δ 1.95 and 2.01 (respectively be s, be total to 3H), and 2.00-2.20 (m, 4H), 3.50-3.65 (m, 4H), 3.87 and 3.89 (they respectively are s, are total to 3H), 4.10-4.20 (m, 2H), 4.50 (m, 1H), 6.85-7.06 (m, 6H), 7.24-7.28 (m, 1H), 7.40-7.44 (m, 3H), 7.89-8.16 (m, 2H), and 8.17-8.18 (m, 1H).
To 4-[1-[4-[N '-(2-bromo phenyl) urea groups]-3-aminomethyl phenyl acetyl group]-the 2S-pyrrolidinyl] methoxyl methyl benzoate (290mg; 0.5mmol) THF (8.0ml) and the agitating solution of methanol (4.0ml) in add the 1N sodium hydroxide (1.0ml, 1.0mmol).Stirred this mixture 24 hours in 70 ℃.This mixture of vacuum concentration adds water wherein, with 1N HCl neutralization.Collect the solid that produces, wash with water, vacuum drying obtains 82 (240mg, 85%), is white crystalline solid.MW 566.44 mp 125-130 ℃; IR (KBr) 3340,1604,1529,1434 cm -1 1H-NMR (DMSO-d 6) δ 1.80-2.10 (m, 4H), 2.18 and 2.20 (they respectively are s, are total to 3H), 3.45-3.80 (m, 4H), and 3.95-4.05 (m, 1H), 4.15-4.20 (m, 1H), 4.25-4.30 (m, 1H), 6.94-7.06 (m, 5H), 7.30-7.34 (m, 1H), and 7.59-7.62 (m, 2H), 7.85-7.90 (m, 2H), 8.01 (d, J=8.1Hz, 1H), 8.44 (s, 1H), 8.62 (s, 1H); MS (FAB) m/z566 (M +); C 28H 28N 3O 5Br0.5H 2The analytical calculation value of O: C, 58.44; H, 5.08; N, 7.30.Measured value: C, 58.57; H, 4.99; N, 7.18.Embodiment 774-[1-[3-methyl-4-[N '-(2-aminomethyl phenyl) urea groups] phenyl acetyl]-(2S)-and the pyrrolidinyl methoxyl group] benzoic acid
Under room temperature, with 3-methyl-4-[N '-(2-aminomethyl phenyl) urea groups] phenylacetic acid (438mg, 1.47mmol), 4-[(2S)-the pyrrolidinyl methoxyl group] essence of Niobe (420mg, 1.79mmol), EDCHCl (410mg, 2.14mmol), HOBt (228mg, 1.69mmol) and triethylamine (240ml, 1.72mmol) mixture in DMF (5ml) stirs and to spend the night.This mixture of dilute with water is used ethyl acetate extraction.With salt water washing extract, through dried over sodium sulfate and evaporation.Residue is through purification by silica gel column chromatography; with chloroform-methanol (50: 1-25: 1,, obtain 4-[1-[3-methyl-4-[N '-(2-aminomethyl phenyl) urea groups v/v) as eluant] phenyl acetyl]-(2S)-the pyrrolidinyl methoxyl group] essence of Niobe (760mg; quantitatively), be the white foam thing. 1H-NMR(CDCl 3)δ1.89(s，3H)，1.94-2.14(m，4H)，2.16(s，3H)，3.50-3.69(m，4H)，3.87(s，3H)，4.09-4.17(m，2H)，4.42-4.45(m，1H)，6.85-7.02(m，6H)，7.10-7.16(m，3H)，7.51-7.53(m，1H)，7.62-7.64(m，1H)，7.91-7.94(m，2H)；MS(FAB)m/z?516(M ++1)。
To 4-[1-[3-methyl-4-[N '-(2-aminomethyl phenyl) urea groups] phenyl acetyl]-(2S)-and the pyrrolidinyl methoxyl group] essence of Niobe (420mg; 0.71mmol) the agitating solution of THF (7ml) in add 0.5N sodium hydroxide (7ml), with the heating 2 hours under refluxing of this reactant mixture.After being cooled to room temperature, in this mixture impouring ice-1N HCl, the precipitation that produces is collected in decompression.Crude product solid recrystallization purifying from chloroform-IPE obtains 83 (526mg, 69%), is white crystalline powder.191-193 ℃ of MW 501.57mp; 1H-NMR (DMSO-d 6) δ 1.87-2.10 (m, 4H), 2.20 (s, 3H), 2.26 (s, 3H), 3.44-3.79 (a plurality of m, 4H altogether), (3.99-4.45 a plurality of m, 3H altogether), 6.91-7.17 (a plurality of m, be total to 7H), and 7.66-7.68 (m, 1H), 7.80-7.90 (m, 3H), 8.19-8.21 (m, 2H), 12.62 (br s, 1H); MS (FAB) m/z 502 (M ++ 1).C 29H 31N 3O 51/4H 2The analytical calculation value of O: C, 68.83; H, 6.27; N, 8.30.Measured value: C, 68.81; H, 6.17; N, 8.23.Embodiment 784-[(4S)-fluoro-1-[4-[N '-(2-methoxyphenyl) urea groups]-3-aminomethyl phenyl acetyl group]-(2S)-and the pyrrolidinyl methoxyl group] benzoic acid
Figure A0080951002661
To 4-amino-3-aminomethyl phenyl tert-butyl acetate (1.36g, 6.15mmol) and triethylamine (170ml, 1.23mmol) the agitating solution of THF (30ml) in add Carbimide. 2-methoxyl group phenyl ester (820ml 6.15mmol), stir the mixture that obtains 27 hours.This mixture vacuum concentration to small size, is added generation precipitation in the residue with hexane, filters and collect this precipitation, obtain 4-[N '-(2-methoxyphenyl) urea groups]-3-aminomethyl phenyl tert-butyl acetate (1.74g, 76%), be white crystalline material.mp?157-158℃； 1H-NMR(CDCl 3)δ1.46(s，9H)，2.30(s，3H)，3.50(s，2H)，3.76(s，3H)，6.43(s，1H)，6.83(br?d，J＝8.4Hz，1H)，6.95(br?d，J＝8.0Hz，1H)，6.98-6.99(m，2H)，7.13(brs，1H)，7.23(brs，1H)，7.48(d，J＝8.8Hz，1H)，8.14(d，J＝8.4Hz，1H)；MS(ESI)m/z371(M ++H)。
To 4-[N '-(2-methoxyphenyl) urea groups]-(1.32g adds trifluoroacetic acid (10ml) to 3-aminomethyl phenyl tert-butyl acetate in the agitating solution of dichloromethane 3.56mmol) (15ml), the mixture that obtains was heated 30 minutes under refluxing.Vacuum concentrated mixture also adds the water generates precipitation, filters and collects this precipitation.Make crude product solid recrystallization from ethanol/hexane, obtain 4-[N '-(2-methoxyphenyl) urea groups]-3-aminomethyl phenyl acetic acid (932mg, 83%), be white powder.mp?260-264℃； 1H-NMR(CD 3OD)δ2.30(s，3H)，3.55(s，2H)，4.87(s，3H)，6.87-6.92(m，2H)，6.97-6.99(m，2H)，7.10-7.24(m，2H)，7.53-7.58(m，1H)，8.04(d，J＝7.2Hz，1H)；MS(ESI)m/z?314(M +)。
Under room temperature, to 4-[N '-(2-methoxyphenyl) urea groups]-3-aminomethyl phenyl acetic acid (336mg, 1.07mmol), 44 (4S)-fluoro-(2S)-pyrrolidinyl methoxyl groups] essence of Niobe (271mg, 1.07mmol) and N, N-dimethyl aminopyridine (130mg, 1.07mmol) the agitating solution of DMF (10ml) in add EDCHCl (226mg 1.18mmol), stir the mixture that obtains 20 hours.In this mixture impouring 1N HCl aqueous solution, use ethyl acetate extraction.With salt water washing organic layer, through anhydrous sodium sulfate drying, vacuum concentration then.Residue is through silica gel column chromatography; with chloroform-methanol (10: 1) as eluant; obtain 4-[(4S)-fluoro-1-[4-[N '-(2-methoxyphenyl) urea groups]-3-aminomethyl phenyl acetyl group]-(2S)-and the pyrrolidinyl methoxyl group] essence of Niobe (583mg, 99%), be colourless amorphous solid. 1H-NMR (CDCl 3) mixture of rotamer, δ 2.05 and 2.12 (s, 3H altogether), 2.05-2.61 (m, 2H), 3.55-4.73 (a plurality of m, 13H), 4.5 1-4.66 (m, 2H), 5.26-5.40 (m, 1H), 6.72-7.01 (a plurality of m, 8H), 7.38-8.13 (a plurality of m, 3H); MS (ESI) m/z550 (M ++ H).
Under room temperature; to 4-[(4S)-fluoro-1-[4-[N '-(2-methoxyphenyl) urea groups]-3-aminomethyl phenyl acetyl group]-(2S)-and the pyrrolidinyl methoxyl group] essence of Niobe (557mg; 1.01mmol) methanol-THF (1: 1; add 1.0N sodium hydrate aqueous solution (4.05ml in agitating solution 10ml); 4.05mmol), the mixture that obtains was heated 2 hours in 60 ℃.In this mixture impouring 1NHCl aqueous solution, use ethyl acetate extraction.With salt water washing organic layer, through anhydrous sodium sulfate drying, vacuum concentration then.Residue as eluant, obtains 84 (492mg, 91%) with chloroform-methanol (10: 1) through silica gel column chromatography, is colourless amorphous solid.MW 535.56 1H-NMR (CD 3OD), the mixture of rotamer, δ 2.96 (s, 3H), 2.11-2.45 (m, 2H), 3.64-4.15 (a plurality of m, 5H), 3.91 (s, 3H), 4.41-4.45 (m, 1H), 4.52-4.61 (m, 1H), 5.25-5.38 (m, 1H), 6.84-7.10 (a plurality of m, 7H), 7.54-7.58 (m, 1H), 7.93 (d, J=8.8Hz, 2H), 8.02 (d, J=8.8Hz, 2H); MS (ESI) m/z536 (M ++ H), 538 (M ++ Na +).Embodiment 794-[(4S)-and fluoro-1-[4-[N '-(2-methoxyphenyl) urea groups] phenyl acetyl]-(2S)-and the pyrrolidinyl methoxyl group] benzoic acid
To 4-amino-3-aminomethyl phenyl ethyl acetate (1.32g, 7.37mmol) and triethylamine (205ml, (980ml 7.37mmol), stirs the mixture that obtains 23 hours to add Carbimide. 2-methoxyl group phenyl ester in the agitating solution of THF 1.47mmol) (20ml).This mixture vacuum concentration to small size, being added generation precipitation in the residue with hexane, collects this precipitation, obtain 4-[N '-(2-methoxyphenyl) urea groups] (2.44g quantitatively), is white crystalline material to the phenylacetic acid ethyl ester.mp107-109℃； 1H-NMR(CDCl 3)δ1.26(t，J＝7.1Hz，3H)，3.56(s，3H)，3.79(s，3H)，4.15(q，J＝7.1Hz，2H)，6.82-6.85(m，1H)，6.91-7.00(m，2H)，7.08(s，1H)，7.17(d，J＝8.5Hz，2H)，7.27(d，J＝8.6Hz，2H)，7.33(s，1H)，8.07-8.10(m，1H)；MS(ESI)m/z?329(M ++H)。
To 4-[N '-(2-methoxyphenyl) urea groups] (2.22g, (10.2ml's phenylacetic acid ethyl ester 10.2mmol), spends the night the mixture stirring that obtains to add the 1.0M-sodium hydrate aqueous solution in the agitating solution of methanol 6.78mmol) (30ml).Add 1N HCl (aqueous solution), this mixture of vacuum concentration.Water is added in the residue, obtain precipitation, filter and collect this precipitation.Make crude product solid recrystallization from ethanol/hexane, obtain 4-[N '-(2-methoxyphenyl) urea groups] phenylacetic acid, be white powder (1.87g, 92%).mp?165-168℃； 1H-NMR(CD 3OD)δ2.30(s，3H)，3.55(s，2H)，4.87(s，3H)，6.87-6.92(m，2H)，6.97-6.99(m，2H)，7.10-7.24(m，2H)，7.53-7.58(m，1H)，8.04(d，J＝7.2Hz，1H)；MS(ESI)m/z?300(M +)。
Under room temperature, to 4-[N '-(2-methoxyphenyl) urea groups] phenylacetic acid (353mg, 1.18mmol), 4-[(4S)-fluoro-(2S)-pyrrolidinyl methoxyl group] essence of Niobe (298mg, 1.18mmol) and N, N-dimethyl aminopyridine (144mg, 1.18mmol) the agitating solution of DMF (10ml) in add EDCHCl (226mg 1.18mmol), stir the mixture that obtains 22 hours.In this mixture impouring 1N HCl aqueous solution, use ethyl acetate extraction.With salt water washing organic layer, through anhydrous sodium sulfate drying, vacuum concentration then.Residue is through silica gel column chromatography; with chloroform-methanol (10: 1) as eluant; obtain 4-[(4S)-fluoro-1-[4-[N '-(2-methoxyphenyl) urea groups] phenyl acetyl]-(2S)-and the pyrrolidinyl methoxyl group] essence of Niobe (594mg, 94%), be colourless amorphous solid. 1H-NMR (CDCl 3) mixture of rotamer, and δ 2.05-2.58 (a plurality of m, 2H), 3.55-4.25 (a plurality of m, 5H), 3.77 (s, 3H), 3.87-3.90 (m, 3H), 4.50-4.63 (m, 2H), and 5.23-5.37 (m, 1H), 6.81-6.84 (m, 1H), 6.91-6.99 (m, 4H), 7.09-7.12 (m, 2H), 7.18-7.26 (m, 2H), 7.45-7.53 (m, 2H), 7.91-8.03 (m, 2H), 8.10-8.12 (m, 1H); MS (ESI) m/z 536 (M ++ H).
Under room temperature; to 4-[(4S)-fluoro-1-[4-[N '-(2-methoxyphenyl) urea groups] phenyl acetyl]-(2S)-and the pyrrolidinyl methoxyl group] essence of Niobe (568mg; 1.06mmol) methanol-THF (1: 1; add 1.0N sodium hydrate aqueous solution (4.24ml in agitating solution 10ml); 4.24mmol), the mixture that obtains was heated 1 hour in 60 ℃.In this mixture impouring 1NHCl aqueous solution, use ethyl acetate extraction.With salt water washing organic layer, through anhydrous sodium sulfate drying, vacuum concentration then.Residue as eluant, obtains 85 (516mg, 93%) with chloroform-methanol (10: 1) through silica gel column chromatography, is colourless amorphous solid.MW 521.54 1H-NMR (CD 3OD), the mixture of rotamer, δ 2.12-2.46 (m, 2H), 3.654.19 (a plurality of m, 5H), 3.88 (s, 3H), 4.42-4.45 (m, 1H), 4.52-4.62 (m, 1H), 5.24-5.39 (m, 1H), 6.85-6.91 (m, 1H), 6.94-7.03 (a plurality of m, 4H), 7.14-7.19 (m, 2H), 7.35-7.40 (m, 2H), 7.92-7.96 (m, 2H), 8.02-8.04 (m, 1H); MS (ESI) m/z 521 (M ++ H), 544 (M ++ Na +).Embodiment 804-[(4S)-fluoro-1-[4-[N '-(2-methoxyphenyl) urea groups]-3-methoxyphenyl acetyl group]-(2S)-and the pyrrolidinyl methoxyl group] benzoic acid
Figure A0080951002691
To 4-amino-3-methoxyphenyl tert-butyl acetate (1.41g, 5.94mmol) and triethylamine (165ml, 1.19mmol) the agitating solution of THF (20ml) in add Carbimide. 2-methoxyl group phenyl ester (790ml 5.94mmol), stir the mixture that obtains 4 days.This mixture vacuum concentration to small size, is added in the residue hexane to produce precipitation, collects this precipitation, obtain 4-[N '-(2-methoxyphenyl) urea groups]-3-methoxyphenyl tert-butyl acetate (2.06g, 90%), be white crystalline material.mp?132-134℃； 1H-NMR(CDCl 3)δ1.46(s，9H)，3.50(s，2H)，3.87(s，3H)，3.88(s，3H)，6.84(s，1H)，6.87-6.90(m，2H)，6.98-7.03(m，2H)，7.12(s，1H)，7.16(s，1H)，8.06(d，J＝8.4Hz，1H)，8.13(dd，J＝7.2，2.0Hz，1H)；MS(ESI)m/z?387(M ++H)。
To 4-[N '-(2-methoxyphenyl) urea groups]-(2.01g adds trifluoroacetic acid (10ml) to 3-methoxyphenyl tert-butyl acetate in the agitating solution of dichloromethane 5.20mmol) (15ml), the mixture that obtains was heated 30 minutes under refluxing.Vacuum concentrated mixture also adds in the residue water to produce precipitation, to filter and collects this precipitation.Make crude product solid recrystallization from ethanol/hexane, obtain 4-[N '-(2-methoxyphenyl) urea groups]-3-methoxybenzene guanidine-acetic acid, be white powder (1.40g, 82%).mp?182-185℃； 1H-NMR(CD 3OD)δ3.55(s，2H)，3.88(s，3H)，3.89(s，3H)，6.80-6.99(m，5H)，7.94(d，J＝8.4Hz，1H)，8.00(d，J＝7.2Hz，1H)；MS(ESI)m/z?330(M +)。
Under room temperature, to 4-[N '-(2-methoxyphenyl) urea groups]-3-methoxybenzene guanidine-acetic acid (353mg, 1.07mmol), 4-[(4S)-fluoro-(2S)-pyrrolidinyl methoxyl group] essence of Niobe (271mg, 1.07mmol) and N, N-dimethyl aminopyridine (131mg, 1.07mmol) the agitating solution of DMF (10ml) in add EDCHCl (224mg 1.18mmol), stir the mixture that obtains 14 hours.In this mixture impouring 1N HCl aqueous solution, use ethyl acetate extraction.With salt water washing organic layer, through anhydrous sodium sulfate drying, vacuum concentration then.Residue is through silica gel column chromatography; with chloroform-methanol (10: 1) as eluant; obtain 4-[(4S)-fluoro-1-[4-[N '-(2-methoxyphenyl) urea groups]-3-methoxyphenyl acetyl group]-(2S)-and the pyrrolidinyl methoxyl group] essence of Niobe (372mg, 61%), be colourless amorphous solid. 1H-NMR (CDCl 3), the mixture of rotamer, and δ 2.04-2.57 (a plurality of m, 2H), 3.58-4.18 (a plurality of m, 5H), 3.79 and 3.83 (s, 3H altogether), 3.86 (s, 3H), 3.87 (s, 3H), 4.51-4.63 (m, 2H), 5.22-5.36 (m, 1H), 6.80-6.89 (m, 3H), 6.94-7.03 (m, 4H), 7.15-7.25 (m, 2H), 7.94-8.01 (m, 2H), 8.04-8.11 (m, 2H); MS (ESI) m/z566 (M ++ H).
Under room temperature; to 4-[(4S)-fluoro-1-[4-[N '-(2-methoxyphenyl) urea groups]-3-methoxyphenyl acetyl group]-(2S)-and the pyrrolidinyl methoxyl group] essence of Niobe (356mg; 0.63mmol) methanol-THF (1: 1; add 1.0N sodium hydrate aqueous solution (1.88ml in agitating solution 10ml); 1.88mmol), the mixture that obtains was heated 2 hours in 60 ℃.In this mixture impouring 1NHCl aqueous solution, use ethyl acetate extraction.With salt water washing organic layer, through anhydrous sodium sulfate drying, vacuum concentration then.Residue as eluant, obtains 86 (335mg, 97%) with chloroform-methanol (10: 1) through silica gel column chromatography, is colourless amorphous solid.MW 551.56 1H-NMR (CD 3OD) mixture of rotamer, δ 2.14-2.48 (m, 2H), 3.69-4.20 (a plurality of m, 5H), 3.88 (s, 3H), 3.89 (s, 3H), 4.46-4.57 (m, 2H), 5.27-5.41 (m, 1H), 6.79-7.04 (m, 7H), 7.90-8.02 (m, 4H); MS (ESI) m/z 552 (M ++ H).Embodiment 814-[1-[4-[N '-(2, the 6-Dichlorobenzene base) urea groups] phenyl acetyl]-(4S)-and fluoro-(2S)-pyrrolidinyl] methoxybenzoic acid
Under room temperature, to 4-aminophenyl ethyl acetate (1.62g, 9.04mmol) and Carbimide. 2, the 6-Dichlorfop (1.70g, 9.04mmol) add in the mixture in THF (40ml) triethylamine (0.25ml, 1.81mmol).Stir after 2 hours the vacuum concentration reactant mixture.Grind residue by adding normal hexane, obtain 4-[N '-(2, the 6-Dichlorobenzene base) urea groups] phenylacetic acid ethyl ester (3.19g, 96%), be colourless powder.Mp 168-170 ℃ (decomposition); 1H-NMR (CDCl 3) δ 1.25 (t, J=7.1Hz, 3H), 3.56 (s, 2H), 4.14 (q, J=7.1Hz, 2H), 6.50 (br, 1H), 6.67 (br, 1H), 7.12-7.52 (m, 7H).
To 4-[N '-(2, the 6-Dichlorobenzene base) urea groups] (3.19g adds 0.25N sodium hydroxide (70ml) to the phenylacetic acid ethyl ester in the agitating solution of THF 8.69mmol) (70ml).After stirring 17 hours under the room temperature, the vacuum concentration solvent.Grind residue by adding entry, obtain 4-[N '-(2, the 6-Dichlorobenzene base) urea groups] phenylacetic acid (2.44g, 82%), be colourless powder.Mp 262-263 ℃ (decomposition); 1H-NMR (DMSO-d 6) δ 3.48 (s, 2H), 7.14 (d, J=8.3Hz, 2H), 7.31 (t, J=8.3Hz, 1H), 7.37 (d, J=8.3Hz, 2H), 7.52 (d, J=8.0Hz, 2H), 8.18 (s, 1H), 8.90 (s, 1H), 12.22 (br, 1H); MS (ESI) m/z339 (M ++ 1), 341 (M ++ 3), 343 (M ++ 5).
Under room temperature, with 4-[N '-(2, the 6-Dichlorobenzene base) urea groups] phenylacetic acid (268mg, 0.79mmol), 4-[(2S, 4S)-4-fluoro-2-pyrrolidinyl] methoxyl methyl benzoate (200mg, 0.79mmol), EDCHCl (227mg, 1.19mmol), HOBT (161mg, 1.19mmol) and triethylamine (0.55ml, 3.95mmol) mixture in DMF (4ml) stirred 18 hours.In this mixture impouring frozen water, use ethyl acetate extraction.Extract with frozen water and salt water washing merging.After dried over sodium sulfate, the vacuum concentration extract.Residue is through TLC[chloroform/methanol (10/1)] purification, obtain 4-[1-[4-[N '-(2, the 6-Dichlorobenzene base) urea groups] phenyl acetyl]-(4S)-and fluoro-(2S)-pyrrolidinyl] methoxyl methyl benzoate (465mg, 100%), be colourless amorphous solid. 1H-NMR (CDCl 3) δ 2.05-2.57 (m, 2H), 3.60 (d, 2H, J=3.4Hz), 3.64-3.84 (m, 2H), 3.88 and 3.89 (they respectively being s, 3H, amide isomers), 3.92-4.63 (m, 3H), 5.22-5.38 (m, 1H), 6.87 and 6.89 (respectively is d, each J=7.9Hz, 2H, amide isomers), 7.01-7.17 (m, 6H), 7.28 (m, 2H), 7.36 (br, 1H), 7.92 (d, J=8.8Hz, 1H), 7.79 (d, J=8.8Hz, 1H); MS (ESI) m/z 574 (M ++ 1), 576 (M ++ 3), 578 (M ++ 5).
To 4-[(2S, 4S)-1-[4-[N '-(2, the 6-Dichlorobenzene base) urea groups] phenyl acetyl]-4-fluoro-2-pyrrolidinyl] (465mg adds 0.25N sodium hydroxide (40ml) to methoxyl methyl benzoate in THF 0.809mmol) (40ml) solution.After stirring 11 hours under the room temperature,, extract with chloroform-methanol (10/1) with this mixture of 1N HCl acidify.Extract and vacuum concentration through the dried over sodium sulfate merging.Residue is through TLC[chloroform/methanol (10/1)] purification, obtain 87 (340mg, 75%), be colourless powder.168-172 ℃ of MW 560.40mp (decomposition); IR (KBr) 3340,1711,1685,1604,1240,773 cm -1 1H-NMR (DMSO-d 6) δ 2.22-2.30 (m, 2H), 3.61 (d, J=7.4Hz, 2H), 3.70-4.75 (m, 6H), 5.30-5.49 (m, 1H), 7.02-7.18 (m, 5H), 7.28-7.41 (m, 4H), 7.52 (dd, J=8.0,2.9Hz, 2H), 7.86 (m, 2H), 8.29 (br, 1H), 9.01 (br, 1H), 12.66 (br, 1H); MS (ESI) m/z560 (M ++ 1), 562 (M ++ 3), 564 (M ++ 5); C 27H 24Cl 2FN 3O 50.5H 2The analytical calculation value of O: C, 56.95; H, 4.43; Cl, 12.45; F, 3.34; N, 7.38.Measured value: C, 57.04; H, 4.34; Cl, 12.98; F, 3.27; N, 7.21.Embodiment 824-[1-[4-[N '-(2, the 6-Dichlorobenzene base) urea groups]-3-methoxyphenyl acetyl group]-(4S)-and fluoro-(2S)-pyrrolidinyl] methoxybenzoic acid
Figure A0080951002731
Under room temperature, to 4-amino-3-methoxyphenyl tert-butyl acetate (2.15g, 9.04mmol) and Carbimide. 2, the 6-Dichlorfop (1.70g, 9.04mmol) add in the mixture in THF (40ml) triethylamine (0.25ml, 9.04mmol).Stir after 18 hours the vacuum concentration reactant mixture.Grind residue by adding normal hexane, obtain 4-[N '-(2, the 6-Dichlorobenzene base) urea groups]-3-methoxyphenyl tert-butyl acetate (2.27g, 59%), be colourless powder.Mp 177-181 ℃ (decomposition); 1H-NMR (CDCl 3) δ 1.43 (s, 9H), 3.74 (s, 2H), 3.83 (s, 3H), 6.34 (s, 1H), 6.81 (s, 1H), 6.84 (d, J=8.3Hz, 1H), 7.06 (br, 1H), 7.27 (t, J=8.1Hz, 1H), 7.39 (d, J=8.1Hz, 2H), 8.04 (d, J=8.3Hz, 1H).
Under room temperature, to 4-[N '-(2, the 6-Dichlorobenzene base) urea groups]-(2.27g adds TFA (20ml) to 3-methoxyphenyl tert-butyl acetate in the agitating solution of dichloromethane 5.34mmol) (50ml).Stir after 2 hours this mixture of vacuum concentration.Grind residue by adding entry, obtain 4-[N '-(2, the 6-Dichlorobenzene base) urea groups]-3-methoxybenzene guanidine-acetic acid (1.50g, 76%), be colourless powder.Mp 246-249 ℃ (decomposition); 1H-NMR (DMSO-d 6) δ 3.49 (s, 2H), 3.88 (s, 3H), 6.75 (d, J=8.3Hz, 1H), 6.93 (s, 1H), 7.30 (t, J=7.8Hz, 1H), 7.52 (d, J=8.0Hz, 2H), 7.97 (d, J=8.0Hz, 1H), 8.40 (s, 1H), 8.86 (s, 1H), 12.23 (br, 1H); MS (ESI) m/z 369 (M ++ 1), 371 (M ++ 3), 373 (M ++ 5).
Under room temperature, with 4-[N '-(2, the 6-Dichlorobenzene base) urea groups]-3-methoxybenzene guanidine-acetic acid (288mg, 0.78mmol), 4-[(2S, 4S)-4-fluoro-2-pyrrolidinyl] methoxyl methyl benzoate (200mg, 0.79mmol), EDCHCl (227mg, 1.19mmol), HOBT (161mg, 1.19mmol) and triethylamine (0.55ml, 3.95mmol) mixture in DMF (4ml) stirred 18 hours.In this mixture impouring frozen water, use ethyl acetate extraction.Extract with frozen water and salt water washing merging.After dried over sodium sulfate, the vacuum concentration extract.Residue is through silica gel column chromatography [50g; chloroform/methanol (40/1)], obtain 4-[1-[4-[N '-(2, the 6-Dichlorobenzene base) urea groups]-3-methoxyphenyl acetyl group]-(4S)-and fluoro-(2S)-pyrrolidinyl] methoxyl methyl benzoate (530mg; 100%), is colourless amorphous solid. 1H-NMR (CDCl 3) δ 2.03-2.62 (m, 2H), 3.61 (d, 2H, J=4.7Hz), 3.62-3.66 (m, 2H), 3.73 and 3.77 (they respectively being s, 3H, amide isomers), 3.78-3.85 (m, 1H), 3.87 with 3.88 (they respectively being s, 3H, amide isomers), and 3.95-4.63 (m, 4H), 5.22-5.36 (m, 1H), 6.82 (s, 1H), 6.88 (d, J=8.8Hz, 1H), 6.95 (d, J=8.8Hz, 2H), 7.14-7.25 (m, 1H), 7.38 (d, J=8.1Hz, 2H), 7.94-8.10 (m, 3H); MS (ESI) m/z 604 (M ++ 1), 606 (M ++ 3), 608 (M ++ 5).
To 4-[1-[4-[N '-(2, the 6-Dichlorobenzene base) urea groups]-3-methoxyphenyl acetyl group]-(4S)-and fluoro-(2S)-pyrrolidinyl] (530mg adds 0.25N sodium hydroxide (40ml) to methoxyl methyl benzoate in THF 0.78mmol) (40ml) solution.After stirring 11 hours under the room temperature,, extract with chloroform-methanol (10/1) with this mixture of 1N HCl acidify.Extract and vacuum concentration through the dried over sodium sulfate merging.Residue is through TLC[chloroform/methanol (10/1)] purification, obtain 88 (420mg, 75%), be colourless amorphous solid.MW 590.43 mp 162-168 ℃ (decomposition); IR (KBr) 3346,2974,1709,1604,1533,1254 cm -1 1H-NMR (DMSO-d 6) δ 1.98-2.36 (m, 2H), 3.58 (s, 2H), 3.78-3.95 (m, 6H), and 4.02-4.68 (m, 2H), 5.31-5.50 (m, 1H), 6.71-7.09 (m, 4H), 7.31 (t, J=7.8Hz, 1H), 7.53 (d, J=8.1Hz, 2H), 7.87 (d, J=8.1Hz, 2H), 7.88-8.00 (m, 1H), 8.30-840 (m, 1H), 8.89 (s, 1H); MS (ESI) m/z 590 (M ++ 1), 592 (M ++ 3), 594 (M ++ 5); C 28H 26Cl 2FN 3O 61.5H 2The analytical calculation value of O: C, 54.47; H, 4.73; F, 3.08; N, 6.81.Measured value: C, 54.53; H, 4.49; F, 2.93; N, 6.65.Embodiment 834-[(2S, 4S)-1-[4-[N '-(2, the 6-Dichlorobenzene base) urea groups]-3-aminomethyl phenyl acetyl group]-4-fluoro-2-pyrrolidinyl] methoxybenzoic acid Under room temperature, to 4-amino-3-aminomethyl phenyl tert-butyl acetate (1.88g, 8.51mmol), Carbimide. 2, the 6-Dichlorfop (1.60g, 8.51mmol) add in the mixture in THF (40ml) triethylamine (0.24ml, 1.70mmol).Stir after 3 hours the vacuum concentration reactant mixture.Grind residue by adding normal hexane, obtain 4-[N '-(2, the 6-Dichlorobenzene base) urea groups]-3-aminomethyl phenyl tert-butyl acetate (2.58g, 74%), be colourless powder.Mp 243-244 ℃ (decomposition); 1H-NMR (CDCl 3) δ 1.45 (s, 9H), 2.30 (s, 3H), 3.49 (s, 2H), 6.24 (s, 2H), 7.12-7.16 (m, 3H), 7.35 (d, J=8.3Hz, 2H), 7.51 (d, J=7.8Hz, 1H).
Under room temperature, to 4-[N '-(2, the 6-Dichlorobenzene base) urea groups]-(2.58g adds TFA (20ml) to 3-aminomethyl phenyl tert-butyl acetate in the agitating solution of dichloromethane 6.30mmol) (50ml).Stir after 2 hours this mixture of vacuum concentration.Grind residue by adding entry, obtain 4-[N '-(2, the 6-Dichlorobenzene base) urea groups]-3-aminomethyl phenyl acetic acid (2.12g, 95%), be colourless powder.Mp 274-283 ℃ (decomposition); 1H-NMR (DMSO-d 6) δ 2.24 (s, 3H), 3.46 (s, 2H), 7.00 (d, J=8.6Hz, 1H), 7.06 (s, 1H), 7.30 (t, J=7.8Hz, 1H), 7.52 (d, J=8.3Hz, 2H), 7.65 (d, J=8.2Hz, 1H), 8.12 (s, 1H), 8.50 (s, 1H), 12.22 (br, 1H); MS (ESI) m/z353 (M ++ 1), 355 (M ++ 3), 357 (M ++ 5).
Under room temperature, with 4-[N '-(2, the 6-Dichlorobenzene base) urea groups]-3-aminomethyl phenyl acetic acid (181mg, 0.51mmol), 4-[(2S, 4S)-4-fluoro-2-pyrrolidinyl] methoxyl methyl benzoate (130mg, 0.51mmol), EDCHCl (147mg, 0.77mmol), HOBT (104mg, 0.77mmol) and triethylamine (0.35ml, 2.55mmol) mixture in DMF (4ml) stirred 18 hours.In this mixture impouring frozen water, use ethyl acetate extraction.Extract with frozen water and salt water washing merging.After dried over sodium sulfate, the vacuum concentration extract.Residue is through TLC[chloroform/methanol (20/1)] purification; obtain 4-[1-[4-[N '-(2; the 6-Dichlorobenzene base) urea groups]-3-aminomethyl phenyl acetyl group]-(4S)-and fluoro-(2S)-pyrrolidinyl] methoxyl methyl benzoate (283mg, 94%), be colourless amorphous solid. 1H-NMR (CDCl 3) δ 1.95-2.61 (m, 3H), 3.55 (br, 2H), 3.67-3.81 (m, 2H), 3.87 (s, 6H), 3.89-4.68 (m, 2H), 5.23-5.43 (m, 1H), 6.81-7.10 (m, 6H), 7.13-7.43 (m, 3H), (7.56 br, 1H, one of isomer), 7.73 (br, 1H, one of isomer), 7.89-8.00 (m, 2H); MS (ESI) m/z 588 (M ++ 1), 590 (M ++ 3), 592 (M ++ 5).
To 4-[1-[4-[N '-(2, the 6-Dichlorobenzene base) urea groups]-3-aminomethyl phenyl acetyl group]-(4S)-and fluoro-(2S)-pyrrolidinyl] (283mg adds 0.25N sodium hydroxide (20ml) to methoxyl methyl benzoate in THF 0.48mmol) (20ml) solution.After stirring 11 hours under the room temperature, with this mixture of ethyl acetate extraction.With 1N HCl acidify remaining aqueous layer and use ethyl acetate extraction.Extract and vacuum concentration through the dried over sodium sulfate merging.Residue is through TLC[chloroform/methanol (20/1)], obtain 89 (450mg, 67%), be the light brown amorphous solid.174-180 ℃ of MW 574.43mp (decomposition); IR (KBr) 3330,3288,1711,1685,1604,1512,1242cm -1 1H-NMR (DMSO-d 6) δ 2.24 (m, 3H), 3.61 (d, 2H, J=6.1Hz), 3.72-4.68 (m, 7H), 5.30-5.50 (m, 1H), 6.97-7.20 (m, 4H), 7.29-7.68 (m, 5H), 7.87 (m, 2H), 8.10-8.95 (m, 1H), 12.65 (br, 1H); MS (ESI) m/z574 (M ++ 1), 576 (M ++ 3), 578 (M ++ 5).C 28H 26Cl 2FN 3O 50.5H 2The analytical calculation value of O: C, 57.64; H, 4.66; Cl, 12.15; F, 3.26; N, 7.20.Measured value: C, 57.37; H, 4.44; Cl, 12.64; F, 3.23; N, 7.25.Embodiment 844-[1-[3-chloro-4-[N '-(2-aminomethyl phenyl) urea groups] phenyl acetyl]-(4S)-and fluoro-(2S)-pyrrolidinyl] methoxybenzoic acid
Figure A0080951002761
Under room temperature, to 3-chlorophenyl acetic acid (21.76g, add in the agitating solution of dichloromethane 127.6mmol) (100ml) methanol (15.6ml, 383mmol) and sulphuric acid (1ml).Stir after 20 minutes, this mixture was heated 2 hours in 80 ℃.In reactant mixture impouring frozen water, use chloroform extraction.Extract with sodium bicarbonate aqueous solution and salt water washing merging.After dried over sodium sulfate, the vacuum concentration extract obtains 3-chlorophenyl methyl acetate (25.4g, 100%), is colorless oil. 1H-NMR(CDCl 3)δ3.60(s，2H)，3.70(s，3H)，7.15-7.26(m，4H)。
In 0 ℃, to 3-chlorophenyl methyl acetate (25.4g, the middle nitric acid (5.5ml.138mmol) that adds that stirs the mixture of sulphuric acid 128mmol) (44ml).This reactant mixture was warming up to room temperature 4 hours gradually.In reactant mixture impouring frozen water, use ethyl acetate extraction.Extract with sodium bicarbonate aqueous solution and salt water washing merging.After dried over sodium sulfate, the vacuum concentration extract.Through silica gel column chromatography purification residue [1kg, n-hexane/ethyl acetate (40/1)], obtain 3-chloro-4-nitrobenzophenone methyl acetate (11.4g, 36%), be yellow oil. 1H-NMR(CDCl 3)δ3.69(s，2H)，3.74(s，3H)，7.33(dd，J＝8.3，1.5Hz，1H)，7.49(d，J＝1.5Hz，1H)，7.87(d，J＝8.3Hz，1H)。
With 3-chloro-4-nitrobenzophenone methyl acetate (10.9g, 47.5mmol), reduced iron powder (8.58g, 153.6mmol), AcONa3H 2O (6.05g, 44.5mmol) and the mixture of acetic acid (17.6ml) in methanol (100/400ml) in 110 ℃ the heating 1 hour.After being cooled to room temperature,, use the methanol wash filter cake by the Celite filter reaction mixture.The filtrate that evaporation merges is also used ethyl acetate extraction.With the extract of salt water washing merging, through dried over sodium sulfate and vacuum concentration.Residue obtains 4-amino-3-chlorophenyl methyl acetate (4.58g, 48%) through silica gel column chromatography [150g, chloroform/ethyl acetate (10/1)], is red oil. 1H-NMR(CDCl 3)δ3.49(s，2H)，3.68(s，3H)，4.01(br，2H)，6.70(d，J＝7.4Hz，1H)，6.96(dd，J＝8.1，2.0Hz，1H)，7.17(d，J＝2.0Hz，1H)。
Under room temperature, to 4-amino-3-chlorophenyl methyl acetate (1.00g, 5.01mmol) and Carbimide. 2-methyl phenyl ester (0.60ml, 5.01mmol) add in the mixture in THF (20ml) triethylamine (0.14ml, 1.00mmol).Stir after 1 day, (0.60ml 5.01mmol) adds in this reactant mixture and stirred 17 hours with Carbimide. 2-methyl phenyl ester.The vacuum concentration reactant mixture.Grind residue by adding normal hexane, obtain 3-chloro-4-[N '-(2-aminomethyl phenyl) urea groups] phenylacetic acid methyl ester (1.23g, 74%), be colourless powder. 1H-NMR(CDCl 3)δ2.34(s，3H)，3.54(s，2H)，3.68(s，3H)，6.24(br，1H)，6.99(br，1H)，7.15(dd，J＝8.3，2.0Hz，1H)，7.21-7.31(m，5H)，7.44(d，J＝7.6Hz，1H)，8.20(d，J＝8.5Hz，1H)；MS(ESI)m/z?333(M ++1)，335(M ++3)。
To 3-chloro-4-[N '-(2-aminomethyl phenyl) urea groups] (1.23g adds 0.25N sodium hydroxide (30ml) to the phenylacetic acid methyl ester in the agitating solution of THF 3.70mmol) (30ml).After stirring 14 hours under the room temperature, this solvent of vacuum concentration.Grind residue by adding 1N HCl,, obtain 3-chloro-4-[N '-(2-aminomethyl phenyl) urea groups in 60 ℃ of decompressions dry 2 days down] phenylacetic acid (1.22g, 100%), be colourless powder. 1H-NMR(DMSO-d 6)δ.26(s，3H)，3.40(s，2H)，6.95(t，J＝7.3Hz，1H)，7.11(d，J＝7.6Hz，2H)，7.16(d，J＝7.3Hz，1H)，7.32(s，1H)，7.76(d，J＝8.0Hz，1H)，7.94(dd，J＝9.3，1.0Hz，1H)，8.72(s，2H)；MS(ESI)m/z319(M ++1)，321(M ++3)，341(M ++Na)。
Under room temperature, with 3-chloro-4-[N '-(2-aminomethyl phenyl) urea groups] phenylacetic acid (319mg, 1.00mmol), 4-[(4S)-fluoro-(2S)-pyrrolidinyl] methoxyl methyl benzoate (253mg, 1.00mmol), EDCHCl (288mg, 1.50mmol), HOBT (203mg, 1.50mmol) and triethylamine (0.70ml, 5.00mmol) mixture in DMF (4ml) stirred 15 hours.In this mixture impouring frozen water, use ethyl acetate extraction.Extract with frozen water and salt water washing merging.After dried over sodium sulfate, the vacuum concentration extract.Residue is through TLC[chloroform/acetone (5/1)] purification, obtain 4-[1-[3-chloro-4-[N '-(2-aminomethyl phenyl) urea groups] phenyl acetyl]-(4S)-and fluoro-(2S)-pyrrolidinyl] methoxyl methyl benzoate (480mg, 87%), be colourless amorphous solid. 1H-NMR (CDCl 3) δ 2.10-2.60 (m, 2H), 2.29 (s, 3H), 3.56 (d, J=6.8Hz, 1H), 3.71-3.84 (m, 1H), 3.87 and 3.89 (respectively being s, 3H, amide isomers), 3.91-4.20 (m, 3H), 4.49-4.60 (m, 2H), 5.32 (dt, J=53.0,4.2Hz, 1H), 6.80 (br, 1H), 6.89 and 6.95 (respectively be d, J=8.8Hz, 2H, amide isomers), 7.09-7.26 (m, 6H), 7.50 (1H), 7.94 and 8.00 (respectively is d for d, J=7.3Hz, J=8.8Hz, 2H, amide isomers), 8.10 and 8.15 (respectively is d, J=8.3Hz, 1H, amide isomers); MS (FAB) m/z554 (M ++ 1), 556 (M ++ 3).
To 4-[1-[3-chloro-4-[N '-(2-aminomethyl phenyl) urea groups] phenyl acetyl]-(4S)-and fluoro-(2S)-pyrrolidinyl] (480mg adds 0.25N sodium hydroxide (30ml) to methoxyl methyl benzoate in THF 0.866mmol) (30ml) solution.After stirring 2 days under the room temperature, this mixture of concentrating under reduced pressure is with 1N HCl acidify.Collecting precipitation washes with water, and drying under reduced pressure obtains 90 (374mg, 80%), is colourless powder.MW 539.98 IR (KBr) 3354,3060,2976,1709,1604,1244 cm -1 1H-NMR (DMSO-d 6) δ 2.27 (s, 3H), 2.31 (s, 2H), 3.66 (d, J=7.2Hz, 2H), 3.71-4.67 (m, 5H), and 5.32-5.53 (m, 1H), 6.97 (t, J=7.3Hz, 1H), 7.04-7.22 (m, 5H), 7.32 and 7.35 (respectively being d, J=1.7Hz, 1H, amide isomers), 7.77 (1H), 7.87 and 7.90 (respectively is d for d, J=7.6Hz, J=9.0Hz, 2H, amide isomers), 8.01 and 8.03 (respectively is d, J=8.5Hz, 1H, amide isomers), 8.57 and 8.59 (respectively is s, 1H, amide isomers), 8.63 and 8.65 (respectively be s, 1H, amide isomers), 12.63 (s, 1H); MS (ESI) m/z 540 (M ++ 1), 542 (M ++ 3).Embodiment 854-[1-[3-chloro-4-[N '-(2-chlorophenyl) urea groups] phenyl acetyl]-(4S)-and fluoro-(2S)-pyrrolidinyl] methoxybenzoic acid
Under room temperature, to 4-amino-3-chlorophenyl methyl acetate (1.00g, 5.01mmol) and Carbimide. 2-chloro phenyl ester (0.60ml, 5.01mmol) add in the mixture in THF (20ml) triethylamine (0.14ml, 1.00mmol).Stir after 1 day, (0.60ml 5.01mmol) adds in the reactant mixture and stirred 17 hours with Carbimide. 2-chloro phenyl ester.The vacuum concentration reactant mixture.Grind residue by adding normal hexane, obtain 3-chloro-4-[N '-(2-chlorophenyl) urea groups] phenylacetic acid methyl ester (1.35g, 76%), be colourless powder. 1H-NMR(CDCl 3)δ3.58(s，3H)，3.71(s，2H)，7.04(m，3H)，7.18(dd，J＝8.5，2.0Hz，1H)，7.27-7.39(m，3H)，8.07(m，2H)；MS(ESI)m/z?353(M ++1)，355(M ++3)，357(M ++5)。
To 3-chloro-4-[N '-(2-chlorophenyl) urea groups] (1.35g adds 0.25N sodium hydroxide (30ml) to the phenylacetic acid methyl ester in the agitating solution of THF 3.82mmol) (30ml).After stirring 14 hours under the room temperature, this solution of vacuum concentration.Grind residue by adding 1N HCl,, obtain 3-chloro-4-[N '-(2-chlorophenyl) urea groups in 60 ℃ of drying under reduced pressure 2 days] phenylacetic acid (1.12g, 86%), be colourless powder. 1H-NMR(DMSO-d 6)δ3.52(s，2H)，7.05(m，1H)，7.17(d，J＝8.5Hz，1H)，7.30(d，J＝7.6Hz，1H)，7.37(s，1H)，7.46(dd，J＝8.0，1.5Hz，1H)，7.95(dd，J＝8.3，1.2Hz，1H)，8.07(d，J＝8.3Hz，1H)，9.00(d，J＝8.0Hz，2H)；MS(FAB)m/z339(M ++1)，34?1(M ++3)，343(M ++5)。
Under room temperature, with 3-chloro-4-[N '-(2-chlorophenyl) urea groups] phenylacetic acid (339mg, 1.00mmol), 4-[(2S, 4S)-and 4-fluoro-2-pyrrolidinyl] methoxyl methyl benzoate (253mg, 1.00mmol), EDCHCl (288mg, 1.50mmol), HOBT (203mg, 1.50mmol) and triethylamine (0.70ml, 5.00mmol) mixture in DMF (4ml) stirred 15 hours.In this mixture impouring frozen water, use ethyl acetate extraction.Extract with frozen water and salt water washing merging.After dried over sodium sulfate, the vacuum concentration extract.Residue is through silica gel column chromatography [50g; chloroform/acetone (10/1)] purification; obtain 4-[1-[3-chloro-4-[N '-(2-chlorophenyl) urea groups] phenyl acetyl]-(4S)-and fluoro-(2S)-pyrrolidinyl] methoxyl methyl benzoate (550mg, 96%), be colourless amorphous solid. 1H-NMR (CDCl 3) δ 2.14-2.64 (m, 2H), 3.59 (d, J=11.2Hz, 2H), and 3.78-3.82 (m, 1H), 3.86 and 3.89 (respectively is s, 3H, amide isomers), 3.91-4.28 (m, 2H), 4.50-4.79 (m, 2H), 5.34 and 5.39 (respectively be dt, J=52.5,4.4Hz, 1H, amide isomers), 6.89-6.98 (m, 3H), and 7.09-7.13 (m, 2H), 7.22 (dt, J=7.3,2.2Hz, 1H), 7.29 (dd, J=8.1,2.0Hz, 1H), 7.79 and 7.86 (respectively is s, 1H, amide isomers), 7.86-8.03 (m, 4H), 8.11 (dd, J=8.3,1.0Hz, 1H); MS (FAB) m/z574 (M ++ 1), 576 (M ++ 3), 578 (M ++ 5).
To 4-[1-[3-chloro-4-[N '-(2-chlorophenyl) urea groups] phenyl acetyl]-(4S)-and fluoro-(2S)-pyrrolidinyl] (550mg adds 0.25N sodium hydroxide (30ml) to methoxyl methyl benzoate in THF 0.957mmol) (30ml) solution.After stirring 2 days under the room temperature, this mixture of concentrating under reduced pressure is with 1N HCl acidify.Collecting precipitation washes with water, and drying under reduced pressure obtains 91 (437mg, 82%), is colourless powder.MW 560.40 IR (KBr) 3348,3072,2954,1703,1604,1529,1439 cm -1 1H-NMR (DMSO-d 6) δ 2.25-2.42 (m, 2H), 3.67 (d, J=8.3Hz, 2H), 3.81-4.68 (m, 5H), 5.39 and 5.46 (respectively be d, J=54.4Hz, 1H, amide isomers), and 7.04-7.10 (m, 3H), 7.18 (d, J=8.3Hz, 1H), 7.31 (1H), 7.33 and 7.37 (respectively is s for t, J=8.3Hz, 1H, amide isomers), 7.47 (d, J=8.1Hz, 1H), 7.88 (dd, J=9.0,3.2Hz, 2H), 7.98 (dd, J=8.5,3.0Hz, 1H) m, 1H), 8.09 (d, J=8.3Hz, 1H), 8.99 (d, J=2.9Hz, 1H), 9.02 (s, 1H), 12.64 (s, 1H); MS (ESI) m/z560 (M ++ 1), 562 (M ++ 3), 564 (m ++ 5).C 27H 24Cl 2FN 3O 50.2H 2The analytical calculation value of O: C, 57.50; H, 4.36; N, 7.45; Cl, 12.57; F, 3.37.Measured value: C, 57.72; H, 4.47; N, 7.14; Cl, 12.44; F, 3.44.Embodiment 864-[1-[4-[N '-(2-bromo phenyl) urea groups]-3-chlorophenyl acetyl group]-(4S)-and fluoro-(2S)-pyrrolidinyl] methoxybenzoic acid
Under room temperature, to 4-amino-3-chlorophenyl methyl acetate (1.00g, 5.01mmol) and Carbimide. 2-bromo phenyl ester (0.62ml, 5.01mmol) add in the mixture in THF (20ml) triethylamine (0.14ml, 1.00mmol).Stir after 1 day, (0.60ml 5.01mmol) adds in the reactant mixture and stirred 24 hours with Carbimide. 2-bromo phenyl ester.The vacuum concentration reactant mixture.Grind residue by adding normal hexane, obtain 4-[N '-(2-bromo phenyl) urea groups]-3-chlorophenyl methyl acetate (1.34g, 67%), be colourless powder. 1H-NMR(CDCl 3)δ3.58(s，3H)，3.70(s，2H)，6.98(m，3H)，7.19(dd，J＝8.3，1.9Hz，1H)，7.32(m，1H)，7.51(m，2H)，8.05(m，1H)；MS(ESI)m/z?398(M ++1)，400(M ++3)，402(M ++5)。
To 4-[N '-(2-bromo phenyl) urea groups]-(1.34g adds 0.25N sodium hydroxide (30ml) to 3-chlorophenyl methyl acetate in the agitating solution of THF 3.37mmol) (30ml).After stirring 14 hours under the room temperature, the vacuum concentration solvent.Grind residue by adding 1N HCl,, obtain 4-[N '-(2-bromo phenyl) urea groups in 60 ℃ of drying under reduced pressure 2 days]-3-chlorophenyl acetic acid (1.03g, 80%), be colourless powder. 1H-NMR(DMSO-d 6)δ3.56(s，2H)，7.00(m，1H)，7.17(dd，J＝9.0，1.7Hz，1H)，7.32-7.40(m，2H)，7.62(dd，J＝8.0，1.2Hz，1H)，7.95(m，2H)，8.83(s，1)，9.01(s，H)，12.41(br，1H)；MS(FAB)m/z385(M ++2)，386(M ++4)，388(M ++6)。
Under room temperature, with 4-[N '-(2-bromo phenyl) urea groups]-3-chlorophenyl acetic acid (384mg, 1.00mmol), 4-[(4S)-fluoro-(2S)-pyrrolidinyl] methoxyl methyl benzoate (253mg, 1.00mmol), EDCHCl (288mg, 1.50mmol), HOBT (203mg, 1.50mmol) and triethylamine (0.70ml, 5.00mmol) mixture in DMF (4ml) stirred 15 hours.In this mixture impouring frozen water, use ethyl acetate extraction.Extract with frozen water and salt water washing merging.After dried over sodium sulfate, the vacuum concentration extract.Residue is through silica gel column chromatography [50g; chloroform/acetone (10/1)] purification; obtain 4-[1-[4-[N '-(2-bromo phenyl) urea groups]-3-chlorophenyl acetyl group]-(4S)-and fluoro-(2S)-pyrrolidinyl] methoxyl methyl benzoate (530mg, 86%), be colourless amorphous solid. 1H-NMR (CDCl 3) δ 2.14-2.63 (m, 2H), 3.58 (d, J=10.0Hz, 1H), and 3.73-3.83 (m, 1H), 3.86 and 3.89 (respectively is s, 3H, amide isomers), 3.90-4.29 (m, 3H), 4.50-4.69 (m, 2H), 5.33 and 5.37 (respectively be m, 1H, amide isomers), and 6.88-6.93 (m, 3H), 7.11-7.14 (m, 2H), 7.26 (m, 1H), 7.46 (d, J=8.1Hz, 1H), 7.62-7.78 (m, 2H), 7.89 and 7.93 (respectively is m, 2H, amide isomers), 8.01 (dd, J=8.8,1.7Hz, 2H); MS (FAB) m/z 618 (M +), 620 (M ++ 2), 622 (M ++ 4).
To 4-[1-[4-[N '-(2-bromo phenyl) urea groups]-3-chlorophenyl acetyl group]-(4S)-and fluoro-(2S)-pyrrolidinyl] (530mg adds 0.25N sodium hydroxide (30ml) to methoxyl methyl benzoate in THF 0.856mmol) (30ml) solution.After stirring 2 days under the room temperature, this mixture of concentrating under reduced pressure is with 1N HCl acidify.Extract this mixture with chloroform/methanol (10/1).Extract with frozen water and salt water washing merging.After dried over sodium sulfate, the vacuum concentration extract.Residue obtains 92 (59mg, 11%) through silica gel column chromatography [20g, chloroform/acetone (10/1)-chloroform/methanol (10/1)], is colourless amorphous solid.MW?604.85?IR(KBr)3329，3060，2976，1712，1526，1435?cm -1； 1H-NMR(DMSO-d 6)δ2.31(m，H)，3.48-4.68(m，7H)，5.32-5.53(m，1H)，6.99-7.19(m，4H)，7.36(s，1H)，7.63(dd，J＝6.7，1.2Hz，1H)，7.86-8.1?8(m，4H)，8.83(s，1H)，9.02(s，1H)，12.67(br，1H)；MS(ESI)m/z604(M ++1)，606(M ++3)，608(M ++5)。C 27H 24BrClFN 3O 50.5H 2The analytical calculation value of O: C, 52.83; H, 4.10; N, 6.85; Cl, 5.78; F, 3.09.Measured value: C, 53.24; H, 4.32; N, 6.43; Cl, 6.01; F, 3.07.Embodiment 874-[1-[3-chloro-4-(N '-the phenyl urea groups) phenyl acetyl]-(4S)-and fluoro-(2S)-pyrrolidinyl] methoxybenzoic acid
Under room temperature, to 4-amino-3-chlorophenyl methyl acetate (1.31g, 6.56mmol) and carbanil (0.71ml, 6.56mmol) add in the mixture in THF (20ml) triethylamine (0.19ml, 1.33mmol).Stir after 15 hours the vacuum concentration reactant mixture.Grind residue by adding normal hexane, obtain 3-chloro-4-(N '-phenyl urea groups) phenylacetic acid methyl ester (1.79g, 86%), be the light brown solid. 1H-NMR(CDCl 3)δ3.56(s，2H)，3.70(s，3H)，6.70(m，1H)，7.06(s，1H)，7.14-7.18(m，2H)，7.26(dd，J＝7.8，1.9Hz，1H)，7.33-7.38(m，4H)，8.14(dd，J＝8.3，3.0Hz，1H)；MS(ESI)m/z?319(M ++1)，321(M ++3)。
(1.79g adds 0.25N sodium hydroxide (30ml) in the agitating solution of THF 5.62mmol) (30ml) to 3-chloro-4-(N '-phenyl urea groups) phenylacetic acid methyl ester.After stirring 20 hours under the room temperature, the vacuum concentration solvent.Grind residue by adding 1N HCl,, obtain 3-chloro-4-(N '-phenyl urea groups) phenylacetic acid (1.58g, 92%), be the light brown solid in 60 ℃ of drying under reduced pressure 2 days. 1H-NMR(DMSO-d 6)δ3.55(s，2H)，6.99(t，J＝7.3Hz，1H)，7.17(d，J＝8.3Hz，1H)，7.29(t，J＝7.6Hz，2H)，7.36(s，1H)，7.46(d，J＝8.0Hz，2H)，8.07(d，J＝8.3Hz，1H)，8.28(s，1H)，9.37(s，1H)，12.37(br?1H)。
Under room temperature, with 3-chloro-4-(N '-phenyl urea groups) phenylacetic acid (305mg, 1.00mmol), 4-[(2S, 4S)-and 4-fluoro-2-pyrrolidinyl] methoxyl methyl benzoate (253mg, 1.00mmol), EDCHCl (288mg, 1.50mmol), HOBT (203mg, 1.50mmol) and triethylamine (0.70ml, 5.00mmol) mixture in DMF (4ml) stirred 17 hours.In this mixture impouring frozen water, use ethyl acetate extraction.Extract with frozen water and salt water washing merging.After dried over sodium sulfate, the vacuum concentration extract.Residue is through silica gel column chromatography [30g; chloroform/acetone (20/1)] purification; obtain 4-[1-[3-chloro-4-(N '-phenyl urea groups) phenyl acetyl]-(4S)-and fluoro-(2S)-pyrrolidinyl] methoxyl methyl benzoate (720mg, 100%), be colourless amorphous solid.MS(FAB)m/z?540(M ++1)，542(M ++3)。
To 4-[1-[3-chloro-4-(N '-phenyl urea groups) phenyl acetyl]-(4S)-and fluoro-(2S)-pyrrolidinyl] (720mg adds 0.25N sodium hydroxide (30ml) to methoxyl methyl benzoate in THF/ methanol (30/30ml) solution 1.00mmol).After stirring 2 hours under the room temperature, this reactant mixture was heated 22 hours in 50 ℃.Remove desolvate after, the residue that obtains with 1N HCl acidify.Collecting precipitation washes with water, and drying under reduced pressure obtains 93[412mg, 78% (two steps)], be colourless powder.MW 525.96 IR (KBr) 3346,3302,2976,1712,1604,1240 cm -1 1H-NMR (DMSO-d 6) δ 2.25-2.31 (m, 2H), 3.66 (d, J=7.8Hz, 2H), 3.71-4.67 (m, 5H), 5.31-5.52 (m, 1H), 6.99 (t, J=7.3Hz, 1H), 7.04 and 7.07 (respectively be d, J=8.7Hz, 2H, amide isomers), 7.14-7.18 (m, 1H), 7.29 (t, J=7.3Hz, 2H), 7.35 (d, J=1.7Hz, 1H), 7.46 (d, J=7.8Hz, 2H), 7.87 and 7.90 (respectively be d, J=9.0Hz, 2H, amide isomers), 8.04 and 8.06 (respectively be d, J=8.5Hz, 1H, amide isomers), 8.26 and 8.28 (respectively being s, 1H, amide isomers), 9.36 (s, 1H), 12.63 (s, 1H); MS (ESI) m/z526 (M ++ 1), 528 (M ++ 3); C 27H 25ClFN 3O 50.5H 2The analytical calculation value of O: C, 60.62; H, 4.90; N, 7.85; Cl, 6.63; F, 3.55.Measured value: C, 61.00; H, 5.19; N, 7.40; Cl, 6.66; F, 3.39.Embodiment 884-[1-[3-bromo-4-[N '-(2-aminomethyl phenyl) urea groups] phenyl acetyl]-(4S)-and fluoro-(2S)-pyrrolidinyl] methoxybenzoic acid
Figure A0080951002841
Under room temperature, to 3-bromobenzene guanidine-acetic acid (10.2g, add in the agitating solution of dichloroethanes 47.4mmol) (50ml) methanol (5.8ml, 142mmol) and sulphuric acid (0.5ml).Stir after 20 minutes, this mixture was heated 7 hours in 80 ℃.In reactant mixture impouring frozen water, use chloroform extraction.Extract with sodium bicarbonate aqueous solution and salt water washing merging.After dried over sodium sulfate, the vacuum concentration extract obtains 3-bromobenzene guanidine-acetic acid methyl ester (10.8g, 99%), is colorless oil. 1H-NMR(CDCl 3)δ3.60(s，2H)，3.71(d，J＝1.0Hz，3H)，7.18-7.44(m，4H)。
In 0 ℃, to 3-chlorophenyl methyl acetate (10.8g, 47.1mmol) the middle nitric acid (2.8ml.70.7mmol) that adds that stirs the mixture in sulphuric acid (15.1ml).This reactant mixture was warming up to room temperature 5.5 hours gradually.In reactant mixture impouring frozen water, use chloroform extraction.Extract with sodium bicarbonate aqueous solution and salt water washing merging.After dried over sodium sulfate, the vacuum concentration extract.Residue obtains 3-bromo-4-nitrobenzophenone methyl acetate (3.69g, 29%) through silica gel column chromatography [500g, n-hexane/ethyl acetate (10/1)], is yellow oil. 1H-NMR(CDCl 3)δ3.68(s，2H)，3.73(s，3H)，7.38(dd，J＝8.3，1.2Hz，1H)，7.67(d，J＝1.3Hz，1H)，7.83(d，J＝8.3Hz，1H)。
With 3-bromo-4-nitrobenzophenone methyl acetate (14.8g, 53.8mmol), reduced iron powder (9.62g, 172mmol), AcONa3H 2O (7.32g, 53.8mmol) and the mixture of acetic acid (20.0ml) in methanol (150/600ml) in 90 ℃ the heating 1 hour.After being cooled to room temperature,, use the methanol wash filter cake by the Celite filter reaction mixture.The filtrate that evaporation merges is also used ethyl acetate extraction.With salt water washing extract, through dried over sodium sulfate and vacuum concentration.Residue obtains 4-amino-3-bromobenzene guanidine-acetic acid methyl ester (9.01g, 69%) through silica gel column chromatography [400g, chloroform/ethyl acetate (20/1)], is brown oil. 1H-NMR(CDCl 3)δ3.48(s，2H)，3.68(s，3H)，4.05(br，2H)，6.69(d，J＝8.3Hz，1H)，7.00(dd，J＝8.1，2.0Hz，1H)，7.32(d，J＝2.0Hz，1H)。
Under room temperature, to 4-amino-3-bromobenzene guanidine-acetic acid methyl ester (587mg, 2.40mmol) and Carbimide. 2-methyl phenyl ester (0.287ml, 2.40mmol) add in the mixture in THF (2ml) triethylamine (33ml, 0.24mmol).Stir 21 hours final vacuum concentrated reaction mixtures.Grind residue by adding normal hexane, obtain 3-bromo-4-[N '-(2-aminomethyl phenyl) urea groups] phenylacetic acid methyl ester (650mg, 72%), be light brown powder. 1H-NMR(CDCl 3)δ2.34(s，3H)，3.53(s，2H)，3.68(s，3H)，6.18(br，1H)，6.96(br，1H)，7.18-7.33(m，4H)，7.29(d，J＝4.4Hz，1H)，7.30(d，J＝7.3Hz，1H)，8.19(d，J＝8.3Hz，1H)；MS(ESI)m/z377(M +)，379(M ++2)。
To 3-bromo-4-[N '-(2-aminomethyl phenyl) urea groups] (650mg adds 0.25N sodium hydroxide (10ml) to the phenylacetic acid methyl ester in the agitating solution of THF 1.72mmol) (10ml).After stirring 14 hours under the room temperature, this solvent of vacuum concentration.Grind residue by adding 1N HCl,, obtain 3-bromo-4-[N '-(2-aminomethyl phenyl) urea groups in 60 ℃ of drying under reduced pressure 2 days] phenylacetic acid (1.22g, 100%), be colourless powder. 1H-NMR(DMSO-d 6)δ2.26(s，3H)，3.32(s，2H)，6.93(m，2H)，7.10-7.17(m，4H)，7.76(d，J＝8.1Hz，2H)，8.52(s，1H)；MS(ESI)m/z?385(M ++Na)，387(M ++2+Na)。
Under room temperature, with 3-bromo-4-[N '-(2-aminomethyl phenyl) urea groups] phenylacetic acid (80mg, 0.22mmol), 4-[(4S)-fluoro-(2S)-pyrrolidinyl] methoxyl methyl benzoate (56mg, 0.22mmol), EDCHCl (63mg, 0.33mmol), HOBT (45mg, 0.33mmol) and triethylamine (0.15ml, 1.10mmol) mixture in DMF (1ml) stirred 18 hours.In this mixture impouring frozen water, use ethyl acetate extraction.Extract with frozen water and salt water washing merging.After dried over sodium sulfate, the vacuum concentration extract.Residue is through TLC[chloroform/acetone (5/1)] purification, obtain 4-[1-[3-bromo-4-[N '-(2-aminomethyl phenyl) urea groups] phenyl acetyl]-(4S)-and fluoro-(2S)-pyrrolidinyl] methoxyl methyl benzoate (140mg, 100%), be yellow oil. 1H-NMR(CDCl 3)δ2.30(s，3H)，2.55(m，1H)，3.56(d，J＝6.4Hz，2H)，3.70-3.84(m，3H)，3.87(s，3H)，3.99-4.59(m，3H)，5.23-5.38(m，1H)，6.83-6.94(m，2H)，6.95(d，J＝8.8Hz，1H)，7.07-7.26(m，5H)，7.36-7.63(m，2H)，7.94-8.15(m，3H)；MS(ESI)m/z?598(M ++1)，600(M ++3)。
To 4-[1-[3-bromo-4-[N '-(2-aminomethyl phenyl) urea groups] phenyl acetyl]-(4S)-and fluoro-(2S)-pyrrolidinyl] (140mg adds 0.25N sodium hydroxide (10ml) to methoxyl methyl benzoate in THF 0.22mmol) (10ml) solution.After stirring 14 hours under the room temperature, this mixture of concentrating under reduced pressure is with 1N HCl acidify.Collecting precipitation washes with water, and drying under reduced pressure obtains 94 (109mg, 85%), is colourless powder.MW 584.43IR (KBr) 3313,3060,2976,1687,1604,1525,1244 cm -1 1H-NMR (DMSO-d 6) δ 2.27 (s, 3H), 2.29 (m, 2H), 3.66 (d, J=8.1Hz, 2H), 3.72-4.68 (m, 5H), and 5.31-5.53 (m, 1H), 6.92-6.99 (m, 1H), 7.04 and 7.07 (respectively be d, J=8.3Hz, 2H, amide isomers), and 7.11-7.21 (m, 3H), 7.48 and 7.51 (s, 1H, amide isomers), 7.75 and 7.79 (respectively is d, J=8.1Hz, 1H, amide isomers), 7.86-7.92 (m, 3H), 8.45 and 8.47 (respectively is s, 1H, amide isomers), 8.59 (s, 1H), 12.64 (s, 1H); MS (FAB) m/z584 (M ++ 1), 586 (M ++ 3); C 28H 27BrFN 3O 5The analytical calculation value: C, 57.54; H, 4.66; N, 7.19; Br, 13.67; F, 3.25.Measured value: C, 57.93; H, 4.97; N, 7.04; Br, 13.35; F, 2.89.Embodiment 894-[1-[3-bromo-4-[N '-(2-chlorophenyl) urea groups] phenyl acetyl]-(4S)-and fluoro-(2S)-pyrrolidinyl] methoxybenzoic acid
Under room temperature, to 4-amino-3-bromobenzene guanidine-acetic acid methyl ester (587mg, 2.40mmol) and Carbimide. 2-chloro phenyl ester (0.29ml, 2.40mmol) add in the mixture in THF (2m1) triethylamine (33ml, 0.24mmol).Stir after 21 hours the vacuum concentration reactant mixture.Grind residue by adding normal hexane, obtain 3-bromo-4-[N '-(2-chlorophenyl) urea groups] phenylacetic acid methyl ester (710mg, 74%), be light brown powder. 1H-NMR(CDCl 3)δ3.57(s，2H)，3.70(s，3H)，7.02-7.28(m，2H)，7.36(d，J＝6.8Hz，1H)，7.48(s，1H)，8.00-8.11(m，2H)；MS(ESI)m/z?397(M +)，399(M ++2)，401(M ++4)。
To 3-bromo-4-[N '-(2-chlorophenyl) urea groups] (710mg adds 0.25N sodium hydroxide (10ml) to the phenylacetic acid methyl ester in the agitating solution of THF 1.79mmol) (10ml).After stirring 14 hours under the room temperature, the vacuum concentration solvent.Grind residue by adding 1N HCl,, obtain 3-bromo-4-[N '-(2-chlorophenyl) urea groups in 60 ℃ of drying under reduced pressure 2 days] phenylacetic acid (643mg, 94%), be colourless powder. 1H-NMR(DMSO-d 6)δ3.56(s，2H)，7.05(m，1H)，7.21(dd，J＝8.6，1.7Hz，1H)，7.29(t，J＝7.8Hz，1H)，7.46(d，J＝8.1Hz，1H)，7.46(d，J＝8.1Hz，1H)，7.53(d，J＝1.7Hz，1H)，7.83(d，J＝8.3Hz，1H)，8.06(d，J＝7.6Hz，1H)，8.86(s，1H)，8.89(s，1H)，12.40(s，1H)；MS(ESI)m/z382(M ++1)，384(M ++3)。
Under room temperature, with 3-bromo-4-[N '-(2-chlorophenyl) urea groups] phenylacetic acid (384mg, 1.00mmol), 4-[(4S)-fluoro-(2S)-pyrrolidinyl] methoxyl methyl benzoate (253mg, 1.00mmo1), EDCHCl (288mg, 1.50mmol), HOBT (203mg, 1.50mmol) and triethylamine (0.70ml, 5.00mmol) mixture in DMF (4ml) stirred 18 hours.In this mixture impouring frozen water, use ethyl acetate extraction.Extract with frozen water and salt water washing merging.After dried over sodium sulfate, the vacuum concentration extract.Residue is through silica gel column chromatography [30g; chloroform/acetone (10/1)] purification; obtain 4-[1-[3-bromo-4-[N '-(2-chlorophenyl) urea groups] phenyl acetyl]-(4S)-and fluoro-(2S)-pyrrolidinyl] methoxyl methyl benzoate (640mg, 100%), be colourless amorphous solid. 1H-NMR(CDCl 3)δ2.07-2.46(m，2H)，2.59(t，J＝18.4Hz，1H)，3.57(d，J＝10.5Hz，2H)，3.63-4.67(m，7H)，5.26-5.44(m，1H)，6.89-6.96(m，3H)，7.13(d，J＝7.6Hz，1H)，7.1(t，J＝7.3Hz，1H)，7.26-7.29(m，2H)，7.52-7.94(m，4H)，8.01(d，J＝8.5Hz，1H)，8.09(d，J＝8.5Hz，1H)；MS(FAB)m/z618(M +)，620(M ++3)，622(M ++5)。
To 4-[1-[3-bromo-4-[N '-(2-chlorophenyl) urea groups] phenyl acetyl]-(4S)-and fluoro-(2S)-pyrrolidinyl] (640mg adds 0.25N sodium hydroxide (40ml) to methoxyl methyl benzoate in THF 1.00mmol) (40ml) solution.After stirring 14 hours under the room temperature, this mixture of concentrating under reduced pressure is with 1N HCl acidify.Collecting precipitation washes with water, and drying under reduced pressure obtains 95 (522mg, 86%), is pale yellow powder.MW 604.85IR (KBr) 3317,3072,1709,1685,1604,1529,1290 cm -1 1H-NMR (DMSO-d 6) δ 2.24-2.50 (m, 2H), 3.67 (d, J=8.3Hz, 2H), and 3.73-4.68 (m, 5H), 5.31-5.52 (m, 1H), and 7.03-7.09 (m, 3H), 7.22 (dt, J=8.3,1.7Hz, 1H), 7.30 (d, J=7.3Hz, 1H), 7.46 (dd, J=8.0,1.4Hz, 1H), 7.49 and 7.52 (respectively be d, J=2.0Hz, 1H, amide isomers), 7.80-7.91 (m, 3H), 8.07 (dd, J=8.3,1.2Hz, 1H), 8.85 with 8.86 (respectively being s, 1H, amide isomers), 8.96 and 8.97 (respectively is s, 1H, amide isomers), 12.62 (s, 1H); MS (FAB) m/z605 (M ++ 1), 607 (M ++ 3), 609 (M ++ 3), 626 (M ++ 1+Na); C 27H 24BrClFN 3O 50.8H 2The analytical calculation value of O: C, 52.37; H, 4.17; N, 6.79; F, 3.07.Measured value: C, 52.63; H, 4.12; N, 6.62; F, 2.97.Embodiment 904-[1-[3-bromo-4-[N '-(2-bromo phenyl) urea groups] phenyl acetyl]-(4S)-and fluoro-(2S)-pyrrolidinyl] methoxybenzoic acid
Under room temperature, to 4-amino-3-bromobenzene guanidine-acetic acid methyl ester (587mg, 2.40mmol) and Carbimide. 2-bromo phenyl ester (0.30ml, 2.40mmol) add in the mixture in THF (2ml) triethylamine (33ml, 0.24mmol).Stir after 4 hours the vacuum concentration reactant mixture.Grind residue by adding normal hexane, obtain 3-bromo-4-[N '-(2-bromo phenyl) urea groups] phenylacetic acid methyl ester (770mg, 73%), be light brown powder. 1H-NMR(CDCl 3)δ3.55(s，2H)，3.70(s，3H)，6.97(dd，J＝7.3，1.5Hz，1H)，7.22(dd，J＝8.5，2.2Hz，1H)，7.29-7.33(m，2H)，7.48(d，J＝1.0，2.2Hz，1H)，7.54(dd，J＝8.0，1.2Hz，1H)，8.01(m，2H)；MS(ESI)m/z?443(M ++1)，445(M ++3)，447(M ++5)。
To 3-bromo-4-[N '-(2-bromo phenyl) urea groups] (770mg adds 0.25N sodium hydroxide (10ml) to the phenylacetic acid methyl ester in the agitating solution of THF 1.74mmol) (10ml).After stirring 14 hours under the room temperature, the vacuum concentration solvent.Grind residue by adding 1N HCl,, obtain 3-bromo-4-[N '-(2-bromo phenyl) urea groups in 60 ℃ of drying under reduced pressure 2 days] phenylacetic acid (702mg, 94%), be colourless powder. 1H-NMR(DMSO-d 6)δ3.56(s，2H)，6.99(dt，J＝7.8，1.5Hz，1H)，7.21(dd，J＝8.3，1.7Hz，1H)，7.33(dt，J＝7.1，1.5Hz，1H)，7.53(d，J＝1.7Hz，1H)，7.62(dd，J＝8.1，1.5Hz，1H)，7.82(d，J＝8.3Hz，1H)，7.93(dd，J＝8.1，1.5Hz，1H)，8.82(s，1H)，8.86(s，1H)，12.39(s，1H)；MS(ESI)m/z428(M ++1)，430(M ++3)。
Under room temperature, with 3-bromo-4-[N '-(2-chlorophenyl) urea groups] phenylacetic acid (428mg, 1.00mmol), 4-[(2S, 4S)-and 4-fluoro-2-pyrrolidinyl] methoxyl methyl benzoate (253mg, 1.00mmol), EDCHCl (288mg, 1.50mmol), HOBT (203mg, 1.50mmol) and triethylamine (0.70ml, 5.00mmol) mixture in DMF (4ml) stirred 18 hours.In this mixture impouring frozen water, use ethyl acetate extraction.Extract with frozen water and salt water washing merging.After dried over sodium sulfate, the vacuum concentration extract.Residue is through silica gel column chromatography [30g; chloroform/acetone (10/1)]; obtain 4-[1-[3-bromo-4-[N '-(2-bromo phenyl) urea groups] phenyl acetyl]-(4S)-and fluoro-(2S)-pyrrolidinyl] methoxyl methyl benzoate (720mg, 100%), be colourless amorphous solid. 1H-NMR(CDCl 3)δ2.07-2.45(m，2H)，2.58(m，1H)，3.58(d，J＝9.0Hz，2H)，3.63-4.69(m，9H)，5.26-543(m，1H)，6.88-6.99(m，3H)，7.16(d，J＝8.3Hz，1H)，7.23-7.32(m，2H)，7.46(dd，J＝8.1，1.5Hz，1H)，7.5?1-8.20(m，5H)；MS(FAB)m/z664(M +)，666(M ++3)，668(M ++5)。
To 4-[1-[3-bromo-4-[N '-(2-bromo phenyl) urea groups] phenyl acetyl]-(4S)-and fluoro-(2S)-pyrrolidinyl] (720mg adds 0.25N sodium hydroxide (40ml) to methoxyl methyl benzoate in THF 1.00mmol) (40ml) solution.After stirring 14 hours under the room temperature, this mixture of concentrating under reduced pressure is with 1N HCl acidify.Extract this mixture with chloroform/methanol (10/1).Extract with frozen water and salt water washing merging.After dried over sodium sulfate, the vacuum concentration extract.Residue grinds through silica gel column chromatography [20g, chloroform/acetone (10/1)-chloroform/methanol (20/1)] and by adding ether, obtains 96 (489mg, 75%), is colourless amorphous solid.MW 649.30 IR (KBr) 3450,3313,3070,1709,1684,1525,1435 cm -1 1H-NMR (DMSO-d 6) δ 2.25-2.50 (m, 2H), 3.67 (d, J=8.3Hz, 2H), 3.73-4.68 (m, 5H), and 5.31-5.53 (m, 1H), 6.98-7.08 (m, 3H), 7.21 (d, J=8.2Hz, 1H), 7.34 (t, J=8.8Hz, 1H), 7.50 with 7.53 (they respectively being s, 1H, amide isomers), 7.62 (d, J=8.0Hz, 1H), 7.80-7.96 (m, 4H), 8.82 (s, 1H), 8.85 with 8.86 (they respectively being s, 1H, amide isomers), 12.63 (br, 1H); MS (FAB) m/z 650 (M ++ 1), 652 (M ++ 3), 654 (M ++ 3), 672 (M ++ Na); C 27H 24Br 2FN 3O 50.9H 2The analytical calculation value of O: C, 48.73; H, 3.91; N, 6.31; F, 2.85.Measured value: C, 48.96; H, 3.98; N, 5.92; F, 2.77.Embodiment 914-[1-[4-[N '-(2-aminomethyl phenyl) urea groups]-2,3-difluorophenyl acetyl group]-(4S)-and fluoro-(2S)-pyrrolidinyl methoxyl group] benzoic acid
Figure A0080951002901
Under room temperature, to tert-butyl group ethyl malonic ester (5.35ml, 28.2mmol) in the agitating solution of DMF (150ml), add sodium hydride (60% in oil, 3.38g, 84.7mmol).After 20 minutes, by Dropping funnel drip in DMF (50ml) 2,3-two fluoro Nitrobenzol (5g, 28.2mmol).After the adding, under room temperature, stirred this mixture 3 hours.In this mixture impouring frozen water and saturated ammonium chloride (100ml).With this mixture of ethyl acetate extraction, with the organic layer of 1M HCl and salt water washing merging.Through dried over mgso, filter and concentrate.Residue is dissolved in the dichloromethane (20ml), under room temperature, adds TFA (20ml).This mixture was refluxed 18 hours.This mixture of vacuum evaporation is with toluene (20ml * 2) coevaporation.Residue through silica gel column chromatography (the medium pressure chromatography system: YAMAZEN YFLC-5404-FC, linear gradient hexane-ethyl acetate 10: 0-1: 1, φ 50mm * 300mm, 15ml/min), obtain 2,3-two fluoro-4-nitrobenzophenone ethyl acetate (5.85g, 85%), is yellow oil. 1H-NMR(CDCl 3)δ1.30(m，3H)，3.78(s，2H)，4.22(m，2H)，7.22(m，1H)，7.84(m，1H)；MS(FAB)m/z?246(M ++1)。
Under room temperature, to 2, (5.85g adds SnCl in the agitating solution of ethanol 23.9mol) (100ml) to 3-two fluoro-4-nitrobenzophenone ethyl acetate 2(16.1g, 71.6mmol).Under refluxing, continue to stir 18 hours.Remove desolvate after, make residue be dissolved in chloroform (100ml) and be poured into frozen water-4M sodium hydroxide (the 4M sodium hydroxide of 40ml is in the 300ml frozen water), extract with chloroform (100ml * 2), through anhydrous magnesium sulfate drying and concentrating under reduced pressure.Residue is through silica gel column chromatography (medium pressure chromatography system: YAMAZEN YFLC-5404, hexane-ethyl acetate 9: 1-7: 3 linear gradient liquid, φ 50mm * 500mm, 15ml/min), obtain 4-amino-2,3-difluorophenylacetic acid ethyl ester (1.94g, 38%), is colorless oil. 1H-NMR(CDCl 3)δ1.25(t，J＝7.3Hz，3H)，3.55(d，J＝1.0Hz，2H)，3.78(brs，2H)，4.15(dd，J＝7.2Hz，14.2Hz，2H)，6.49(dt，J＝1.8，8.2Hz，1H)，6.78(m，1H)；MS(FAB)m/z216(M ++1)。
Under room temperature, to 4-amino-2,3-difluorophenylacetic acid ethyl ester (323mg, 1.5mmol) add in the agitating solution in DMF (8ml) triethylamine (0.209ml, 1.5mmol) and Carbimide. 2-methyl phenyl ester (0.372ml, 3.0mmol).Continue to stir 48 hours in 80 ℃.The vacuum evaporation reactant mixture, with solid suspension in normal hexane.Pass through solid collected by filtration.(1: 1, v/v 20ml), added 4M sodium hydroxide (10ml) under room temperature to make this solid be dissolved in THF-methanol.Under room temperature, continue to stir 18 hours.In this reactant impouring 1M HCl, collect the precipitation that produces by filtering.Make this solid recrystallization with chloroform-normal hexane, obtain the 4-[(2-aminomethyl phenyl) urea groups]-2,3-difluorophenylacetic acid (200mg, 42%) is white solid. 1H-NMR(CDCl 3)δ2.30(s，3H)，3.35(s，2H)，6.98(m，1H)，7.04(m，1H)，7.18(d，J＝7.3Hz，2H)，7.69(d，J＝8.1Hz，1H)，7.90(m，1H)；MS(FAB)m/z321(M ++1)。
To 4-(4-S-4-fluoro-2-pyrrolidinyl) methoxyl methyl benzoate (63mg, 0.25mmol) and 4-[N '-(2-aminomethyl phenyl) urea groups]-2,3-difluorophenylacetic acid (82mg, 0.25mmol) add EDCHCl (72mg in the agitating solution in DMF (5ml), 0.38mmol), HOBt (69mg, 0.48mmol) and DMAP (cat.), under room temperature, continue to stir to spend the night.(50ml) dilutes this mixture with ethyl acetate, with 1M sodium hydroxide, 1M HCl and salt water washing, through anhydrous magnesium sulfate drying and concentrating under reduced pressure.Make residue be dissolved in the THF-methanol-water (21ml, 1: 1: 1, v/v/v) in, under room temperature, continue to stir 6 hours.In this mixture impouring 1M HCl, and the usefulness chloroform-methanol (9: 1, v/v) extract.Organic facies and concentrating under reduced pressure through the anhydrous magnesium sulfate drying merging.Residue is through TLC[Whatman, PLK-5F, chloroform/methanol, 20/1, v/v] purification, obtain 97 (69mg, 51%), be white powder.MW 541.52 IR (KBr) 3340,1604,1540,1251,1168,754 cm -1 1H-NMR (DMSO-d 6) δ 2.25 (s, 3H), 2.32 (m, 2H), 3.68-4.40 (m, 7H), 5.32-5.55 (m, 1H), 6.98 (m, 2H), 7.05 (d, J=8.8Hz, 2H), 7.83 (d, J=8.8Hz, 2H), 7.82-7.92 (m, 2H), 8.40 (s, 1H), 9.14 (s, 1H); MS (ESI) m/z564 (M ++ Na); C 28H 26F 3N 3O 52.0H 2The analytical calculation value of O: C, 58.23; H, 5.24; N, 7.28.Measured value: C, 58.07; H, 4.84; N, 7.03.Embodiment 924-[1-[4-[N '-(2-aminomethyl phenyl) urea groups]-2,5-difluorophenyl acetyl group]-(4S)-and fluoro-(2S)-pyrrolidinyl methoxyl group] benzoic acid
Figure A0080951002921
Under room temperature, to the ethyl malonic acid di tert butyl carbonate (6.32ml, 28.2mmol) in the agitating solution of DMF (150ml), add sodium hydride (60% in oil, 3.38g, 84.7mmol).After 20 minutes, by Dropping funnel drip in DMF (50ml) 2, the 5-difluoro nitrobenzene (5g, 28.2mmol).After the adding, under room temperature, stirred this mixture 3 hours.In this mixture impouring frozen water and saturated ammonium chloride (100ml).With this mixture of ethyl acetate extraction, with the organic layer of 1M HCl and salt water washing merging.Through dried over mgso, filter and concentrate.Residue is dissolved in the dichloromethane (20ml), under room temperature, adds TFA (20ml).This mixture was refluxed 18 hours.This mixture of vacuum evaporation is with toluene (20ml * 2) coevaporation.Residue is dissolved in the methanol (150ml), adds concentrated sulphuric acid (5ml).This mixture was refluxed 18 hours.(300ml) dilutes this mixture with ethyl acetate, and water, 1M HCl and salt water washing are through anhydrous magnesium sulfate drying and concentrating under reduced pressure.Residue through silica gel column chromatography (the medium pressure chromatography system: YAMAZEN YFLC-5404-FC, linear gradient hexane-ethyl acetate 10: 0-1: 1, φ 50mm * 300mm, 15ml/min), obtain 2,5-two fluoro-4-nitrobenzophenone ethyl acetate (6.53g, 90%), is yellow oil. 1H-NMR(CDCl 3)δ3.75(s，2H)，3.76(s，3H)，7.29(dd，J＝5.8Hz，10.5Hz，1H)，7.81(dd，J＝6.0Hz，8.4Hz，1H)；MS(ESI)m/z232(M ++1)。
Under room temperature, to 2, (5.88g adds SnCl in the agitating solution of ethanol 25.4mol) (100ml) to 5-two fluoro-4-nitrobenzophenone ethyl acetate 2(17.2g, 76.3mmol).Under refluxing, continue to stir 18 hours.Remove desolvate after, make residue be dissolved in chloroform (100ml) and be poured into frozen water-4M sodium hydroxide (the 4M sodium hydroxide of 40ml is in the 300ml frozen water), extract with chloroform (100ml * 2), through anhydrous magnesium sulfate drying and concentrating under reduced pressure.Residue is through silica gel column chromatography (medium pressure chromatography system: YAMAZEN YFLC-5404, hexane-ethyl acetate 9: 1-7: 3 linear gradient liquid, φ 50mm * 500mm, 15ml/min), obtain 4-amino-2,5-difluorophenylacetic acid ethyl ester (2.85g, 52%), is colorless oil. 1H-NMR(CDCl 3)δ1.28(t，J＝7.3Hz，3H)，3.51(s，2H)，3.78(brs，2H)，4.15(dd，J＝7.2Hz，14.2Hz，2H)，6.47(dd，J＝7.5，10.4Hz，1H)，6.88(dd，J＝6.7，11.0Hz，1H)；MS(FAB)m/z216(M ++1)。
Under room temperature, to 4-amino-2,5-difluorophenylacetic acid ethyl ester (323mg, 1.5mmol) add in the agitating solution in DMF (8ml) triethylamine (0.209ml, 1.5mmol) and Carbimide. 2-methyl phenyl ester (0.372ml, 3.0mmol).Continue to stir 48 hours in 80 ℃.The vacuum evaporation reactant mixture, with solid suspension in normal hexane.Pass through solid collected by filtration.(1: 1, v/v 20ml), added 4M sodium hydroxide (10ml) under room temperature to make this solid be dissolved in THF-methanol.Under room temperature, continue to stir 18 hours.In this reactant impouring 1M HCl, collect the precipitation that produces by filtering.Make this solid recrystallization with chloroform-normal hexane, obtain the 4-[(2-aminomethyl phenyl) urea groups]-2,5-difluorophenylacetic acid (214mg, 46%) is white solid. 1H-NMR(CDCl 3)δ2.30(s，3H)，3.35(m，2H)，7.02(m，2H)，7.18(d，J＝7.3Hz，2H)，7.69(d，J＝7.8Hz，1H)，8.03(m，1H)；MS(FAB)m/z321(M ++1)。
To 4-[(4S)-fluoro-(2S)-pyrrolidinyl] methoxyl methyl benzoate (63mg, 0.25mmol) and 2,5-two fluoro-4-[N '-(2-aminomethyl phenyl) urea groups] phenylacetic acid (82mg, 0.25mmol) add EDCHCl (72mg in the agitating solution in DMF (5ml), 0.38mmol), HOBt (69mg, 0.48mmol) and DMAP (cat.), under room temperature, continue to stir to spend the night.(50ml) dilutes this mixture with ethyl acetate, with 1M sodium hydroxide, 1M HCl and salt water washing, through anhydrous magnesium sulfate drying and concentrating under reduced pressure.Make residue be dissolved in the THF-methanol-water (21ml, 1: 1: 1, v/v/v) in, under room temperature, continue to stir 6 hours.In this mixture impouring 1M HCl, and the usefulness chloroform-methanol (9: 1, v/v) extract.Organic facies and concentrating under reduced pressure through the anhydrous magnesium sulfate drying merging.Residue is through TLC[Whatman, PLK-5F, chloroform/methanol, 20/1, v/v] purification, obtain 98 (69mg, 51%), be white powder.MW 541.52 IR-ATR:3351,1604,1537,1167,754 (cm -1); 1H-NMR (DMSO) δ 2.25 (s, 3H), 2.32 (m, 2H), 3.68-4.70 (m, 7H), and 5.32-5.55 (m, 1H), 6.97 (t, J=7.6Hz, 1H), 7.06 (d, J=8.5Hz, 2H), 7.20 (m, 3H), 7.87 (d, J=8.8Hz, 2H), 7.83-8.04 (m, 2H), 8.45 (s, 1H), 9.1 8 (s, 1H); MS (ESI) m/z 564 (M ++ Na); C 28H 26F 3N 3O 51.75H 2The analytical calculation value of O: C, 58.69; H, 5.19; N, 7.33.Measured value: C, 58.54; H, 4.85; N, 6.98.Embodiment 934-[1-[4-[N '-(2-chlorophenyl) urea groups]-3-methoxyphenyl acetyl group]-4-fluoro-2-pyrrolidinyl] the methylamino benzoic acid 4-[1-[4-[N '-(2-bromo phenyl) urea groups]-3-methoxyphenyl acetyl group]-4-fluoro-2-pyrrolidinyl] the methylamino benzoic acid
Under room temperature, (1.2g adds 1N sodium hydroxide (5ml) in the agitating solution of methanol 4.85mmol) (5ml), stirred this mixture 1 hour under room temperature to 1-tert-butoxycarbonyl-4-pyrrolidines-2-methyl formate.This mixture of vacuum concentration adds water wherein, with 1N HCl neutralization.With this mixture of ethyl acetate extraction.Wash extract with water, then through dried over sodium sulfate, vacuum concentration obtains 1-tert-butoxycarbonyl-4-pyrrolidines-2-formic acid, and (1.1g quantitatively), is colorless oil. 1H-NMR(CDCl 3)δ1.47(br?s，9H)，2.78-2.83(br?s，3H)，4.37(s，2H)，6.73-6.76(m，3H)，7.17(m，1H)。
In 0 ℃, (1.1g adds BH in the agitating solution of THF 4.7mmol) (10.0ml) to 1-tert-butoxycarbonyl-4-pyrrolidines-2-formic acid 3THF (the THF solution of 1.0M, 10.0ml.10.0mmol)。After this under room temperature, stirred 1.0 hours.After the cooling, this mixture of vacuum concentration.In 0 ℃ water is added wherein, use ethyl acetate extraction.Wash extract with water, then through dried over sodium sulfate, vacuum concentration obtains 1-tert-butoxycarbonyl-4-fluoro-2-pyrrolidinyl methanol, and (1.0g quantitatively), is colorless oil. 1H-NMR(CDCl 3)δ1.48(s，9H)，2.29-2.39(m，1H)，3.38-3.59(m，2H)，3.74-3.88(m，2H)，4.09-4.14(m，2H)，4.85(m，1H)，5.03(br?s，1H)，5.16(br?s，1H)。
In-78 ℃, (0.28ml adds DMSO (0.39ml) in the agitating solution of dichloromethane 2.3mmol) (20.0ml) to oxalyl chloride.After 5 minutes, in this mixture, be added in 1-tert-butoxycarbonyl-4-fluoro-2-pyrrolidinyl methanol in the dichloromethane (5.0ml) (500mg, 2.28mmol).In-78 ℃ this mixture was stirred 30 minutes, add triethylamine (l.6ml).In-78 ℃, mixture was stirred 30 minutes.Under room temperature, stirred 30 minutes again.Water is added in this mixture, use dichloromethane extraction.Through dried over sodium sulfate organic layer and vacuum concentration.Crude product can need not to be further purified and be used for subsequent reaction.In 0 ℃, (302mg 2.0mmol) and in the agitating solution of the DCE (10ml) of acetic acid (0.13ml) adds NaBH (OAc) to crude product, 4-Methyl anthranilate 3(656mg, 3.09mmol).Under room temperature, stirred this reactant mixture 18 hours.This mixture of vacuum concentration adds saturated sodium bicarbonate in the residue, uses dichloromethane extraction.With salt water washing extract, through dried over sodium sulfate and vacuum concentration.Residue is through purification by silica gel column chromatography, with ethyl acetate-normal hexane (1: 3, v/v) as eluant, obtain 4-(1-tert-butoxycarbonyl-4-fluoro-2-pyrrolidinyl) methylamino essence of Niobe (541mg, 77%), be faint yellow oily thing. 1H-NMR (CDCl 3) δ 1.55-1.59 (m, 1H), 2.16-2.27 (m, 1H), 2.89-3.03 (m, 2H), 3.19-3.28 (m, 2H), 3.69-3.73 (m, 1H), 3.84 (s, 3H), 5.15 and 5.29 (they respectively are s, are total to 1H), 6.55-6.58 (m, 2H), 7.84-7.86 (m, 2H).
In 0 ℃, (541mg adds TFA (4.0ml) to the methylamino essence of Niobe in the agitating solution of dichloromethane 1.53mmol) (8.0ml) to 4-(1-tert-butoxycarbonyl-4-fluoro-2-pyrrolidinyl).Under room temperature, stirred this reactant mixture 0.5 hour.This mixture of vacuum concentration adds saturated sodium bicarbonate in the residue, uses dichloromethane extraction.With salt water washing extract, through dried over sodium sulfate and vacuum concentration.Crude product can need not to be further purified and be used for subsequent reaction.In 0 ℃, to crude product (151mg, 0.6mmol), 4-[N '-(2-chlorophenyl) urea groups]-3-methoxybenzene guanidine-acetic acid (201mg, 0.6mmol), HOBt (94mg, 0.7mmol) and triethylamine (167 μ l, 1, add in the agitating solution of THF 1.2mmol) (10.0ml) and MeCN (10.0ml) EDCHCl (173mg, 0.9mmol).Under room temperature, stirred this reactant mixture 16 hours, vacuum concentration.Water is added in the residue, use ethyl acetate extraction.With saturated sodium bicarbonate, 2-M citric acid and saturated sodium bicarbonate washing extract, then through dried over sodium sulfate and vacuum concentration.Residue is through purification by silica gel column chromatography; with normal hexane-ethyl acetate (1: 2; v/v) as eluant; obtain 4-[1-[4-[N '-(2-chlorophenyl) urea groups]-3-methoxyphenyl acetyl group]-4-fluoro-2-pyrrolidinyl] methylamino essence of Niobe (320mg; 94%), is amorphous solid. 1H-NMR (CDCl 3) δ 1.80-1.95 (m, 1H), 2.42-2.58 (m, 1H), 3.20-3.51 (m, 3H), 3.51-3.76 (m, 5H), 3.84 (s, 3H), 3.85-3.98 (m, 1H), 4.67-4.70 (m, 1H), 5.10 and 5.23 (they respectively are s, are total to 1H), 5.50 (br s, 1H), 6.49-6.52 (m, 2H), 6.78-6.81 (m, 2H), 6.97-7.01 (m, 1H), 7.14-7.18 (m, 2H), 7.24-7.36 (m, 2H), 7.80-7.82 (m, 2H), and 7.99-8.01 (m, 1H), 8.15-8.18 (m, 1H).
To 4-[1-[4-[N '-(2-chlorophenyl) urea groups]-3-methoxyphenyl acetyl group]-4-fluoro-2-pyrrolidinyl] methylamino essence of Niobe (320mg; 0.56mmol) THF (5.0ml) and the agitating solution of methanol (3.0ml) in add the 1N sodium hydroxide (0.8ml, 0.8mmol).This mixture was stirred 24 hours in 70 ℃.This mixture of vacuum concentration adds water wherein, with 1N HCl neutralization.Collect the solid that produces, wash with water, vacuum drying obtains 99 (280mg, 90%), is white crystalline solid.132-136 ℃ of MW 555.00mp; IR (KBr) 3332,2937,1602,1531,1174,752 cm -1 1H-NMR (DMSO-d 6) δ 2.00-2.40 (m, 2H), 3.50-3.90 (m, 4H), 3.75-3.85 (m, 5H), 4.27 (m, 1H), 5.23 with 5.37 (they respectively are s, are total to 1H), and 6.51-7.03 (m, 5H), 7.25-7.29 (m, 1H), 7.41-7.44 (m, 1H), 7.64-7.68 (m, 2H), 7.92-8.10 (m, 2H), and 8.87-8.94 (m, 2H); C 28H 28N 4O 5FCl0.6H 2The analytical calculation value of O: C, 59.44; H, 5.20; N, 9.90.Measured value: C, 59.41; H, 5.19; N, 9.72.
In 0 ℃, (541mg adds TFA (4.0ml) to the methylamino essence of Niobe in the agitating solution of dichloromethane 1.53mmol) (8.0ml) to 4-(1-tert-butoxycarbonyl-4-fluoro-2-pyrrolidinyl).Under room temperature, stirred this reactant mixture 0.5 hour.This mixture of vacuum concentration adds saturated sodium bicarbonate in the residue, uses dichloromethane extraction.With salt water washing extract, through dried over sodium sulfate and vacuum concentration.Crude product can need not to be further purified and be used for subsequent reaction.In 0 ℃, to crude product (151mg, 0.6mmol), 4-[N '-(2-bromo phenyl) urea groups]-3-methoxybenzene guanidine-acetic acid (227mg, 0.6mmol), HOBt (94mg, 0.7mmol) and triethylamine (167 μ l, 1.2mmol) THF (10.0ml) and the agitating solution of MeCN (10.0ml) in add EDCHCl (173mg, 0.9mmol).Under room temperature, stirred this reactant mixture 16 hours, vacuum concentration.Water is added in the residue, use ethyl acetate extraction.With saturated sodium bicarbonate, 2-M citric acid and saturated sodium bicarbonate washing extract, then through dried over sodium sulfate and vacuum concentration.Residue is through purification by silica gel column chromatography; with normal hexane-ethyl acetate (2: 3; v/v) as eluant; obtain 4-[1-[4-[N '-(2-bromo phenyl) urea groups]-3-methoxyphenyl acetyl group]-4-fluoro-2-pyrrolidinyl] methylamino essence of Niobe (280mg; 76%), is colorless oil. 1H-NMR (CDCl 3) δ 1.80-1.98 (m, 1H), 1.42-1.58 (m, 1H), 3.20-3.52 (m, 3H), 3.67-3.79 (m, 5H), 3.84 (s, 3H), 3.94-3.97 (m, 1H), 4.68-4.71 (m, 1H), 5.10 and 5.23 (they respectively are s, are total to 1H), 5.5 1 (br s, 1H), and 6.50-6.52 (m, 2H), 6.79-7.07 (m, 5H), 7.25-7.33 (m, 1H), 7.51-7.53 (m, 1H), 7.80-7.83 (m, 2H), and 7.98-8.00 (m, 1H), 8.11-8.14 (m, 1H).
To 4-[1-[4-[N '-(2-bromo phenyl) urea groups]-3-methoxyphenyl acetyl group]-4-fluoro-2-pyrrolidinyl] methylamino essence of Niobe (280mg; 0.46mmol) THF (8.0ml) and the agitating solution of methanol (8.0ml) in add the 1N sodium hydroxide (2.8ml, 2.8mmol).This mixture was stirred 18 hours in 70 ℃.This mixture of vacuum concentration adds water wherein, with 1N HCl neutralization.Collect the solid that produces, wash with water, vacuum drying obtains 100 (260mg, 95%), is white crystalline solid.131-135 ℃ of MW 599.45mp; IR (KBr) 3332,2935,1602,1529,1174 cm -1 1H-NMR (DMSO-d 6) δ 1.95-2.01 (m, 1H), 2.20-2.35 (m, 1H), 3.10-3.20 (m, 1H), and 3.50-3.70 (m, 3H), 3.80-3.85 (m, 5H), 4.27 (m, 1H), 5.24 and 5.37 (they respectively are s, are total to 1H), 6.54-6.99 (m, 5H), 7.30-7.33 (m, 1H), 7.58-7.94 (m, 3H), 7.94-7.98 (m, 2H), 8.73 (m, 1H), 8.93 (m, 1H); C 28H 28N 4O 5BrF0.7H 2The analytical calculation value of O: C, 54.95; H, 4.84; N, 9.15.Measured value: C, 54.98; H, 4.81; N, 8.93.Embodiment 944-[1-[3-methoxyl group-4-[N '-(2-aminomethyl phenyl) urea groups] phenyl acetyl]-(4R)-and fluoro-(2S)-pyrrolidinyl methoxyl group] benzoic acid
Figure A0080951002981
With 4-[(4R)-fluoro-(2S)-pyrrolidinyl methoxyl group] essence of Niobe (634mg, 2.50mmol), 3-methoxyl group-4-[N '-(2-aminomethyl phenyl) urea groups] phenylacetic acid (787mg, 2.50mmol), EDCHCl (718mg, 3.75mmol), the mixture of HOBt (cat.), DMAP (cat.) and DMF (10ml) stirs and spend the night.(300m1) dilutes this mixture with ethyl acetate.(2 * 100ml) wash this solution, through dried over mgso and vacuum concentration with saline.Residue is through silica gel column chromatography; with chloroform-ethyl acetate (4: 1) as eluant; obtain 4-[1-[3-methoxyl group-4-[N '-(2-aminomethyl phenyl) urea groups] phenyl acetyl]-(4R)-and fluoro-(2S)-pyrrolidinyl methoxyl group] (1.37g quantitatively), is faint yellow viscous solid to essence of Niobe. 1H-NMR (CDCl 3) δ 2.24 (s, 3H), 2.26-2.47 (m, 2H), 3.46 (s, 3H), 3.49-3.64 (m, 4H), 3.87 (s, 3H), 4.06 (dd, J=9.5,2.0Hz, 1H), 4.51-4.62 (m, 2H), 5.20 and 5.33 (they respectively are br s, are total to 1H), 6.63 (s, 1H), 6.72 (d, J=8.3Hz, 1H), 6.77 (d, J=9.0Hz, 2H), 7.05 (t, J=7.6Hz, 1H), 7.16-7.20 (m, 3H), 7.53 (s, 1H), 7.63 (d, J=7.8Hz, 1H), 7.91 (d, J=9.0Hz, 2H), 8.07 (d, J=8.1Hz, 1H).
With 4-[1-[3-methoxyl group-4-[N '-(2-aminomethyl phenyl) urea groups] phenyl acetyl]-(4R)-and fluoro-(2S)-pyrrolidinyl methoxyl group] essence of Niobe (1.37g; 2.49mmol), the 0.25N sodium hydroxide (20ml, 5.00mmol) and the mixture of THF (20ml) stirred 3 days.In this mixture impouring 1NHCl (100ml), (5: 1,2 * 200ml) extracted with chloroform-methanol.Extract and vacuum concentration through the dried over mgso merging.Residue is through silica gel column chromatography, with chloroform-methanol (20: 1-4: 1) eluting, obtain 101 (930mg, 70%), be faint yellow amorphous solid.MW 535.56 1H-NMR (DMSO-d 6) δ 2.24-2.41 (m, 5H altogether), and 3.42-4.66 (a plurality of m, 10H), 5.31 and 5.44 (they respectively are br s, are total to 1H), 6.71-7.16 (a plurality of m, 7H), 7.79 (d, J=8.1Hz, 1H), 7.85-7.89 (m, 2H), 7.98-8.00 (m, 1H), 8.47 (s, 1H), 8.55 (s, 1H); MS (FAB) m/z 536 (M ++ 1).Embodiment 954-[(4S)-and chloro-1-[3-methoxyl group-4-[N '-(2-chlorophenyl) urea groups] phenyl acetyl]-(2S)-and pyrrolidinyl] methoxybenzoic acid
Under room temperature, to 4-(anti-form-1-tert-butoxycarbonyl-4-hydroxyl-(2S)-pyrrolidinyl) methoxyl methyl benzoate (351mg, 1.0mmol) and triphenyl phasphine (393mg adds carbon tetrachloride (5.0ml) in the agitating solution of chloroform 1.5mmol) (5.0ml).In 50 ℃ reactant mixture was stirred 24 hours.With this reactant mixture vacuum concentration.Residue is through purification by silica gel column chromatography, with normal hexane to normal hexane-ethyl acetate (4: 1, v/v) as eluant, obtain 4-(cis-1-tert-butoxycarbonyl-4-chloro-(2S)-pyrrolidinyl) methoxyl methyl benzoate (340mg, 92%), is faint yellow oily thing. 1H-NMR(CDCl 3)δ1.48(s，9H)，2.38-2.65(m，2H)，3.50-3.90(m，1H)，3.88(s，3H)，3.89-4.05(m，1H)，4.26-4.41(m，4H)，6.95-6.97(m，2H)，7.98(d，J＝8.5Hz，2H)。
In 0 ℃, (369mg adds TFA (3.0ml) to methoxyl methyl benzoate in the agitating solution of dichloromethane 1.0mmol) (3.0ml) to 4-(1-tert-butoxycarbonyl-(4S)-chloro-(2S)-pyrrolidinyl).The mixture that obtains was stirred under room temperature 0.5 hour.This mixture of vacuum concentration.Saturated sodium bicarbonate is added in the residue, use dichloromethane extraction.With salt water washing extract, through dried over sodium sulfate and vacuum concentration.Crude product can need not to be further purified and be used for subsequent reaction.In 0 ℃, to crude product (185mg, 0.5mmol), 4-[N '-(2-chlorophenyl) urea groups]-3-methoxybenzene guanidine-acetic acid (167mg, 0.5mmol), HOBt (68mg, 0.5mmol) and triethylamine (208ml, 1.5mmol) THF (8.0ml) and the agitating solution of MeCN (8.0ml) in add EDCHCl (144mg, 0.75mmol).Under room temperature, stirred this reactant mixture 16 hours, vacuum concentration.Water is added in the residue, use ethyl acetate extraction.With saturated sodium bicarbonate, 2-M citric acid and saturated sodium bicarbonate washing extract, then through dried over sodium sulfate and vacuum concentration.Residue is through purification by silica gel column chromatography; with normal hexane-ethyl acetate (1: 2; v/v) as eluant; obtain 4-[(4S)-chloro-1-[3-methoxyl group-4-[N '-(2-chlorophenyl) urea groups] phenyl acetyl]-(2S)-and pyrrolidinyl] methoxyl methyl benzoate (210mg; 72%), is colorless oil. 1H-NMR(CDCl 3)δ3.35-3.50(m，1H)，3.55-3.65(m，1H)，3.61-3.66(m，3H)，3.75(s，3H)，3.88(s，3H)，3.99-4.04(m，1H)，4.35-4.40(m，3H)，4.48-4.53(m，1H)，6.77-7.10(m，7H)，7.25-7.36(m，2H)，7.93-8.00(m，2H)，8.18(d，J＝8.0Hz，1H)。
To 4-[(4S)-chloro-1-[3-methoxyl group-4-[N '-(2-chlorophenyl) urea groups] phenyl acetyl]-(2S)-and pyrrolidinyl] methoxyl methyl benzoate (210mg; 0.35mmol) THF (6.0ml) and the agitating solution of methanol (3.0ml) in add the 1N sodium hydroxide (0.7ml, 0.7mmol).This mixture was stirred 18 hours in 70 ℃.This mixture of vacuum concentration adds water wherein, with 1N HCl neutralization.Collect the solid that produces, wash with water, vacuum drying obtains 102 (200mg, 98%), is white crystalline solid.MW 572.44 mp 126-131 ℃; IR (KBr) 3330,1685,1604,1533,1438 cm -1 1H-NMR (DMSO-d 6) δ 2.15-2.25 (m, 1H), 2.58-2.63 (m, 1H), 3.58-3.78 (m, 3H), 3.83 (s, 3H), 4.13-4.42 (m, 4H), 4.73 (m, 1H), 6.75-7.45 (m, 7H), 7.86-8.10 (m, 4H), 8.90 (s, 1H), 8.95 (s, 1H); MS (FAB) m/z 572 (M ++ 1); C 28H 27N 3O 6The analytical calculation value of Cl: C, 58.75; H, 4.75; N, 7.34.Measured value: C, 58.93; H, 4.85; N, 7.15.Embodiment 964-[1-[4-[N '-(2-bromo phenyl) urea groups]-3-methoxyphenyl acetyl group]-(4S)-and chloro-(2S)-pyrrolidinyl] methoxybenzoic acid
Figure A0080951003001
In 0 ℃, to 4-[1-tert-butoxycarbonyl-(4S)-chloro-(2S)-pyrrolidinyl] (369mg adds TFA (3.0ml) to methoxyl methyl benzoate in the agitating solution of dichloromethane 1.0mmol) (3.0ml).Under room temperature, reactant mixture was stirred 0.5 hour.This mixture of vacuum concentration.Saturated sodium bicarbonate is added in the residue, use dichloromethane extraction.With salt water washing extract, through dried over sodium sulfate and vacuum concentration.Crude product can need not to be further purified and be used for subsequent reaction.In 0 ℃, to crude product (185mg, 0.5mmol), 4-[N '-(2-bromo phenyl) urea groups]-3-methoxybenzene guanidine-acetic acid (190mg, 0.5mmol), HOBt (68mg, 0.5mmol) and triethylamine (208ml, 1.5mmol) THF (8.0ml) and the agitating solution of MeCN (8.0ml) in add EDCHCl (144mg, 0.75mmol).Under room temperature, stirred this reactant mixture 16 hours, vacuum concentration.Water is added in the residue, use ethyl acetate extraction.With saturated sodium bicarbonate, 2-M citric acid and saturated sodium bicarbonate washing extract, then through dried over sodium sulfate and vacuum concentration.Residue is through purification by silica gel column chromatography; with normal hexane-ethyl acetate (1: 2; v/v) as eluant; obtain 4-[1-[4-[N '-(2-bromo phenyl) urea groups]-3-methoxyphenyl acetyl group]-(4S)-and chloro-(2S)-pyrrolidinyl] methoxyl methyl benzoate (260mg; 83%), is colorless oil. 1H-NMR(CDCl 3)δ2.32-2.50(m，1H)，2.53-2.65(m，1H)，3.61-3.67(m，3H)，3.75(s，3H)，3.88(s，3H)，3.99-4.03(m，1H)，4.35-4.40(m，3H)，4.45-4.55(m，1H)，6.78-7.10(m，7H)，7.28-7.33(m，1H)，7.52(d，J＝8.0Hz，1H)，7.94-7.99(m，3H)，8.14(d，J＝8.3Hz，1H)。
To 4-[1-[4-[N '-(2-bromo phenyl) urea groups]-3-methoxyphenyl acetyl group]-(4S)-and chloro-(2S)-pyrrolidinyl] methoxyl methyl benzoate (260mg; 0.4mmol) THF (6.0ml) and the agitating solution of methanol (3.0ml) in add the 1N sodium hydroxide (0.8ml, 0.8mmol).This mixture was stirred 24 hours in 70 ℃.This mixture of vacuum concentration adds water wherein, with 1N HCl neutralization.Collect the solid that produces, wash with water, vacuum drying obtains 103 (210mg, 83%), is white crystalline solid.MW 616.89 mp 127-132 ℃; IR (KBr) 3330,1685,1604,1529,1434 cm -1 1H-NMR (DMSO-d 6) δ 2.18-2.28 (m, 1H), 2.60-2.70 (m, 1H), 3.55-3.75 (m, 3H), 3.83 (s, 3H), 4.12-4.42 (m, 4H), and 4.60-4.75 (m, 1H), 6.75-7.06 (m, 5H), and 7.30-7.34 (m, 1H), 7.60 (d, J=7.3Hz, 1H), 7.86-7.94 (m, 5H), 8.75 (s, 1H), 8.94 (s, 1H); MS (FAB) m/z 616 (M +), 618 (M ++ 2); C 28H 27N 3O 6The analytical calculation value of ClBr: C, 54.52; H, 4.41; N, 6.81.Measured value: C, 54.98; H, 4.54; N, 6.66.Embodiment 974-[(4R)-and chloro-1-[3-methoxyl group-4-[N '-(2-aminomethyl phenyl) urea groups] phenyl acetyl]-(2S)-and the pyrrolidinyl methoxyl group] benzoic acid 4-[(4R)-chloro-1-[4-[N '-(2-chlorophenyl) urea groups]-3-methoxyphenyl acetyl group]-(2S)-and the pyrrolidinyl methoxyl group] benzoic acid
Figure A0080951003022
To 1-(tert-butoxycarbonyl)-(4S)-hydroxyl-(2S)-pyrrolidinyl methyl formate (1.81g, 7.34mmol) carbon tetrachloride-dichloromethane (20ml, 1: 1, v/v) add triphenyl phasphine (3.87mmol in the agitating solution, 14.75mmol), under room temperature, reactant mixture was stirred 2 hours.In this mixture, add ethanol (5ml), under room temperature, this reactant mixture stirring is spent the night.Remove desolvate after, residue is through purification by silica gel column chromatography, with normal hexane-ethyl acetate (3: 1, v/v) as eluant, obtain 1-(tert-butoxycarbonyl)-(4R)-chloro-(2S)-pyrrolidinyl methyl formate (1.36g, 70%), be colorless oil. 1H-NMR (CDCl 3) δ 1.42 (s, 9H), 2.32-2.39 (m, 1H), 2.49-2.54 (m, 1H), 3.66-3.92 (a plurality of s and m. be total to 5H), 4.44-4.55 (m, 2H); MS (FAB) m/z 264 (M ++ 1).
(1.35g adds 0.5N sodium hydroxide (10ml) in the agitating solution of THF 5.12mmol) (10ml), this reactant mixture was heated 1.5 hours under refluxing to 1-(tert-butoxycarbonyl)-(4R)-chloro-(2S)-pyrrolidinyl methyl formate.After being cooled to room temperature, in this mixture impouring ice-1N HCl, the usefulness chloroform-methanol (9: 1, v/v) extract this mixture.With salt water washing extract, obtain 1-(tert-butoxycarbonyl)-(4R)-chloro-(2S)-pyrrolidinyl formic acid (1.28g, quantitative) through dried over sodium sulfate and evaporation, be colorless oil. 1H-NMR(CDCl 3)δ1.44(s，9H)，2.37-2.54(m，2H)，3.68-3.88(m，2H)，4.42-4.45(m，2H)。
(1.28g drips BH in the agitating solution of THF 5.13mmol) (20ml) to 1-(tert-butoxycarbonyl)-(4R)-chloro-(2S)-pyrrolidinyl formic acid by syringe 3(0.60ml, 6.33mmol), stirred reaction mixture is 1 hour under room temperature for DMS.Except that after desolvating, residue is dissolved in the dichloromethane.Water, this solution of salt water washing are through dried over sodium sulfate and evaporation.Residue is through purification by silica gel column chromatography, with chloroform-methanol (50: 1, v/v) as eluant, obtain 1-(tert-butoxycarbonyl)-(4R)-chloro dried meat ammonia alcohol (0.88g, 73%), be colorless oil. 1H-NMR(CDCl 3)δ1.48(s，9H)，1.98(m，1H)，2.26-2.32(m，1H)，3.56-3.65(m，2H)，3.77(m，2H)，4.24(m，1H)，4.41-4.46(m，2H)；MS(FAB)m/z236(M ++1)。
To 4-methyl hydroxybenzoate (560mg, 3.68mmol), 1-(tert-butoxycarbonyl)-(4R)-chloro dried meat ammonia alcohol (870mg, 3.69mmol), triphenyl phasphine (1.16g, 4.42mmol) the agitating solution of cold (0 ℃) of THF (15ml) in add DIAD (870ml, 4.42mmol), with heating under this reaction mixture refluxed 10 hours.After being cooled to room temperature, evaporate this mixture.Residue is through purification by silica gel column chromatography, with normal hexane-ethyl acetate (5: 1,, obtain 4-[1-(tert-butoxycarbonyl)-(4R)-chloro-(2S)-pyrrolidinyl methoxyl group v/v) as eluant] essence of Niobe (890mg, 65%), be white solid.Mp 116-120 ℃; 1H-NMR (CDCl 3) δ 1.47 (s, 9H), 2.39-2.53 (m, 2H), 3.69-3.70 and 4.13-4.17 (m, 3H altogether), 3.88 (s, 3H), 4.30-4.41 (m, 2H), 4.50-4.55 (m, 1H), 6.90-6.92 (m, 2H), 7.96-7.98 (m, 2H); MS (FAB) m/z370 (M ++ 1); C 18H 24ClNO 5The analytical calculation value: C, 58.46; H, 6.54; Cl, 9.59; N, 3.79.Measured value: C, 58.35; H, 6.56; Cl, 9.75; N, 3.77.
To 4-[1-(tert-butoxycarbonyl)-(4R)-chloro-(2S)-pyrrolidinyl methoxyl group] (840mg adds TFA (10ml) to essence of Niobe in the agitating solution of dichloromethane 2.27mmol) (10ml).This reactant mixture stirred under room temperature spend the night.This mixture of vacuum concentration, with saturated sodium bicarbonate make be alkalescence.With this mixture of chloroform extraction, use the salt water washing, obtain 4-[(4R through dried over sodium sulfate and evaporation)-chloro-(2S)-pyrrolidinyl methoxyl group] essence of Niobe (580mg, 95%), be white solid.mp?61-64℃； 1H-NMR(CDCl 3)δ1.85(br?s，1H)，2.03-2.10(m，1H)，2.29-2.35(m，1H)，3.19-3.31(m，2H)，3.88(s，3H)，3.92-4.06(m，3H)，4.53-4.56(m，1H)，6.91(d，J＝8.8Hz，2H)，7.98(d，J＝8.8Hz，2H)；MS(FAB)m/z270(M ++1)。
Under room temperature, with 3-methoxyl group-4-[N '-(2-aminomethyl phenyl) urea groups] phenylacetic acid (385mg, 1.22mmol), 4-[(4R)-chloro-(2S)-pyrrolidinyl methoxyl group] essence of Niobe (330mg, 1.22mmol), EDCHCl (281mg, 1.47mmol), HOBt (200mg, 1.48mmol) and triethylamine (205ml, 1.47mmol) mixture in THF (10ml) stirs and to spend the night.This mixture of dilute with water is used ethyl acetate extraction.With salt water washing extract, through dried over sodium sulfate and evaporation.Residue is through purification by silica gel column chromatography; with chloroform-methanol (100: 1-50: 1; v/v) as eluant; obtain 4-[(4R)-chloro-1-[3-methoxyl group-4-[N '-(2-aminomethyl phenyl) urea groups] phenyl acetyl]-(2S)-and the pyrrolidinyl methoxyl group] essence of Niobe (670mg; 97%), is the white foam thing. 1H-NMR(CDCl 3)δ2.28(s，3H)，2.33-2.57(m，2H)，3.50(s，3H)，3.59-3.60(m，2H)，3.75-3.82(m，2H)，3.88(s，3H)，4.06-4.09(m，1H)，4.51-4.63(m，3H)，6.65-6.80(m，5H)，7.09-7.13(m，1H)，7.20-7.27(m，3H)，7.56-7.58(m，1H)，7.91-7.93(m，2H)，8.05-8.07(m，1H)；MS(FAB)m/z?566(M ++1)。
To 4-[(4R)-chloro-1-[3-methoxyl group-4-[N '-(2-aminomethyl phenyl) urea groups] phenyl acetyl]-(2S)-and the pyrrolidinyl methoxyl group] (480mg adds 0.5N sodium hydroxide (5ml) to essence of Niobe in the agitating solution of THF 0.85mmol) (5ml).With this reaction mixture refluxed heating 2 hours.After being cooled to room temperature, in this mixture impouring ice-1N HCl, the precipitation that produces is collected in decompression.Make the crude product solid be dissolved in the chloroform-methanol and evaporation.With ether wash residual thing, obtain 104 (355mg, 76%), be white amorphous solid.128-132 ℃ of MW 552.02mp; 1H-NMR (DMSO-d 6) δ 2.25 (s, 3H), 2.29-2.46 (m, 2H), 3.57-3.73 (m, 2H), 3.78 (s, 3H), 3.81-3.99 (m, 2H), 4.11-4.31 (m, 2H), 4.43-4.45 and 4.64-4.67 (respectively be m, be total to 1H), 4.83-4.85 (m, 1H), and 6.71-7.17 (m, 7H), 7.78-7.80 (m, 1H), 7.87-7.91 (m, 2H), 7.99-8.01 (m, 1H), 8.47 (s, 1H), 8.56 (s, 1H), 12.66 (brs, 1H); MS (FAB) m/z552 (M ++ 1); C 29H 30ClN 3O 63/4H 2The analytical calculation value of O; C, 61.59; H, 5.61; Cl, 6.27; N, 7.43.Measured value: C, 61.56; H, 5.51; Cl, 6.68; N, 7.26.
Under room temperature, with 4-[N '-(2-chlorophenyl) urea groups]-3-methoxybenzene guanidine-acetic acid (400mg, 1.19mmol), 4-[(4R)-chloro-(2S)-pyrrolidinyl methoxyl group] essence of Niobe (320mg, 1.19mmol), EDCHCl (275mg, 1.43mmol), HOBt (195mg, 1.44mmol) and triethylamine (200ml, 1.43mmol) mixture in THF (10ml) stirs and to spend the night.This mixture of dilute with water is used ethyl acetate extraction.With salt water washing extract, through dried over sodium sulfate and evaporation.Residue is through purification by silica gel column chromatography; with chloroform-methanol (100: 1; v/v) as eluant; obtain 4-[(4R)-chloro-1-[4-[N '-(2-chlorophenyl) urea groups]-3-methoxyphenyl acetyl group]-(2S)-and the pyrrolidinyl methoxyl group] essence of Niobe (690mg; 98%), is the weak yellow foam thing. 1H-NMR(CDCl 3)δ2.35-2.41(m，1H)，2.49-2.59(m，1H)，3.55(s，3H)，3.57-3.70(m，2H)，3.73-3.86(m，2H)，3.88(s，3H)，4.06-4.09(m，1H)，4.54-4.66(m，3H)，6.67-6.81(m，4H)，6.95-6.99(m，1H)，7.23-7.25(m，1H)，7.29-7.33(m，1H)，7.47-7.49(m，2H)，7.90-7.99(m，3H)，8.18-8.21(m，1H)；MS(FAB)m/z586(M ++1)。
To 4-[(4R)-chloro-1-[4-[N '-(2-chlorophenyl) urea groups]-3-methoxyphenyl acetyl group]-(2S)-and the pyrrolidinyl methoxyl group] (410mg adds 0.5N sodium hydroxide (5ml) to essence of Niobe in the agitating solution of THF 0.70mmol) (5ml).With this reaction mixture refluxed heating 5 hours.After being cooled to room temperature, in this mixture impouring ice-1N HCl, the precipitation that produces is collected in decompression.Make the crude product solid be dissolved in the chloroform-methanol and evaporation.With ether wash residual thing, obtain 105 (282mg, 70%), be amorphous solid.MW 572.44 mp 131-136 ℃; 1H-NMR (DMSO-d 6) δ 2.29-2.35 (m, 1H), 2.44-2.47 (m, 1H), 3.58-3.74 (m, 2H), 3.78 (s, 3H), 3.81-3.99 (m, 2H), 4.10-4.32 (m, 2H), 4.44-4.46 with 4.66 (they respectively are m, are total to 1H), 4.84 (m, 1H), 6.74-7.04 (m, 5H), 7.26-7.30 (m, 1H), 7.43-7.45 (m, 1H), 7.87-7.91 (m, 2H), 7.96 (d, J=8.3Hz, 1H), 8.09 (d, J=8.3Hz, 1H), 8.90 (s, 1H), 8.94 (s, 1H); C 28H 27Cl 2N 3O 63/4H 2The analytical calculation value of O: C, 57.39; H, 4.90; Cl, 11.66; N, 7.17.Measured value: C, 57.57; H, 4.94; Cl, 11.66; N, 6.89.Embodiment 984-[(4S)-and hydroxyl-1-[4-[N '-(2-aminomethyl phenyl) urea groups] phenyl acetyl]-(2S)-and pyrrolidinyl] methoxybenzoic acid
Figure A0080951003061
Under room temperature, to 4-[(4S)-acetoxyl group-1-tert-butoxycarbonyl-(2S)-pyrrolidinyl] (2.31g adds TFA (10ml) to methoxyl methyl benzoate in the agitating solution of dichloromethane 5.87mmol) (46ml).Stir after 3.5 hours this mixture of vacuum concentration.By adding dichloromethane and 1N sodium hydroxide dilution residue, use dichloromethane extraction.With the extract of salt water washing merging, through dried over sodium sulfate and vacuum concentration.Residue obtains 4-[(4S through silica gel column chromatography [100g, chloroform/methanol (20/1)])-acetoxyl group-(2S)-pyrrolidinyl] methoxyl methyl benzoate (1.89mg, 100%), be the lilac solid. 1H-NMR(CDCl 3)δ2.10(s，3H)，2.14(m，1H)，2.65(m，1H)，3.52-3.63(m，H)，3.89(s，3H)，4.18(m，1H)，4.28(d，J＝5.9Hz，2H)，5.38(m，1H)，6.93(d，J＝8.8Hz，2H)，7.99(d，J＝8.8Hz，2H)。
Under room temperature, with 3-methoxyl group-4-[N '-(2-aminomethyl phenyl) urea groups] phenylacetic acid (343mg, 1.09mmol), 4-[(4S)-acetoxyl group-(2S)-pyrrolidinyl] methoxyl methyl benzoate (320mg, 1.09mmol), EDCHCl (313mg, 1.64mmol), HOBt (222mg, 1.64mmol) and triethylamine (0.76ml, 5.45mmol) mixture in DMF (7ml) stirred 16 hours.In this mixture impouring frozen water, use ethyl acetate extraction.Extract with frozen water and salt water washing merging.After dried over sodium sulfate, the vacuum concentration extract.Residue is through silica gel column chromatography [50g; chloroform/acetone (5/1)]; obtain 4-[(4S)-acetoxyl group-1-[3-methoxyl group-4-[N '-(2-aminomethyl phenyl) urea groups] phenyl acetyl]-(2S)-and pyrrolidinyl] methoxyl methyl benzoate (520mg, 81%), be brown amorphous solid. 1H-NMR (CDCl 3) δ 2.00 (s, 3H, one of isomer), 2.03 (s, 3H, one of isomer), 2.28 (m, 5H), 3.54 (s, 1H), 3.58 (s, 2H), 3.64 (s, 1H), 3.67 and 3.69 (respectively is s, 3H, amide isomers), 3.85 (d, J=5.4Hz, 1H), 3.88 (s, 3H), 4.04 (t, J=9.3Hz, 1H), 5.27-5.34 (m, 1H), 6.51 (m, 1H), 6.76-6.89 (m, 2H), 6.94 (d, J=8.1Hz, 1H), 7.14 (m, 1H), 7.25 (m, 4H), 7.53 (d, J=8.3Hz, 1H), 7.96 (d, J=8.0Hz, 1H), 8.00-8.10 (m, 2H); MS (ESI) m/z590 (M ++ 1).
To 4-[(4S)-acetoxyl group-1-[4-[N '-(2-aminomethyl phenyl) urea groups] phenyl acetyl]-(2S)-and pyrrolidinyl] (520mg adds 0.25N sodium hydroxide (30ml) to methoxyl methyl benzoate in THF 0.882mmol) (30ml) solution.With this reactant mixture after stirring 2 days under the room temperature, with this mixture of ethyl acetate extraction.With 1N HCl acidify water layer, extract with chloroform-methanol (10/1).Extract with salt water washing merging.After dried over sodium sulfate, the vacuum concentration extract.Make the residue crystallization by adding chloroform, ethanol and ether, obtain 106 (68mg, 14%), be colourless powder.MW 533.57 mp 148-152 ℃ (decomposition); IR (KBr) 3356,2939,1687,1604,1533,1454,1255 cm -1 1H-NMR (DMSO-d 6) δ 1.95-2.09 (m, 2H), 2.25 (s, 3H), 3.59 (d, J=5.9Hz, 2H), 3.71 (m, 1H), 3.81 and 3.85 (respectively be s, 3H, amide isomers), 4.13-4.47 (m, 4H), 5.19 (br, 1H), 6.70-7.21 (m, 7H), 7.79 (d, J=7.9Hz, 1H), 7.86 (d, J=8.8Hz, 2H), 8.01 (d, J=8.3Hz, 1H), 8.47 (s, 1H), 8.57 (s, 1H); MS (ESI) m/z 533 (M ++ 1); C 29H 31N 3O 71H 2The analytical calculation value of O: C, 63.15; H, 6.03; N, 7.62.Measured value: C, 63.29; H, 5.76; N, 7.46.Embodiment 994-[1-[4-[N '-(2-chlorophenyl) urea groups]-3-methoxyphenyl acetyl group]-(4S)-hydroxyl-(2S)-pyrrolidinyl] methoxybenzoic acid
Figure A0080951003071
Under room temperature, with 4-[N '-(2-chlorophenyl) urea groups]-3-methoxybenzene guanidine-acetic acid (342mg, 1.02mmol), 4-[(2S, 4S)-and 4-acetoxyl group-2-pyrrolidinyl] methoxyl methyl benzoate (300mg, 1.02mmol), EDCHCl (293mg, 1.53mmol), HOBt (207mg, 1.53mmol) and triethylamine (0.71ml, 5.10mmol) mixture in DMF (6ml) stirred 15 hours.In this mixture impouring frozen water, use ethyl acetate extraction.Extract with frozen water and salt water washing merging.After dried over sodium sulfate, the vacuum concentration extract.Residue is through silica gel column chromatography [50g; chloroform/acetone (5/1)]; obtain 4-[(4S)-acetoxyl group-1-[4-[N '-(2-chlorophenyl) urea groups]-3-methoxyphenyl acetyl group]-(2S)-and pyrrolidinyl] methoxyl methyl benzoate (510mg, 82%), be the light brown amorphous solid. 1H-NMR (CDCl 3) δ 2.01 and 2.04 (respectively being s, 3H, amide isomers), 2.17 (m, 2H), 3.56-3.66 (m, 3H), 3.61 (s, 3H), 3.88 (s, 3H), 3.89 (m, 1H), 4.07 (t, J=9.6Hz, 1H), 4.45 (dd, J=9.2,3.4Hz, 1H), 4.56 (m, 1H), and 5.31-5.39 (m, 1H), 6.80-7.01 (m, 4H), 7.23 (d, J=8.1Hz, 4H), 7.34 (d, J=8.1Hz, 1H), 7.95 (d, J=8.5Hz, 1H), 8.00 (m, 1H), 8.18 (d, J=8.3Hz, 1H); MS (ESI) m/z610 (M ++ 1), 612 (M ++ 3).
To 4-[(4S)-acetoxyl group-1-[4-[N '-(2-chlorophenyl) urea groups]-3-methoxyphenyl acetyl group]-(2S)-and pyrrolidinyl] (510mg adds 0.25N sodium hydroxide (30ml) to methoxyl methyl benzoate in THF 0.836mmol) (30ml) solution.With this reactant mixture after stirring 2 days under the room temperature, with this mixture of ethyl acetate extraction.With 1N HCl acidify water layer, extract with chloroform-methanol (10/1).Extract with salt water washing merging.After dried over sodium sulfate, the vacuum concentration extract.Make the residue crystallization by adding ethanol and ether, obtain 107 (22mg, 5%), be colourless powder.MW 553.99 mp 138-142 ℃ (decomposition); IR (KBr) 3334,2939,1685,1604,1533,1439,1248 cm -1 1H-NMR (DMSO-d 6) δ 1.93-2.14 (m, 2H), 3.60 (d, J=5.7Hz, 2H), 3.71 (m, 1H), 3.81 and 3.85 (they respectively being s, 3H, amide isomers), 4.14-4.50 (m, 4H), 5.19 (br, 1H), 6.72 and 6.76 (respectively is m, 1H, amide isomers), 6.85 and 6.90 (they respectively being s, 1H, amide isomers), 7.00-7.08 (m, 3H), 7.28 (t, J=7.3Hz, 1H), 7.43 (dd, J=8.1,1.2Hz, 1H), and 7.86-7.95 (m, 2H), 7.97 (d, J=8.1Hz, 1H), 8.10 (dd, J=8.3,1.5Hz, 1H), 8.90 (s, 1H), 8.94 (s, 1H), 12.64 (br, 1H); MS (ESI) m/z554 (M ++ 1), 556 (M ++ 3); C 28H 28ClN 3O 7The analytical calculation value: C, 60.71; H, 5.05; Cl, 6.40; N, 7.58.Measured value: C, 60.47; H, 5.37; Cl, 6.31; N, 7.19.Embodiment 1004-[(4S)-and acetoxyl group-1-[4-[N '-(2-bromo phenyl) urea groups] phenyl acetyl]-(2S)-and pyrrolidinyl] methoxybenzoic acid
Under room temperature, with 4-[N '-(2-bromo phenyl) urea groups]-3-methoxybenzene guanidine-acetic acid (387mg, 1.02mmol), 4-[(4S)-acetoxyl group-(2S)-pyrrolidinyl] methoxyl methyl benzoate (300mg, 1.02mmol), EDCHCl (293mg, 1.53mmol), HOBt (207mg, 1.53mmol) and triethylamine (0.71ml, 5.10mmol) mixture in DMF (6ml) stirred 15 hours.In this mixture impouring frozen water, use ethyl acetate extraction.Extract with frozen water and salt water washing merging.After dried over sodium sulfate, the vacuum concentration extract.Residue obtains 4-[(4S through silica gel column chromatography [50g, chloroform/acetone (5/1)])-acetoxyl group-1-[4-[N '-(2-bromo phenyl) urea groups] phenyl acetyl]-(2S)-and pyrrolidinyl] methoxyl methyl benzoate (510mg, 76%), be yellow oil. 1H-NMR (CDCl 3) δ 2.01 and 2.04 (respectively being s, 3H, amide isomers), 2.31 (m, 2H), 3.54-3.68 (m, 3H), 3.76 (s, 2H), 3.88 (s, 3H), and 3.89-4.58 (m, 4H), 5.31-5.36 (m, 1H), 6.81-6.96 (m, 5H), 7.19-7.32 (m, 3H), 7.51 (d, J=8.0Hz, 1H), 7.93-8.00 (m, 3H), 8.13 (d, J=8.3Hz, 1H); MS (ESI) m/z654 (M ++ 1), 656 (M ++ 3).
To 4-[(4S)-acetoxyl group-1-[4-[N '-(2-bromo phenyl) urea groups] phenyl acetyl]-(2S)-and pyrrolidinyl] (510mg adds 0.25N sodium hydroxide (30ml) to methoxyl methyl benzoate in THF 0.779mmol) (30ml) solution.With this reactant mixture after stirring 2 days under the room temperature, with this mixture of ethyl acetate extraction.With 1N HCl acidify water layer, extract with chloroform-methanol (10/1).Extract with salt water washing merging.After dried over sodium sulfate, the vacuum concentration extract.Make the residue crystallization by adding ethanol and ether, obtain 108 (87mg, 19%), be light brown powder.MW 598.44 mp 143-151 ℃ (decomposition); IR (KBr) 3332,2937,1685,1604,1529,1529,1435 cm -1 1H-NMR (DMSO-d 6) δ 1.92-2.14 (m, 2H), 3.60 (d, J=5.9Hz, 2H), 3.72 (m, 1H), 3.81 and 3.85 (they respectively being s, 3H, amide isomers), 4.14-4.49 (m, 4H), 5.19 (br, 1H), 6.72 and 6.75 (respectively is m, 1H, amide isomers), 6.85 and 6.90 (they respectively being m, 1H, amide isomers), 6.97 (t, J=6.1Hz, 1H), 7.06 (d, J=8.8Hz, 2H), 7.32 (t, J=7.1Hz, 1H), 7.60 (dd, J=7.8,1.2Hz, 1H), 7.86 (d, J=8.8Hz, 2H), 7.87-7.97 (m, 3H), 8.74 (s, 1H), 8.93 (s, 1H), 12.60 (br, 1H); MS (ESI) m/z 559 (M ++ 1), 561 (M ++ 3); C 28H 28BrN 3O 70.1H 2The analytical calculation value of O: C, 56.03; H, 4.74; Br, 13.31; N, 7.00.Measured value: C, 55.80; H, 4.84; Br, 13.64; N, 6.66.Embodiment 1014-[1-[4-[N '-(2-chlorophenyl) urea groups]-3-methoxyphenyl acetyl group]-(4R)-hydroxyl-(2S)-the pyrrolidinyl methoxyl group] benzoic acid
Figure A0080951003101
To 4-[(4R)-acetoxyl group-1-(tert-butoxycarbonyl)-(2S)-pyrrolidinyl methoxyl group] essence of Niobe (835mg, 2.12mmol) the agitating solution of dichloromethane (5ml) in add TFA (5ml), this reactant mixture was stirred under room temperature 1 hour.This mixture of vacuum concentration, with saturated sodium bicarbonate make be alkalescence.With this mixture of chloroform extraction.Use the salt water washing, dry and evaporation obtains 4-[(4R through potassium carbonate)-acetoxyl group-(2S)-the pyrrolidinyl methoxyl group] essence of Niobe (580mg, 95%), be brown oil. 1H-NMR (CDCl 3) δ 1.86-1.93 (m, 1H), 2.00-2.12 (a plurality of s and m, 5H altogether), 3.03-3.29 (m, 1H), 3.73-3.80 (m, 1H), 3.88 (s, 3H), 3.93-4.01 (m, 2H), 5.27-5.30 (m, 1H), 6.91 (d, J=9.0Hz, 2H), 7.98 (d, J=9.0Hz, 2H); MS (FAB) m/z 294 (M ++ 1).
Under room temperature, with 4-[N '-(2-chlorophenyl) urea groups]-3-methoxybenzene guanidine-acetic acid (365mg, 1.09mmol), 4-[(4R)-acetoxyl group-(2S)-pyrrolidinyl methoxyl group] essence of Niobe (320mg, 1.09mmol), EDCHCl (250mg, 1.30mmol), HOBt (180mg, 1.33mmol) and triethylamine (182ml, 1.31mmol) mixture in THF (5ml) stirred 2 days.This mixture of dilute with water is also used ethyl acetate extraction.With salt water washing extract, also evaporation after dried over sodium sulfate.Residue is through purification by silica gel column chromatography; with chloroform-methanol (50: 1; v/v) as eluant; obtain 4-[(4R)-acetoxyl group-1-[4-[N '-(2-chlorophenyl) urea groups]-3-methoxyphenyl acetyl group]-(2S)-and the pyrrolidinyl methoxyl group] essence of Niobe (500mg; 75%), is the white foam thing. 1H-NMR(CDCl 3)δ2.01(s，3H)，2.03-2.05(m，1H)，2.20-2.26(m，1H)，2.37-2.43(m，1H)，3.59(s，2H)，3.62(s，3H)，3.66-3.87(m，2H)，3.89(s，3H)，4.07-4.09(m，1H)，4.48-4.51(m，1H)，4.59(m，1H)，6.70-6.82(m，4H)，6.97-7.01(m，1H)，7.24-7.35(m，4H)，7.92-7.98(m，3H)，8.12-8.21(m，1H)；MS(FAB)m/z610(M ++1)。
To 4-[(4R)-acetoxyl group-1-[4-[N '-(2-chlorophenyl) urea groups]-3-methoxyphenyl acetyl group]-(2S)-and the pyrrolidinyl methoxyl group] essence of Niobe (500mg; 0.82mmol) the agitating solution of THF (5ml) in add 0.5N sodium hydroxide (5ml), with this reactant mixture heated overnight under refluxing.After being cooled to room temperature, in this mixture impouring ice-1N HCl, the precipitation that produces is collected in decompression.Make the crude product solid through from chloroform-' recrystallization purifying obtains 109 (223mg, 49%) the E, is white crystalline powder.MW 553.99 mp 137-142 ℃; 1H-NMR (DMSO-d 6) δ 1.95-2.09 (m, 2H), 3.41-3.43 (m, 1H), 3.57 (m, 3H), 3.78 (s, 3H), 4.07-4.40 (a plurality of m, 4H altogether), 5.07 (m, 1H), 6.72-6.74 (m, 1H), 6.85 (m, 1H), 6.99-7.03 (m, 3H), 7.25-7.29 (m, 1H), 7.42-7.43 (m, 1H), 7.85-7.87 (m, 2H), 7.93-7.95 (m, 1H), 8.07-8.09 (m, 1H), 8.88 (s, 1H), 8.92 (s, 1H), 12.65 (br s, 1H); MS (FAB) m/z554 (M ++ 1); C 28H 28ClN 3O 71/2H 2The analytical calculation value of O: C, 59.73; H, 5.19; Cl, 6.30; N, 7.46.Measured value: C, 59.58; H, 5.32; Cl, 6.99; N, 7.21.Embodiment 1024-[(4R)-and hydroxyl-1-[3-methoxyl group-4-[N '-(2-aminomethyl phenyl) urea groups] phenyl acetyl]-(2S)-and the pyrrolidinyl methoxyl group] benzoic acid
Under room temperature, with 3-methoxyl group-4-[N '-(2-aminomethyl phenyl) urea groups] phenylacetic acid (320mg, 1.02mmol), 4-[(4R)-acetoxyl group-(2S)-pyrrolidinyl methoxyl group] essence of Niobe (300mg, 1.02Hmol), EDCHCl (235mg, 1.23mmol), HOBt (166mg, 1.23mmol) and triethylamine (171ml, 1.23mmol) mixture in THF (5ml) stirred 2 days.This mixture of dilute with water is also used ethyl acetate extraction.With salt water washing extract, also evaporation after dried over sodium sulfate.Residue is through purification by silica gel column chromatography; with chloroform-methanol (50: 1; v/v) as eluant; obtain 4-[(4R)-acetoxyl group-1-[3-methoxyl group-4-[N '-(2-aminomethyl phenyl) urea groups] phenyl acetyl]-(2S)-and the pyrrolidinyl methoxyl group] essence of Niobe (420mg; 70%), is the white foam thing. 1H-NMR (CDCl 3) δ 1.99 (s, 3H), 2.02-2.05 (m, 1H), 2.15-2.41 (a plurality of s and m, 5H altogether), 3.55 (s, 3H), 3.57 (s, 2H), 3.63-3.73 (m, 2H), 3.89 (s, 3H), 4.07-4.10 (m, 1H), 4.45-4.48 (m, 1H), 4.57 (m, 1H), 6.56 (s, 1H), 6.66 (m, 1H), 6.75-6.82 (m, 3H), 7.11-7.24 (m, 4H), 7.54-7.56 (m, 1H), and 7.92-7.94 (m, 2H), 8.05-8.07 (m, 1H); MS (FAB) m/z590 (M ++ 1).
To 4-[(4R)-acetoxyl group-1-[3-methoxyl group-4-[N '-(2-aminomethyl phenyl) urea groups] phenyl acetyl]-(2S)-and the pyrrolidinyl methoxyl group] essence of Niobe (420mg; 0.71mmol) add 0.5N sodium hydroxide (5ml) in the agitating solution in THF (5ml), with this reactant mixture heated overnight under refluxing.After being cooled to room temperature, in this mixture impouring ice-1N HCl, the precipitation that produces is collected in decompression.Make the crude product solid obtain 110 (182mg, 48%), be white crystalline powder through recrystallization purifying from chloroform-IPE.MW 533.57 mp 178-182 ℃; 1H-NMR (DMSO-d 6) δ 1.92-2.10 (m, 2H), 2.23 (s, 3H), 3.40-3.44 (m, 1H), 3.56-3.67 (m, 3H), 3.78 (s, 3H), (4.05-4.39 a plurality of m, 4H altogether), 5.06 (m, 1H), 6.71-7.01 (m, 5H), and 7.10-7.16 (m, 2H), 7.77-7.79 (m, 1H), 7.85-7.89 (m, 2H), 7.98-8.00 (m, 1H), 8.45 (s, 1H), 8.54 (s, 1H), 12.59 (br s, 1H); MS (FAB) m/z534 (M ++ 1); C 29H 31N 3O 71/2H 2The analytical calculation value of O: C, 64.20; H, 5.94; N, 7.74.Measured value: C, 64.35; H, 5.83; N, 7.68.Embodiment 1034-[(4S)-and fluoro-1-[3-methoxyl group-4-[N '-(2-aminomethyl phenyl) urea groups] phenyl acetyl]-2-pyrrolidinyl methyl]-1-piperazinyl acetic acid
To N-(tert-butoxycarbonyl) (4S)-fluoro dried meat ammonia alcohol (1.26g, 5.75mmol), triethylamine (4ml, 28.5mmol) and DMSO (4.1ml adds SO in the agitating solution of dichloromethane 57.5mmol) (20ml) 3Pyridine (2.74g, 17.2mmol).Stir after 5 hours, evaporate this mixture to remove dichloromethane, with ether (200ml) dilution.Wash this solution with 1N HCl (200ml) and saline (200ml), through dried over mgso and evaporation.The residue that obtains is through silica gel column chromatography, with hexane-ethyl acetate (4: 1) eluting, obtain N-(tert-butoxycarbonyl) (4S)-fluoro dried meat ammonium aldehyde (628mg, 50%), be yellow oil. 1H-NMR (CDCl 3) δ 1.41-1.47 (m, 9H), 2.02-2.48 (m, 2H), 3.47-3.94 (m, 2H), 4.16 and 4.29 (they respectively being d, J=9.8Hz, 1H altogether), 5.13 and 5.26 (they respectively are s, are total to 1H).
To N-(tert-butoxycarbonyl) (4S)-fluoro dried meat ammonium aldehyde (1.44g, 6.63mmol), 1-piperazinyl ethyl acetate (1.71g, 9.94mmol) and acetic acid (759 μ l add NaBH in the agitating solution of methanol 13.3mmol) (20ml) 3CN (880mg, 13.3mmol).This reactant mixture stirred spend the night and evaporate.With saturated sodium bicarbonate (100ml) quencher residue, with chloroform (2 * 200ml) evaporations.The extract that merges through dried over mgso also evaporates.The oiliness residue with chloroform-methanol (20: 1) eluting, obtains 4-[1-(tert-butoxycarbonyl)-(4S)-fluoro-2-pyrrolidinyl methyl through silica gel column chromatography]-1-piperazinyl ethyl acetate (2.38g, 95%), be yellow oil.
With 4-[1-(tert-butoxycarbonyl)-(4S)-fluoro-2-pyrrolidinyl methyl]-1-piperazinyl ethyl acetate (2.38g, 6.37mmol), the mixture of TFA (5ml) and dichloromethane (5ml) stirred 3 hours.Evaporate this mixture, make residue be alkalescence with saturated sodium bicarbonate (100ml).With chloroform-methanol (4: 1,2 * 150ml) extract this mixture, through dry extract that merges of potassium carbonate and evaporation, obtain 4-[(4S)-fluoro-2-pyrrolidinyl methyl]-1-piperazinyl ethyl acetate (1.44g, 83%), be brown oil. 1H-NMR (CDCl 3) δ 1.27 (dt, J=7.1,2.0Hz, 3H), 1.66-3.35 (a plurality of m, 17H), 4.18 (dq, J=7.1,2.0Hz, 2H), 5.09 and 5.22 (they respectively are m, are total to 1H).
With 4-[(4S)-fluoro-2-pyrrolidinyl methyl]-1-piperazinyl ethyl acetate (1.44g, 5.27mmol), 3-methoxyl group-4-[N '-(2-aminomethyl phenyl) urea groups] phenylacetic acid (1.66g, 5.27mmol), EDCHCl (1.52g, 7.91mmol), HOBt (cat.) and DMAP (cat.) mixture in DMF (10ml) stirs and spend the night.With ethyl acetate-methanol (10: 1,220ml) dilute this mixture.(200ml) washs this solution with saline, through dried over mgso and evaporation.Residue is through silica gel column chromatography; with chloroform-methanol (20: 1) as eluant; obtain 4-[(4S)-fluoro-1-[3-methoxyl group-4-[N '-(2-aminomethyl phenyl) urea groups] phenyl acetyl]-2-pyrrolidinyl methyl]-1-piperazinyl ethyl acetate (2.47g, 82%), be yellow viscous solid. 1H-NMR (CDCl 3) δ 1.24-1.29 (m, 3H), 1.92-4.36 (a plurality of m, 7H), 5.16 and 5.29 (they respectively are m, are total to 1H), 6.43 (s, 1H), 6.74-6.81 (m, 2H), 7.12-7.29 (m, 4H), 7.50 (d, J=7.8Hz, 1H), 8.01-8.07 (m, 1H).
With 4-[(4S)-fluoro-1-[3-methoxyl group-4-[N '-(2-aminomethyl phenyl) urea groups] phenyl acetyl]-2-pyrrolidinyl methyl]-1-piperazinyl ethyl acetate (1.0g; 1.76mmol) and the 0.25N sodium hydroxide (14ml, 3.50mmol) mixture in THF (15ml) stirs and to spend the night.With 1N HCl neutralize this mixture and the evaporation.(DIAION, HP20) purification as eluant, obtain 111 MW 541.61 (400mg, 40%) with water-methanol to residue, are faint yellow amorphous solid through ion exchange resin. 1H-NMR (CD 3OD) δ 2.00-3.95 (a plurality of m, 24H), 4.34-4.40 (m, 1H), 5.23 and 5.36 (respectively are m, altogether 1H), 6.78-6.82 (m, 1H), 6.92 (m, 1H), 7.00-7.04 (m, 1H), 7.09-7.23 (m, 4H), 7.59 (d, J=7.1Hz, 1H), 7.99-8.02 (m, 1H); MS (FAB) m/z542 (M ++ 1).Embodiment 1044-[1-[4-[N '-(2-chlorophenyl) urea groups]-3-methoxyphenyl acetyl group]-4,4-two fluoro-2-pyrrolidinyl methyl]-1-piperazinyl acetic acid
Figure A0080951003141
To 1-(tert-butoxycarbonyl)-4,4-two fluoro-2-pyrrolidinyl methanol (2.11g, 8.89mmol), triethylamine (6.2ml, 44.5mmol), (6.3ml adds SO in the agitating solution of dichloromethane 88.9mmol) (20ml) to DMSO 3Pyridine (4.25g, 26.7mmol).Stir after 3 hours, this mixture of vacuum concentration is with ether (200ml) dilution.Wash the mixture that obtains with 1N HCl (100ml) and saline (100ml), through dried over mgso and vacuum concentration.Residue with hexane-ethyl acetate (4: 1) eluting, obtains 1-(tert-butoxycarbonyl)-4 through silica gel column chromatography, and 4-two fluoro-2-pyrrolidine formaldehyde (1.40g, 67%) are yellow oil. 1H-NMR (CDCl 3) δ 1.45-1.52 (m, 9H), 2.49 (m, 2H), 3.75-3.88 (m, 2H), 4.29-4.42 (m, 1H), 9.54 and 9.60 (they respectively are s, are total to 1H).
In 0 ℃, to 1-(tert-butoxycarbonyl)-4,4-two fluoro-2-pyrrolidine formaldehyde (1.40g, 5.95mmol) and 1-piperazinyl ethyl acetate (1.02g, methanol-acetic acid 5.95mmol) (12: 1, add NaBH in agitating solution 13ml) 3CN (787mg, 11.9mmol).Stir after 3 days, by adding this mixture of saturated sodium bicarbonate (100ml) quencher, (2 * 200ml) extract with chloroform.Extract and vacuum concentration through the dried over mgso merging.Residue as eluant, obtains 4-[1-(tert-butoxycarbonyl)-4,4-two fluoro-2-pyrrolidinyl methyl with chloroform-methanol (20: 1) through silica gel column chromatography]-1-piperazinyl ethyl acetate (822mg, 35%), be yellow oil. 1H-NMR (CDCl 3) δ 1.27 (t, J=7.1Hz, 3H), 1.46 (m, 9H), 1.64 (m, 2H), 2.39-2.64 (m, 10H), 3.19 (s, 2H), 3.42-4.05 (a plurality of m, 3H), 4.18 (q, J=7.1Hz, 2H).
With 4-[1-(tert-butoxycarbonyl)-4,4-two fluoro-2-pyrrolidinyl methyl]-1-piperazinyl ethyl acetate (820mg, 2.09mmol) and the solution stirring in the dichloromethane (5ml) of TFA (5ml) 1 hour.This mixture of vacuum concentration makes residue be alkalescence with saturated sodium bicarbonate.(5: 1,2 * 200ml) extracted the mixture that obtains with chloroform-methanol.Through dry extract and the vacuum concentration that merges of potassium carbonate, obtain 4-(4,4-two fluoro-2-pyrrolidinyl methyl)-1-piperazinyl ethyl acetate (493mg, 81%), be brown oil. 1H-NMR(CDCl 3)δ1.27(t，J＝7.1Hz，3H)，1.91(m，2H)，2.27-2.60(m，10H)，3.09-3.34(m，4H)，3.46-3.53(m，1H)，4.19(q，J＝7.1Hz，2H)。
With 4-(4,4-two fluoro-2-pyrrolidinyl methyl)-1-piperazinyl ethyl acetate (490mg, 1.69mmol), 4-[N '-(2-chlorophenyl) urea groups]-3-methoxybenzene guanidine-acetic acid (567mg, 1.69mmol), EDCHCl (486mg, 2.54mmol), HOBt (cat.) and DMAP (cat.) mixture in DMF (10ml) stirs and spend the night.(250ml) dilutes this mixture with ethyl acetate, and (2 * 200ml) washings are through dried over mgso and vacuum concentration with saline.Residue is through silica gel column chromatography; with chloroform-ethyl acetate (4: 1) to chloroform-methanol (10: 1) as eluant; obtain 4-[1-[4-[N '-(2-chlorophenyl) urea groups]-3-methoxyphenyl acetyl group]-4; 4-two fluoro-2-pyrrolidinyl methyl]-1-piperazinyl ethyl acetate (973mg; 95%), is yellow viscous oil shape thing. 1H-NMR (CDCl 3) δ 1.25 (t, J=7.1Hz, 3H), 2.31-2.68 (m, 12H), 3.17-3.20 (m, 2H), 3.52-3.91 (m, 4H), 4.10-4.48 (a plurality of m, 3H), and 6.75-6.84 (m, 2H), 7.00 (dt, J=7.8,1.5Hz, 1H), and 7.16-7.29 (m, 3H), 7.35 (dd, J=8.3,1.5Hz, 1H), 8.00 (d, J=8.3Hz, 1H), 8.18 (dd, J=8.3,1.5Hz, 1H).
To 4-[1-[4-[N '-(2-chlorophenyl) urea groups]-3-methoxyphenyl acetyl group]-4; 4-two fluoro-2-pyrrolidinyl methyl]-1-piperazinyl ethyl acetate (292mg; 0.480mmol) the agitating solution of THF (4ml) in add the 0.25N sodium hydroxide (3.8ml, 0.960mmol).Stir after 2 days, neutralizing with 1N HCl, also (4: 1,2 * 200ml) extracted this mixture, through the extract and the vacuum concentration of dried over mgso merging with chloroform-methanol.Residue with chloroform-methanol (5: 1) eluting, obtains 112 MW 580.02 (81.7mg, 29%) through purification by silica gel column chromatography, is faint yellow amorphous solid.MW 580.02 1H-NMR (DMSO-d 6) δ 2.24-2.50 (a plurality of m, 12H), 3.40-4.47 (a plurality of m, 10H), 6.76 (d, J=8.1Hz, 1H), 6.88 (s, 1H), 7.02 (t, J=8.1Hz, 1H), 7.28 (t, J=8.1Hz, 1H), 7.44 (d, J=8.1Hz, 1H), 7.97 (d, J=8.1Hz, 1H), 8.08 (d, J=8.1Hz, 1H), 8.96-8.99 (m, 2H); MS (FAB) m/z580 (M ++ 1) embodiment 1054-[1-[3-methoxyl group-4-[N '-(2-aminomethyl phenyl) urea groups] phenyl acetyl]-(4S)-phenoxy group-(2S)-pyrrolidinyl] methyl isophthalic acid-piperazinyl acetic acid
Figure A0080951003161
Under room temperature, blanket of nitrogen, to (2S, 4R)-1-tert-butoxycarbonyl-4-hydroxyl-2-pyrrolidinyl methyl formate (4.69g, 19.1mmol), phenol (1.98g, 21.0mmol) and triphenyl phasphine (5.51g, 21.0mmol) add in the stirred mixture in THF (80ml) DIAD (4.13ml, 21.0mmol).This mixture stirring is spent the night.Remove desolvate after, the residue that obtains is through silica gel column chromatography purification [700g, chloroform/ethyl acetate (10/1)], obtains that (2S 4S)-1-tert-butoxycarbonyl-4-phenoxy group-2-pyrrolidinyl methyl formate (5.31g, 86%), is colorless oil. 1H-NMR (CDCl 3) δ 1.43 (br, 9H, one of isomer), 1.48 (br, 9H, one of isomers), 2.48 (m, 1H), 3.75 (br, 3H), 4.42-4.96 (m, 2H), 6.88-7.35 (m, 5H).
Under room temperature, to (2S, 4S)-1-tert-butoxycarbonyl-4-phenoxy group-2-pyrrolidinyl methyl formate (5.31g, and adding 0.25N sodium hydroxide in the agitating solution of THF 16.5mmol) (132ml) (132ml, 33.0mmol).The mixture stirring that obtains is spent the night.Except that after desolvating,, use chloroform extraction by adding this mixture of 1N HCl acidify.Extract with salt water washing merging.Through dried over sodium sulfate and evaporation.Make residue recrystallization in normal hexane-chloroform, obtain (2S, 4S)-1-tert-butoxycarbonyl-4-phenoxy group-2-pyrrolidinyl formic acid (2.96g, 58%), be white powder. 1H-NMR(DMSO-d 6)δ1.36(s，9H)，2.16(d，J＝13.2Hz，1H)，2.56(m，1H)，3.46(m，1H)，3.71(dt，J＝12.0，5.4Hz，1H)，4.26(dt，J＝9.5，7.1Hz，1H)，4.99(m，1H)，6.85(m，2H)，6.94(t，J＝7.3Hz，1H)，7.28(t，J＝7.3Hz，1H)。
In 0 ℃, to (2S, 4S)-(2.39g adds BH in the agitating solution of THF 7.76mmol) (50ml) to 1-tert-butoxycarbonyl-4-phenoxy group-2-pyrrolidinyl formic acid 3DMS (1.55ml, 15.5mmol).After 10 minutes, stir down, make this mixture rise to room temperature, then in 50 ℃ of heating 2 hours in uniform temp.After being cooled to room temperature, this mixture of vacuum concentration, in 0 ℃ by adding the entry quencher.With this mixture of ethyl acetate extraction.Extract with the salt water washing merges also evaporates through dried over sodium sulfate.Residue is through silica gel column chromatography purification [60g, chloroform/methanol (50/1)], obtain (2S, 4S)-1-tert-butoxycarbonyl-4-phenoxy group-2-pyrrolidinyl methanol (2.83g, 100%), be colorless oil. 1H-NMR(CDCl 3)δ1.47(s，9H)，1.95(br，1H)，2.36(m，1H)，3.56-3.74(m，3H)，3.89-4.52(m，3H)，4.85(br，1H)，6.84(dd，J＝8.8，1.2Hz，2H)，6.97(t，J＝7.2Hz，1H)，7.29(t，2H，J＝7.8Hz)。
In 0 ℃, to (2S, 4S)-1-tert-butoxycarbonyl-4-phenoxy group-2-pyrrolidinyl methanol (2.75g, 9.37mmol), triethylamine (7.84ml, 56.2mmol), (6.66ml 9.37mmol) adds SO to DMSO in the stirring the mixture in dichloromethane (30ml) 3(4.47g 28.1mmol), makes the mixture that obtains be warming up to room temperature to pyridine then.Stir after 2.5 hours this mixture of vacuum concentration.In the mixture that obtains, add entry, use ether extraction.With the extract of salt water washing merging, through dried over sodium sulfate and vacuum concentration.Residue is through silica gel column chromatography [100g, chloroform/acetone (5/1)], obtain (2S, 4S)-1-tert-butoxycarbonyl-4-phenoxy group-2-pyrrolidine formaldehyde (2.54g, 93%), be yellow oil. 1H-NMR (CDCl 3) δ 1.45 (s, 9H, one of isomer), 1.49 (s, 9H, one of isomers), 2.17 (br, 2H), 3.65-4.31 (m, 3H), 4.91 (br, 1H), 6.79 (d, J=7.8Hz, 1H), 6.77 (m, 1H), 7.28 (m, 2H), 9.66 (m, 1H).
Under room temperature, to 1-tert-butoxycarbonyl-(4S)-phenoxy group-(2S)-pyrrolidine formaldehyde (1.36g, 4.67mmol), (1.61g adds NaBH (OAc) in the stirring the mixture of THF 9.37mmol) (30ml) to 1-piperazinyl ethyl acetate 3(1.98g, 9.34mmol).Stir after 3 hours,, use ethyl acetate extraction by adding this mixture of entry quencher.Extract with sodium bicarbonate aqueous solution and salt water washing merging.Through dried over sodium sulfate organic layer and vacuum concentration.Residue is through silica gel column chromatography [50g, chloroform/methanol (10/1)], obtains 4-[1-tert-butoxycarbonyl-(4S)-phenoxy group-(2S)-pyrrolidinyl] methyl isophthalic acid-piperazinyl ethyl acetate (1.05g, 50%), be colorless oil. 1H-NMR(CDCl 3)δ1.27(t，J＝7.3Hz，3H)，1.57(s，9H)，2.1?8(m，1H)，2.33-2.74(m，9H)，3.17(s，2H)，3.52-4.10(m，5H)，4.17(q，J＝7.3Hz，2H)，4.89(br，1H)，6.84(d，J＝6.8Hz，2H)，6.95(m，1H)，7.26(m，3H)。
Under room temperature, to 4-[1-tert-butoxycarbonyl-(4S)-phenoxy group-(2S)-pyrrolidinyl] (1.05g adds TFA (20ml) to methyl isophthalic acid-piperazinyl ethyl acetate in the agitating solution of dichloromethane 2.35mmol) (20ml).Stir after 3 hours, this mixture of vacuum concentration dilutes this mixture with chloroform-methanol (10/1), makes by adding 1N sodium hydroxide to be alkalescence.Extract the reactant mixture that merges with chloroform-methanol (10/1).With salt water washing organic layer, dry and concentrate through sodium carbonate, obtain 44 (4S)-phenoxy groups-(2S)-pyrrolidinyl] (1.12g quantitatively), is brown oil to methyl isophthalic acid-piperazinyl ethyl acetate, and it can use without being further purified. 1H-NMR(CDCl 3)δ1.25(tt，J＝7.1，7.1Hz，3H)，1.91(d，J＝12.0Hz，1H)2.42-2.85(m，10H)，3.22(s，2H)，3.50(s，2H)，3.54-3.82(m，2H)，4.15(q，J＝7.1Hz，2H)，4.98(br，1H)，6.84(d，J＝8.1Hz，2H)，6.76(t，J＝7.1Hz，1H)，7.26-7.31(m，3H)，7.40(br，1H)。
Under room temperature, with 4-[N '-(2-aminomethyl phenyl) urea groups]-3-methoxybenzene guanidine-acetic acid (337mg, 1.07mmol), 4-[(4S)-phenoxy group-(2S)-pyrrolidinyl] methyl isophthalic acid-piperazinyl ethyl acetate (373mg, 1.07mmol), EDCHCl (308mg, 1.61mmol) and DMAP (197mg, 1.61mmol) mixture in DMF (6ml) stirred 18 hours.In this mixture impouring frozen water, use ethyl acetate extraction.Extract with frozen water and salt water washing merging.After dried over sodium sulfate, the vacuum concentration extract.Residue is through silica gel column chromatography [50g; chloroform/methanol (50/1)]; obtain 4-[1-[3-methoxyl group-4-[N '-(2-aminomethyl phenyl) urea groups] phenyl acetyl]-(4S)-phenoxy group-(2S)-pyrrolidinyl] methyl isophthalic acid-piperazinyl ethyl acetate (430mg, 62%), be colourless amorphous solid. 1H-NMR(CDCl 3)δ1.25(t，J＝7.8Hz，3H)，1.98-2.17(m，2H)，2.26(s，3H)，2.36-2.82(m，11H)，3.13(s，1H)，3.17(s，1H)，3.55(d，J＝2.4Hz，1H)，3.66(d，J＝0.9Hz，3H)，3.67-3.84(m，2H)，3.98-4.35(m，3H)，4.85-4.95(m，1H)，6.27-6.89(m，6H)，7.08(t，J＝7.3Hz，1H)，7.19(d，J＝6.8Hz，1H)，7.28(m，2H)，7.41(d，J＝4.9Hz，1H)，7.55(d，J＝7.8Hz，1H)，8.05(dd，J＝7.3，2.2Hz，1H)。For hydrochlorate: be the light brown amorphous solid.IR (KBr) 3265,3059,1747,1533,1225 cm -1MS (FAB) m/z 644 (M ++ 1); C 36H 45N 5O 6HCl2.1H 2The analytical calculation value of O: C, 60.22; H, 7.05; N, 9.75.Measured value: C, 59.97; H, 6.72; N, 9.54.
To 4-[1-[3-methoxyl group-4-[N '-(2-aminomethyl phenyl) urea groups] phenyl acetyl]-(4S)-phenoxy group-(2S)-pyrrolidinyl] (240mg adds 0.25N sodium hydroxide (3.0ml) to methyl isophthalic acid-piperazinyl ethyl acetate in THF 0.373mmol) (3.0ml) solution.After stirring 20 hours under the room temperature, neutralize this mixture also with chloroform-methanol (10/1) extraction, through the extract and the vacuum concentration of dried over sodium sulfate merging with 1N HCl.Grind residue by adding ether, obtain 113 MW 615.72 (143mg, 62%), be white powder.IR (KBr) 3346,2949,1633,1533,1227 cm -1 1H-NMR (DMSO-d 6) δ 1.76 (m, 1H), 2.18 (br, 2H), 2.25 (s, 3H), 2.42-2.83 (m, 9H), 3.17 (s, 1H), 3.20 (s, 1H), 3.38 (m, 1H), 3.70 (s, 2H), and 3.72-3.78 (m, 2H), 3.85 (s, 3H, one of isomer), 3.87 (s, 3H, one of isomers), 3.95 (m, 1H), 4.27 (br, 1H), 5.08 (m, 1H), 6.75 (d, J=8.3Hz, 1H), 6.93 (m, 5H), 7.14 (m, 2H), 7.30 (d, J=7.6Hz, 1H), 7.79 (d, J=9.0Hz, 1H), 8.02 (m, 1H), 8.50 (s, 1H), 8.58 (s, 1H); MS (FAB) m/z 616 (M ++ 1).C 34H 41N 5O 60.1EtOH2H 2The analytical calculation value of O: C, 62.58; H, 7.00; N, 10.67.Measured value: C, 62.73; H, 6.58; N, 10.24.Embodiment 1064-[1-[4-[NN '-(2-chlorophenyl) urea groups]-3-methoxyphenyl acetyl group]-(4S)-phenoxy group-(2S)-pyrrolidinyl] methyl isophthalic acid-piperazinyl acetic acid
Figure A0080951003201
Under room temperature, with 4-[N '-(2-chlorophenyl) urea groups]-3-methoxybenzene guanidine-acetic acid (358mg, 1.07mmol), 4-[(4S)-phenoxy group-(2S)-pyrrolidinyl methyl]-1-piperazinyl ethyl acetate (373mg, 1.07mmol), EDCHCl (308mg, 1.61mmol) and DMAP (197mg, 1.61mmol) mixture in DMF (6ml) stirred 18 hours.In this mixture impouring frozen water, use ethyl acetate extraction.Extract with frozen water and salt water washing merging.After dried over sodium sulfate, the vacuum concentration extract.Residue is through silica gel column chromatography [50g; chloroform/methanol (50/1)]; obtain 4-[1-[4-[N '-(2-chlorophenyl) urea groups]-3-methoxyphenyl acetyl group]-(4S)-phenoxy group-(2S)-pyrrolidinyl] methyl isophthalic acid-piperazinyl ethyl acetate (320mg, 45%), be colourless amorphous solid. 1H-NMR (CDCl 3) δ 1.25 (qq, J=7.3,7.3Hz, 3H), 2.05-2.20 (m, 2H), 2.35-2.80 (m, 1H), 3.12 and 3.18 (they respectively being s, 1H, amide isomers), 3.57 (d, J=5.4Hz, 1H), 3.66 and 3.38 (they respectively being s, 3H, amide isomers), 3.70-3.90 (m, 2H), and 4.02-4.43 (m, 3H), 4.88-4.97 (m, 1H), 6.83-6.99 (m, 6H), 7.18-7.32 (m, 3H), 7.68 (d, J=8.3Hz, 1H), 7.74 (s, 1H), 8.16 (dd, J=8.3,1.4Hz, 1H).For hydrochlorate: be the light brown amorphous solid.IR (KBr) 3300,2978,1745,1533,1225cm -1MS (FAB) m/z 664 (M ++ 1), 666 (M ++ 3); C 35H 42ClN 5O 6HCl2.4H 2The analytical calculation value of O: C, 56.51; H, 6.48; N, 9.41.Measured value: C, 56.51; H, 6.18; N, 9.28.
To 4-[1-[4-[N '-(2-chlorophenyl) urea groups]-3-methoxyphenyl acetyl group]-(4S)-phenoxy group-(2S)-pyrrolidinyl] (181mg adds 0.25N sodium hydroxide (2.2ml) to methyl isophthalic acid-piperazinyl ethyl acetate in THF 0.273mmol) (2.2ml) solution.After stirring 20 hours under the room temperature, neutralize this mixture also with chloroform-methanol (10/1) extraction with 1N HCl.Extract and vacuum concentration through the dried over sodium sulfate merging.Grind residue by adding ether, obtain 114 (133mg, 77%), be white powder.MW 636.14 IR (KBr) 3317,2949,1701,1631,1595,1225 cm -1 1H-NMR (DMSO-d 6) δ 2.13-3.05 (m, 11H), 3.22 and 3.36 (they respectively being s, 2H, amide isomers), 3.38 (m, 1H), 3.60 (s, 2H), 3.71 (m, 1H), 3.85 (s, 3H), 3.95 (m, 1H), 4.28 (br, 1H), 5.06 (m, 1H), 6.76 (d, J=8.3Hz, 1H), 6.91-7.03 (m, 5H), 7.29 (m, 3H), 7.44 (d, J=7.9Hz, 1H), 7.97 (dd, J=8.1,4.1Hz, 1H), 8.08 (d, J=7.0Hz, 1H), 8.91 (s, 1H), 8.95 (s, 1H); MS (FAB) m/z636 (M ++ 1), 638 (M ++ 3).C 33H 38ClN 5O 60.2EtOH1.3H 2The analytical calculation value of O: C, 59.98; H, 6.30; N, 10.47.Measured value: C, 60.25; H, 6.12; N, 10.11.Embodiment 1074-[1-[4-[N '-(2-bromo phenyl) urea groups]-3-methoxyphenyl acetyl group]-(4S)-phenoxy group-(2S)-pyrrolidinyl] methyl isophthalic acid-piperazinyl acetic acid
Figure A0080951003211
Under room temperature, with 4-[N '-(2-bromo phenyl) urea groups]-3-methoxybenzene guanidine-acetic acid (406mg, 1.07mmol), 4-[(4S)-phenoxy group-(2S)-pyrrolidinyl] methyl isophthalic acid-piperazinyl ethyl acetate (373mg, 1.07mmol), EDCHCl (308mg, 1.61mmol) and DMAP (197mg, 1.61mmol) mixture in DMF (6ml) stirred 18 hours.In this mixture impouring frozen water, use ethyl acetate extraction.Extract with frozen water and salt water washing merging.After dried over sodium sulfate, the vacuum concentration extract.Residue is through silica gel column chromatography [50g; chloroform/methanol (50/1)]; obtain 4-[1-[4-[N '-(2-bromo phenyl) urea groups]-3-methoxyphenyl acetyl group]-(4S)-phenoxy group-(2S)-pyrrolidinyl] methyl isophthalic acid-piperazinyl ethyl acetate (560mg, 74%), be colourless amorphous solid. 1H-NMR (CDCl 3) δ 1.25 (tt, J=7.1,7.1Hz, 3H), 2.04-2.84 (m, 13H), 3.12 and 3.18 (respectively be s, 1H, amide isomers), 3.57 (d, J=4.4Hz, 1H), 3.66 with 3.68 (they respectively being s, 3H, amide isomers), 3.68-3.87 (m, 3H), and 4.05-4.41 (m, 2H), 4.87-4.96 (m, 1H), and 6.76-6.98 (m, 6H), 7.20-7.33 (m, 3H), 7.46 (d, J=8.1Hz, 1H), 7.67 with 7.71 (respectively being s, 1H, amide isomers), 7.78 and 7.81 (respectively is s, 1H, amide isomers), 7.95 (d, J=8.3Hz, 1H), 8.09 (d, J=8.1Hz, 1H).For hydrochlorate: be the light brown amorphous solid.IR (KBr) 3384,2978,1745,1340,1120 cm -1MS (FAB) m/z 708 (M ++ 1), 710 (M ++ 3); C 35H 42BrN 5O 6HCl2.5H 2The analytical calculation value of O: C, 53.20; H, 6.12; N, 8.86.Measured value: C, 52.98; H, 5.79; N, 8.66.
To 4-[1-[4-[N '-(2-bromo phenyl) urea groups]-3-methoxyphenyl acetyl group]-(4S)-phenoxy group-(2S)-pyrrolidinyl] (330mg adds 0.25N sodium hydroxide (3.7ml) to methyl isophthalic acid-piperazinyl ethyl acetate in THF 0.466mmol) (3.7ml) solution.After stirring 20 hours under the room temperature, neutralize this mixture also with chloroform-methanol (10/1) extraction with 1N HCl.Extract and vacuum concentration through the dried over sodium sulfate merging.Grind residue by adding ether, obtain 115 (217mg, 68%), be white powder.MW 680.59 IR (KBr) 3315,3095,2941,1631,1529,1435 cm -1 1H-NMR (DMSO-d 6) δ 1.75 (m, 1H), 2.18 and 2.23 (they respectively being s, 2H, amide isomers), 2.30-2.78 (m, 9H), 3.15 (2,2H), 3.47 (m, 1H), 3.58 (s, 2H), 3.60-3.82 (m, 3H), 3.84 (m, 3H), 3.93 (m, 1H), 4.26 (br, 1H), 5.08 (m, 1H), 6.75 (d, J=7.8Hz, 1H), 6.98 (m, 5H), 7.30 (m, 3H), 7.60 (dd, J=8.1,2.2Hz, 1H), 7.94 (m, 2H), 8.75 (s, 1H), 8.93 (s, 1H); MS (FAB) m/z680 (M ++ 1), 82 (M ++ 3).C 33H 38BrN 5O 60.2EtOH2H 2The analytical calculation value of O: C, 55.27; H, 6.00; N, 9.65.Measured value: C, 55.32; H, 5.56; N, 9.25.Embodiment 1084-[(4S)-and (4-carboxyl phenoxy group)-1-[3-methoxyl group-4-[N '-(2-aminomethyl phenyl) urea groups] phenyl acetyl]-(2S)-and the pyrrolidinyl methoxyl group] benzoic acid
Figure A0080951003221
To 1-(tert-butoxycarbonyl)-(4R)-hydroxyl-(2S)-pyrrolidinyl methyl formate (10.4g, 0.04mol) and imidazoles (8.66g, 0.13mol) the agitating solution of DMF (40m) in add TBS-Cl (7.03g 0.05mol), stir this reactant mixture 3 hours in 60 ℃.After being cooled to room temperature, dilute this mixture, use ether extraction with saline.With salt water washing extract, through dried over sodium sulfate and evaporation.Residue is through purification by silica gel column chromatography, with normal hexane-ethyl acetate (5: 1, v/v) as eluant, obtain 1-(tert-butoxycarbonyl)-(4R)-(t-butyldimethylsilyloxy base)-(2S)-pyrrolidinyl methyl formate (15.0g, 98%), is colorless oil. 1H-NMR (CDCl 3) δ 0.06 (s, 6H), 0.87 (s, 9H), 1.41 and 1.46 (respectively be s, 9H), 1.99-2.03 (m, 1H), 2.16-2.18 (m, 1H), 3.31-3.42 (m, 1H), 3.56-3.63 (m, 1H), 3.73 and 3.74 (respectively be s, 3H), 4.31-4.42 (m, 2H); MS (ESI) m/z360 (M ++ 1).
To 1-(tert-butoxycarbonyl)-(4R)-(t-butyldimethylsilyloxy base)-(2S)-pyrrolidinyl methyl formate (15.0g, 0.04mol) the agitating solution of THF (60ml) in add 1N sodium hydroxide (60ml), in 60 ℃ reactant mixture was stirred 2 hours.After being cooled to room temperature, this mixture is concentrated into small size,, uses ethyl acetate extraction with 1N HCl acidify.With salt water washing extract, through dried over sodium sulfate and evaporation, obtain 1-(tert-butoxycarbonyl)-(4R)-(t-butyldimethylsilyloxy base)-(2S)-pyrrolidinyl formic acid (12.8g, 89%), be colorless oil. 1H-NMR (CDCl 3) δ 0.07 and 0.08 (respectively be s, 6H), 0.87 (s, 9H), 1.49 (s, 9H), 2.06-2.11 (m, 1H), 2.41-244 (m, 1H), 3.40-3.59 (m, 2H), 4.36-4.50 (m, 2H); MS (ESI) m/z346 (M ++ 1).
(12.8g is added dropwise to BH in the agitating solution of THF 0.04mol) (150ml) cold (0 ℃) to 1-(tert-butoxycarbonyl)-(4R)-(t-butyldimethylsilyloxy base)-(2S)-pyrrolidinyl formic acid 3DMS (5.30ml.0.06mol) stirs this reactant mixture and to spend the night under room temperature.By adding saturated ammonium chloride quencher reactant mixture, use ethyl acetate extraction.With salt water washing extract, through dried over sodium sulfate and evaporation.Residue is through purification by silica gel column chromatography, with toluene-acetone (5: 1, v/v) as eluant, obtain 1-(tert-butoxycarbonyl)-(4R)-(t-butyldimethylsilyloxy base)-(2S)-dried meat ammonia alcohol (10.5g, 85%), be colorless oil. 1H-NMR(CDCl 3)δ0.06(s，6H)，0.87(s，9H)，1.47(s，9H)，1.58-1.63(m，1H)，1.93-1.98(m，1H)，3.32-344(m，2H)，3.51-3.57(m，1H)，3.67-3.71(m，1H)，4.13-4.15(m，1H)，4.27(m，1H)，4.87-4.89(m，1H)。
To 4-methyl hydroxybenzoate (4.81g, 0.03mol), 1-(tert-butoxycarbonyl)-(4R)-(t-butyldimethylsilyloxy base)-(2S)-dried meat ammonia alcohol (10.5g, 0.03mol) and triphenyl phasphine (9.96g, 0.04mol) the agitating solution of cold (0 ℃) of THF (160ml) in be added dropwise to DIAD (7.48ml.0.04mol), with this reaction mixture refluxed heating 7 hours, after being cooled to room temperature, evaporate this mixture.Residue is through purification by silica gel column chromatography, with normal hexane-ethyl acetate (6: 1, v/v) eluting obtains 4-[1-(tert-butoxycarbonyl)-(4R)-(t-butyldimethylsilyloxy base)-(2S)-pyrrolidinyl methoxyl group] essence of Niobe (9.58g, 65%), is white solid.mp?86-88℃； 1H-NMR(CDCl 3)δ0.08(s，6H)，0.88(s，9H)，1.46(s，9H)，2.04-2.15(m，2H)，3.29-3.48(m，2H)，3.88(s，3H)，4.06-4.30(m，3H)，4.46-4.51(m，1H)，6.91-6.93(m，2H)，7.96-7.98(m，2H)；MS(ESI)m/z466(M ++1)。
To 4-[1-(tert-butoxycarbonyl)-(4R)-(t-butyldimethylsilyloxy base)-(2S)-pyrrolidinyl methoxyl group] essence of Niobe (1.49g, 3.20mmol) the agitating solution of cold (0 ℃) of THF (15ml) in add TBAF (6.40ml, 6.40mmol, the solution of 1M THF), this reactant mixture was stirred under room temperature 2 hours.Dilute this mixture with ethyl acetate, water, salt water washing are through dried over sodium sulfate and evaporation.Residue is through purification by silica gel column chromatography, with toluene-acetone (5: 1,, obtain 4-[1-(tert-butoxycarbonyl)-(4R)-hydroxyl-(2S)-pyrrolidinyl methoxyl group v/v) as eluant] essence of Niobe (1.05g, 93%), be white solid.mp?103-105℃； 1H-NMR(CDCl 3)δ1.46(s，9H)，2.11-2.28(m，2H)，3.49-3.60(m，2H)，3.88(s，3H)，4.15-4.34(m，3H)，4.53-4.57(m，1H)，6.91(d，J＝8.6Hz，2H)，7.97(d，J＝8.6Hz，2H)；MS(ESI)m/z352(M ++1)。
To 4-methyl hydroxybenzoate (0.56g, 3.68mmol), 4-[1-(tert-butoxycarbonyl)-(4R)-hydroxyl-(2S)-pyrrolidinyl methoxyl group] essence of Niobe (1.30g, 3.70mmol) and triphenyl phasphine (1.16g, 4.42mmol) the agitating solution of cold (0 ℃) of THF (20ml) in drip DIAD (0.87ml, 4.42mmol), this reactant mixture was stirred under room temperature 3 hours.Evaporate this mixture, residue is through purification by silica gel column chromatography, with normal hexane-ethyl acetate (6: 1, v/v) as eluant, obtain 4-[1-(tert-butoxycarbonyl)-(4S)-(4-methoxycarbonyl phenoxy group)-(2S)-pyrrolidinyl methoxyl group] essence of Niobe (1.80g, q.y.), be colorless oil. 1H-NMR(CDCl 3)δ1.49(s，9H)，2.3?1-2.38(m，1H)，2.45-2.49(m，1H)，3.64-3.77(m，2H)，3.88(s，6H)，4.07-4.15(m，1H)，4.33-4.44(m，2H)，4.95-5.01(m，1H)，6.85(d，J＝8.8Hz，2H)，6.94(br?s，2H)，7.97(d，J＝8.8Hz，4H)；MS(ESI)m/z486(M ++1)。
To 4-[1-(tert-butoxycarbonyl)-(4S)-(4-methoxycarbonyl phenoxy group)-(2S)-pyrrolidinyl methoxyl group] essence of Niobe (1.80g, 3.71mmol) the agitating solution of dichloromethane (15ml) in add TFA (15ml), under room temperature, this reactant mixture was stirred 1.5 hours.This mixture of vacuum concentration, by saturated sodium bicarbonate make be alkalescence, use chloroform extraction.With salt water washing extract, dry and evaporation obtains 4-[(4S through potassium carbonate)-(4-methoxycarbonyl phenoxy group)-(2S)-pyrrolidinyl methoxyl group] (1.50g q.y.), is faint yellow oily thing to essence of Niobe. 1H-NMR(CDCl 3)δ1.87-1.92(m，1H)，2.41-2.48(m，1H)，3.18-3.23(m，1H)，3.34-3.37(m，1H)，3.60-3.66(m，1H)，3.88(s，3H)，3.89(s，3H)，4.04-4.13(m，2H)，4.94-5.00(m，1H)，6.87-6.93(m，4H)，7.96-8.00(m，4H)；MS(ESI)m/z?386(M ++1)。
Under room temperature, with 3-methoxyl group-4-[N '-(2-aminomethyl phenyl) urea groups] phenylacetic acid (400mg, 1.27mmol), 4-[(4S)-(4-methoxycarbonyl phenoxy group)-(2S)-pyrrolidinyl methoxyl group] essence of Niobe (491mg, 1.27mmol), EDCHCl (293mg, 1.53mmol), HOBt (207mg, 1.53mmol) and triethylamine (215 μ l, 1.54mmol) mixture in THF (10ml) stirs and to spend the night.This mixture of dilute with water is used ethyl acetate extraction.With salt water washing extract, through dried over sodium sulfate and evaporation.Residue is through purification by silica gel column chromatography; with chloroform-methanol (60: 1-50: 1; v/v) as eluant; obtain 4-[1-[3-methoxyl group-4-[N '-(2-aminomethyl phenyl) urea groups] phenyl acetyl]-(4S)-and (4-methoxycarbonyl phenoxy group)-(2S)-pyrrolidinyl methoxyl group] essence of Niobe (532mg; 61%), is the white foam thing. 1H-NMR (CDCl 3) δ 2.26-2.49 (a plurality of s and m, 5H altogether), 3.56-3.93 (a plurality of s and m, 13H altogether), (4.07-4.59 a plurality of m, 3H altogether), 5.01 (m, 1H), 6.69-6.94 (m, 7H), 7.09-7.13 (m, 1H), and 7.20-7.31 (m, 3H), 7.52-7.57 (m, 1H), 7.92-8.00 (m, 4H), and 8.06-8.09 (m, 1H); MS (ESI) m/z 682 (M ++ 1).
To 4-[1-[3-methoxyl group-4-[N '-(2-aminomethyl phenyl) urea groups] phenyl acetyl]-(4S)-and (4-methoxycarbonyl phenoxy group)-(2S)-pyrrolidinyl methoxyl group] essence of Niobe (532mg; 0.78mmol) the agitating solution of THF (5ml) in add 0.5N sodium hydroxide (5ml), with the heating 5 hours under refluxing of this reactant mixture.After being cooled to room temperature, in this mixture impouring ice-1N HCl, collect the precipitation that generates.The crude product solid is recrystallization from methanol-chloroform-ether, obtains 116 (125mg, 25%), is light yellow crystalline powder.MW 653.68 mp 154-159 ℃; 1H-NMR (DMSO-d 6) δ 2.24 (s, 3H), 2.38-2.49 (m, 2H), 3.63 (s, 2H), 3.67-3.88 (a plurality of s and m, 4H altogether), 4.01-4.06 and 4.15-4.19 (respectively be m, be total to 2H), 4.27-4.31 and 4.38-4.42 (respectively be m, altogether 2H), 5.18-5.25 (m, 1H), 6.72-6.77 (m, 1H), 6.85-7.16 (a plurality of m, 8H altogether), 7.78-7.89 (m, 5H), 7.99-8.02 (m, 1H), 8.46 (s, 1H), 8.57 (s, 1H), 12.65 (br s, 2H); MS (ESI) m/z 654 (M ++ 1); C 36H 35N 3O 91/2H 2The analytical calculation value of O: C, 65.25; H, 5.48; N, 6.34.Measured value: C, C, 65.29; H, 5.54; N, 6.20.Embodiment 1094-[(4S)-(4-carboxyl phenoxy group)-1-[4-[N '-(2-chlorophenyl) urea groups]-3-methoxyphenyl acetyl group]-(2S)-and the pyrrolidinyl methoxyl group] benzoic acid
Figure A0080951003261
Under room temperature, with 4-[N '-(2-chlorophenyl) urea groups]-3-methoxybenzene guanidine-acetic acid (420mg, 1.25mmol), 4-[(4S)-(4-methoxycarbonyl phenoxy group)-(2S)-pyrrolidinyl methoxyl group] essence of Niobe (483mg, 1.25mmol), EDCHCl (288mg, 1.50mmol), HOBt (203mg, 1.50mmol) and triethylamine (210 μ l, 1.51mmol) mixture in THF (10ml) stirs and to spend the night.This mixture of dilute with water is used ethyl acetate extraction.With salt water washing extract, through dried over sodium sulfate and evaporation.Residue is through purification by silica gel column chromatography; with chloroform-methanol (50: 1; v/v) as eluant; obtain 4-[1-[4-[N '-(2-chlorophenyl) urea groups]-3-methoxyphenyl acetyl group]-(4S)-and (4-methoxycarbonyl phenoxy group)-(2S)-pyrrolidinyl methoxyl group] essence of Niobe (488mg; 55%), is the white foam thing. 1H-NMR (CDCl 3) δ 2.28-2.51 (m, 2H), 3.62-3.94 (a plurality of s and m, 13H altogether), 4.07-4.62 (a plurality of m, 3H altogether), 4.99-5.03 (m, 1H), 6.78-6.99 (m, 7H), 7.23-7.34 (m, 2H), 7.42-7.52 (m, 2H), 7.92-8.01 (m, 5H), 8.17-8.20 (m, 1H); MS (ESI) m/z 702 (M ++ 1).
To 4-[1-[4-[N '-(2-chlorophenyl) urea groups]-3-methoxyphenyl acetyl group]-(4S)-and (4-methoxycarbonyl phenoxy group)-(2S)-pyrrolidinyl methoxyl group] essence of Niobe (488mg; 0.70mmol) THF (5ml) agitating solution in add 0.5N sodium hydroxide (5ml), with the heating 3 hours under refluxing of this reactant mixture.After being cooled to room temperature, in this mixture impouring ice-1N HCl, collect the precipitation that generates.The crude product solid is recrystallization from methanol-chloroform-ether, obtains 117 (137mg, 29%), is white crystalline powder.150-153 ℃ of MW674.10 mp; 1H-NMR (DMSO-d 6) δ 4.17-4.21 (respectively be m, be total to 2H), 4.30-4.34 and 4.40-4.45 (respectively be m, be total to 2H), 5.20-5.27 (m, 1H), and 6.77-6.81 (m, 1H), 6.89-6.92 (m, 1H), 7.01-7.08 (m, 5H), 7.27-7.3 1 (m, 1H), 7.43-7.46 (m, 1H), and 7.86-7.92 (m, 4H), 7.97-8.00 (m, 1H), 8.10-8.12 (m, 1H), 8.91 (s, 1H), 8.96 (s, 1H), 12.65 (br s, 2H); MS (ESI) m/z 674 (M ++ 1).C 35H 32ClN 3O 91/4H 2The analytical calculation value of O: C, 61.95; H, 4.83; N, 6.19; Cl, 5.22.Measured value: C, 61.77; H, 4.86; N, 6.13; Cl, 5.49.Embodiment 1104-[1-[4-[N '-(2-bromo phenyl) urea groups]-3-methoxyphenyl acetyl group]-(4S)-and (4-carboxyl phenoxy group)-(2S)-pyrrolidinyl methoxyl group] benzoic acid
Figure A0080951003271
Under room temperature, with 4-[N '-(2-bromo phenyl) urea groups]-3-methoxybenzene guanidine-acetic acid (464mg, 1.22mmol), 4-[(4S)-(4-methoxycarbonyl phenoxy group)-(2S)-pyrrolidinyl methoxyl group] essence of Niobe (472mg, 1.22mmol), EDCHCl (282mg, 1.47mmol), HOBt (200mg, 1.48mmol) and triethylamine (205 μ l, 1.47mmol) mixture in THF (10ml) stirs and to spend the night.This mixture of dilute with water is used ethyl acetate extraction.With salt water washing extract, through dried over sodium sulfate and evaporation.Residue is through purification by silica gel column chromatography; with chloroform-methanol (60: 1-50: 1; v/v) as eluant; obtain 4-[1-[4-[N '-(2-bromo phenyl) urea groups]-3-methoxyphenyl acetyl group]-(4S)-and (4-methoxycarbonyl phenoxy group)-(2S)-pyrrolidinyl methoxyl group] essence of Niobe (379mg; 41%), is the white foam thing. 1H-NMR (CDCl 3) δ 2.28-2.51 (m, 2H), 3.59-3.95 (a plurality of s and m, 13H altogether), 4.07-4.62 (a plurality of m, 3H altogether), 4.99-5.03 (m, 1H), 6.79-6.95 (m, 7H), 7.27-7.36 (m, 3H), 7.49-7.51 (m, 1H), 7.93-8.01 (m, 5H), 8.11-8.14 (m, 1H); MS (ESI) m/z 747 (M ++ 1).
To 4-[1-[4-[N '-(2-bromo phenyl) urea groups]-3-methoxyphenyl acetyl group]-(4S)-and (4-methoxycarbonyl phenoxy group)-(2S)-pyrrolidinyl methoxyl group] essence of Niobe (379mg; 0.51mmol) THF (5ml) agitating solution in add 0.5N sodium hydroxide (5ml), with the heating 5 hours under refluxing of this reactant mixture.After being cooled to room temperature, in this mixture impouring ice-1N HCl, collect the precipitation that generates.The crude product solid obtains 118 (51mg, 14%) through preparation property TLC purification, is faint yellow amorphous solid.MW 718.55 1H-NMR (DMSO-d 6) δ 2.20-2.40 (m, 2H), 3.65-3.89 (a plurality of m, 6H altogether), 4.02-4.63 (a plurality of m, 4H altogether), 5.19-5.26 (m, 1H), and 6.74-7.06 (m, 7H), 7.30-7.34 (m, 1H), 7.59-7.61 (m, 1H), 7.83-7.96 (m, 6H), 8.74 (s, 1H), 8.93 (s, 1H); C 35H 32BrN 3O 92H 2The analytical calculation value of O: C, 55.71; H, 4.81; N, 5.57.Measured value: C, 55.92; H, 4.80; N, 5.30.Embodiment 1114-[(4S)-and (4-carboxyl phenoxy group)-1-[4-[N '-(2-aminomethyl phenyl) urea groups] phenyl acetyl]-(2S)-and the pyrrolidinyl methoxyl group] benzoic acid
Figure A0080951003281
Under room temperature, with 4-[N '-(2-aminomethyl phenyl) urea groups] phenylacetic acid (328mg, 1.15mmol), 4-[(4S)-(4-methoxycarbonyl phenoxy group)-(2S)-pyrrolidinyl methoxyl group] essence of Niobe (444mg, 1.15mmol), EDCHCl (265mg, 1.38mmol), HOBt (187mg, 1.38mmol) and triethylamine (195 μ l, 1.40mmol) mixture in THF (10ml) stirs and to spend the night.This mixture of dilute with water is used ethyl acetate extraction.With salt water washing extract.Through dried over sodium sulfate and evaporation.Residue is through purification by silica gel column chromatography; with chloroform-methanol (60: 1-50: 1; v/v) as eluant; obtain 4-[(4S)-(4-methoxycarbonyl phenoxy group)-1-[4-[N '-(2-aminomethyl phenyl) urea groups] phenyl acetyl]-(2S)-and the pyrrolidinyl methoxyl group] essence of Niobe (332mg; 44%), is the white foam thing. 1H-NMR (CDCl 3) δ 2.13 (s, 3H), 2.24-2.48 (m, 2H), 3.52-3.90 (a plurality of s and m, 10H altogether), (4.05-4.58 a plurality of m, 3H altogether), 5.01 (m, 1H), 6.78-6.90 (m, 4H), 6.98-7.20 (m, 8H), 7.51-7.56 (m, 2H), 7.90-8.00 (m, 4H); MS (ESI) m/z652 (M ++ 1).
To 4-[(4S)-(4-methoxycarbonyl phenoxy group)-1-[4-[N '-(2-aminomethyl phenyl) urea groups] phenyl acetyl]-(2S)-and the pyrrolidinyl methoxyl group] essence of Niobe (332mg; 0.5 add 0.5N sodium hydroxide (5ml) in the agitating solution of THF 1mmol) (5ml), this reactant mixture heated 3 hours under refluxing.After being cooled to room temperature, in this mixture impouring ice-1N HCl, collect the precipitation that generates.The crude product solid obtains 119 (118mg, 37%) through recrystallization from methanol-chloroform-ether, is white crystalline powder.MW 623.65 mp 157-160 ℃; 1H-NMR (DMSO-d 6) δ 2.20-2.25 (a plurality of s and m, 4H altogether), and 2.39-2.47 (m, 1H), 3.64 (s, 2H), and 3.68-3.89 (m, 1H), 4.02-4.08 and 4.16-4.20 (respectively be m, be total to 2H), 4.29-4.33 and 4.39-4.43 (respectively be m, altogether 2H), 5.20-5.26 (m, 1H), 6.92-6.96 (m, 1H), and 7.02-7.08 (m, 4H), 7.12-7.18 (m, 4H), 7.39-7.41 (m, 2H), 7.84-7.92 (m, 6H), 9.01 (s, 1H), 12.65 (br s, 2H); MS (ESI) m/z 624 (M ++ 1).C 35H 33N 3O 81H 2The analytical calculation value of O: C, 65.51; H, 5.50; N, 6.55.Measured value: C, 65.48; H, 5.36; N, 6.52.Embodiment 1124-[4-(2,4 difluorobenzene oxygen base)-1-[3-methoxyl group-4-[N '-(2-aminomethyl phenyl) urea groups] phenyl acetyl]-2-pyrrolidinyl methoxybenzoic acid
To 1-(tert-butoxycarbonyl)-4-L-Hydroxyproline methyl ester (4.0g, 16.3mmol), triphenyl phasphine (5.14g, 19.6mmol) and 2,4-difluorophenol (2.55g, 19.6mmol) the agitating solution of THF (50ml) in add DIAD (3.9ml, 19.6mmol), with this mixture reflux 3 hours.After being cooled to room temperature, this mixture of vacuum concentration, residue is through silica gel column chromatography, with chloroform-ethyl acetate (4: 1) eluting, obtain 1-(tert-butoxycarbonyl)-4-(2,4 difluorobenzene oxygen base) pyrrolidine-2-methyl formate (5.82g, quantitatively), be yellow oil.
To 1-(tert-butoxycarbonyl)-4-(2,4 difluorobenzene oxygen base) pyrrolidine-2-methyl formate (5.82g, add in the agitating solution of THF 16.3mmol) (130ml) the 0.25N sodium hydroxide (130ml, 32.6mmol).The mixture stirring that obtains is spent the night.In this mixture impouring 1N HCl (100ml), (2 * 200ml) extract with chloroform.Through dried over mgso extract and evaporation.Residue with chloroform-ethyl acetate (4: 1) eluting, obtains 1-(tert-butoxycarbonyl)-4-(2,4 difluorobenzene oxygen base) pyrrolidine-2-formic acid (2.55g, 46%) through silica gel column chromatography, is the colourless foam thing. 1H-NMR (CDCl 3) δ 1.42-1.47 (m, 9H), 2.29-2.74 (a plurality of m, 2H), 3.66-3.71 (m, 2H), 4.46-4.51 (m, 1H), 4.83 (m, 1H), 6.73-6.95 (m, 3H).
(2.55g adds BH in the agitating solution of THF 7.43mmol) (50ml) to 1-(tert-butoxycarbonyl)-4-(2,4 difluorobenzene oxygen base) pyrrolidine-2-formic acid 3DMS (452 μ l, 7.43mmol).This mixture reflux is spent the night.After being cooled to room temperature, this mixture of vacuum concentration is by adding entry (100ml) quencher.(2 * 200ml) extract this mixture, through dried over mgso and evaporation with chloroform.Residue as eluant, obtains 1-(tert-butoxycarbonyl)-4-(2,4 difluorobenzene oxygen base)-2-pyrrolidinyl methanol (1.76g, 72%) with chloroform-ethyl acetate (4: 1) through silica gel column chromatography, is colorless oil. 1H-NMR (CDCl 3) δ 1.45 (s, 9H), 2.28-2.36 (m, 2H), 3.58-4.99 (a plurality of m, 8H), 6.74-6.90 (m, 3H).
To 1-(tert-butoxycarbonyl)-4-(2,4-two fluorophenoxies)-2-pyrrolidinyl methanol (500mg, 1.52mmol), 4-methyl hydroxybenzoate (277mg, 1.82mmol) and triphenyl phasphine (477mg, 1.82mmol) the agitating solution of THF (10ml) in add DIAD (358 μ l, 1.82mmol), with this mixture reflux 5 hours.After being cooled to room temperature, this mixture of vacuum concentration.Residue as eluant, obtains 4-[1-(tert-butoxycarbonyl)-4-(2,4 difluorobenzene oxygen base)-2-pyrrolidinyl methoxyl group with chloroform-ethyl acetate (20: 1) through silica gel column chromatography] essence of Niobe (529mg, 75%), be colorless oil. 1H-NMR(CDCl 3)δ1.46(s，9H)，2.20-2.47(m，2H)，3.64(m，2H)，3.86(s，3H)，4.07-4.43(m，3H)，4.86(m，1H)，6.74-6.87(m，3H)，6.94(d，2H，J＝8.5Hz)，7.95(d，2H，J＝8.5Hz)。
To 4-[1-(tert-butoxycarbonyl)-4-(2,4 difluorobenzene oxygen base)-2-pyrrolidinyl methoxyl group] (529mg adds TFA (5ml) to essence of Niobe in the agitating solution of dichloromethane 1.15mmol) (5ml).This mixture stirring is spent the night.This mixture of vacuum concentration makes residue be alkalescence by adding saturated sodium bicarbonate.(2 * 100ml) extract this mixture with chloroform.Through dry extract of potassium carbonate and evaporation, obtain 4-[4-(2,4 difluorobenzene oxygen base)-2-pyrrolidinyl methoxyl group] essence of Niobe (385mg, 92%), be yellow oil. 1H-NMR(CDCl 3)δ1.89-1.95(m，1H)，2.28-2.35(m，1H)，3.09(dd，J＝12.5，4.9Hz，1H)，3.33(d，J＝12.5Hz，1H)，3.60(m，1H)，3.86(s，3H)，4.10(d，J＝5.6Hz，2H)，4.84(m，1H)，6.73-6.89(m，3H)，6.91(d，J＝8.5Hz，2H)，7.96(d，J＝8.5Hz，2H)。
With 4-[4-(2,4-two fluorophenoxies)-and 2-pyrrolidinyl methoxyl group] essence of Niobe (380mg, 1.05mmol), 3-methoxyl group-4-[N '-(2-aminomethyl phenyl) urea groups] phenylacetic acid (329mg, 1.05mmol), EDCHCl (302mg, 1.58mmol) and the HOBt of catalytic amount and the mixture of DMAP in DMF (10ml) stirred 3 days.(200ml) dilutes this mixture with ethyl acetate, with saline (2 * 200ml) washings.Except that after desolvating; residue is through silica gel column chromatography; with chloroform-ethyl acetate (4: 1) to chloroform-methanol (10: 1) as eluant; obtain 4-[4-(2; 4-two fluorophenoxies)-and 1-[3-methoxyl group-4-[N '-(2-aminomethyl phenyl) urea groups] phenyl acetyl]-2-pyrrolidinyl methoxyl methyl benzoate (693mg, quantitatively). 1H-NMR (CDCl 3) δ 2.16-2.53 (m, 5H), 3.61-4.93 (a plurality of m, 14H), 6.48-8.12 (a plurality of m, 16H).To 4-[4-(2; 4-two fluorophenoxies)-and 1-[3-methoxyl group-4-[N '-(2-aminomethyl phenyl) urea groups] phenyl acetyl]-2-pyrrolidinyl methoxyl methyl benzoate (693mg; 1.05mmol) THF (8ml) agitating solution in add 0.25N sodium hydroxide (8.4ml; 2.10mmol), this mixture stirring is spent the night.In this mixture impouring 1NHCl (200ml), sucking filtration is collected the precipitation that generates.Solid is through silica gel column chromatography, with chloroform-methanol (50: 1-10: 1) as eluant, obtain 120 (323mg, 48%), be colourless amorphous solid.MW 645.65 1H-NMR (DMSO-d 6) δ 2.25 (s, 3H), 2.35 (m, 2H), 3.33-5.18 (a plurality of m, 11H), 6.75 (dd, 1H, J=8.3,1.7Hz), 6.87-7.30 (a plurality of m, 8H), 7.79 (d, 1H, J=8.3Hz), 7.85-7.90 (m, 3H), 8.01 (d, 1H, J=8.3Hz), 8.49 (s, 1H), 8.57 (s, 1H); MS (FAB) m/z646 (M ++ 1).Embodiment 1134-[1-[4-[N '-(2-chlorophenyl) urea groups]-3-methoxyphenyl acetyl group]-4-(6-quinolyl oxygen base-2S-pyrrolidinyl] methoxybenzoic acid
In 0 ℃, to (anti-form-1-tert-butoxycarbonyl-4-hydroxyl-2-pyrrolidinyl) methoxyl methyl benzoate (1.0g, 3.0mmol), 6-hydroxyquinoline (435mg, 3.0mmol) and triphenyl phasphine (943mg, 3.6mmol) the agitating solution of THF (10ml) in add DIAD (727mg, 3.6mmol).This reactant mixture was stirred under room temperature 18 hours.This mixture of vacuum concentration, residue be through purification by silica gel column chromatography, with normal hexane-ethyl acetate (1: 2, v/v) eluting.In 0 ℃, in the agitating solution of the dichloromethane (6.0ml) of this product, add TFA (6.0ml).Stirred reaction mixture is 0.5 hour under room temperature.This mixture of vacuum concentration.Saturated sodium bicarbonate is added in the residue, use dichloromethane extraction.With salt water washing extract, through dried over sodium sulfate and vacuum concentration.Residue with methanol-dichloromethane (1%-10% v/v) as eluant, obtains 4-[(4S)-[(6-quinolyl oxygen base-2S-pyrrolidinyl)] methoxyl methyl benzoate (900mg, 82%), is faint yellow oily thing through purification by silica gel column chromatography. 1H-NMR(CDCl 3)δ1.92-2.10(m，1H)，2.45-2.55(m，1H)，3.20-3.30(m，1H)，3.38-3.50(m，1H)，3.60-3.70(m，1H)，3.88(s，3H)，4.05-4.18(m，2H)，5.03(m，1H)，6.91(d，J＝8.5Hz，1H)，7.02(d，J＝2.7Hz，1H)，7.35-7.38(m，2H)，7.96(d，J＝8.5Hz，1H)，8.00-8.05(m，2H)，8.76(d，J＝3.2Hz，1H)。
In 0 ℃, to 4-(4S-(6-quinolyl oxygen base-2S-pyrrolidinyl) methoxyl methyl benzoate (300mg, 0.79mmo1), 4-[N '-(2-chlorophenyl) urea groups]-3-methoxybenzene guanidine-acetic acid (264mg, 0.79mmol), HOBt (107mg, 0.79mmol) and triethylamine (330m1,2.37mmol) THF (10.0ml) and the agitating solution among the MeCN (10.0ml) in add EDCHCl (228mg, 1.2mmol).Under room temperature, stirred this reactant mixture 16 hours, vacuum concentration.Water is added in the residue, use ethyl acetate extraction.Wash extract with saturated sodium bicarbonate, then through dried over sodium sulfate and vacuum concentration.Residue is through purification by silica gel column chromatography; with ethyl acetate to ethanol-ethyl acetate (10%; v/v) as eluant; obtain 4-[1-[4-[N '-(2-chlorophenyl) urea groups]-3-methoxyphenyl acetyl group]-4-(6-quinolyl oxygen base-(2 S)-pyrrolidinyl] methoxyl methyl benzoate (520mg; 95%), is colorless oil. 1H-NMR(CDCl 3)δ2.30-2.60(m，3H)，3.64(s，2H)，3.73(s，3H)，3.80-3.95(m，1H)，3.87(s，3H)，4.15-4.30(m，1H)，4.50-4.70(m，2H)，5.11(br?s，1H)，6.81-7.01(m，6H)，7.26-7.39(m，6H)，7.93-8.03(m，5H)，8.19(d，J＝8.3Hz，1H)，8.80(s，1H)。
To 4-[1-[4-[N '-(2-chlorophenyl) urea groups]-3-methoxyphenyl acetyl group]-4-(6-quinolyl oxygen base-2S-pyrrolidinyl] methoxyl methyl benzoate (520mg; 0.75mmol) THF (10.0ml) and the agitating solution of methanol (5.0ml) in add the 1N sodium hydroxide (1.5ml, 1.5mmol).This mixture was stirred 18 hours in 60 ℃.This mixture of vacuum concentration adds water wherein, with 1N HCl neutralization.Collect the solid that produces, wash with water, vacuum drying obtains 121 (450mg, 88%), is white crystalline solid.681.13 mp 129-133 ℃; IR (KBr) 3332,1704,1604,1531,1419,1222,1166 cm -1 1H-NMR (DMSO-d 6) δ 2.25-2.55 (m, 2H), 3.67 (s, 2H), 3.82 (s, 3H), and 3.81-3.92 (m, 1H), 4.02-4.15 (m, 2H), 4.40-4.50 (m, 2H), 5.25-5.40 (m, 1H), 5.33-7.07 (m, 5H), 7.26-7.49 (m, 5H), 7.83-8.23 (m, 6H), 8.73-8.74 (m, 1H), 8.90 (s, 1H), 8.94 (s, 1H); MS (FAB) m/z681 (M ++ 1); C 37H 33N 4O 7Cl0.5H 2The analytical calculation value of O: C, 64.39; H, 4.97; N, 8.12.Measured value: C, 64.22; H, 4.90; N, 7.96.Embodiment 144-[1-[4-[N '-(2-bromo phenyl) urea groups]-3-methoxyphenyl acetyl group]-(4S)-(6-quinolyl oxygen base-(2S)-pyrrolidinyl] methoxybenzoic acid
In 0 ℃, to 4-(4S-(6-quinolyl oxygen base-2S-pyrrolidinyl) methoxyl methyl benzoate (300mg, 0.79mmol), 4-[N '-(2-bromo phenyl) urea groups]-3-methoxybenzene guanidine-acetic acid (299mg, 0.79mmol), HOBt (107mg, 0.79mmol) and triethylamine (330ml, 2.37mmol) THF (10.0ml) and the agitating solution of MeCN (10.0ml) in add EDCHCl (228mg, 1.2mmol).Under room temperature, stirred this reactant mixture 16 hours, vacuum concentration.Water is added in the residue, use ethyl acetate extraction.Wash extract with saturated sodium bicarbonate, then through dried over sodium sulfate and vacuum concentration.Residue is through purification by silica gel column chromatography; with ethyl acetate to ethanol-ethyl acetate (10%; v/v) as eluant; obtain 4-[1-[4-[N '-(2-bromo phenyl) urea groups]-3-methoxyphenyl acetyl group]-(4S)-(6-quinolyl oxygen base-(2S)-pyrrolidinyl] methoxyl methyl benzoate (530mg; 91%), is colorless oil. 1H-NMR(CDCl 3)δ2.30-2.62(m，3H)，3.65(s，2H)，3.75(s，3H)，3.80-3.95(m，1H)，3.93(s，3H)，4.10-4.30(m，1H)，4.50-4.70(m，2H)，5.11(br?s，1H)，6.82-6.98(m，6H)，7.15-7.39(m，5H)，7.52(d，J＝8.0Hz，1H)，7.93-8.03(m，5H)，8.14(d，J＝8.3Hz，1H)，8.80(s，1H)。
To 4-[1-[4-[N '-(2-bromo phenyl) urea groups]-3-methoxyphenyl acetyl group]-4-(6-quinolyl oxygen base-2S-pyrrolidinyl] methoxyl methyl benzoate (530mg; 0.72mmol) THF (10.0ml) and the agitating solution of methanol (5.0ml) in add the 1N sodium hydroxide (1.4ml, 1.4mmol).This mixture was stirred 24 hours in 70 ℃.This mixture of vacuum concentration adds water wherein, with 1N HCl neutralization.Collect the solid that produces, wash with water, vacuum drying obtains 122 (460mg, 88%), is white crystalline solid.MW 725.59 mp 149-153 ℃; IR (KBr) 3332,1704,1604,1527,1222,1164 cm -1 1H-NMR (DMSO-d 6) δ 2.28-2.58 (m, 2H), 3.67 (s, 2H), 3.82 (s, 3H), and 3.85-3.90 (m, 1H), 4.05-4.15 (m, 2H), 4.40-4.50 (m, 2H), 5.20-5.32 (m, 1H), 6.77-7.07 (m, 5H), 7.31-7.61 (m, 5H), 7.83-7.97 (m, 5H), 8.21-8.22 (m, 1H), 8.73-8.74 (m, 2H), 8.92 (s, 1H); MS (FAB) m/z725 (M +), 727 (M ++ 2); C 37H 33N 4O 6Br0.5H 2The analytical calculation value of O: C, 60.50; H, 4.67; N, 7.63; Br, 10.88.Measured value: C, 60.51; H, 4.60; N, 7.52; Br, 11.06.Embodiment 1154-[(2S, 4S)-1-[3-methoxyl group-4-[N '-(2-aminomethyl phenyl) urea groups] phenyl acetyl]-4-(2-naphthoxy)-2-pyrrolidinyl] methoxybenzoic acid
Under room temperature, blanket of nitrogen, to (2S, 4R)-1-tert-butoxycarbonyl-4-hydroxyl-2-pyrrolidinyl methyl formate (4.22g, 17.2mmol), beta naphthal (2.73g, 18.9mmol) and triphenyl phasphine (4.96g, the middle DIAD (3.72ml.18.9mmol) of adding that stirs the mixture of THF 18.9mmol) (80ml).After stirring is spent the night, this mixture of vacuum concentration.Residue is through silica gel column chromatography [600g, chloroform/ethyl acetate (10/1)], obtain (2S, 4S)-1-tert-butoxycarbonyl-4-(2-naphthoxy)-2-pyrrolidinyl methyl formate (5.37g), it need not to be further purified and uses.
Under room temperature, to (2S, 4S)-add in the agitating solution of the THF (116ml) of 1-tert-butoxycarbonyl-4-(2-naphthoxy)-2-pyrrolidinyl methyl formate (5.37g) the 0.25N sodium hydroxide (116ml, 29.0mmol).The mixture stirring that obtains is spent the night.Except that after desolvating,, use chloroform extraction by adding this mixture of 1N HCl acidify.Extract with the salt water washing merges also evaporates through dried over sodium sulfate.Residue is recrystallization from normal hexane-chloroform, obtain (2S, 4S)-1-tert-butoxycarbonyl-4-(2-naphthoxy)-2-pyrrolidinyl formic acid (4.44g, 85% (two steps)), be white powder. 1H-NMR (DMSO-d 6) δ 1.37 and 1.41 (s, 9H, amide isomers), 2.26 (d, J=13.9Hz, 1H), 2.65 (m, 1H), 3.47 (d, J=11.5Hz, 1H), 3.81 (m, 1H), 4.30 (m, 1H), 5.14 (m, 1H), 7.02-7.86 (m, 7H).
In 0 ℃, to (2S, 4S)-(1.12g adds BH in THF 3.13mmol) (30ml) agitating solution to 1-tert-butoxycarbonyl-4-(2-naphthoxy)-2-pyrrolidinyl formic acid 3DMS (0.63ml, 6.3mmol).This mixture is warming up to room temperature immediately, then in 50 heating 1.5 hours.After being cooled to room temperature, in 0 ℃, by adding this mixture of entry quencher, with this mixture of ethyl acetate extraction.Extract with the salt water washing merges also evaporates through dried over sodium sulfate.Residue is through silica gel column chromatography [50g, chloroform/methanol (50/1)], obtain (2S, 4S)-1-tert-butoxycarbonyl-4-(2-naphthoxy)-2-pyrrolidinyl methanol (1.10g, 100%), be faint yellow oily thing. 1H-NMR(CDCl 3)δ1.48(s，9H)，2.45(m，1H)，3.58-4.80(m，4H)，5.01(br，1H)，7.04-7.99(m，7H)。
Under room temperature, blanket of nitrogen, to (2S, 4S)-1-tert-butoxycarbonyl-4-(2-naphthoxy)-2-pyrrolidinyl methanol (640mg, 1.86mmol), 4-methyl hydroxybenzoate (283mg, 1.86mmol) and triphenyl phasphine (488mg, the middle DIAD (0.37ml.1.86mmol) of adding that stirs the mixture of THF 1.86mmol) (18ml).This mixture stirring is spent the night.Except that after desolvating, the residue that obtains is through silica gel column chromatography [100g, n-hexane/ethyl acetate (2/1)], obtain 4-[(2S, 4S)-and 1-tert-butoxycarbonyl-4-(2-naphthoxy)-2-pyrrolidinyl] methoxyl methyl benzoate (830mg, 93%), be colorless oil. 1H-NMR (CDCl 3) δ 1.50 (d, J=8.3Hz, 9H), 2.34 (m, 1H), 2.53 (d, J=14.2Hz, 1H), 3.72-3.85 (m, 1H), 3.86 and 3.87 (s, 3H, amide isomers), 4.17 (m, 1H), 4.26-4.52 (m, 2H), 5.06 (br, 1H), 6.87 (d, J=8.8Hz, 1H), 6.94 (d, J=8.8Hz, 2H), 7.04 (br, 2H), 7.33 (t, J=7.3Hz, 1H), 7.42 (t, J=7.3Hz, 1H), 7.64-8.02 (m, 5H).
Under room temperature, to 4-[(2S, 4S)-and 1-tert-butoxycarbonyl-4-(2-naphthoxy)-2-pyrrolidinyl] (870mg adds TFA (6ml) to methoxyl methyl benzoate in the agitating solution of dichloromethane 1.74mmol) (24ml).This mixture stirring is spent the night.This mixture of vacuum concentration dilutes residue with dichloromethane, makes by adding 1N sodium hydroxide to be alkalescence, and it is used dichloromethane extraction.With salt water washing organic layer, through dried over sodium sulfate and concentrated.Residue obtains 4-[(2S through silica gel column chromatography [100g, n-hexane/ethyl acetate (2/1)], 4S)-4-(2-naphthoxy)-2-pyrrolidinyl] methoxyl methyl benzoate (750mg, 100%), be the dark oil thing. 1H-NMR (CDCl 3) δ 1.99 (dd, J=14.2,5.6Hz, 1H), 2.48 (m, 1H), 3.22 (dd, J=12.2,4.6Hz, 1H), 3.43 (d, J=12.5Hz, 1H), 3.67 (m, 1H), 3.86 and 3.87 (s, 3H, amide isomers), 4.11 (m, 2H), 5.04 (m, 1H), 6.83 (d, J=8.5Hz, 1H), 6.89 (d, J=8.8Hz, 2H), 7.07 (d, J=2.0Hz, 1H), 7.12 (d, J=9.0Hz, 1H), 7.33 (dt, J=8.1,1.2Hz, 1H), 7.44 (dt, J=6.8,1.2Hz, 1H), 7.70 (d, J=8.1Hz, 1H), 7.75 (dd, J=9.0,5.1Hz, 2H), 7.90 (d, J=8.5Hz, 1H), 7.96 (dd, J=6.8,2.0Hz, 2H).
Under room temperature, with 3-methoxyl group-4-[N '-(2-aminomethyl phenyl) urea groups] phenylacetic acid (333mg, 0.106mmol), 4-[(2S, 4S)-and 4-(2-naphthoxy)-2-pyrrolidinyl] methoxyl methyl benzoate (400mg, 106mmol), EDCHCl (305mg, 1.59mmol) and DMAP (194mg, 1.59mmol) mixture in DMF (10ml) stirred 3 days.In this mixture impouring frozen water, use ethyl acetate extraction, with the extract of frozen water and salt water washing merging.After dried over sodium sulfate, the vacuum concentration extract.Residue is through silica gel column chromatography [100g; n-hexane/ethyl acetate (1/1) chloroform/methanol (50/1)]; obtain 4-[(2S; 4S)-and 1-[3-methoxyl group-4-[N '-(2-aminomethyl phenyl) urea groups] phenyl acetyl]-4-(2-naphthoxy)-2-pyrrolidinyl] methoxyl methyl benzoate (520mg; 73%), is the light brown amorphous substance. 1H-NMR(CDCl 3)δ2.28(s，3H)，2.29(m，1H)，2.55(d，J＝14.2Hz，1H)，3.60(d，J＝3.4Hz，2H)，3.66(d，J＝3.7Hz，3H)，3.68-4.00(m，5H)，4.05-4.67(m，3H)，5.09(br，1H)，6.61(s，1H)，6.77(m，2H)，6.87(d，J＝8.8Hz，1H)，6.94-7.54(m，8H)，7.68-8.09(m，8H)；MS(ESI)m/z?674(M ++1)。
To 4-[(2S; 4S)-and 1-[3-methoxyl group-4-[N '-(2-aminomethyl phenyl) urea groups] phenyl acetyl]-4-(2-naphthoxy)-2-pyrrolidinyl] (415mg adds 0.25N sodium hydroxide (4.9ml) to methoxyl methyl benzoate in the solution of THF 0.616mmol) (4.9ml).After stirring 3 days under the room temperature,, extract with chloroform-methanol (10/1) with this mixture of 1N HCl acidify.Extract and vacuum concentration through the dried over sodium sulfate merging.Residue is through TLC[chloroform/methanol (10/1)] purification, obtain 123 (180mg, 44%), be colorless amorphous substance.MW?659.73?IR(KBr)3354，2937，1685，1601，1533，1255?cm -1； 1H-NMR(DMSO-d 6)δ2.24(s，3H)，2.25-2.43(m，2H)，3.65(s，2H)，3.81(s，3H)，3.83(m，1H)，4.05-4.70(m，4H)，5.21-5.33(br，1H)，6.76(d，J＝7.3Hz，1H)，6.86-7.35(m，9H)，7.44(t，J＝7.3Hz，1H)，7.76-7.89(m，6H)，8.01(d，J＝8.3Hz，1H)，8.48(s，1H)，8.56(s，1H)；MS(FAB)m/z660(M ++1)。Embodiment 1164-[(2S, 4S)-1-[4-[N '-(2-chlorophenyl) urea groups]-3-methoxyphenyl acetyl group]-4-(2-naphthoxy)-2-pyrrolidinyl] methoxybenzoic acid
Figure A0080951003381
Under room temperature, to 4-[N '-(2-chlorophenyl) urea groups]-3-methoxybenzene guanidine-acetic acid (310mg, 0.93mmol), 4-[(2S, 4S)-and 4-(2-naphthoxy)-2-pyrrolidinyl] methoxyl methyl benzoate (350mg, 0.93mmol), EDCHCl (267mg, 1.40mmol) and DMAP (171mg, 1.40mmol) mixture in DMF (10ml) stirred 3 days.In this mixture impouring frozen water, use ethyl acetate extraction.Extract with frozen water and salt water washing merging.After dried over sodium sulfate, the vacuum concentration extract.Residue is through silica gel column chromatography [100g; n-hexane/ethyl acetate (1/1) chloroform/methanol (50/1)]; obtain 4-[(2S; 4S)-1-[4-[N '-(2-chlorophenyl) urea groups]-3-methoxyphenyl acetyl group]-4-(2-naphthoxy)-2-pyrrolidinyl] methoxyl methyl benzoate (450mg; 68%), is the light brown amorphous substance. 1H-NMR(CDCl 3)δ2.32(m，1H)，2.58(d，J＝14.5Hz，1H)，3.63(d，J＝2.7Hz，1H)，3.70(s，3H)，3.86(s，3H)，3.84-3.95(m，2H)，4.15-4.64(m，4H)，5.11(br，1H)，6.79-7.06(m，7H)，7.21-7.46(m，7H)，7.66-7.77(m，3H)，7.92(d，J＝8.8Hz，1H)，7.97(m，1H)，8.17(d，J＝8.4Hz，1H)；MS(ESI)m/z694(M ++1)，696(M ++3)。
To 4-[(2S; 4S)-1-[4-[N '-(2-chlorophenyl) urea groups]-3-methoxyphenyl acetyl group]-4-(2-naphthoxy)-2-pyrrolidinyl] (381mg adds 0.25N sodium hydroxide (4.3ml) to methoxyl methyl benzoate in the solution of THF 0.535mmol) (4.3ml).After stirring 3 days under the room temperature,, extract with chloroform-methanol (10/1) with this mixture of 1N HCl acidify.Extract and vacuum concentration through the dried over sodium sulfate merging.Residue is through TLC[chloroform/methanol (10/1)] purification, obtain 124 (140mg, 39%), be colorless amorphous substance.MW 680.15 IR (KBr) 3323,2935,1704,1601,1529,1529,1508 cm -1 1H-NMR (DMSO-d 6) δ 2.27-2.49 (m, 2H), 3.65 (s, 2H), 3.81 (s, 3H), 3.83 (m, 1H), 4.05-4.71 (m, 4H), 5.30 (br, 1H), 6.77 (d, J=7.8Hz, 1H), 6.87-7.16 (m, 4H), 7.14 (dd, J=8.8,2.2Hz, 1H), 7.27 (t, J=7.3Hz, 1H), 7.29-7.46 (m, 4H), 7.76-7.86 (m, 5H), 7.96 (d, J=8.3Hz, 1H), 8.08 (dd, J=8.3,1.2Hz, 1H), 8.90 (s, 1H), 8.93 (s, 1H); MS (FAB) m/z 680 (M ++ 1), 682 (M ++ 3); C 38H 34ClN 3O 71H 2The analytical calculation value of O: C, 65.37; H, 5.20; N, 6.02.Measured value: C, 65.43; H, 5.11; N, 5.93.Embodiment 1172-[(2S, 4S)-1-[3-methoxyl group-4-[N '-(2-aminomethyl phenyl) urea groups] phenyl acetyl]-4-(2-naphthoxy)-2-pyrrolidinyl] methoxyl group-5-pyridine carboxylic acid
Under room temperature, blanket of nitrogen, to (2S, 4R)-1-tert-butoxycarbonyl-4-hydroxyl-2-pyrrolidinyl methyl formate (4.22g, 17.2mmol), beta naphthal (2.73g, 18.9mmol) and triphenyl phasphine (4.96g, the middle DIAD (3.72ml.18.9mmol) of adding that stirs the mixture of THF 18.9mmol) (80ml).After stirring is spent the night, this mixture of vacuum concentration.Residue is through silica gel column chromatography [600g, chloroform/ethyl acetate (10/1)], obtains that (2S, 4S)-1-tert-butoxycarbonyl-4-(2-naphthoxy)-2-pyrrolidinyl methyl formate (5.37g), it need not to be further purified and is used for next step reaction.
Under room temperature, to (2S, 4S)-add in the agitating solution of the THF (116ml) of 1-tert-butoxycarbonyl-4-(2-naphthoxy)-2-pyrrolidinyl methyl formate (5.37g) the 0.25N sodium hydroxide (116ml, 29.0mmol).The mixture stirring that obtains is spent the night.Except that after desolvating,, use chloroform extraction by adding this mixture of 1N HCl acidify.Extract with the salt water washing merges also evaporates through dried over sodium sulfate.Residue is recrystallization from normal hexane-chloroform, obtain (2S, 4S)-1-tert-butoxycarbonyl-4-(2-naphthoxy)-2-pyrrolidinyl formic acid (4.44g, 85% (two steps)), be white powder. 1H-NMR (DMSO-d 6) δ 1.37 and 1.41 (s, 9H, amide isomers), 2.26 (d, J=13.9Hz, 1H), 2.65 (m, 1H), 3.47 (d, J=11.5Hz, 1H), 3.81 (m, 1H), 4.30 (m, 1H), 5.14 (m, 1H), 7.02-7.86 (m, 7H).
In 0 ℃, to (2S, 4S)-(1.12g adds BH in THF 3.13mmol) (30ml) agitating solution to 1-tert-butoxycarbonyl-4-(2-naphthoxy)-2-pyrrolidinyl formic acid 3DMS (0.63ml, 6.3mmol).This mixture is warming up to room temperature immediately, then in 50 heating 1.5 hours.After being cooled to room temperature, in 0 ℃, by adding this mixture of entry quencher.Use ethyl acetate extraction.Extract with the salt water washing merges also evaporates through dried over sodium sulfate.Residue is through silica gel column chromatography [50g, chloroform/methanol (50/1)], obtain (2S, 4S)-1-tert-butoxycarbonyl-4-(2-naphthoxy)-2-pyrrolidinyl methanol (1.10g, 100%), be faint yellow oily thing. 1H-NMR(CDCl 3)δ1.48(s，9H)，2.45(m，1H)，3.58-4.80(m，4H)，5.01(br，1H)，7.04-7.99(m，7H)。
Under room temperature, blanket of nitrogen, to (2S, 4S)-1-tert-butoxycarbonyl-4-(2-naphthoxy)-2-pyrrolidinyl methanol (484mg, 1.41mmol), 2-hydroxyl-5-pyridine carboxylic acid methyl ester (216mg, 1.41mmol) and triphenyl phasphine (370mg, 1.41mmol) the stirring the mixture of THF (15ml) in add DIAD (0.28ml, 1.41mmol).This mixture stirring is spent the night.Except that after desolvating, the residue that obtains is through silica gel column chromatography [50g, n-hexane/ethyl acetate (2/1)], obtain 2-[(2S, 4S)-and 1-tert-butoxycarbonyl-4-(2-naphthoxy)-2-pyrrolidinyl] methoxypyridine-5-methyl formate (170mg, 25%), be colorless oil. 1H-NMR(CDCl 3)δ1.47(s，9H)，2.37(m，1H)，2.46(d，J＝14.2Hz，1H)，3.71-4.00(m，2H)，3.89(s，3H)，4.30-4.56(m，2H)，4.74(dd，J＝9.8，4.6Hz，1H)，5.06(br，1H)，6.70(d，J＝8.8Hz，2H)，7.05-7.09(m，2H)，7.33(t，J＝6.9Hz，1H)，7.42(t，J＝6.9Hz，1H)，7.67-7.75(m，3H)，8.09(d，J＝8.8Hz，1H)，8.77(d，J＝2.2Hz，1H)。
Under room temperature, to 5-carboxymethyl-2-[(2S, 4S)-and 1-tert-butoxycarbonyl-4-(2-naphthoxy)-2-pyrrolidinyl] (170mg adds TFA (2ml) to methoxypyridine in the agitating solution of dichloromethane 0.36mmol) (5ml).Stir after 2 hours, this mixture of vacuum concentration dilutes it with dichloromethane, by adding the alkalization of 1N sodium hydroxide.Reactant mixture with the dichloromethane extraction merging.With salt water washing organic layer, through dried over sodium sulfate and concentrated.Residue is through TLC[chloroform/methanol (10/1)] purification, obtain 2-[(2S, 4S)-and 4-(2-naphthoxy)-2-pyrrolidinyl] methoxypyridine-5-methyl formate (107mg, 80%), be colorless oil. 1H-NMR(CDCl 3)δ1.95(m，1H)，2.27(br，1H)，2.46(m，1H)，3.19(dd，J＝12.2，4.9Hz，1H)，3.41(d，J＝12.2Hz，1H)，3.65(m，1H)，3.89(s，3H)，4.58(m，2H)，5.00(br，1H)，6.77(d，J＝8.8Hz，1H)，7.06(br，1H)，7.11(dd，J＝8.8，2.7Hz，1H)，7.31-7.45(m，2H)，7.69-7.76(m，3H)，8.13(dd，J＝8.8，2.4Hz，1H)，8.78(d，J＝2.2Hz，1H)。
Under room temperature, with 3-methoxyl group-4-[N '-(2-aminomethyl phenyl) urea groups] phenylacetic acid (89mg, 0.283mmol), 2-[(2S, 4S)-and 4-(2-naphthoxy)-2-pyrrolidinyl] methoxypyridine-5-methyl formate (107mg, 0.78mmol), EDCHCl (81mg, 0.425mmol) and DMAP (52mg, 0.425mmol) mixture in DMF (3ml) stirred 18 hours.In this mixture impouring frozen water, use ethyl acetate extraction, with the extract of frozen water and salt water washing merging.After dried over sodium sulfate, the vacuum concentration extract.Residue is through TLC[chloroform/methanol (10/1)] purification; obtain 2-[(2S; 4S)-and 1-[3-methoxyl group-4-[N '-(2-aminomethyl phenyl) urea groups] phenyl acetyl]-4-(2-naphthoxy)-2-pyrrolidinyl] methoxyl group-5-pyridine carboxylic acid methyl ester (193mg, 100%), be colorless amorphous substance. 1H-NMR (CDCl 3) δ 2.27 (d, J=3.2Hz, 3H), 2.30 (m, 1H), 2.49 (dd, J=14.2,2.0Hz, 1H), 3.60 (d, J=3.9Hz, 1H), 3.67 (d, J=5.9Hz, 3H), 3.81 (s, 1H), 3.85 (s, 1H), 3.88 and 3.91 (s, 3H, amide isomers), 3.95 (m, 1H), 4.02-5.09 (m, 4H), 6.67 (d, J=8.8Hz, 1H), and 6.73-7.13 (m, 3H), 7.20-7.45 (m, 7H), 7.53 (t, J=7.8Hz, 1H), 7.67-7.77 (m, 3H), 8.02-8.84 (m, 3H).For hydrochlorate: a kind of light brown amorphous substance.IR(KBr)3346，2951，1720，1601，1533，1281?cm -1；MS(FAB)m/z?675?(M ++1)。
To 2-[(2S; 4S)-and 1-[3-methoxyl group-4-[N '-(2-aminomethyl phenyl) urea groups] phenyl acetyl]-4-(2-naphthoxy)-2-pyrrolidinyl] (158mg adds 0.25N sodium hydroxide (1.8ml) to methoxyl group-5-pyridine carboxylic acid methyl ester in the solution of THF 0.23mmol) (1.8ml).After stirring 22 hours under the room temperature, with 1N HCl this mixture that neutralizes, with chloroform-methanol (10/1) extraction.Extract and vacuum concentration through the dried over sodium sulfate merging.Residue is through TLC[chloroform/methanol (5/1)] purification, obtain 125 (51mg, 34%), be colourless amorphous solid.MW 660.72 IR (KBr) 3354,2956,1601,1533,1255,1022 cm -1 1H-NMR (DMSO-d 6) δ 2.24 (s, 3H), 3.30-5.32 (m, 13H), 6.72-8.82 (m, 19H); MS (FAB) m/z 661 (M ++ 1); C 38H 36N 4O 70.5EtOH1H 2The analytical calculation value of O: C, 66.75; H, 5.89; N, 7.98.Measured value: C, 66.39; H, 5.55; N, 7.66.Embodiment 1184-[5-(R)-benzyloxymethyl-1-[4-[N '-(2-aminomethyl phenyl) urea groups] phenyl acetyl]-2-(S)-pyrrolidinyl methoxyl group] benzoic acid
In-78 ℃, (5.0g adds chlorination pi-allyl magnesium and (1.0M in ether, 30.5ml.30.5mmol), stirs down and makes the mixture that obtains be warmed to room temperature gradually in the agitating solution of THF 30.5mmol) (100ml) to benzyl-(S)-glycidyl ether.In this mixture impouring water, vacuum concentration is used chloroform extraction then.Through anhydrous sodium sulfate drying organic layer and vacuum concentration.Residue as eluant, obtains 1-benzyloxy-2-(R)-hydroxyl-5-hexene (2.18g, 35%) with hexane-ethyl acetate (5: 1) through silica gel column chromatography, is colorless oil. 1H-NMR (CDCl 3) δ 1.52-1.60 (m, 2H), 2.11-2.25 (m, 2H), 2.34 (d, J=3.2Hz, 1H), 3.35 (dd, J=9.6,8.0Hz, 1H), 3.52 (dd, J=9.6,3.2Hz, 1H), and 3.84-3.86 (m, 1H), 4.57 (s, 2H), 4.96-5.07 (a plurality of m, 2H), 5.78-5.88 (m, 1H), 7.29-7.38 (m, 5H); MS (ESI) m/z224 (M ++ NH 4 +).
Under room temperature, to 1-benzyloxy-2-(R)-hydroxyl-5-hexene (2.18g, 10.5mmol), triphenyl phasphine (3.32g, 12.7mmol) and phthalimide (1.86g, 12.7mmol) agitating solution in add diisopropyl azo-2-carboxylic acid (2.62ml, 12.7mmol), the mixture that obtains stirred under room temperature spend the night.This mixture of vacuum concentration is used ethyl acetate extraction.Wash organic layer with water, through anhydrous sodium sulfate drying and vacuum concentration.Residue as eluant, obtains the adjacent diformazan acylimino of 1-benzyloxy-2-(S)-benzene-5-hexene (2.95g, 83%) with hexane-ethyl acetate (5: 1) through silica gel column chromatography, is colorless oil. 1H-NMR (CDCl 3) δ 1.76-1.84 (m, 2H), 2.06 (dd, J=14.4,6.8Hz, 2H), 2.12-2.22 (m, 1H), 3.69 (dd, J=10.0,5.6Hz, 1H), 4.00 (t, J=9.6Hz, 1H), 4.46 (d, J=12.0Hz, 1H), 4.53 (d, J=12.0Hz, 1H), 4.51-4.58 (m, 1H), 4.91-4.99 (a plurality of m, 2H), 5.72-5.79 (m, 1H), and 7.21-7.26 (m, 5H), 7.71-7.83 (a plurality of m, 2H) MS (ESI) m/z 336 (M ++ H).
Under room temperature, to the adjacent diformazan acylimino of 1-benzyloxy-2-(S)-benzene-5-hexene (2.95g, (80% in water to add hydrazine hydrate in the agitating solution of ethanol 8.80mmol) (30ml), 460ml, 11.4mmol), stir down, the mixture that obtains was heated 7.5 hours under refluxing.Filter this solution, vacuum concentrated filtrate.In residue impouring sodium bicarbonate aqueous solution, use chloroform extraction.Through the anhydrous sodium sulfate drying organic layer, vacuum concentration obtains 2-(S)-amino-1-benzyloxy-5-hexene (1.90mg, quantitative) then, is colorless oil. 1H-NMR (CDCl 3) δ 1.37-1.55 (a plurality of m, 4H), 2.08-2.19 (m, 2H), 2.99-3.03 (m, 1H), 3.25 (dd, J=9.2,7.6Hz, 1H), 3.45 (dd, J=9.2,4.0Hz, 1H), 4.53 (s, 2H), and 4.94-5.06 (a plurality of m, 2H), 5.76-5.85 (m, 1H), 7.27-7.37 (m, 5H); MS (ESI) m/z 206 (M ++ H), 247 (M ++ H+CH 3CN).
Under room temperature, to 2-(S)-amino-1-benzyloxy-5-hexene (1.89g, 9.21mmol) and triethylamine (1.28ml adds Benzenecarbonyl chloride. (1.07ml in the agitating solution of dichloromethane 9.21mmol) (20ml), 9.21mmol), the mixture that obtains was stirred 23 hours.In this mixture impouring water, use dichloromethane extraction.Wash organic layer with water, through anhydrous sodium sulfate drying, vacuum concentration then.Residue as eluant, obtains N-[2-(S)-(1-benzyloxy)-5-hexenyl with hexane-ethyl acetate (5: 1) through silica gel column chromatography] Benzoylamide (2.67g, 94%), be colourless spicule.Mp 78-79 ℃; 1H-NMR (CDCl 3) δ 1.76-1.82 (m, 2H), 2.11-2.17 (m, 2H), 3.59 (brs, 2H), 4.29-4.35 (m, 1H), 4.54 (dd, J=19.2,12.0Hz, 2H), 4.96-5.05 (a plurality of m, 2H), 5.78-5.89 (m, 1H), 6.39 (d, J=8.0Hz, 1H), 7.27-7.51 (m, 8H), 7.74 (d, J=7.2Hz, 2H); MS (ESI) m/z 310 (M ++ H).
To N-[2-(S)-(1-benzyloxy)-5-hexenyl] Benzoylamide (2.41g, CH 7.79mmol) 3CN-H 2(3: 1, (2.97g 23.4mmol), stirred the mixture that obtains 20 hours disposable adding iodine in agitating solution 40ml) O.With in this mixture impouring sodium thiosulfate solution and vacuum concentration.Use chloroform extraction then.With salt water washing organic layer, through anhydrous sodium sulfate drying, vacuum concentration then.Residue is dissolved in the dichloromethane (30ml), add Bis(tert-butoxycarbonyl)oxide (2.55g, 11.7mmol), triethylamine (1.63ml, 11.7mmol) and N, (180mg's N-dimethyl aminopyridine 1.47mmol), spends the night the mixture stirring that obtains under room temperature.In this mixture impouring water, use dichloromethane extraction.Wash organic layer with water, through dried over sodium sulfate and vacuum concentration.Residue as eluant, obtains N-Boc-2-(S)-benzoyloxy methyl-5-(S)-benzyloxymethyl pyrrolidine (1.27g, 38%) with hexane-ethyl acetate (5: 1) through silica gel column chromatography, is colorless oil. 1H-NMR (CDCl 3) δ 1.41 and 1.49 (s, 9H altogether), 1.85-2.00 (a plurality of m, 4H), 3.33-4.59 (a plurality of m, 8H), 7.26-7.32 (m, 5H), 7.41-7.46 (m, 2H), 7.54-7.57 (m, 1H), 8.02 (d, J=7.6Hz, 2H); MS (ESI) m/z426 (M ++ H), 448 (M ++ Na +).
Under room temperature, to N-Boc-2-(S)-benzoyloxy methyl-5-(S)-benzyloxymethyl pyrrolidine (1.23g, add in the agitating solution of methanol 2.89mmol) (30ml) sodium hydroxide (1.0M in water, 3.47ml, 3.47mmol), the mixture that obtains was stirred 4 hours.With 1N HCl aqueous solution neutralize this mixture and vacuum concentration.Use chloroform extraction then.With salt water washing organic layer, through anhydrous sodium sulfate drying, vacuum concentration then.Residue as eluant, obtains N-Boc-5-(S)-methylol-2-(S)-benzyloxymethyl pyrrolidine (847mg, 91%) with hexane-ethyl acetate (3: 1) through silica gel column chromatography, is colorless oil. 1H-NMR(CDCl 3)δ1.41(s，9H)，1.57(brs，1H)，1.95-1.97(m，2H)，2.05-2.18(m，1H)，3.36(t，J＝8.4Hz，1H)，3.56-3.62(m，2H)，3.67-3.72(m，2H)，3.95(brs，1H)，4.03(brs，1H)，4.51(s，1H)，7.28-7.37(m，5H)；MS(FAB)m/z?322(M ++H)。
Under room temperature, to N-Boc-2-(S)-hydroxymethyl-5-(S)-benzyloxymethyl pyrrolidine (388mg, 1.21mmol), triphenyl phasphine (380mg, 1.45mmol) and 4-methyl hydroxybenzoate (220mg, 1.45mmol) agitating solution in add diisopropyl azo-2-carboxylic acid (200ml, 1.45mmol), the mixture stirring that obtains is spent the night.This mixture of vacuum concentration is used ethyl acetate extraction.Wash organic layer with water, through anhydrous sodium sulfate drying, vacuum concentration then.Residue as eluant, obtains 4-[2-(S)-(N-Boc-5-(S)-benzyloxymethyl) pyrrolidinyl methoxyl group with hexane-ethyl acetate (3: 1) through silica gel column chromatography] essence of Niobe (462mg, 84%), be colorless oil. 1H-NMR (CDCl 3) δ 1.40 and 1.48 (s, 9H altogether), 1.98-2.13 (m, 4H), 3.83 and 3.85 (s, 3H), 3.33-4.25 (a plurality of m, 4H), 4.47-4.59 (m, 2H), 6.91-6.96 (m, 2H), 7.26-7.34 (m, 5H), 7.95-7.98 (m, 2H); MS (FAB) m/z 456 (M ++ H), 478 (M ++ Na +).
Under room temperature, to 4-[2-(S)-(N-Boc-5-(S)-benzyloxymethyl) pyrrolidinyl methoxyl group] essence of Niobe (446mg, 0.98mmol) the agitating solution of dichloromethane (10ml) in add trifluoroacetic acid (10ml), the mixture that obtains was stirred 1 hour.This mixture of vacuum concentration in this mixture impouring sodium bicarbonate aqueous solution, is used chloroform extraction then.Wash organic layer with water, through anhydrous sodium sulfate drying, vacuum concentration obtains 4-[2-(S)-(5-(S)-benzyloxymethyl) pyrrolidinyl methoxyl group then] (363mg quantitatively), is yellow oil to essence of Niobe.This product is used for subsequent reaction without being further purified. 1H-NMR (CDCl 3) δ 1.43-1.65 (m, 2H), 1.93-2.07 (m, 3H), 3.36-3.68 (a plurality of m, 4H), 3.89 (s, 3H), 3.86-3.93 (overlapping, 2H), 4.55 (s, 2H), 6.90 (d, J=8.4Hz, 2H), 7.26-7.37 (m, 5H), 7.97 (d, J=8.4Hz, 2H); MS (FAB) m/z 356 (M ++ H).
Under room temperature, to 4-[2-(S)-(5-(S)-benzyloxymethyl) pyrrolidinyl methoxyl group] essence of Niobe (115mg, 0.32mmol), 4-[N '-(2-aminomethyl phenyl) urea groups] phenylacetic acid (92.0mg, 0.32mmol) and N, N-dimethyl aminopyridine (52.0mg, 0.42mmol) the agitating solution of DMF (10ml) in add EDCHCl (81.0mg 0.42mmol), stir the mixture that obtains and spends the night.In reactant mixture impouring water, use chloroform extraction.With salt water washing organic layer, through anhydrous sodium sulfate drying, vacuum concentration then.Residue is through silica gel column chromatography, with chloroform-methanol (20/1) as eluant, obtain 4-[5-(R)-benzyloxymethyl-1-[4-[N '-(2-aminomethyl phenyl) urea groups] the phenyl acetylamino]-2-(S)-pyrrolidinyl methoxyl group] essence of Niobe (169mg, 84%), be colourless amorphous solid. 1H-NMR (CDCl 3) rotamer mixture δ 1.92-2.18 (m, 3H), 2.24 and 2.25 (s, altogether 3H), 2.20-2.31 (overlapping, 1H), 3.39-3.70 (a plurality of m, 4H), 3.87 with 3.89 (s, 3H altogether), 4.17 and 4.18 (s, 2H altogether), 4.30-4.45 (a plurality of m, 2H), 4.53 (s, 2H), 6.43-7.13 (a plurality of m, 9H), 7.20-7.36 (a plurality of m, 7H), and 7.58-7.99 (a plurality of m, 3H); MS (FAB) m/z, 622 (M ++ H).
Under room temperature; to 4-[5-(R)-benzyloxymethyl-1-[4-[N '-(2-aminomethyl phenyl) urea groups] phenyl acetyl]-2-(S)-pyrrolidinyl methoxyl group] essence of Niobe (156mg; 0.24mmol) methanol-THF (1: 5; adding 1.0M sodium hydroxide in agitating solution 12ml) (1.2ml, 1.20mmol).Stir down, the mixture that obtains was heated 7 hours in 80 ℃.In this mixture impouring 1N HCl, use chloroform extraction then.With salt water washing organic layer, through anhydrous sodium sulfate drying, vacuum concentration then.Residue as eluant, obtains 126 (94.0mg, 65%) with chloroform-methanol (5: 1) through silica gel column chromatography, is colourless amorphous solid.MW607.70 1H-NMR (CD 3OD), the mixture δ 1.85-2.35 of rotamer (a plurality of m, 4H), 2.43-2.92 (a plurality of m, 5H), 2.28 (s, 3H), 3.55-4.55 (a plurality of m, 10H), 6.85-7.95 (a plurality of m, 17H); MS (ESI) m/z 630 (M ++ Na).Embodiment 1194-[5-(R)-benzyloxymethyl-1-[4-[N '-(2-chlorophenyl) urea groups]-3-methoxyphenyl acetyl group]-2-(S)-pyrrolidinyl methoxyl group] benzoic acid
Under room temperature, to 4-[2-(S)-(5-(S)-benzyloxymethyl) pyrrolidinyl methoxyl group] essence of Niobe (117mg, 0.33mmol), 4-[N '-(2-chlorophenyl) urea groups]-3-methoxybenzene guanidine-acetic acid (110mg, 0.33mmol) and N, N-dimethyl aminopyridine (50.0mg, 0.40mmol) the agitating solution of DMF (10ml) in add EDCHCl (76.0mg 0.40mmol), stir the mixture that obtains and spends the night.In reactant mixture impouring water, use chloroform extraction.With salt water washing organic layer, through anhydrous sodium sulfate drying, vacuum concentration then.Residue is through silica gel column chromatography; with chloroform-methanol (20: 1) as eluant; obtain 4-[5-(R)-benzyloxymethyl-1-[4-[N '-(2-chlorophenyl) urea groups]-3-methoxyphenyl acetyl group]-2-(S)-pyrrolidinyl methoxyl group] essence of Niobe (189mg; 85%), is colourless amorphous solid. 1H-NMR (CDCl 3), the mixture δ 1.91-2.30 of rotamer (a plurality of m, 4H), 3.39-3.74 (a plurality of m, 3H), 3.73 (s, 2H), 4.15-4.02 (m, 2H), and 4.32-4.44 (m, 1H), 4.54 (s, 2H), and 6.71-7.02 (a plurality of m, 5H), 7.06 (s, 1H), 7.17 (s, 1H), 7.24-7.40 (a plurality of m, 7H), and 7.89-8.22 (a plurality of m, 4H); MS (FAB) m/z 672 (M ++ H).
Under room temperature; to 4-[5-(R)-benzyloxymethyl-1-[4-[N '-(2-chlorophenyl) urea groups]-3-methoxyphenyl acetyl group]-2-(S)-pyrrolidinyl methoxyl group] essence of Niobe (169mg; 0.25mmol) methanol-THF (2: 5; adding 1.0M sodium hydroxide in agitating solution 7ml) (750ml, 0.75mmol).Stir down, the mixture that obtains was heated 2 hours in 80 ℃.In this mixture impouring 1N HCl, use chloroform extraction then.With salt water washing organic layer, through anhydrous sodium sulfate drying, vacuum concentration then.Residue as eluant, obtains 127 (114mg, 69%) with chloroform-methanol (15: 1) through silica gel column chromatography, is colourless amorphous solid.MW 658.14 1H-NMR (CD 3OD), the mixture δ 1.88-2.37 of rotamer (a plurality of m, 4H), 3.51-4.49 (a plurality of m, 8H), 3.64 and 3.73 (s, 3H altogether), 4.86 (s, 2H), 6.85-7.95 (a plurality of m, 16H); MS (ESI) m/z658 (M ++ H), 680 (M ++ Na +); C 36H 36ClN 3O 7H 2The analytical calculation value of O: C, 63.95; H, 5.66; N, 6.21.Measured value: C, 63.65; H, 5.40; N, 5.95.Embodiment 1204-[5-(R)-benzyloxymethyl-1-[4-[N '-(2-bromo phenyl) urea groups]-3-methoxyphenyl acetyl group]-2-(S)-pyrrolidinyl methoxyl group] benzoic acid
Under room temperature, to 4-[2-(S)-(5-(S)-benzyloxymethyl) pyrrolidinyl methoxyl group] essence of Niobe (119mg, 0.34mmol), 4-[N '-(2-bromo phenyl) urea groups]-3-methoxybenzene guanidine-acetic acid (127mg, 0.34mmol) and N, N-dimethyl aminopyridine (50.0mg, 0.40mmol) the agitating solution of DMF (10ml) in add EDCHCl (77.0mg 0.40mmol), stir the mixture that obtains and spends the night.In reactant mixture impouring water, use chloroform extraction.With salt water washing organic layer, through anhydrous sodium sulfate drying, vacuum concentration then.Residue is through silica gel column chromatography; with chloroform-methanol (20: 1) as eluant; obtain 4-[5-(R)-benzyloxymethyl-1-[4-[N '-(2-bromo phenyl) urea groups]-3-methoxyphenyl acetyl group]-2-(S)-pyrrolidinyl methoxyl group] essence of Niobe (217mg; 90%), is colourless amorphous solid. 1H-NMR (CDCl 3) rotamer mixture δ 1.92-2.31 (a plurality of m, 4H), 3.39-3.73 (a plurality of m, 3H), 3.65 (s, 2H), 4.15-4.02 (m, 2H), 4.32-4.44 (m, 1H), 4.54 (s, 2H), 6.71-6.99 (a plurality of m, 5H), 7.04 (s, 1H), 7.11 (s, 1H), 7.22-7.39 (a plurality of m, 7H), and 7.51-8.17 (a plurality of m, 4H); MS (FAB) m/z 716 (M +), 718 (M ++ 2).
Under room temperature; to 4-[5-(R)-benzyloxymethyl-1-[4-[N '-(2-bromo phenyl) urea groups]-3-methoxyphenyl acetyl group]-2-(S)-pyrrolidinyl methoxyl group] essence of Niobe (178mg; 0.25mmol) methanol-THF (2: 5; adding 1.0M sodium hydroxide in agitating solution 7ml) (750ml, 0.75mmol).Stir down, the mixture that obtains was heated 1.5 hours in 80 ℃.In this reactant mixture impouring 1N HCl, use chloroform extraction then.With salt water washing organic layer, through anhydrous sodium sulfate drying, vacuum concentration then.Residue as eluant, obtains 128 (159mg, 91%) with chloroform-methanol (15: 1) through silica gel column chromatography, is colourless amorphous solid.MW 702.59 1H-NMR (CD 3OD), the mixture δ 1.88-2.35 of rotamer (a plurality of m, 4H), 3.51-4.49 (a plurality of m, 8H), 3.64 and 3.72 (s, 3H altogether), 4.87 (s, 2H), 6.65-8.05 (a plurality of m, 16H); MS (ESI) m/z 702 (M +), 704 (M ++ 2); C 36H 36BrN 3O 7H 2The analytical calculation value of O: C, 60.00; H, 5.32; N, 5.83.Measured value: C, 59.66; H, 5.04; N, 5.65.Embodiment 1213-[5-(R)-benzyloxymethyl-1-[4-[N '-(2-aminomethyl phenyl) urea groups]-3-methoxyphenyl acetyl group]-2-(S)-pyrrolidinyl methoxyl group] benzoic acid
Under room temperature, to N-Boc-2-(S)-methylol-5-(S)-benzyloxymethyl pyrrolidine (415mg, 1.29mmol), triphenyl phasphine (410mg, 1.55mmol) and 3-methyl hydroxybenzoate (240mg, 1.55mmol) agitating solution in add diisopropyl azo-2-carboxylic acid (320ml, 1.55mmol), the mixture stirring that obtains is spent the night.This mixture of vacuum concentration, residue as eluant, obtains 3-[2-(S)-(N-Boc-5-(S)-benzyloxymethyl) pyrrolidinyl methoxyl group with hexane-ethyl acetate (3: 1) through silica gel column chromatography then] essence of Niobe (513mg, 87%), is colorless oil. 1H-NMR (CDCl 3) δ 1.40 and 1.46 (s, 9H altogether), and 1.95-2.20 (a plurality of m, 4H), 3.33-3.72 (a plurality of m, 2H), 3.82-4.00 (m, 1H), 3.90 with 3.91 (s, 3H altogether), 4.09-4.21 (m, 3H), and 4.21-4.57 (m, 2H), 7.11-7.15 (m, 1H), 7.26-7.37 (m, 6H), and 7.53-7.65 (m, 2H); MS (ESI) m/z 456 (M ++ H).
Under room temperature, to 3-[2-(S)-(N-Boc-5-(S)-benzyloxymethyl) pyrrolidinyl-methoxyl group] essence of Niobe (501mg, 1.10mmol) the agitating solution of dichloromethane (10ml) in add trifluoroacetic acid (10ml), the mixture that obtains was stirred 1 hour.In this mixture of vacuum concentration and the impouring sodium bicarbonate aqueous solution, use chloroform extraction.Washing organic layer with water,, obtain 3-[2-(S)-(5-(S)-benzyloxymethyl) pyrrolidinyl-methoxyl group through anhydrous sodium sulfate drying and vacuum concentration] (387mg quantitatively), is yellow oil to essence of Niobe.This product is used for subsequent reaction without being further purified. 1H-NMR(CDCl 3)δ1.26-1.65(m，2H)，1.94-2.04(m，3H)，3.37-3.52(m，2H)，3.63-3.66(m，1H)，3.85-3.93(m，1H)，3.91(s，3H)，4.55(s，2H)，7.09-7.11(m，1H)，7.27-7.54(m，6H)，7.54-7.55(m，1H)，7.61-7.63(m，2H)；MS(ESI)m/z?35(M ++H)。
Under room temperature, to 3-[2-(S)-(5-(S)-benzyloxymethyl) pyrrolidinyl methoxyl group] essence of Niobe (140mg, 0.39mmol), 4-[N '-(2-aminomethyl phenyl) urea groups]-3-methoxybenzene guanidine-acetic acid (125mg, 0.39mmol) and N, N-dimethyl aminopyridine (58.0mg, 0.47mmol) the agitating solution of THF (15ml) in add EDCHCl (90.0mg 0.47mmol), stir the mixture that obtains and spends the night.In reactant mixture impouring water, use chloroform extraction.With salt water washing organic layer, through anhydrous sodium sulfate drying, vacuum concentration then.Residue is through silica gel column chromatography; with chloroform-methanol (10: 1) as eluant; obtain 3-[5-(R)-benzyloxymethyl-1-[4-[N '-(2-aminomethyl phenyl) urea groups]-3-methoxyphenyl acetyl group]-2-(S)-pyrrolidinyl methoxyl group] essence of Niobe (256mg; quantitatively), be colourless amorphous solid. 1H-NMR (CDCl 3), the mixture δ 1.67 of rotamer (s, 3H), 1.97-2.40 (a plurality of m, 4H), 3.42-3.85 (a plurality of m, 5H), 3.60 (s, 3H), 3.95 and 3.97 (s, be total to 3H), and 4.15-4.26 (m, 2H), 4.36-4.49 (m, 1H), 4.59 (s, 2H), 632 and 6.36 (s, 1H altogether), 6.75-6.87 (a plurality of m, 2H), 7.20 (brs, 2H), 7.16-7.72 (a plurality of m, 10H), 8.03-8.09 (m, 1H); MS (ESI) m/z652 (M ++ H).
Under room temperature; to 3-[5-(R)-benzyloxymethyl-1-[4-[N '-(2-aminomethyl phenyl) urea groups]-3-methoxyphenyl acetyl group]-2-(S)-pyrrolidinyl methoxyl group] essence of Niobe (185mg; 0.28mmol) methanol-THF (2: 5; adding 1.0M sodium hydroxide in agitating solution 7ml) (860ml, 0.86mmol).Stir down, the mixture that obtains was heated 1 hour in 60 ℃.In reactant mixture impouring 1N HCl, use chloroform extraction then.With salt water washing organic layer, through anhydrous sodium sulfate drying, vacuum concentration then.Residue as eluant, obtains 129 (171mg, 94%) with chloroform-methanol (5: 1) through silica gel column chromatography, is colourless amorphous solid.MW 637.72 1H-NMR (CD 3OD), the mixture δ 1.89-2.37 of rotamer (a plurality of m, 4H), 2.29 (s, 3H), 3.52-4.53 (a plurality of m, 8H), 3.66 and 3.74 (s, 3H altogether), 4.85 (s, 2H), 6.66-7.98 (a plurality of m, 16H); MS (ESI) m/z 638 (M ++ H), 660 (M ++ Na +); C 37H 39N 3O 7H 2The analytical calculation value of O: C, 67.77; H, 6.30; N, 6.41.Measured value: C, 67.40; H, 5.95; N, 6.14.Embodiment 1223-[5-(R)-benzyloxymethyl-1-[4-[N '-(2-chlorophenyl) urea groups]-3-methoxyphenyl acetyl group]-2-(S)-pyrrolidinyl methoxyl group] benzoic acid
Figure A0080951003501
Under room temperature, to 3-[2-(S)-(5-(S)-benzyloxymethyl) pyrrolidinyl methoxyl group] essence of Niobe (118mg, 0.33mmol), 4-[N '-(2-chlorophenyl) urea groups]-3-methoxybenzene guanidine-acetic acid (112mg, 0.33mmol) and N, N-dimethyl aminopyridine (50.0mg, 0.40mmol) the agitating solution of THF (15ml) in add EDCHCl (80.0mg 0.40mmol), stir the mixture that obtains and spends the night.In reactant mixture impouring water, use chloroform extraction.With salt water washing organic layer, through anhydrous sodium sulfate drying, vacuum concentration then.Residue is through silica gel column chromatography, with chloroform-methanol (10: 1) as eluant, obtain 3-[5-(R)-benzyloxymethyl-1-[4-[N '-(2-chlorophenyl) urea groups]-3-methoxyphenyl acetylamino]-2-(S)-pyrrolidinyl methoxyl group] essence of Niobe (226mg, quantitatively), be colourless amorphous solid. 1H-NMR (CDCl 3), the mixture δ 1.99-2.40 of rotamer (a plurality of m, 4H), 3.43-3.92 (a plurality of m, 5H), 3.67 (s, 3H), 3.98 and 4.02 (s, be total to 3H), and 4.18-4.29 (m, 2H), 4.36-4.51 (m, 1H), 4.60 (s, 2H), 6.75-6.92 (a plurality of m, 2H), and 7.01-7.22 (a plurality of m, 4H), 7.29-7.53 (a plurality of m, 9H), and 7.62-8.26 (a plurality of m, 3H); MS (ESI) m/z 672 (M ++ H).
Under room temperature; to 3-[5-(R)-benzyloxymethyl-1-[4-[N '-(2-chlorophenyl) urea groups]-3-methoxyphenyl acetyl group]-2-(S)-pyrrolidinyl methoxyl group] essence of Niobe (169mg; 0.25mmol) methanol-THF (2: 5; adding 1.0M sodium hydroxide in agitating solution 7ml) (760ml, 0.76mmol).Stir down, the mixture that obtains was heated 1.5 hours in 60 ℃.In reactant mixture impouring 1N HCl, use chloroform extraction then.With salt water washing organic layer, through anhydrous sodium sulfate drying, vacuum concentration then.Residue as eluant, obtains 130 (155mg, 94%) with chloroform-methanol (5: 1) through silica gel column chromatography, is colourless amorphous solid.MW658.14 1H-NMR (CD 3OD), the mixture δ 1.89-2.35 of rotamer (a plurality of m, 4H), 3.52-4.53 (a plurality of m, 8H), 3.68 and 3.75 (s, 3H altogether), 4.85 (s, 2H), 6.68-8.03 (a plurality of m, 1 6H); MS (ESI) m/z 658 (M ++ H), 680 (M ++ Na +); C 36H 36ClN 3O 7H 2The analytical calculation value of O: C, 63.95; H, 5.66; N, 6.21.Measured value: C, 64.01; H, 5.38; N, 5.96.Embodiment 1233-[5-(R)-benzyloxymethyl-1-[4-[N '-(2-bromo phenyl) urea groups]-3-methoxyphenyl acetyl group]-2-(S)-pyrrolidinyl methoxyl group] benzoic acid
Under room temperature, to 3-[2-(S)-(5-(S)-benzyloxymethyl) pyrrolidinyl methoxyl group] essence of Niobe (116mg, 0.33mmol), 4-[N '-(2-bromo phenyl) urea groups]-3-methoxybenzene guanidine-acetic acid (124mg, 0.33mmol) and N, N-dimethyl aminopyridine (48.0mg, 0.39mmol) the agitating solution of THF (15ml) in add EDCHCl (75.0mg 0.39mmol), stir the mixture that obtains and spends the night.In reactant mixture impouring water, use chloroform extraction, with salt water washing organic layer, through anhydrous sodium sulfate drying, vacuum concentration then.Residue is through silica gel column chromatography; with chloroform-methanol (10: 1) as eluant; obtain 3-[5-(R)-benzyloxymethyl-1-[4-[N '-(2-bromo phenyl) urea groups]-3-methoxyphenyl acetyl group]-2-(S)-pyrrolidinyl methoxyl group] essence of Niobe (209mg; 89%), is colourless amorphous solid. 1H-NMR (CDCl 3), the mixture δ 1.99-2.37 of rotamer (a plurality of m, 4H), 3.43-3.91 (a plurality of m, 5H), 3.70 (s, 3H), 3.93 and 3.96 (s, be total to 3H), and 4.19-4.28 (m, 2H), 4.37-4.51 (m, 1H), 4.60 (s, 2H), 6.77-7.11 (a plurality of m, 6H), and 7.28-7.74 (a plurality of m, 10H), 7.91-7.95 (a plurality of m, 1H), and 8.20-8.23 (a plurality of m, 1H); MS (ESI) m/z 716 (M +), 718 (M ++ 2).
Under room temperature; to 3-[5-(R)-benzyloxymethyl-1-[4-[N '-(2-bromo phenyl) urea groups]-3-methoxyphenyl acetyl group]-2-(S)-pyrrolidinyl methoxyl group] essence of Niobe (176mg; 0.25mmol) methanol-THF (2: 5; adding 1.0M sodium hydroxide in agitating solution 7ml) (760ml, 0.76mmol).Stir down, the mixture that obtains was heated 1.5 hours in 60 ℃.In reactant mixture impouring 1N HCl, use chloroform extraction then.With salt water washing organic layer, through anhydrous sodium sulfate drying, vacuum concentration then.Residue as eluant, obtains 131 (156mg, 88%) with chloroform-methanol (5: 1) through silica gel column chromatography, is colourless amorphous solid.MW702.59 1H-NMR (CD 3OD), the mixture δ 1.89-2.37 of rotamer (a plurality of m, 4H), 2.29 (s, 3H), 3.52-4.53 (a plurality of m, 8H), 3.68 and 3.75 (s, 3H altogether), 4.85 (s, 2H), 6.67-7.95 (a plurality of m, 16H); MS (ESI) m/z702 (M ++ H), 704 (M ++ Na); C 36H 36BrN 3O 7H 2The analytical calculation value of O: C, 60.00; H, 5.32; N, 5.83.Measured value: C: 59.65; H, 5.02; N, 5.65.Embodiment 1244-[(2S, 4S)-4-methoxyl group-1-[3-methoxyl group-4-[N '-(2-aminomethyl phenyl) urea groups] phenyl acetyl]-the 2-pyrrolidinyl] methoxybenzoic acid
Under room temperature, blanket of nitrogen, to (2S, 4R)-1-tert-butoxycarbonyl-2-tert-butyl diphenyl siloxy methyl-4-hydroxyl pyrrolidine (21.7g, 47.6mmol), acetic acid (3.0ml, 52.4mmol) and triphenyl phasphine (12.5g, the middle DIAD (9.4ml.47.6mmol) of adding that stirs the mixture of THF 52.4mmol) (330ml).After uniform temp stirs 2 hours down, heated these mixture 2 hours in 50 ℃.After being cooled to room temperature, this reactant mixture of vacuum concentration.The residue that obtains is through silica gel column chromatography [1Kg, n-hexane/ethyl acetate (5/1)], obtain (2S, 4S)-4-acetoxyl group-1-tert-butoxycarbonyl-2-tert-butyl diphenyl siloxy crassitude (23.3g, 99%), be colorless oil. 1H-NMR (CDCl 3) δ 1.06 (s, 9H), 1.35 and 1.43 (s, 9H, amide isomers), 1.92 (br, 3H), 2.20-2.45 (m, 2H), 3.31-4.07 (m, 5H), 5.17-5.30 (m, 1H), 7.36-7.44 (m, 6H), 7.65-7.71 (m, 4H).
In 0 ℃, to (2S, 4S)-4-acetoxyl group-1-tert-butoxycarbonyl-2-tert-butyl diphenyl siloxy crassitude (23.3g, 46.9mmol) and acetic acid (6.0ml, 104.8mmol) the stirring the mixture of THF (470ml) in add TBAF (93.8ml, 93.8mmol).Stir after 24 hours this mixture of vacuum concentration.With the residue that ethyl acetate and aqueous ammonium chloride solution dilution obtain, use ethyl acetate extraction.The extract that merges with the salt water washing, with it with dried over sodium sulfate and vacuum concentration.Residue is through silica gel column chromatography [700g, chloroform/ethyl acetate (4/1)], obtain (2S, 4S)-4-acetoxyl group-1-tert-butoxycarbonyl-2-pyrrolidine carbinol (9.70g, 8%), be colorless oil. 1H-NMR(CDCl 3)δ1.47(s，9H)，1.63(m，1H)，1.81(m，1H)，2.07(s，3H)，2.34(m，1H)，3.42(dd，J＝12.7，0.9Hz，1H)，3.62-3.85(m，3H)，4.48(br，1H)，5.20(br，1H)。
Under room temperature, to (2S, 4S)-4-acetoxyl group-1-tert-butoxycarbonyl-2-pyrrolidine carbinol (9.70g, 37.4mmol), right-methyl hydroxybenzoate (5.69g, 37.4mmol) and triphenyl phasphine (10.8g, 41.1mmol) the stirring the mixture of THF (200ml) in add DIAD (8.10ml, 41.1mmol).Stir after 1.5 hours this mixture of vacuum concentration.The residue that obtains obtains 4-[(2S through silica gel column chromatography [700g, chloroform/ethyl acetate (10/1)], 4S)-4-acetoxyl group-1-tert-butoxycarbonyl-2-pyrrolidinyl] methoxyl methyl benzoate (11.8g, 81%), be faint yellow oily thing. 1H-NMR(CDCl 3)δ1.48(s，9H)，2.03(s，3H)，2.27(m，2H)，3.46(m，1H)，3.72(m，1H)，3.88(s，3H)，3.98(t，J＝9.0Hz，1H)，4.21-4.47(m，2H)，5.31(br，1H)，6.96(br，2H)，7.98(d，J＝8.8Hz，2H)。
Under room temperature, to 4-[(2S, 4S)-and 4-acetoxyl group-1-tert-butoxycarbonyl-4-hydroxyl-2-pyrrolidinyl] methoxyl methyl benzoate (7.43g, the potassium carbonate of adding catalytic amount in the agitating solution of methanol 18.9mmol) (150ml).Stir after 1 day this mixture of vacuum concentration.Make the residue recrystallization that obtains by adding chloroform-normal hexane, obtain 4-[(2S, 4S)-1-tert-butoxycarbonyl-4-hydroxyl-2-pyrrolidinyl] methoxyl methyl benzoate (5.76g, 87%), be colorless solid. 1H-NMR(CDCl 3)δ1.46(s，9H)，2.11(m，1H)，2.35(br，1H)，3.27-3.65(m，2H)，3.89(s，3H)，4.07-4.54(m，4H)，6.96(d，J＝6.9Hz，2H)，7.99(d，J＝6.9Hz，2H)。
In 0 ℃, to 4-[(2S, 4S)-1-tert-butoxycarbonyl-4-hydroxyl-2-pyrrolidinyl] methoxyl methyl benzoate (2.10g, and the sodium hydride in the agitating solution of THF 5.98mmol) (60ml) in the oil of adding 60% (359mg, 8.97mmol).Stir after 15 minutes, under uniform temp, (1.20ml 8.97mmol) adds in this mixture, makes the mixture that obtains rise to room temperature with 1 hour with MeI.Then, under room temperature, with the sodium hydride in 60% the oil (359mg, 8.97mmol) and MeI (1.20ml 8.97mmol) adds in this reactant mixture and stirred 14 hours.In this reactant mixture impouring frozen water, use chloroform extraction.Extract with sodium bicarbonate aqueous solution and salt water washing merging.After dried over sodium sulfate, the vacuum concentration extract.Residue obtains 4-[(2S through silica gel column chromatography [50g, n-hexane/ethyl acetate (4/1)], 4S)-1-tert-butoxycarbonyl-4-methoxyl group-2-pyrrolidinyl] methoxyl methyl benzoate (1.32g, 60%), be colorless oil. 1H-NMR(CDCl 3)δ1.48(s，9H)，2.05(m，1H)，2.29(d，J＝14.2Hz，1H)，3.30(s，3H)，3.36-4.38(m，4H)，6.76(br，2H)，7.97(d，J＝8.8Hz，2H)。
Under room temperature, to 4-[(2S, 4S)-and 1-tert-butoxycarbonyl-4-methoxyl group-2-pyrrolidinyl] (2.38g adds TFA (23ml) to methoxyl methyl benzoate in the agitating solution of dichloromethane 3.61mmol) (46ml).Stir after 14 hours this mixture of vacuum concentration.By adding dichloromethane and 1N sodium hydroxide dilution residue and using dichloromethane extraction.With the extract of salt water washing merging, through dried over sodium sulfate, with its vacuum concentration.Residue obtains 4-[(2S through silica gel column chromatography [50g, chloroform/methanol (20/1)], 4S)-4-methoxyl group-2-pyrrolidinyl] methoxyl methyl benzoate (950mg, 99%), be yellow oil. 1H-NMR(CDCl 3)δ2.16(t，J＝5.3Hz，1H)，2.72(s，1H)，2.95(d，J＝6.8Hz，1H)，3.11(d，J＝11.0Hz，1H)，3.26(t，J＝1.9Hz，3H)，3.52(br，1H)，3.84(d，J＝1.7Hz，3H)，3.92(s，1H)，4.00(d，J＝4.1Hz，2H)，6.88(m，2H)，7.94(m，2H)。
Under room temperature, with 3-methoxyl group-4-[N '-(2-aminomethyl phenyl) urea groups] phenylacetic acid (375mg, 1.19mmol), 4-[(2S, 4S)-and 4-methoxyl group-2-pyrrolidinyl] methoxyl methyl benzoate (317mg, 1.19mmol), EDCHCl (342mg, 1.79mmol), HOBt (242mg, 1.79mmol) and triethylamine (0.83ml, 5.95mmol) mixture in DMF (5ml) stirred 13 hours.In this mixture impouring frozen water, use ethyl acetate extraction.Extract with frozen water and salt water washing merging.After dried over sodium sulfate, the vacuum concentration extract.Residue is through silica gel column chromatography [50g; chloroform/acetone (10/1) chloroform/methanol (20/1)]; obtain 4-[(2S; 4S)-and 4-methoxyl group-1-[3-methoxyl group-4-[N '-(2-aminomethyl phenyl) urea groups] phenyl acetyl]-the 2-pyrrolidinyl] methoxyl methyl benzoate (650mg; 98%), is the light brown amorphous solid. 1H-NMR(CDCl 3)δ2.03(m，1H)，2.31(s，3H)，2.32(m，1H)，3.29(d，J＝1.0Hz，3H)，3.57-3.68(m，5H)，3.88(d，J＝1.0Hz，3H)，3.99-4.06(m，2H)，4.46(m，1H)，6.19(m，1H)，6.80(s，1H)，6.81(d，J＝9.0Hz，1H)，6.96-7.19(m，4H)，7.29(m，2H)，7.50(d，J＝6.7Hz，1H)，7.95-7-8.10(m，3H)；MS(ESI)m/z562(M ++1)。
To 4-[(2S; 4S)-and 4-methoxyl group-1-[3-methoxyl group-4-[N '-(2-aminomethyl phenyl) urea groups] phenyl acetyl]-the 2-pyrrolidinyl] (650mg adds 0.25N sodium hydroxide (18.5ml) to methoxyl methyl benzoate in the solution of THF 1.16mmol) (18.5ml).After stirring 12 hours under the room temperature,, extract with chloroform-methanol (10/1) with this mixture of 1N HCl acidify.Extract and vacuum concentration through the dried over sodium sulfate merging.Residue obtains 132 (540mg, 85%) through silica gel column chromatography [50g, chloroform/methanol (20/1)], is colourless amorphous solid.MW 547.60 IR (KBr) 3354,2937,1709,1685,1604,1533,1454 cm -1 1H-NMR (DMSO-d 6) δ 2.11 (m, 2H), 2.25 (s, 3H), 3.22 (s, 3H), 3.49-3.78 (m, 4H), 3.82 with 3.86 (s, 3H, amide isomers), 3.87-4.52 (m, 4H), and 6.71-7.17 (m, 7H), 7.79 (d, J=8.1Hz, 1H), 7.86-8.03 (m, 3H), 8.45-8.57 (m, 2H), 12.64 (br, 1H); MS (ESI) m/z 548 (M ++ 1); C 30H 33N 3O 71Na1.5H 2The analytical calculation value of O: C, 60.29; H, 6.07; N, 7.03.Measured value: C, 59.90; H, 5.59; N, 6.69.Embodiment 1254-[(2S, 4S)-1-[4-[N '-(2-chlorophenyl) urea groups]-3-methoxyphenyl acetyl group]-4-methoxyl group-2-pyrrolidinyl] methoxybenzoic acid
Figure A0080951003551
Under room temperature, with 4-[N '-(2-chlorophenyl) urea groups]-3-methoxybenzene guanidine-acetic acid (398mg, 1.19mmol), 4-[(2S, 4S)-and 4-methoxyl group-2-pyrrolidinyl] methoxyl methyl benzoate (317mg, 1.19mmol), EDCHCl (342mg, 1.79mmol), HOBt (242mg, 1.79mmol) and triethylamine (0.83ml, 5.95mmol) mixture in DMF (5ml) stirred 13 hours.In this mixture impouring frozen water, use ethyl acetate extraction.Extract with frozen water and salt water washing merging.After dried over sodium sulfate, the vacuum concentration extract.Residue is through silica gel column chromatography [50g; chloroform/acetone (10/1) chloroform/methanol (20/1)]; obtain 4-[(2S; 4S)-1-[4-[N '-(2-chlorophenyl) urea groups]-3-methoxyphenyl acetyl group]-4-methoxyl group-2-pyrrolidinyl] methoxyl methyl benzoate (600mg; 87%), is colourless amorphous solid. 1H-NMR(CDCl 3)δ1.99-2.06(m，1H)，2.34(d，J＝13.9Hz，1H)，3.30(s，3H)，3.59(d，J＝7.4Hz，1H)，3.62(d，J＝3.2Hz，2H)，3.83(s，3H)，3.88(s，3H)，4.00-4.18(m，3H)，4.42-4.51(m，2H)，6.82-7.07(m，7H)，7.28(d，J＝8.3Hz，1H)，7.35(dd，J＝7.9，1.5Hz，1H)，7.94-8.00(m，3H)，8.18(d，J＝8.3Hz，1H)；MS(ESI)m/z?582(M ++1)，584(M ++3)。
To 4-[(2S; 4S)-1-[4-[N '-(2-chlorophenyl) urea groups]-3-methoxyphenyl acetyl group]-4-methoxyl group-2-pyrrolidinyl] (600mg adds 0.25N sodium hydroxide (16ml) to methoxyl methyl benzoate in the solution of THF 1.03mmol) (16ml).After stirring 12 hours under the room temperature,, extract with chloroform-methanol (10/1) with this mixture of 1NHCl acidify.Extract and vacuum concentration through the dried over sodium sulfate merging.Residue is through TLC[chloroform/methanol (10/1)] purification, obtain 133 (495mg, 75%), be colourless amorphous solid.MW 568.02 IR (KBr) 3330,3070,2937,1709,1685,1604,1533 cm -1 1H-NMR (DMSO-d 6) δ 2.11 (m, 2H), 3.22 (s, 3H), 3.56-3.78 (m, 4H), 3.81 with 3.85 (s, 3H, amide isomers), and 3.88-4.56 (m, 4H), 6.73 and 6.77 (d, J=8.1Hz, 1H, amide isomers), 6.85 and 6.91 (s, 1H, amide isomers), 7.01-7.07 (s, 3H), 7.28 (t, J=8.1Hz, 1H), 7.43 (d, J=8.1Hz, 1H), 7.85-7.94 (m, 2H), 7.97 (d, J=8.6Hz, 1H), 8.09 (d, J=8.3Hz, 1H), 8.90-8.95 (m, 2H); MS (FAB) m/z 570 (M ++ 1), 572 (M ++ 3).Embodiment 1264-[(2S, 4S)-1-[4-[N '-(2-bromo phenyl) urea groups]-3-methoxyphenyl acetyl group]-4-methoxyl group-2-pyrrolidinyl] methoxybenzoic acid
Under room temperature, with 4-[N '-(2-bromo phenyl) urea groups]-3-methoxybenzene guanidine-acetic acid (451mg, 1.19mmol), 4-[(2S, 4S)-and 4-methoxyl group-2-pyrrolidinyl] methoxyl methyl benzoate (317mg, 1.19mmol), EDCHCl (342mg, 1.79mmol), HOBt (242mg, 1.79mmol) and triethylamine (0.83ml, 5.95mmol) mixture in DMF (5ml) stirred 13 hours.In this mixture impouring frozen water, use ethyl acetate extraction.Extract with frozen water and salt water washing merging.After dried over sodium sulfate, the vacuum concentration extract.Residue is through silica gel column chromatography [50g; chloroform/acetone (10/1)], obtain 4-[(2S, 4S)-1-[4-[N '-(2-bromo phenyl) urea groups]-3-methoxyphenyl acetyl group]-4-methoxyl group-2-pyrrolidinyl] methoxyl methyl benzoate (760mg; 100%), is yellow oil. 1H-NMR(CDCl 3)δ1.99-2.32(m，1H)，2.34(d，J＝13.4Hz，1H)，3.30(s，3H)，3.59(m，1H)，3.63(d，J＝3.2Hz，2H)，3.68(dd，J＝12.2，5.1Hz，1H)，3.81(br，3H)，3.88(s，3H)，3.91-4.16(m，2H)，4.49-4.51(m，2H)，6.82-7.15(m，7H)，7.31(t，J＝8.1Hz，1H)，7.52(d，J＝8.1Hz，1H)，7.93-8.00(m，3H)，8.14(d，J＝8.3Hz，1H)；MS(ESI)m/z626(M ++1)，628(M ++3)。
To 4-[(2S, 4S)-1-[4-[N '-(2-bromo phenyl) urea groups] phenyl acetyl]-4-methoxyl group-2-pyrrolidinyl] (760mg adds 0.25N sodium hydroxide (19ml) to methoxyl methyl benzoate in the solution of THF 1.19mmol) (19ml).After stirring 12 hours under the room temperature,, extract with chloroform-methanol (10/1) with this mixture of 1N HCl acidify.Extract and vacuum concentration through the dried over sodium sulfate merging.Residue obtains 134 (580mg, 78%) through silica gel column chromatography [50g, chloroform/methanol (20/1)], is colourless amorphous solid.MW 612.47 IR (KBr) 3330,2935,1709,1685,1604,1529,1434 cm -1 1H-NMR (DMSO-d 6) δ 2.11 (m, 2H), 3.22 (s, 3H), 3.58-3.78 (m, 4H), 3.81 and 3.86 (s, 3H, amide isomers), 3.92-4.52 (m, 4H), 6.72 (d, J=8.6Hz, 1H), 6.77 (d, J=8.3Hz, 1H), 6.85 and 6.91 (s, 1H, amide isomers), 6.97 (t, J=7.1Hz, 1H), 7.02 and 7.06 (d, J=8.6Hz, 2H, amide isomers), 7.32 (t, J=7.3Hz, 1H), 7.59 (dd, J=8.1,1.0Hz, 1H), 7.94 (dd, J=8.1,1.2Hz, 2H), 7.95-7.98 (m, 2H), 8.74 (s, 1H), 8.94 (s, 1H), 12.63 (br, 1H); MS (FAB) m/z 612 (M ++ 1), 614 (M ++ 3).Embodiment 1274-[(4R)-and methoxyl group-1-[3-methoxyl group-4-[N '-(2-aminomethyl phenyl) urea groups] phenyl acetyl]-(2S)-and the pyrrolidinyl methoxyl group] benzoic acid
Under room temperature, to 1-(tert-butoxycarbonyl)-(4R)-methoxyl group-(2S)-(2.87g adds BH in the agitating solution of THF 11.7mmol) (25ml) to pyrrolidinyl formic acid 3(1.66ml's DMS 17.5mmol), spends the night this reactant mixture stirring under room temperature.Evaporate this mixture, residue is dissolved in the dichloromethane.Water, this solution of salt water washing are through dried over sodium sulfate and evaporation.Residue is through purification by silica gel column chromatography, and (50/1, v/v) eluting obtains 1-(tert-butoxycarbonyl)-(4R)-methoxyl group-(2S)-dried meat ammonia alcohol (1.79g, 66%), is colorless oil with chloroform-methanol. 1H-NMR(CDCl 3)δ1.47(s，9H)，1.69-1.73(m，1H)，2.12-2.17(m，1H)，3.31(s，3H)，3.37-3.40(m，1H)，3.53-3.62(m，2H)，3.68-3.73(m，1H)，3.83-3.87(m，1H)，4.04-4.07(m，1H)，4.90-4.92(m，1H)；MS(FAB)m/z?232(M ++1)。
To 4-methyl hydroxybenzoate (1.18g, 7.76mmol), 1-(tert-butoxycarbonyl)-(4R)-methoxyl group-(2S)-dried meat ammonia alcohol (1.79g, 7.74mmol) and triphenyl phasphine (2.44g, 9.30mmol) the agitating solution of THF (30ml) in add DIAD (1.83ml, 9.29mmol), with this reaction mixture refluxed heating 5 hours.After being cooled to room temperature, evaporate this mixture.Through the filtered through silica gel residue, and usefulness toluene-acetone (5: 1, v/v) as eluant, obtain crude product.This crude product is dissolved in the dichloromethane (20ml).In this solution, add TFA (20ml), under room temperature, this reactant mixture stirring is spent the night.This mixture of vacuum concentration, by saturated sodium bicarbonate make be alkalescence.With this mixture of chloroform extraction, use the salt water washing, dry and evaporation through potassium carbonate.Residue is through purification by silica gel column chromatography, with chloroform-methanol (30: 1-30: 2, v/v) as eluant, obtain 4-[(4R)-methoxyl group-(2S)-pyrrolidinyl methoxyl group] essence of Niobe (1.67g, 81% (two steps)), be rufous grease. 1H-NMR(CDCl 3)δ1.65-1.72(m，1H)，1.89(bs，1H)，2.05-2.22(m，1H)，2.95-3.15(m，2H)，3.31(s，3H)，3.69-3.76(m，1H)，3.88(s，3H)，3.91-4.06(m，3H)，6.89-6.92(m，2H)，7.96-7.98(m，2H)；MS(FAB)m/z266(M ++1)。
Under room temperature, with 3-methoxyl group-4-[N '-(2-aminomethyl phenyl) urea groups] phenylacetic acid (470mg, 1.50mmol), 4-[(4R)-methoxyl group-(2S)-pyrrolidinyl methoxyl group] essence of Niobe (396mg, 1.49mmol), EDCHCl (343mg, 1.79mmol), HOBt (242mg, 1.79mmol) and triethylamine (250ml, 1.79mmol) mixture in THF (10ml) stirs and to spend the night.This mixture of dilute with water is used ethyl acetate extraction.With salt water washing extract, through dried over sodium sulfate and evaporation.Residue is through purification by silica gel column chromatography; with chloroform-methanol (100: 1; v/v) as eluant; obtain 4-[(4R)-methoxyl group-1-[3-methoxyl group-4-[N '-(2-aminomethyl phenyl) urea groups] phenyl acetyl]-(2S)-and the pyrrolidinyl methoxyl group] essence of Niobe (822mg; 98%), is the white foam thing. 1H-NMR (CDCl 3) δ 2.14-2.24 (m, 2H), 2.27 (s, 3H), 3.25 (s, 3H), 3.51 (s, 3H), 3.58-3.73 (m, 4H), 3.88 (s, 3H), 3.98-4.09 (m, 2H), 4.40-4.53 (m, 2H), (6.67-7.29 a plurality of m, 9H altogether), and 7.57-7.59 (m, 1H), 7.91-7.93 (m, 2H), 8.04-8.06 (m, 1H); MS (FAB) m/z 562 (M ++ 1).
To 4-[(4R)-methoxyl group-1-[3-methoxyl group-4-[N '-(2-aminomethyl phenyl) urea groups] phenyl acetyl]-(2S)-and the pyrrolidinyl methoxyl group] essence of Niobe (517mg; 0.92mmol) the agitating solution of THF (5ml) in add 0.5N sodium hydroxide (5ml), with this reaction mixture refluxed heating 3 hours.After being cooled to room temperature, in this mixture impouring ice-1N HCl, the precipitation that produces is collected in decompression.The crude product solid is recrystallization purifying from methanol-chloroform-IPE, obtains 135 (144mg, 29%), is white crystalline powder.MW 547.60 mp 112-115 ℃; 1H-NMR (DMSO-d 6) δ 2.04-2.17 (m, 2H), 2.25 (s, 3H), 3.21 (s, 3H), 3.56-3.75 (m, 4H), 3.79 (s, 3H), 4.04-4.35 (m, 4H), 6.73-7.17 (a plurality of m, 7H altogether), 7.79-7.81 (m, 1H), 7.87-7.89 (m, 2H), 7.99-8.01 (m, 1H), 8.47 (s, 1H), 8.55 (s, 1H), 12.63 (bs, 1H); MS (FAB) m/z 548 (M ++ 1); C 30H 33N 3O 71/4H 2The analytical calculation value of O: C, 65.26; H, 6.12; N, 7.61.Measured value: C, 65.36; H, 6.45; N, 7.24.Embodiment 1284-[1-[4-[N '-(2-fluoro phenyl) urea groups]-3-methoxyphenyl acetyl group]-(4R)-methoxyl group-(2S)-the pyrrolidinyl methoxyl group] benzoic acid
Figure A0080951003601
Under room temperature, with 4-[N '-(2-fluoro phenyl) urea groups]-3-methoxybenzene guanidine-acetic acid (476mg, 1.50mmol), 4-[(4R)-methoxyl group-(2S)-pyrrolidinyl methoxyl group] essence of Niobe (397mg, 1.50mmol), EDCHCl (344mg, 1.79mmol), HOBt (243mg, 1.80mmol) and triethylamine (250ml, 1.79mmol) mixture in THF (10ml) stirs and to spend the night.This mixture of dilute with water is used ethyl acetate extraction.With salt water washing extract, through dried over sodium sulfate and evaporation.Residue is through purification by silica gel column chromatography; with chloroform-methanol (100: 1; v/v) as eluant; obtain 4-[1-[4-[N '-(2-fluoro phenyl) urea groups]-3-methoxyphenyl acetyl group]-(4R)-methoxyl group-(2S)-the pyrrolidinyl methoxyl group] essence of Niobe (806mg; 95%), is the weak yellow foam thing. 1H-NMR (CDCl 3) δ 2.14-2.37 (m, 2H), 3.28 (s, 3H), 3.44 (s, 3H), 3.48-3.74 (m, 4H), 3.88 (s, 3H), 4.02-4.15 (m, 2H), 4.43-4.58 (m, 2H), 6.63-7.10 (a plurality of m, 7H altogether), 7.68-7.73 (m, 1H), 7.89-8.02 (m, 4H), and 8.16-8.20 (m, 1H); MS (FAB) m/z 566 (M ++ 1).
To 4-[1-[4-[N '-(2-fluoro phenyl) urea groups]-3-methoxyphenyl acetyl group]-(4R)-methoxyl group-(2S)-the pyrrolidinyl methoxyl group] essence of Niobe (491mg; 0.87mmol) the agitating solution of THF (5ml) in add 0.5N sodium hydroxide (5ml), with this reaction mixture refluxed heating 3 hours.After being cooled to room temperature, in this mixture impouring ice-1N HCl, the precipitation that produces is collected in decompression.The crude product solid is recrystallization purifying from methanol-chloroform-IPE, obtains 136 (173mg, 36%), is white crystalline powder.MW 551.56 mp 111-116 ℃; 1H-NMR (DMSO-d 6) δ 2.08-2.17 (m, 2H), 3.21 (s, 3H), 3.56-3.73 (m, 4H), 3.78 (s, 3H), 4.04-4.33 (m, 4H), 6.74-7.22 (a plurality of m, 7H altogether), 7.87-7.89 (m, 2H), 7.99-8.01 (m, 1H), 8.16-8.20 (m, 1H), 8.70 (s, 1H), 9.18 (s, 1H), 12.64 (brs, 1H); MS (FAB) m/z 552 (M ++ 1); C 29H 30FN 3O 70.15H 2The analytical calculation value of O: C, 62.84; H, 5.51; F, 3.43; N, 7.58.Measured value: C, 63.08; H, 5.83; F, 3.30; N, 7.15.Embodiment 1294-[1-[4-[N '-(2-chlorophenyl) urea groups]-3-methoxyphenyl acetyl group]-(4R)-methoxyl group-(2S)-the pyrrolidinyl methoxyl group] benzoic acid
Under room temperature, with 4-[N '-(2-chlorophenyl) urea groups]-3-methoxybenzene guanidine-acetic acid (460mg, 1.37mmol), 4-[(4R)-methoxyl group-(2S)-pyrrolidinyl methoxyl group] essence of Niobe (365mg, 1.38mmol), EDCHCl (316mg, 1.65mmol), HOBt (223mg, 1.65mmol) and triethylamine (230ml, 1.65mmol) mixture in THF (10ml) stirs and to spend the night.This mixture of dilute with water is used ethyl acetate extraction.With salt water washing extract, through dried over sodium sulfate and evaporation.Residue is through purification by silica gel column chromatography; with chloroform-methanol (100: 1; v/v) as eluant; obtain 4-[1-[4-[N '-(2-chlorophenyl) urea groups]-3-methoxyphenyl acetyl group]-(4R)-methoxyl group-(2S)-the pyrrolidinyl methoxyl group] essence of Niobe (801mg; q.y.), be the white foam thing. 1H-NMR (CDCl 3) δ 2.13-2.36 (m, 2H), 3.27 (s, 3H), 3.58 (s, 3H), 3.61-3.73 (m, 4H), 3.88 (s, 3H), 4.06-4.14 (m, 2H), 4.43-4.56 (m, 2H), 6.70-6.99 (a plurality of m, 5H altogether), 7.23-7.42 (m, 4H), 7.90-8.00 (m, 3H), and 8.17-8.20 (m, 1H); MS (FAB) m/z 582 (M ++ 1).
To 4-[1-[4-[N '-(2-chlorophenyl) urea groups]-3-methoxyphenyl acetyl group]-(4R)-methoxyl group-(2S)-the pyrrolidinyl methoxyl group] essence of Niobe (541mg; 0.93mmol) the agitating solution of THF (5ml) in add 0.5N sodium hydroxide (5ml), with this reaction mixture refluxed heating 3 hours.After being cooled to room temperature, in this mixture impouring ice-1N HCl, the precipitation that produces is collected in decompression.The crude product solid is recrystallization purifying from methanol-chloroform-IPE, obtains 137 (281mg, 53%), is white crystalline powder.MW 568.02 mp 116-119 ℃; 1H-NMR (DMSO-d 6) δ 2.08-2.17 (m, 2H), 3.21 (s, 3H), 3.56-3.73 (m, 4H), 3.79 (s, 3H), 4.04-4.33 (m, 4H), 6.75 (d, J=8.3Hz, 1H), 6.87 (s, 1H), 7.02 (d, J=8.3Hz, 3H), 7.28 (t, J=7.8Hz, 1H), 7.44 (d, J=7.8Hz, 1H), 7.87-7.89 (m, 2H), 7.96 (d, J=8.3Hz, 1H), 8.10 (d, J=8.3Hz, 1H), 8.89 (s, 1H), 8.93 (s, 1H), 12.63 (br s, 1H); MS (FAB) m/z 568 (M ++ 1); C 29H 30ClN 3O 71/4H 2The analytical calculation value of O: C, 60.84; H, 5.37; Cl, 6.19; N, 7.34.Measured value: C, 61.03; H, 5.56; Cl, 6.27; N, 7.03.Embodiment 1304-[1-[4-[N '-(2-bromo phenyl) urea groups]-3-methoxyphenyl acetyl group]-(4R)-methoxyl group-(2S)-the pyrrolidinyl methoxyl group] benzoic acid
Under room temperature, with 4-[N '-(2-bromo phenyl) urea groups]-3-methoxybenzene guanidine-acetic acid (600mg, 1.58mmol), 4-[(4R)-methoxyl group-(2S)-pyrrolidinyl methoxyl group] essence of Niobe (420mg, 1.58mmol), EDCHCl (3 64mg, 1.90mmol), HOBt (214mg, 1.58mmol) and triethylamine (265ml, 1.90mmol) mixture in THF (15ml) stirs and to spend the night.This mixture of dilute with water is used ethyl acetate extraction.With salt water washing extract, through dried over sodium sulfate and evaporation.Residue is through purification by silica gel column chromatography; with chloroform-methanol (100: 1; v/v) as eluant; obtain 4-[1-[4-[N '-(2-bromo phenyl) urea groups]-3-methoxyphenyl acetyl group]-(4R)-methoxyl group-(2S)-the pyrrolidinyl methoxyl group] essence of Niobe (1.01g; q.y.), be the weak yellow foam thing. 1H-NMR (CDCl 3) δ 2.13-2.33 (m, 2H), 3.27 (s, 3H), 3.57 (s, 3H), 3.61-3.72 (m, 4H), 3.88 (s, 3H), 4.05-4.14 (m, 2H), 4.43-4.57 (m, 2H), 6.70-7.00 (a plurality of m, 5H altogether), 7.29-7.52 (m, 4H), 7.92-8.01 (m, 3H), and 8.12-8.15 (m, 1H); MS (FAB) m/z 626 (M ++ 1).
To 4-[1-[4-[N '-(2-bromo phenyl) urea groups]-3-methoxyphenyl acetyl group]-(4R)-methoxyl group-(2S)-the pyrrolidinyl methoxyl group] essence of Niobe (697mg; 1.11mmol) the agitating solution of THF (8ml) in add 0.5N sodium hydroxide (8ml), with this reaction mixture refluxed heating 2 hours.After being cooled to room temperature, in this mixture impouring ice-1N HCl, the precipitation that produces is collected in decompression.The crude product solid is recrystallization purifying from methanol-chloroform-IPE, obtains 138 (252mg, 37%), is white crystalline powder.MW 612.47 mp 125-130 ℃; 1H-NMR (DMSO-d 6) δ 2.08-2.17 (m, 2H), 3.21 (s, 3H), 3.60-3.72 (m, 4H), 3.79 (s, 3H), 3.95-4.33 (m, 4H), (6.75-7.08 a plurality of m, 5H altogether), and 7.31-7.34 (m, 1H), 7.59-7.61 (m, 1H), 7.87-7.89 (m, 2H), 7.93-7.96 (m, 2H), 8.73 (s, 1H), 8.91 (s, 1H), 12.63 (br s, 1H); MS (FAB) m/z 612 (M ++ 1); C 29H 30BrN 3O 7The analytical calculation value: C, 56.87; H, 4.94; Br, 13.05; N, 6.86.Measured value: C, 56.67; H, 4.97; Br, 13.07; N, 6.68.Embodiment 1314-[4,4-two fluoro-1-[3-methoxyl group-4-[N '-(2-aminomethyl phenyl) urea groups] phenyl acetyl]-2-pyrrolidinyl methoxyl group] benzoic acid
To the N-Boc proline methyl ester (2.0g, add in the agitating solution of dichloromethane 8.15mmol) 3A molecular sieve (2g) and PDC (4.60g, 12.2mmol).This mixture was stirred 3 days.Filter this mixture, evaporated filtrate by the Celiete pad.Residue as eluant, obtains 1-(tert-butoxycarbonyl)-4-oxo-pyrrolidine-2-methyl formate (1.13g, 57%) with chloroform-methanol (10: 1) through silica gel column chromatography, is colorless oil. 1H-NMR(CDCl 3)δ1.46-1.48(m，9H)，2.56-2.61(m，1H)，2.88-3.00(m，1H)，3.77(s，3H)，3.82-3.88(m，2H)，4.71-4.83(m，1H)。
To 1-(tert-butoxycarbonyl)-4-oxo-pyrrolidine-2-methyl formate (1.13g, 4.65mmol) add in cold (78 ℃) solution of the stirring in dichloromethane (20ml) methyl DAST (1.1ml, 11.6mmol).This mixture is warmed to room temperature.Stir after 15 hours, in this mixture impouring water (50ml), extract with ethyl acetate (200ml).(2 * 200ml) washing extracts are through dried over mgso and evaporation with saline.Residue as eluant, obtains 1-(tert-butoxycarbonyl)-4 with chloroform-ethyl acetate (20: 1) through silica gel column chromatography, and 4-two fluoropyrrolidines-2-methyl formate (885mg, 72%) are yellow oil. 1H-NMR (CDCl 3) δ 1.42 and 147 (respectively be s, be total to 9H), 2.46 (ddd, J=26.9,13.7,5.1Hz, 1H), 2.62-2.78 (m, 1H), 3.75-3.95 (m, 5H), 4.43-4.57 (m, 1H).
To 1-(tert-butoxycarbonyl)-4, (885mg, (26.7ml 6.67mmol) also continues to stir 1 hour 4-two fluoropyrrolidines-2-methyl formate to add the 0.25N sodium hydroxide in the agitating solution of THF 3.34mmol) (25ml).In this mixture impouring 1N HCl (100ml), (2 * 200ml) extract with chloroform.Extract with saline (100ml) washing merges through dried over mgso and evaporation, obtains 1-(tert-butoxycarbonyl)-4, and 4-two fluoropyrrolidines-2-formic acid (775mg, 92%) are the yellow crystal solid.Mp 113-117 ℃; 1H-NMR (CDCl 3) δ 1.44 and 1.49 (respectively be s, be total to 9H), and 2.53-2.80 (m, 2H), 3.71-3.90 (m, 2H), 4.20-4.61 (m, 1H); MS (FAB) m/z 252 (M ++ 1); C 10H 15F 2O 4The analytical calculation value: C, 47.81; H, 6.02; N, 5.58.Measured value: C, 48.06; H, 6.05; N, 5.45.
Under room temperature, to N-(tert-butoxycarbonyl) 4, (3.00g adds BH in the agitating solution of THF 11.9mmol) (20ml) to 4-difluoro proline 3DMS (1.1ml.11.9mmol) was with this mixture reflux 2 hours.After being cooled to room temperature, this mixture of vacuum concentration.By adding entry (100ml) quencher residue, (2 * 200ml) extract with chloroform.The extract that merges through dried over mgso also evaporates.Residue as eluant, obtains 1-(tert-butoxycarbonyl)-4 with chloroform-ethyl acetate (4: 1) through silica gel column chromatography, and 4-two fluoro-2-pyrrolidinyl methanol (2.11g, 75%) are colorless oil. 1H-NMR(CDCl 3)δ1.48(s，9H)，2.04-2.55(m，2H)，3.59-4.17(m，5H)。
Under room temperature, to 1-(tert-butoxycarbonyl)-4,4-two fluoro-2-pyrrolidinyl methanol (600mg, 2.53mmol), 4-methyl hydroxybenzoate (462mg, 3.03mmol), triphenyl phasphine (795mg, 3.03mmol) the stirring the mixture of THF (10ml) in add DIAD (597 μ l, 3.03mmol).Stir down, with this mixture reflux 3 hours.After being cooled to room temperature, this mixture of vacuum concentration.Residue as eluant, obtains 4-[1-(tert-butoxycarbonyl)-4,4-two fluoro-2-pyrrolidinyl methoxyl groups with hexane-ethyl acetate (4: 1) through silica gel column chromatography] essence of Niobe (831mg, 88%), be colorless oil. 1H-NMR (CDCl 3) δ 1.48 (s, 9H), 2.53-2.61 (m, 2H), 3.63-4.41 (a plurality of m, 8H altogether), 6.94 (d, J=8.8Hz, 2H), 7.99 (d, J=8.8Hz, 2H).
With 4-[1-(tert-butoxycarbonyl)-4,4-two fluoro-2-pyrrolidinyl methoxyl groups] essence of Niobe (830mg, 2.23mmol) and the mixture of TFA (5ml) in dichloromethane (5ml) stirred 3 hours and vacuum concentration.Make residue be alkalescence with saturated sodium bicarbonate, (2 * 200ml) extract with chloroform.Through dry extract and the vacuum concentration that merges of potassium carbonate, obtain 4-(4,4-two fluoro-2-pyrrolidinyl methoxyl groups) essence of Niobe (550mg, 91%), be faint yellow solid. 1H-NMR (CDCl 3) δ 2.19 (m, 1H), 2.43 (m, 1H), 3.19-3.41 (m, 2H), 3.77 (m, 1H), 3.89 (s, 3H), 4.00-4.09 (m, 2H), 6.92 (d, J=9.0Hz, 2H), 7.99 (d, J=9.0Hz, 2H); MS (FAB) m/z 272 (M ++ 1); C 13H 15F 2N0 3The analytical calculation value: C, 57.56; H, 5.57; N, 5.16.Measured value: C, 57.65; H, 5.67; N, 5.16.
With 4-(4,4-two fluoro-2-pyrrolidinyl methoxyl groups) essence of Niobe (540mg, 1.99mmol), 3-methoxyl group-4-[N '-(2-aminomethyl phenyl) urea groups] phenylacetic acid (626mg, 1.99mmol), EDCHCl (572mg, 2.99mmol), HOBt (cat.) and DMAP (cat.) mixture in DMF (10ml) stirs and spend the night.(300ml) dilutes this mixture with ethyl acetate, and (2 * 100ml) washings are through dried over mgso and vacuum concentration with saline.Residue is through silica gel column chromatography; with chloroform-methanol (20: 1) as eluant, obtain 4-(4,4-two fluoro-1-[3-methoxyl group-4-[N '-(2-aminomethyl phenyl) urea groups] phenyl acetyl]-2-pyrrolidinyl methoxyl group] essence of Niobe (1.00g; 89%), is the colourless foam thing. 1H-NMR (CDCl 3) δ 2.31 (s, 3H), 2.47-2.63 (m, 2H), 3.52-3.97 (a plurality of s and m, 10H altogether), 4.07-4.30 (m, 2H), 4.67-4.69 (m, 1H), 6.45 (s, 1H), 6.65 (d, J=1.7Hz, 1H), 6.74-6.76 (m, 1H), 6.84 (d, J=8.8Hz, 2H), 7.14 (m, 2H), 7.24 (m, 2H), 7.52-7.54 (m, 1H), 7.94 (d, J=8.8Hz, 2H), 8.09 (d, J=8.1Hz, 1H).
With 4-(4; 4-two fluoro-1-[3-methoxyl group-4-[N '-(2-aminomethyl phenyl) urea groups] phenyl acetyl]-2-pyrrolidinyl methoxyl group] essence of Niobe (1.00g; 1.76mmol) and the 0.25N sodium hydroxide (14ml, 3.50mmol) mixture in THF (14ml) stirs and to spend the night.With this mixture of 1N HCl acidify, (10: 1,2 * 200ml) extracted with chloroform-methanol.The extract that merges through dried over mgso also evaporates.Residue is through silica gel column chromatography, with chloroform-methanol (20: 1-10: 1) as eluant, obtain 139 (658mg, 68%), be the colourless crystallization powder.MW 553.55 mp 135-140 ℃; 1H-NMR (DMSO-d 6) δ 2.23 (s, 3H), 2.49-2.73 (m, 2H), 3.36-4.55 (a plurality of m, 10H), 6.73 (d, J=8.3Hz, 1H), 6.84 (s, 1H), 6.93 (t, J=7.3Hz, 1H), 7.00 (d, J=8.3Hz, 2H), 7.10-7.16 (m, 2H), 7.78 (d, J=8.3Hz, 1H), 7.86 (d, J=8.3Hz, 2H), 8.00 (d, J=8.3Hz, 1H), 8.47 (s, 1H), 8.56 (s, 1H); MS (FAB) m/z 554 (M ++ 1); C 29H 29F 2N 3O 63/4H 2The analytical calculation value of O: C, 61.42; H, 5.44; N, 7.06.Measured value: C, 61.30; H, 5.44; N, 7.06.Embodiment 1324-[1-[4-[N '-(2-chlorophenyl) urea groups]-3-methoxyphenyl acetyl group]-4,4-two fluoro-2-pyrrolidinyl methoxyl groups] benzoic acid
With 4-(4,4-two fluoro-2-pyrrolidinyl methoxyl groups) essence of Niobe (229mg, 0.845mmol), 4-[N '-(2-chlorophenyl) urea groups]-3-methoxybenzene guanidine-acetic acid (283mg, 0.845mmol), EDCHCl (243mg, 1.27mmol), the mixture of HOBt (cat.), DMAP (cat.) and DMF (10ml) stirs and spend the night.(300ml) dilutes this mixture with ethyl acetate, and (2 * 100ml) wash this solution, through dried over mgso and vacuum concentration with saline.Residue is through silica gel column chromatography; with chloroform-ethyl acetate (20: 1-4:, obtain 4-[1-[4-[N '-(2-chlorophenyl) urea groups 1) as eluant]-3-methoxyphenyl acetyl group]-4,4-two fluoro-2-pyrrolidinyl methoxyl groups] essence of Niobe (482mg; 97%), is colourless viscous solid. 1H-NMR (CDCl 3) δ 2.50-2.67 (m, 2H), 3.54-4.71 (a plurality of m, 13H), 6.69 (d, J=1.5Hz, 1H), 6.76 (d, J=8.3Hz, 1H), 6.84 (d, J=8.8Hz, 2H), 6.98 (dt, J=7.8,1.5Hz, 1H), 7.23-7.27 (m, 1H), 7.33 (d, J=8.3Hz, 1H), 7.39 (m, 2H), 7.94 (d, J=8.8Hz, 2H), 8.00 (d, J=8.3Hz, 1H), 8.19 (dd, J=8.3,1.5Hz, 1H).
With 4-[1-[4-[N '-(2-chlorophenyl) urea groups]-3-methoxyphenyl acetyl group]-4; 4-two fluoro-2-pyrrolidinyl methoxyl groups] essence of Niobe (480mg; 0.816mmol), the 0.25N sodium hydroxide (6.5ml, 1.65mmol) and the mixture of THF (20ml) stirred 3 days.With (5: 1,2 * 200ml) extracted with chloroform-methanol among this mixture impouring 1N HCl (100ml).Extract and vacuum concentration through the dried over mgso merging.Residue is through silica gel column chromatography, with chloroform-methanol (20: 1-5: 1) as eluant, obtain 140 (270mg, 58%), be faint yellow amorphous solid.MW573.97 1H-NMR (DMSO-d 6) δ 2.45-2.74 (m, 2H), 3.63-4.83 (a plurality of m, 10H), 6.76 (d, J=8.3Hz, 1H), 6.87 (s, 1H), 7.00-7.05 (m, 3H), and 7.26-7.30 (m, 1H), 7.44 (dd, J=8.3,1.2Hz, 1H), and 7.88-7.93 (m, 2H), 7.98 (d, J=8.3Hz, 1H), 8.10 (d, J=8.3Hz, 1H), 8.92 (s, 1H), 8.96 (s, 1H); MS (FAB) m/z 574 (M ++ 1); C 28H 26ClF 2N 3O 6H 2The analytical calculation value of O: C, 56.81; H, 4.77; N, 7.10.Measured value: C, 56.75; H, 4.69; N, 6.79.Embodiment 1334-[(2R, 3R, 4S)-3,4-isopropylidene dioxy base-1-[4-[N '-(2-aminomethyl phenyl) urea groups]-3-methoxyphenyl acetyl group]-the 2-pyrrolidinyl] methoxybenzoic acid
To (2S, 3R, 4S)-1-benzyloxycarbonyl-3,4-isopropylidene dioxy base-2-pyrrolidinyl methyl formate (10.7g, 31.9mmol) the solution of THF (250ml) in add 0.25N sodium hydroxide (255ml), after stirring 24 hours under the room temperature,, use ethyl acetate extraction with this mixture of 1N HCl acidify.The extract that merges with the salt water washing through dried over sodium sulfate and vacuum concentration, obtains that (4S)-1-benzyloxycarbonyl-3,4-isopropylidene dioxy base-2-pyrrolidinyl formic acid (9.87g, 96%) is colorless oil for 2S, 3R with it. 1H-NMR (CDCl 3) δ 1.32 (s, 3H), 1.46 (d, J=2.7Hz, 3H), 3.61 (m, 1H), 3.82 and 3.92 (d, J=12.7Hz, 1H, amide isomers), 4.58 and 4.64 (s, 1H, amide isomers), 4.77 (t, J=5.1Hz, 1H), 4.83 and 4.89 (d, J=5.9Hz, 1H, amide isomers), 5.15 and 5.19 (m, 2H, amide isomers), 7.31-7.37 (m, 5H).
In 0 ℃, to (2S, 3R, 4S)-and 1-benzyloxycarbonyl-3, (9.87g adds BH in the agitating solution of THF 30.7mmol) (200ml) to 4-isopropylidene dioxy base-2-pyrrolidinyl formic acid 3DMS (6.14ml, 61.4mmol).Make this mixture be back to room temperature, reflux is 2 hours then.After being cooled to room temperature, this mixture of vacuum concentration, in 0 ℃ by adding the entry quencher, with this mixture of ethyl acetate extraction.The extract that water and salt water washing merge, with it with dried over sodium sulfate and vacuum concentration.Residue is through silica gel column chromatography [200g, chloroform/methanol (20: 1)], obtain (2R, 3R, 4S)-and 1-benzyloxycarbonyl-3,4-isopropylidene dioxy base-2-pyrrolidinyl methanol (10.1g, 100%) is colorless oil. 1H-NMR(CDCl 3)δ1.31(s，3H)，1.45(s，3H)，3.56-4.74(m，7H)，5.14(s，2H)，7.34(m，5H)。
Under 0 ℃, blanket of nitrogen, to (2R, 3R, 4S)-1-benzyloxycarbonyl-3,4-isopropylidene dioxy base-2-pyrrolidinyl methanol (312mg, 0.64mmol), right-methyl hydroxybenzoate (67ml, 0.70mmol), triphenyl phasphine (184mg, 0.70mmol) the stirring the mixture of THF (7ml) in add DIAD (138ml, 0.70mmol).Make this mixture rise to room temperature and stirred 3 hours, except that after desolvating, the residue that obtains is through silica gel column chromatography [10g, n-hexane/ethyl acetate (4: 1)], obtain 4-[(2R, 3R, 4S)-benzyloxycarbonyl-3,4-isopropylidene dioxy base-2-pyrrolidinyl] methoxyl methyl benzoate (321mg, 83%), be colorless oil. 1H-NMR (CDCl 3) δ 1.01 (s, 6H), 1.03 (s, 3H), 2.23 (m, 1H), 2.63 (m, 1H), 3.61 (d, J=12.5Hz, 1H), 3.80-4.27 (m, 4H), 4.84 (br, 1H), 5.01 and 5.08 (ABq, J=12.2Hz, 1H, amide isomers), 6.75-6.87 (m, 3H), and 7.19-7.63 (m, 15H).
Under room temperature, nitrogen atmosphere, stir 4-[(2R, 3R, 4S)-and 1-benzyloxycarbonyl-3,4-isopropylidene dioxy base-2-pyrrolidinyl] (2.37g is 5.76mmol) with the suspension of 10% Pd/C (240mg) in ethanol (170ml) for methoxyl methyl benzoate.Stir after 1 day, change this catalyst and solvent into 10%Pd/C (500mg) and THF (50ml).Under room temperature, nitrogen atmosphere, stirred this suspension 5 days.After removing by filter catalyst, vacuum concentrated filtrate.Residue obtains 4-[(2R through silica gel column chromatography [100g, chloroform/acetone (20/1)], 3R, 4S)-3, and 4-isopropylidene dioxy base-2-pyrrolidinyl] methoxyl methyl benzoate (930mg, 53%), be brown oil. 1H-NMR(CDCl 3)δ1.35(s，3H)，1.50(s，3H)，3.02(dd，J＝13.7，4.1Hz，1H)，3.13(d，J＝13.7Hz，1H)，3.58(t，J＝6.3Hz，1H)，3.88(s，3H)，3.90(dd，J＝9.3，6.6Hz，1H)，4.02(dd?J＝9.5，3.9Hz，1H)，4.74(d，J＝5.6Hz，1H)，4.79(m，1H)，6.90(d，J＝9.0Hz，2H)，7.98(d，J＝9.0Hz，2H)。
Under room temperature, with 3-methoxyl group-4-[N '-(2-aminomethyl phenyl) urea groups] phenylacetic acid (437mg, 1.39mmol), 4-[(2R, 3R, 4S)-3,4-isopropylidene dioxy base-2-pyrrolidinyl] and methoxyl methyl benzoate (428mg, 1.39mmol), EDCHCl (400mg, 2.09mmol) and DMAP (170mg, 1.39mmol) mixture in DMF (12ml) stirred 20 hours.In this mixture impouring frozen water, use ethyl acetate extraction.Extract with frozen water and salt water washing merging.After dried over sodium sulfate, the vacuum concentration extract.Residue is through silica gel column chromatography [70g; chloroform/acetone (10/1)]; obtain 4-[(2R; 3R; 4S)-3; 4-isopropylidene dioxy base-1-[4-[N '-(2-aminomethyl phenyl) urea groups]-3-methoxyphenyl acetyl group]-the 2-pyrrolidinyl] methoxyl methyl benzoate (840mg, 100%), be colourless amorphous solid.IR (KBr) 3354,2985,2939,1716,1533,1254 cm -1 1H-NMR (CDCl 3) δ 1.31 (s, 3H), 1.42 (s, 3H), 2.05 (s, 3H), 3.50 (s, 3H), 3.55-3.88 (m, 4H), 3.89 (s, 3H), 4.13 (m, 1H), 4.67 (br, 1H), 4.78 (d, J=6.1Hz, 1H), 4.88 (d, J=5.6Hz, 1H), 6.46 (s, 1H), 6.62 (d, J=1.5Hz, 1H), 6.74 (m, 3H), 7.05 (s, 1H), 7.14 (d, J=7.3Hz, 1H), 7.23 (m, 2H), 7.57 (d, J=7.8Hz, 1H), 7.91-8.08 (m, 3H); MS (ESI) m/z 604 (M ++ 1); C 33H 37N 3O 80.6H 2The analytical calculation value of O: C, 64.50; H, 6.27; N, 6.84.Measured value: C, 64.38; H, 6.18; N, 6.66.
Under room temperature; with 4-[(2R, 3R, 4S)-3; 4-isopropylidene dioxy base-1-[4-[N '-(2-aminomethyl phenyl) urea groups]-3-methoxyphenyl acetyl group]-the 2-pyrrolidinyl] methoxyl methyl benzoate (183mg, 0.303mmol) and the mixture of g.HCl-MeOH (6ml) stirred 17 hours.This mixture of vacuum concentration.Residue is through TLC[chloroform/methanol (10/1)] purification, obtain 4-[(2R, 3R; 4S)-3; 4-dihydroxy-1-[4-[N '-(2-aminomethyl phenyl) urea groups]-3-methoxyphenyl acetyl group]-the 2-pyrrolidinyl] methoxyl methyl benzoate (162mg, 95%), be colourless amorphous solid.IR(KBr)3342，1716，1604，1535，1255?cm -1； 1H-NMR(CDCl 3)δ2.25(br，3H)，3.33-3.75(m，7H)，3.87(s，3H)，4.10(d，J＝8.3Hz，1H)，4.24(s，2H)，4.37(m，2H)，6.62-7.94(m，13H)；MS(ESI)m/z?564(M ++1)。
To 4-[(2R; 3R; 4S)-3,4-isopropylidene dioxy base-1-[4-[N '-(2-aminomethyl phenyl) urea groups]-3-methoxyphenyl acetyl group]-the 2-pyrrolidinyl] (490mg adds 0.25N sodium hydroxide (9.8ml) to methoxyl methyl benzoate in the solution of THF 0.812mmol) (9.8ml).After stirring 4 days under the room temperature,, extract with chloroform-methanol (10/1) with this mixture of 1N HCl acidify.Extract and vacuum concentration through dried over sodium sulfate merges obtain 141 (445mg, 93%), are colourless amorphous solid.MW 689.64 IR (KBr) 3354,2983,2937,1707,1604,1533 cm -1 1H-NMR (DMSO-d 6) δ .24 and 1.26 (s, 3H, amide isomers), 1.26 and 1.32 (s, 3H, amide isomers), 2.24 (s, 3H), 3.40 (dd, J=14.0,5.1Hz, 1H), 3.60 (m, 2H), 3.71 (m, 1H), 3.76 (s, 3H), 3.82 (s, 3H), 3.92-4.96 (m, 5H), 6.74 and 6.78 (m, 1H, amide isomers), 6.83-7.16 (m, 6H), 7.79 (d, J=8.3Hz, 1H), 7.87 (t, J=9.1Hz, 2H), 8.01 (m, 1H), 8.49 (d, J=3.4Hz, 1H), 8.57 (s, 1H); MS (FAB) m/z 590 (M ++ 1); C 32H 35N 3O 82.3H 2The analytical calculation value of O: C, 60.90; H, 6.32; N, 6.66.Measured value: C, 61.00; H, 6.00; N, 6.27.Embodiment 1344-[(2R, 3R, 4S)-3,4-dihydroxy-1-[4-[N '-(2-aminomethyl phenyl) urea groups]-3-methoxyphenyl acetyl group]-the 2-pyrrolidinyl] methoxybenzoic acid
To 4-[(2R; 3R; 4S)-3,4-dihydroxy-1-[4-[N '-(2-aminomethyl phenyl) urea groups]-3-methoxyphenyl acetyl group]-the 2-pyrrolidinyl] (63mg adds 0.25N sodium hydroxide (0.89ml) to methoxyl methyl benzoate in the solution of THF 0.112mmol) (0.89ml).After stirring 3 days under the room temperature,, extract with chloroform-methanol (10/1) with this mixture of 1N HCl acidify.Extract and vacuum concentration through dried over sodium sulfate merges obtain 142 (54mg, 88%), are colourless amorphous solid.MW 549.57 IR (KBr) 3356,2958,2927,1685,1604,1535,1255 cm -1 1H-NMR (DMSO-d 6) δ 2.24 (s, 3H), 3.40 (m, 1H), 3.58 (s, 2H), 3.66 (dd, J=9.8,6.6Hz, 1H), 3.80 (s, 3H), 3.99-4.30 (m, 5H), 5.10 (br, 1H), 6.72 (d, J=8.1Hz, 1H), 6.85 (s, 1H), 6.93 (t, J=7.3Hz, 1H), 7.03 (d, J=8.8Hz, 2H), 7.14 (t, J=8.8Hz, 2H), 7.79 (d, J=8.1Hz, 2H), 7.86 (d, J=8.8Hz, 2H), 7.99 (d, J=8.3Hz, 1H), 8.46 (s, 1H), 8.56 (s, 1H); MS (ESI) m/z 550 (M ++ 1); C 29H 31N 3O 80.85H 2The analytical calculation value of O: C, 61.66; H, 5.83; N, 7.44.Measured value: C, 62.09; H, 5.93; N, 6.95.Embodiment 1354-[(2R, 3R, 4S)-1-[4-[N '-(2-chlorophenyl) urea groups]-3-methoxyphenyl acetyl group]-3,4-isopropylidene dioxy base-2-pyrrolidinyl] methoxybenzoic acid
Under room temperature, with 4-[N '-(2-chlorophenyl) urea groups]-3-methoxybenzene guanidine-acetic acid (487mg, 1.45mmol), 4-[(2R, 3R, 4S)-3,4-isopropylidene dioxy base-2-pyrrolidinyl] and methoxyl methyl benzoate (447mg, 1.45mmol), EDCHCl (418mg, 2.18mmol) and DMAP (177mg, 1.45mmol) mixture in DMF (12ml) stirred 19 hours.In this mixture impouring frozen water, use ethyl acetate extraction.Extract with frozen water and salt water washing merging.After dried over sodium sulfate, the vacuum concentration extract.Residue is through silica gel column chromatography [70g; chloroform/acetone (10/1)]; obtain 4-[(2R; 3R; 4S)-1-[4-[N '-(2-chlorophenyl) urea groups]-3-methoxyphenyl acetyl group]-3; 4-isopropylidene dioxy base-2-pyrrolidinyl] methoxyl methyl benzoate (850mg, 94%), be colourless amorphous solid.IR (KBr) 3329,2939,1716,1627,1531,1254cm -1 1H-NMR (CDCl 3) δ 1.33 (s, 3H), 1.43 (s, 3H), 3.56 (s, 3H), 3.61 (s, 1H), 3.64 (s, 1H), 3.70 (m, 1H), 3.79 (d, J=10.4Hz, 1H), 3.88 (s, 3H), 4.14 (dd, J=9.8,2.2Hz, 1H), 4.40 (dd, J=9.8,3.4Hz, 1H), 4.67 (s, 1H), 4.80 (d, J=6.1Hz, 1H), 4.90 (t, J=4.6Hz, 1H), 6.65 (d, J=1.7Hz, 1H), 6.71-6.84 (m, 3H), 6.98 (dt, J=7.6,1.5Hz, 1H), 7.27 (m, 2H), 7.33 (dd, J=8.0,1.2Hz, 2H), 7.90-8.01 (m, 3H), 8.20 (dd, J=8.3,1.5Hz, 1H); MS (ESI) m/z 624 (M ++ 1), 626 (M ++ 3); C 32H 34N 3O 81.4H 2The analytical calculation value of O: C, 59.19; H, 5.71; N, 6.47.Measured value: C, 58.85; H, 5.35; N, 6.21.
Under room temperature; with 4-[(2R, 3R, 4S)-1-[4-[N '-(2-chlorophenyl) urea groups]-3-methoxyphenyl acetyl group]-3; 4-isopropylidene dioxy base-2-pyrrolidinyl] methoxyl methyl benzoate (177mg, 0.284mmol) and the mixture of g.HCl-MeOH (4ml) stirred 2 days.This mixture of vacuum concentration.Residue is through TLC[chloroform/methanol (15/1)] purification; obtain 4-[(2R; 3R; 4S)-1-[4-[N '-(2-chlorophenyl) urea groups]-3-methoxyphenyl acetyl group]-3; 4-isopropylidene dioxy base-2-pyrrolidinyl] methoxyl methyl benzoate (140mg; 85%), is colourless amorphous solid.IR(KBr)3338，2949，1712，1623，1604，1533?cm -1； 1H-NMR(CDCl 3)δ2.27(m，1H)，2.79(m，1H)，3.53(dd，J＝10.5，5.9Hz，1H)，3.63(s，3H)，3.88(s，3H)，4.21(d，J＝7.8Hz，1H)，4.31(m，2H)，4.43(dd，J＝9.8，4.4Hz，1H)，4.52(t，J＝4.6Hz，1H)，6.71(s，1H)，6.80(m，3H)，6.99(t，J＝7.3Hz，1H)，7.16(s，1H)，7.21(s，1H)，7.39(d，J＝8.1Hz，1H)，7.91(d，J＝8.6Hz，2H)，8.16(d，J＝8.3Hz，1H)；MS(ESI)m/z?584(M ++1)，586(M ++3)。
To 4-[(2R; 3R; 4S)-1-[4-[N '-(2-chlorophenyl) urea groups]-3-methoxyphenyl acetyl group]-3,4-isopropylidene dioxy base-2-pyrrolidinyl] (511mg adds 0.25N sodium hydroxide (9.8ml) to methoxyl methyl benzoate in the solution of THF 0.819mmol) (9.8ml).After stirring 20 hours under the room temperature,, extract with chloroform-methanol (10/1) with this mixture of 1N HCl acidify.Extract and vacuum concentration through dried over sodium sulfate merges obtain 143 (504mg, 100%), are colourless amorphous solid.MW 610.05 IR (KBr) 3330,2983,2937,1711,1689,1604,1533,1252 cm -1 1H-NMR (DMSO-d 6) δ 1.26 (s, 3H), 1.32 (s, 3H), 3.40 (m, 1H), 3.60 and 3.61 (d, J=2.5Hz, 3H, amide isomers), 3.62 (m, 1H), 3.78 and 3.83 (s, 3H, amide isomers), 4.16 (m, 2H), 4.42-4.98 (m, 3H), 6.74-7.15 (m, 6H), 7.28 (t, J=7.3Hz, 1H), 7.43 (d, J=8.1Hz, 1H), 7.78-7.97 (m, 4H), 8.08 (d, J=8.3Hz, 1H), 8.89 (s, 1H), 8.92 (s, 1H), 12.68 (br, 1H); MS (ESI) m/z 610 (M ++ 1), 612 (M ++ 3).Embodiment 1364-[(2R, 3R, 4S)-1-[4-[N '-(2-chlorophenyl) urea groups]-3-methoxyphenyl acetyl group]-3,4-dihydroxy-2-pyrrolidinyl] methoxybenzoic acid
Figure A0080951003721
To 4-[1-[4-[N '-(2-chlorophenyl) urea groups]-3-methoxyphenyl acetyl group]-3,4-dihydroxy-2-pyrrolidinyl methoxyl group] (63mg adds 0.25N sodium hydroxide (0.80ml) to essence of Niobe in the solution of THF 0.108mmol) (0.80ml).After stirring 3 days under the room temperature,, extract with chloroform-methanol (10/1) with this mixture of 1N HCl acidify.Extract and vacuum concentration through dried over sodium sulfate merges obtain 144 (61mg, 100%), are colourless amorphous solid.MW 569.99IR (KBr) 3338,1687,1604,1533,1255,1169,1036 cm -1 1H-NMR (DMSO-d 6) δ 3.59 (d, J=5.5Hz, 2H), 3.61 (m, 1H), 3.66 (dd, J=10.0,7.1Hz, 1H), 3.80 (s, 3H), 4.00-4.33 (m, 5H), 5.10 (br, 1H), 6.74 (d, J=8.3Hz, 1H), 6.87 (s, 1H), 7.03 (m, 3H), 7.28 (t, J=8.3Hz, 1H), 7.43 (d, J=6.6Hz, 1H), 7.87 (d, J=8.5Hz, 2H), 7.95 (d, J=8.3Hz, 1H), 8.09 (d, J=8.3Hz, 1H), 8.32 (s, 1H), 8.89 (s, 1H), 8.93 (s, 1H); MS (ESI) m/z 570 (M ++ 1), 572 (M ++ 3); C 28H 28ClN 3O 81.4H 2The analytical calculation value of O: C, 57.19; H, 5.14; N, 7.15.Measured value: C, 57.52; H, 5.22; N, 6.76.Embodiment 1374-[1-[4-[N '-(2-aminomethyl phenyl) urea groups]-3-methoxyphenyl acetyl group]-5-(R)-phenyl-2-(S)-pyrrolidinyl methoxyl group] benzoic acid
Figure A0080951003731
In-78 ℃, to N-Boc-pyrroles's 1,3-propanedicarboxylic acid (pyrroglutarate) benzyl ester (8.93g, 28.0mmol) the agitating solution of THF (100ml) in add phenyl lithium (1.0M be in ether-cyclohexane extraction, 33.5ml, 33.5mmol), make the mixture that obtains be warmed to-40 ℃ gradually, stir then and spend the night.Aqueous ammonium chloride solution is added in the reactant mixture, and vacuum is removed THF, uses ethyl acetate extraction then.Wash organic layer with water, through anhydrous sodium sulfate drying and vacuum concentration.Residue is through silica gel column chromatography, and as eluant, recrystallization from hexane-ethyl acetate obtains [2-(S)-(N-Boc-amino)-5-oxo-6-phenyl] benzyl valerianate (5.02g, 45%) then, is colourless spicule with hexane-ethyl acetate (4: 1).Mp 85-87 ℃; 1H-NMR (CDCl 3) δ 1.43 (s, 9H), 2.07-2.19 (m, 1H), 2.27-2.36 (m, 1H), 2.97-3.13 (m, 2H), 4.44 (brs, 1H), 5.19 (dd, J=25.2,12.0Hz, 2H), 5.19 (overlapping, 1H), 7.28-7.98 (a plurality of m, 10H); MS (ESI) m/z 322 (M ++ H).
Under room temperature, (2.20g adds trifluoroacetic acid (15ml) in the agitating solution of dichloromethane 5.54mmol) (50ml), the mixture that obtains was stirred 2 hours to [2-(S)-(N-Boc-amino)-5-oxo-6-phenyl] benzyl valerianate.This mixture of vacuum concentration also is poured in the sodium bicarbonate aqueous solution, uses ethyl acetate extraction then.Through anhydrous sodium sulfate drying organic layer and vacuum concentration, obtain 5-phenyl-5-pyrrolin-2-(S)-benzyl formate and (l.60g, quantitatively), be yellow solid.This product can need not to be further purified and be used for next step reaction: 1H-NMR (CDCl 3) δ 2.20-2.29 (m, 1H), 2.32-2.42 (m, 1H), 2.96-3.05 (m, 1H), 3.12-3.21 (m, 1H), 4.96-5.00 (m, 1H), 5.24 (s, 2H), 7.31-7.49 (m, 8H), 7.88-7.91 (m, 2H); MS (ESI) m/z 280 (M ++ H).
Under room temperature, with 5-phenyl-5-pyrrolin-2-(S)-benzyl formate (1.59g, 5.69mmol) and Pd/C (10%, 128mg) mixture in methanol (30ml) stirred 28 hours under nitrogen atmosphere.Filter this mixture, vacuum concentrated filtrate.Make residue be dissolved in acetonitrile-water (3: 2,25ml) in, add Bis(tert-butoxycarbonyl)oxide (1.86g, 8.54mmol) and the 1.0M sodium hydroxide (8.54ml 8.54mmol), stirs the mixture that obtains 30 minutes.This mixture of vacuum concentration also is poured in the sodium bicarbonate aqueous solution, uses ethyl acetate extraction then.Water, saturated brine washing organic layer are through anhydrous sodium sulfate drying and vacuum concentration.Residue is through silica gel column chromatography, with chloroform-methanol (9: 1) eluting, recrystallization from hexane-ethyl acetate, obtains N-Boc-5-(R)-phenyl-(S)-proline (810mg, 49%), is colorless solid.Mp?113-117℃； 1H-NMR(CDCl 3)δ1.13(s，9H)，1.43(brs，1H)，1.96(brs，1H)，2.09(brs，1H)，2.31-2.34(m，1H)，2.46(brs，1H)，4.52(brs，1H)，4.69(brs，1H)，7.22-7.37(m，5H)。
Under room temperature, (1.14g adds 10M-BH in the agitating solution of THF 3.91mmol) (20ml) to N-Boc-5-(R)-phenyl-2-(S)-proline 3Me 2(780ml 7.82mmol), heats the mixture that obtains 30 minutes under refluxing S.In this mixture impouring 1N HCl aqueous solution, use ethyl acetate extraction.Through anhydrous sodium sulfate drying organic layer, vacuum concentration then.Residue is through silica gel column chromatography, and as eluant, (1.11g quantitatively), is colorless oil to obtain N-Boc-2-(S)-methylol-5-(R)-Phenylpyrrolidine with chloroform-methanol (10: 1). 1H-NMR(CDCl 3)δ1.19(brs，9H)，1.65(brs，1H)，1.83-1.90(m，1H)，1.98-2.06(m，1H)，2.22-2.31(m，1H)，3.75-3.86(m，2H)，4.16-4.19(m，1H)，4.83(t，J＝6.8Hz，1H)，4.89(brs，1H)，7.19-7.31(m，5H)；MS(ESI)?m/z?278(M ++H)。
Under room temperature, to N-Boc-2-(S)-methylol-5-(R)-Phenylpyrrolidine (1.10g, 3.97mmol), triphenyl phasphine (1.25g, 4.76mmol) and 4-methyl hydroxybenzoate (724mg, 4.76mmol) agitating solution in add diisopropyl azo-2-carboxylic acid (955ml, 4.76mmol), the mixture that obtains was stirred 45 minutes in 60 ℃.This mixture of vacuum concentration, residue as eluant, obtain 4-[N-Boc-5-(R)-phenyl-2-(S)-pyrrolidinyl methoxyl group with hexane-ethyl acetate (4: 1) through silica gel column chromatography] essence of Niobe (1.31g, 80%), be colorless oil. 1H-NMR (CDCl 3) δ 1.19 and 1.47 (brs, 9H altogether), and 2.09-2.15 (m, 3H), 2.33-2.37 (m, 1H), 3.94 (s, 3H), 4.30 (brs, 1H), 4.41 (brs, 2H), 4.77 (brs, 1H), 7.03 (d, J=8.8Hz, 2H), 7.24-7.36 (m, 5H), 8.03-8.06 (m, 2H); MS (ESI) m/z 412 (M ++ H).
Under room temperature, to 4-[N-Boc-5-(R)-phenyl-2-(S)-pyrrolidinyl methoxyl group] (1.28g adds trifluoroacetic acid (10ml) to essence of Niobe in the agitating solution of dichloromethane 3.11mmol) (30ml), the mixture that obtains was stirred 45 minutes.In this mixture of vacuum concentration and the impouring sodium bicarbonate aqueous solution, use chloroform extraction then.Wash organic layer with water, through anhydrous sodium sulfate drying, vacuum concentration obtains 4-[5-(R)-phenyl-2-(S)-pyrrolidinyl methoxyl group then] (363mg quantitatively), is yellow oil to essence of Niobe.This product is used for subsequent reaction without being further purified. 1H-NMR(CDCl 3)δ1.71-1.83(m，2H)，2.03-2.10(m，1H)，2.15-2.24(m，1H)，3.68-3.74(m，1H)，3.89(s，3H)，4.01-4.09(m，2H)，4.28(t，J＝7.2Hz，1H)，6.95(d，J＝8.8Hz，2H)，7.22-7.27(m，1H)，7.33(t，J＝8.0Hz，2H)，7.42(d，J＝7.6Hz，2H)，8.00(d，J＝8.8Hz，2H)；MS(ESI)m/z?312(M ++H)，353(M ++CH 3CN)。
Under room temperature, to 4-[5-(R)-phenyl-2-(S)-pyrrolidinyl methoxyl group] essence of Niobe (135mg, 0.43mmol), 4-[N '-(2-aminomethyl phenyl) urea groups]-3-methoxybenzene guanidine-acetic acid (136mg, 0.43mmol) and N, (52.9mg adds EDCHCl (90.8mg in the agitating solution of DMF 0.43mmol) (10ml) to N-two-methylamino pyridine, 0.48mmol), the mixture stirring that obtains is spent the night.In reactant mixture impouring water, use ethyl acetate extraction.With salt water washing organic layer, through anhydrous sodium sulfate drying, vacuum concentration then.Residue is through silica gel column chromatography; with hexane-ethyl acetate (1: 5) as eluant; obtain 4-[1-[4-[N '-(2-aminomethyl phenyl) urea groups]-3-methoxyphenyl acetyl group]-5-(R)-phenyl-2-(S)-pyrrolidinyl methoxyl group] essence of Niobe (271mg; quantitatively), be colourless amorphous solid. 1H-NMR (CDCl 3) δ 2.00-2.18 (m, 3H), 2.31-2.41 (m, 1H), 2.27 (s, 3H), 3.31 (s, 2H), 3.67 (s, 3H), 3.89 (s, 3H), 4.35-4.48 (m, 2H), 4.60 (brs, 1H), 4.92 (t, J=6.8Hz, 1H), 6.51 (d, J=8.4Hz, 1H), 6.62 (s, 1H), 6.96 (d, J=8.8Hz, 1H), 7.11-7.40 (a plurality of m, 8H), 7.51 (d, J=8.0Hz, 1H), 7.97-8.00 (m, 2H); MS (ESI) m/z 608 (M ++ H).
Under room temperature; to 4-[1-[4-[N '-(2-aminomethyl phenyl) urea groups]-3-methoxyphenyl acetyl group]-5-(R)-phenyl-2-(S)-pyrrolidinyl methoxyl group] essence of Niobe (243mg; 0.40mmol) methanol-THF (1: 1; add 1.0M sodium hydroxide (2.4ml in agitating solution 10ml); 2.40mmol); stir down, the mixture that obtains was heated 1.5 hours in 60 ℃.In this reactant mixture impouring 1N HCl, use chloroform extraction then.With salt water washing organic layer, through anhydrous sodium sulfate drying, vacuum concentration then.Residue with chloroform-methanol (10: 1) eluting, obtains 145 (224mg, 94%) through silica gel column chromatography, is colourless amorphous solid.MW 593.67 1H-NMR (CD 3OD), and the mixture δ 2.00-2.19 of rotamer (m, 3H), 2.28 and 2.30 (s, 3H altogether), 2.45-2.49 (m, 1H), 3.37 (dd, J=39,16Hz, 2H), 3.77 and 3.80 (s, be total to 3H), and 3.92-5.18 (a plurality of m, 4H), 6.48-8.03 (a plurality of m, 16H); MS (FAB) m/z594 (M ++ H).Embodiment 1384-[1-[4-[N '-(2-chlorophenyl) urea groups]-3-methoxyphenyl acetyl group]-5-(R)-phenyl-2-(S)-pyrrolidinyl methoxyl group] benzoic acid
Under room temperature, to 4-[5-(R)-phenyl-2-(S)-pyrrolidinyl methoxyl group] essence of Niobe (142mg, 0.46mmol), 4-[N '-(2-chlorophenyl) urea groups]-3-methoxybenzene guanidine-acetic acid (153mg, 0.46mmol) and N, (55.7mg adds EDCHCl (95.7mg in the agitating solution of DMF 0.46mmol) (10ml) to the N-dimethyl aminopyridine, 0.50mmol), the mixture stirring that obtains is spent the night.In reactant mixture impouring water, use ethyl acetate extraction.With salt water washing organic layer, through anhydrous sodium sulfate drying, vacuum concentration then.Residue is through silica gel column chromatography; with hexane-ethyl acetate (1: 5) as eluant; obtain 4-[1-[4-[N '-(2-chlorophenyl) urea groups]-3-methoxyphenyl acetyl group]-5-(R)-phenyl-2-(S)-pyrrolidinyl methoxyl group] essence of Niobe (260mg; 90%), is colourless amorphous solid. 1H-NMR (CDCl 3) δ 2.00-2.19 (m, 3H), 2.35-2.44 (m, 1H), 3.35 (s, 2H), 3.76 (s, 3H), 3.89 (s, 3H), 4.38-4.48 (m, 2H), 4.63 (brs, 1H), 4.94 (t, J=7.2Hz, 1H), 6.53 (d, J=8.4Hz, 1H), 6.66 (s, 1H), 6.96-7.01 (m, 3H), 7.12-7.42 (a plurality of m, 8H), 7.87 (d, J=8.0Hz, 1H), 7.98 (d, J=8.8Hz, 2H), 8.17-8.19 (m, 1H); MS (ESI) m/z 627 (M +), 628 (M ++ H).
Under room temperature; to 4-[1-[4-[N '-(2-chlorophenyl) urea groups]-3-methoxyphenyl acetyl group]-5-(R)-phenyl-2-(S)-pyrrolidinyl methoxyl group] essence of Niobe (251mg; 0.40mmol) methanol-THF (1: 1; add 1.0M sodium hydroxide (2.4ml in agitating solution 10ml); 2.40mmol); stir down, the mixture that obtains was heated 1.5 hours in 60 ℃.In this reactant mixture impouring 1N HCl, use chloroform extraction then.With salt water washing organic layer, through anhydrous sodium sulfate drying, vacuum concentration then.Residue with chloroform-methanol (10: 1) eluting, obtains 146 (181mg, 74%) through silica gel column chromatography, is colourless amorphous solid.MW 614.09 1H-NMR (CD 3OD), the mixture δ 1.99-2.19 of rotamer (m, 3H), 2.42-2.53 (m, 1H), 3.38 (dd, J=39,15Hz, 2H), 3.79 and 3.80 (s, 3H altogether), 3.94-5.19 (a plurality of m, 4H), 6.49-8.05 (a plurality of m, 16H); MS (FAB) m/z 614 (M ++ H); C 35H 34ClN 3O 6H 2The analytical calculation value of O: C, 65.06; H, 5.62; N, 6.50.Measured value: C, 65.03; H, 5.75; N, 6.45.Embodiment 1394-[1-[4-[N '-(2-bromo phenyl) urea groups]-3-methoxyphenyl acetyl group]-5-(R)-phenyl-2-(S)-pyrrolidinyl ethyoxyl] benzoic acid
Under room temperature, to 4-[5-(R)-phenyl-2-(S)-pyrrolidinyl methoxyl group] essence of Niobe (146mg, 0.47mmol), 4-[N '-(2-bromo phenyl) urea groups]-3-methoxybenzene guanidine-acetic acid (178mg, 0.47mmol) and N, (57.4mg adds EDCHCl (99.0mg in the agitating solution of DMF 0.47mmol) (10ml) to the N-dimethyl aminopyridine, 0.52mmol), the mixture stirring that obtains is spent the night.In reactant mixture impouring water, use ethyl acetate extraction.With salt water washing organic layer, through anhydrous sodium sulfate drying, vacuum concentration then.Residue is through silica gel column chromatography; with hexane-ethyl acetate (1: 5) as eluant; obtain 4-[1-[4-[N '-(2-bromo phenyl) urea groups]-3-methoxyphenyl acetyl group]-5-(R)-phenyl-2-(S)-pyrrolidinyl methoxyl group] essence of Niobe (288mg; 91%), is colourless amorphous solid. 1H-NMR (CDCl 3) δ 2.00-2.20 (m, 3H), 2.34-2.43 (m, 1H), 3.35 (s, 2H), 3.72 (s, 3H), 3.89 (s, 3H), 4.38-4.49 (m, 2H), 4.62 (brs, 1H), 4.94 (t, J=7.2Hz, 1H), 6.54 (d, J=8.4Hz, 1H), 6.67 (s, 1H), 6.91-7.05 (m, 4H), 7.28-7.42 (a plurality of m, 7H), 7.51 (d, J=8.0Hz, 2H), 7.87 (d, J=8.4Hz, 1H), 7.99 (d, J=8.8Hz, 2H), 8.14 (d, J=8.4Hz, 2H); MS (ESI) m/z 672 (M +), 674 (M ++ 2).
Under room temperature; to 4-[1-[4-[N '-(2-bromo phenyl) urea groups]-3-methoxyphenyl acetyl group]-5-(R)-phenyl-2-(S)-pyrrolidinyl methoxyl group] essence of Niobe (270mg; 0.40mmol) methanol-THF (1: 1; add 1.0M sodium hydroxide (2.0ml in agitating solution 10ml); 2.0mmol); stir down, the mixture that obtains was heated 1 hour in 60 ℃.In this reactant mixture impouring 1N HCl, use chloroform extraction then.With salt water washing organic layer, through anhydrous sodium sulfate drying, vacuum concentration then.Residue with chloroform-methanol (10: 1) eluting, obtains 147 (212mg, 80%) through silica gel column chromatography, is colourless amorphous solid.MW 658.54 1H-NMR (CD 3OD), the mixture δ 1.99-2.19 of rotamer (m, 3H), 2.42-2.53 (m, 1H), 3.38 (dd, J=39,16Hz, 2H), 3.79 and 3.80 (s, 3H altogether), 3.94-5.19 (a plurality of m, 4H), 6.49-8.00 (a plurality of m, 16H); MS (FAB) m/z 658 (M +), 660 (m ++ 2); Embodiment 1404-[1-[4-[N '-(2, the 6-Dichlorobenzene base) urea groups]-3-methoxyphenyl acetyl group]-5-(R)-phenyl-2-(S)-pyrrolidinyl methoxyl group] benzoic acid
Figure A0080951003791
Under room temperature, to 4-[5-(R)-phenyl-2-(S)-pyrrolidinyl methoxyl group] essence of Niobe (109mg, 0.35mmol), 4-[N '-(2, the 4-Dichlorobenzene base) urea groups]-3-methoxybenzene guanidine-acetic acid (129mg, 0.35mmol) and N, (42.8mg adds EDCHCl (73.4mg in the agitating solution of DMF 0.35mmol) (10ml) to the N-dimethyl aminopyridine, 0.39mmol), the mixture that obtains was stirred 6 hours.In reactant mixture impouring water, use ethyl acetate extraction.With salt water washing organic layer, through anhydrous sodium sulfate drying, vacuum concentration then.Residue is through silica gel column chromatography; with hexane-ethyl acetate (1: 4) as eluant; obtain 4-[1-[4-[N '-(2; 6-dichloro-phenyl) urea groups]-3-methoxyphenyl acetyl group]-5-(R)-phenyl-2-(S)-pyrrolidinyl methoxyl group] essence of Niobe (208mg; 90%), is colourless amorphous solid. 1H-NMR (CDCl 3) δ 2.00-2.21 (m, 3H), 2.33-2.39 (m, 1H), 3.31 (s, 2H), 3.69 (s, 3H), 3.88 (s, 3H), 4.34-4.45 (m, 2H), 4.59 (brs, 1H), 4.93 (t, J=6.8Hz, 1H), 6.47 (d, J=8.0Hz, 1H), 6.63 (s, 1H), 6.68 (s, 1H), 6.92-6.95 (m, 2H), and 7.12-7.41 (a plurality of m, 9H), 7.96-8.01 (m, 4H); MS (FAB) m/z 662 (M ++ H).
Under room temperature; to 4-[1-[4-[N '-(2; 6-dichloro-phenyl) urea groups]-3-methoxyphenyl acetyl group]-5-(R)-phenyl-2-(S)-pyrrolidinyl methoxyl group] essence of Niobe (186mg; 0.28mmol) methanol-THF (1: 1; add 1.0M sodium hydroxide (1.4ml in agitating solution 10ml); 1.4mmol), stir down, the mixture that obtains was heated 2.5 hours in 60 ℃.In this reactant mixture impouring 1N HCl, use chloroform extraction then.With salt water washing organic layer, through anhydrous sodium sulfate drying, vacuum concentration then.Residue with chloroform-methanol (10: 1) eluting, obtains 148 (166mg, 91%) through silica gel column chromatography, is colourless amorphous solid.MW 648.53 1H-NMR (CDCl 3) δ 2.00-2.18 (m, 3H), 2.34-2.40 (m, 1H), 3.33 (s, 2H), 3.68 (s, 3H), 4.37-4.47 (m, 2H), 4.61 (brs, 1H), 4.94 (t, J=6.8Hz, 1H), 6.48 (d, J=8.0Hz, 1H), 6.63 (s, 1H), 6.96 (d, J=8.4Hz, 3H), 7.12-7.38 (a plurality of m, 9H), 7.95 (d, J=8.0Hz, 1H), 8.01 (d, J=8.8Hz, 2H); MS (FAB) m/z 648 (M ++ H).Embodiment 1414-[1-[4-[N '-(2-bromo phenyl) urea groups]-3-aminomethyl phenyl acetyl group]-5-(R)-phenyl-2-(S)-pyrrolidinyl methoxyl group] benzoic acid
Under room temperature, to 4-[5-(R)-phenyl-2-(S)-pyrrolidinyl methoxyl group] essence of Niobe (125mg, 0.40mmol), 4-[N '-(2-bromo phenyl) urea groups]-3-aminomethyl phenyl acetic acid (146mg, 0.40mmol) and N, N-dimethyl aminopyridine (49.0mg, 0.40mmol) the agitating solution of DMF (10ml) in add EDCHCl (84.1mg 0.44mmol), stir the mixture that obtains 6 hours.In reactant mixture impouring water, use ethyl acetate extraction.With salt water washing organic layer, through anhydrous sodium sulfate drying, vacuum concentration then.Residue is through silica gel column chromatography; with hexane-ethyl acetate (1: 4) as eluant; obtain 4-[1-[4-[N '-(2-bromo phenyl) urea groups]-3-methoxyphenyl acetyl group]-5-(R)-phenyl-2-(S)-pyrrolidinyl methoxyl group] essence of Niobe (238mg; 90%), is colourless amorphous solid. 1H-NMR (CDCl 3) δ 1.92 (s, 3H), 2.09-2.27 (m, 3H), 2.42-2.50 (m, 1H), 3.22-3.41 (m, 2H), 3.88 (s, 3H), 4.39 (d, J=4.4Hz, 1H), 4.64 (brs, 1H), 5.00 (t, J=6.8Hz, 1H), 6.72 (s, 1H), 6.81-6.93 (a plurality of m, 8H), 7.22-7.42 (a plurality of m, 6H), 8.01 (d, J=8.4Hz, 2H), 8.13 (d, J=8.0Hz, 1H); MS (FAB) m/z 656 (M +), 658 (M ++ 2).
Under room temperature; to 4-[1-[4-[N '-(2-bromo phenyl) urea groups]-3-aminomethyl phenyl acetyl group]-5-(R)-phenyl-2-(S)-pyrrolidinyl methoxyl group] essence of Niobe (216mg; 0.33mmol) methanol-THF (1: 1; add 1.0M sodium hydroxide (1.7ml in agitating solution 10ml); 1.7mmol); stir down, the mixture that obtains was heated 2.5 hours in 60 ℃.In this reactant mixture impouring 1N HCl, use chloroform extraction then.With salt water washing organic layer, through anhydrous sodium sulfate drying, vacuum concentration then.Residue with chloroform-methanol (10: 1) eluting, obtains 149 (166mg, 91%) through silica gel column chromatography, is colourless amorphous solid.MW 642.54 1H-NMR (CDCl 3) δ 2.01 (s, 3H), 2.05-2.25 (m, 3H), 2.43-2.48 (m, 1H), 3.34 (dd, J=45,16Hz, 2H), 4.38-4.45 (m, 2H), 4.66 (brs, 1H), 4.99 (t, J=6.8Hz, 1H), 6.77 (s, 1H), 6.82-6.88 (m, 2H), 6.94 (d, J=8.8Hz, 2H), 7.15-7.55 (a plurality of m, 10H), 8.00 (d, J=8.8Hz, 2H), 8.14 (d, J=7.2Hz, 1H); MS (FAB) m/z 642 (M +), 644 (M ++ 2).Embodiment 1424-[1-[4-[N '-(2-aminomethyl phenyl) urea groups]-3-methoxyphenyl acetyl group]-5-(R)-methyl-2-(S)-pyrrolidinyl methoxyl group] benzoic acid
Figure A0080951003811
In-78 ℃, (7.55g, (1.1M is in ether, and 28.4ml 32.4mmol), makes the mixture that obtains be warmed to room temperature gradually, stirs then and spends the night to add lithium methide in the agitating solution of THF 23.6mmol) (100ml) to N-Boc-pyrroles's 1,3-propanedicarboxylic acid benzyl ester.Aqueous ammonium chloride solution is added in the reactant mixture, and vacuum is removed THF, uses ethyl acetate extraction then.Wash organic layer with water, through anhydrous sodium sulfate drying and vacuum concentration.Residue as eluant, obtains [2-(S)-(N-Boc-amino)-5-oxo-6-methyl] benzyl valerianate (5.02g, 45%) with hexane-ethyl acetate (3: 1) through silica gel column chromatography, is colourless spicule.Mp 85-87 ℃; 1H-NMR (CDCl 3) δ 1.43 (s, 9H), 1.61-2.15 (a plurality of m, 3H), 2.09 (s, 3H), 2.41-2.55 (m, 2H), 4.30 (brs, 1H), 4.70 (d, J=5.6Hz, 1H), 5.12-5.21 (m, 2H), 7.29-7.37 (m, 5H); MS (ESI) m/z 336 (M ++ H).
Under room temperature, (4.46g adds trifluoroacetic acid (20ml) in the agitating solution of dichloromethane 13.3mmol) (50ml), the mixture that obtains was stirred 1.5 hours to [2-(S)-(N-Boc-amino)-5-oxo-6-methyl] benzyl valerianate.This mixture of vacuum concentration is dissolved in the toluene it, and evaporation obtains 5-methyl-5-pyrrolin-2-(S)-benzyl formate trifluoroacetate (5.74g, quantitative) then, is the crude product brown oil.To this chemical compound in methanol (30ml) (1.97g, add in 5.94mmol) Pd/C (10%, 153mg), under nitrogen atmosphere, the mixture that obtains was stirred 3 days.Filter this mixture, vacuum concentrated filtrate obtains 5-methyl-5-pyrrolidine-2-(S)-methanoic acid trifluoro acetate (956mg, 66%), is white crude product solid.Under room temperature, to this chemical compound (939mg, 3.86mmol) and the methanol-water of Bis(tert-butoxycarbonyl)oxide (15: 1, (8.49ml 8.49mmol), stirred the mixture that obtains 1 hour 16ml) to add the 1.0M sodium hydroxide in the solution.The mixture that evaporation obtains also is poured in the 1N HCl aqueous solution, uses chloroform/methanol (5: 1) to extract then.Through anhydrous sodium sulfate drying organic layer, vacuum concentration then.Residue is through silica gel column chromatography, with chloroform-methanol (7: 1) eluting, obtains N-Boc-5-(R)-methyl-(S)-proline (711mg, 80%), is colorless oil. 1H-NMR(CD 3OD)δ1.27(d，J＝6.0Hz，3H)，1.41-1.46(m，9H)，1.62-1.64(m，1H)，1.96-2.01(m，2H)，2.22(brs，1H)，3.94(brs，1H)，4.17(brs，1H)；MS(ESI)m/z?230(M ++H)。
Under room temperature, (1.03g adds 10M-BH in the agitating solution of THF 4.49mmol) (20ml) to N-Boc-5-(R)-methyl-2-(S)-proline 3Me 2(1.57ml 15.7mmol), heats the mixture that obtains 5 hours under refluxing S.In this mixture impouring 1N HCl aqueous solution, use ethyl acetate extraction.Through anhydrous sodium sulfate drying organic layer, vacuum concentration then.Residue as eluant, obtains N-Boc-2-(S)-methylol-5-(R)-crassitude (838mg, 87%) with hexane-ethyl acetate (1: 3) through silica gel column chromatography, is colorless oil. 1H-NMR(CDCl 3)δ1.17(d，J＝6.0Hz，3H)，1.48(s，9H)，1.48-1.64(m，2H)，1.90-2.11(m，2H)，3.52-3.57(m，1H)，3.68-3.70(m，1H)，3.94-4.13(m，1H)。
Under room temperature, to N-Boc-2-(S)-methylol-5-(R)-crassitude (820mg, 3.81mmol), triphenyl phasphine (1.10g, 4.19mmol) and 4-methyl hydroxybenzoate (580mg, 3.81mmol) agitating solution in add diisopropyl azo-2-carboxylic acid (841ml, 4.19mmol), the mixture that obtains was stirred 1 hour in 60 ℃.This mixture of vacuum concentration, residue as eluant, obtain 4-[N-Boc-5-(R)-methyl-2-(S)-pyrrolidinyl methoxyl group with hexane-ethyl acetate (5: 1) through silica gel column chromatography] essence of Niobe (1.32g, 80%), be colorless oil. 1H-NMR (CDCl 3) δ 1.24 (brs, 3H), 1.49 (s, 9H), 1.55-1.70 (m, 2H), 1.94-2.11 (m, 2H), 3.88 (s, 3H), 3.88 (overlapping, 1H), 4.06-4.20 (m, 2H), 6.93-6.96 (m, 2H), 7.97 (d, J=8.8Hz, 2H); MS (ESI) m/z 350 (M ++ H).
Under room temperature, to 4-[N-Boc-5-(R)-methyl-2-(S)-pyrrolidinyl methoxyl group] (1.29g adds trifluoroacetic acid (10ml) to essence of Niobe in the agitating solution of dichloromethane 3.70mmol) (30ml), the mixture that obtains was stirred 35 minutes.In this mixture of vacuum concentration and the impouring sodium bicarbonate aqueous solution, use chloroform extraction then.Wash organic layer with water, through anhydrous sodium sulfate drying, vacuum concentration obtains 4-[5-(R)-methyl-2-(S)-pyrrolidinyl methoxyl group then] essence of Niobe (871mg, 95%), be colorless oil.This product is used for subsequent reaction without being further purified. 1H-NMR(CDCl 3)δ1.18(d，J＝6.4Hz，3H)，1.30-1.40(m，1H)，1.59-1.67(m，1H)，1.87-1.97(m，2H)，3.19-3.27(m，1H)，3.49-3.55(m，1H)，3.87(s，3H)，3.89-4.05(m，2H)，6.89(d，J＝8.8Hz，2H)，7.96(d，J＝8.8Hz，2H)；MS(ESI)m/z?250(M ++H)。
Under room temperature, to 4-[5-(R)-methyl-2-(S)-pyrrolidinyl methoxyl group] essence of Niobe (141mg, 0.57mmol), 4-[N '-(2-aminomethyl phenyl) urea groups]-3-methoxybenzene guanidine-acetic acid (178mg, 0.57mmol) and N, N-dimethyl aminopyridine (69.0mg, 0.57mmol) the agitating solution of DMF (10ml) in add EDCHCl (120mg 0.62mmol), stir the mixture that obtains and spends the night.In reactant mixture impouring water, use ethyl acetate extraction.With salt water washing organic layer, through anhydrous sodium sulfate drying, vacuum concentration then.Residue is through silica gel column chromatography; with ethyl acetate as eluant; obtain 4-[1-[4-[N '-(2-aminomethyl phenyl) urea groups]-3-methoxyphenyl acetyl group]-5-(R)-methyl-2-(S)-pyrrolidinyl methoxyl group] essence of Niobe (297mg, 96%), be colourless amorphous solid. 1H-NMR (CDCl 3) δ 1.24-1.34 (m, 3H), 1.93-2.18 (a plurality of m, 4H), 2.28 (s, 3H), 3.65 (s, 3H), 3.88 (s, 3H), 3.62-3.87 (m, 3H), 4.11-4.38 (a plurality of m, 3H), 6.42-8.06 (a plurality of m, 13H); MS (ESI) m/z546 (M ++ H).
Under room temperature; to 4-[1-[4-[N '-(2-aminomethyl phenyl) urea groups]-3-methoxyphenyl acetyl group]-5-(R)-methyl-2-(S)-pyrrolidinyl methoxyl group] essence of Niobe (279mg; 0.51mmol) methanol-THF (1: 1; add 1.0M sodium hydroxide (2.56ml in agitating solution 10ml); 2.56mmol); stir down, the mixture that obtains was heated 2 hours in 60 ℃.In this reactant mixture impouring 1N HCl, use chloroform extraction then.With salt water washing organic layer, through anhydrous sodium sulfate drying, vacuum concentration then.Residue with chloroform-methanol (15: 1) eluting, obtains 150 (269mg, 99%) through silica gel column chromatography, is colourless amorphous solid.MW 531.60 1H-NMR (CD 3OD), the mixture δ 1.28-1.35 of rotamer (m, 3H), 1.74-2.21 (a plurality of m, 4H), 2.28 (s, 3H), 3.71-4.37 (a plurality of m, 6H), 6.76-7.99 (a plurality of m, 11H); MS (ESI) m/z 532 (M ++ H).Embodiment 1434-[is trans-4-amino-1-[3-methoxyl group-4-[N '-(2-aminomethyl phenyl) urea groups] and phenyl acetyl]-(2S)-and the pyrrolidinyl methoxyl group] benzoic acid
Figure A0080951003841
In 0 ℃, to 4-(trans-4-amino-1-tert-butoxycarbonyl-(2S)-pyrrolidinyl) methoxyl methyl benzoate (1.0g, 2.86mmol) and TEA (1.2ml, add in dichloromethane 8.6mmol) (20.0ml) solution trifluoroacetic anhydride (720mg, 3.43mmol).After stirring 2.5 hours under the room temperature, water is added in this solution, use dichloromethane extraction.Wash extract with water, through dried over sodium sulfate and vacuum concentration.Residue is through purification by silica gel column chromatography, with ethyl acetate-normal hexane (1: 3, v/v) as eluant, obtain 4-(anti-form-1-tert-butoxycarbonyl-4-trifluoroacetamido-(2S)-pyrrolidinyl) methoxyl methyl benzoate (940mg, 74%), is colorless oil. 1H-NMR(CDCl 3)δ1.46(s，9H)，2.02-2.18(m，1H)，2.41-2.52(m，1H)，3.30-3.45(m，1H)，3.80-3.90(m，1H)，3.88(s，3H)，4.00-4.30(m，3H)，4.65-4.75(m，1H)，6.50(brs，1H)，6.91-6.94(m，2H)，7.96-7.99(m，2H)。
In 0 ℃, (470mg adds TFA (5.0ml) to methoxyl methyl benzoate in the agitating solution of dichloromethane 1.05mmol) (10.0ml) to 4-(anti-form-1-tert-butoxycarbonyl-4-trifluoroacetamido-(2S)-pyrrolidinyl).Under room temperature, this reactant mixture was stirred 0.5 hour.This mixture of vacuum concentration.Saturated sodium bicarbonate is added in this residue, use dichloromethane extraction.With salt water washing extract, through dried over sodium sulfate and vacuum concentration.Crude product is used for subsequent reaction without being further purified.In 0 ℃, to crude product, 3-methoxyl group-4-[N '-(2-aminomethyl phenyl) urea groups] phenylacetic acid (314mg, 1.0mmol), HOBt (162mg, 1.2mmol) and triethylamine (417ml, 3.0mmol) in the agitating solution of THF (10.0ml) and MeCN (10.0ml), add EDCHCl (288mg, 1.5mmol).Under room temperature, this reactant mixture was stirred 16 hours vacuum concentration.Water is added in the residue, use ethyl acetate extraction.With saturated sodium bicarbonate, 2M citric acid and saturated sodium bicarbonate washing organic layer, through dried over sodium sulfate and vacuum concentration.Residue is through purification by silica gel column chromatography; with ethyl acetate-normal hexane (3: 1; v/v) as eluant; obtain 4-[anti-form-1-[3-methoxyl group-4-[N '-(2-aminomethyl phenyl) urea groups] phenyl acetyl]-the 4-trifluoroacetamido-(2S)-pyrrolidinyl] methoxyl methyl benzoate (350mg; 52%), is colorless oil. 1H-NMR(CDCl 3)δ2.01-2.10(m，1H)，2.31(s，3H)，2.42-2.48(m，1H)，3.45-3.50(m，1H)，3.56-3.59(m，5H)，3.89(s，3H)，4.07-4.14(m，2H)，4.38-4.42(m，1H)，4.50-4.60(m，1H)，4.72-4.80(m，1H)，6.33(s，1H)，6.60-6.85(m，3H)，7.06-7.26(m，3H)，7.45-7.52(m，1H)，7.93-8.05(m，3H)。
To 4-[anti-form-1-[3-methoxyl group-4-[N '-(2-aminomethyl phenyl) urea groups] phenyl acetyl]-the 4-trifluoroacetamido-(2S)-pyrrolidinyl] methoxyl methyl benzoate (150mg; 0.23mmol) THF (3.0ml) and the agitating solution of methanol (2.0ml) in add the 1N sodium hydroxide (0.70ml, 0.70mmol).Stirred this mixture 18 hours in 60 ℃.This mixture of vacuum concentration adds water wherein, with 1N HCl neutralization.Collect the solid that produces, wash with water, vacuum drying obtains 151 (100mg, 81%), is white crystalline solid.MW 532.59 mp 170-171 ℃; IR (KBr) 3264,2937,1604,1535,1415,1376,1255,1224,1033 cm -1 1H-NMR (DMSO-d 6) δ 1.80-1.90 (m, 1H), 2.10-2.20 (m, 1H), 2.24 (s, 3H), 3.55-3.80 (m, 3H), 3.57 (s, 2H), 4.08-4.18 (m, 2H), 4.36-4.60 (m, 1H), 6.72-7.16 (m, 7H), 7.77-8.01 (m, 4H), 8.46 (s, 1H), 8.54 (s, 1H); MS (FAB) m/z 532 (M ++ 1); C 29H 32N 40 62.0H 20 analytical calculation value: C, 61.26; H, 6.38; N, 9.85.Measured value: C, 61.07; H, 6.32; N, 9.58.Embodiment 1444-[is trans-4-amino-1-[3-methoxyl group-4-[N '-(2-aminomethyl phenyl) urea groups] and phenyl acetyl]-(2S)-and pyrrolidinyl] the methoxyl methyl benzoate hydrochlorate
Under room temperature; to 4-[anti-form-1-[3-methoxyl group-4-[N '-(2-aminomethyl phenyl) urea groups] phenyl acetyl]-the 4-trifluoroacetamido-(2S)-pyrrolidinyl] methoxyl methyl benzoate (200mg; 0.31mmol) the agitating solution of methanol (4.0mml) in add entry (2.0ml) and potassium carbonate (138mg, 1.0mmol).After stirring 18 hours under the room temperature, water is added in this mixture, use dichloromethane extraction.Wash extract with water, through dried over sodium sulfate and vacuum concentration.Residue is through purification by silica gel column chromatography, with methanol-dichloromethane (5: 95-15: 85, v/v) as eluant.This product is dissolved in the ethanol (5.0ml), to wherein add 1N HCl (in ethanol) (1.0ml, 1.0mmol).This mixture of vacuum concentration obtains 152 (120mg, 63%), is amorphous solid.MW 546.61IR (KBr) 3382,2948,2879,1604,1533,1286,1255,771 cm -1 1H-NMR (DMSO-d 6) δ 2.25 (s, 3H), 2.10-2.30 (m, 2H), 3.59-3.70 (m, 3H), 3.77-3.80 (m, 8H), 4.00-4.24 (m, 2H), 4.47-4.67 (m, 1H), 6.70-7.16 (m, 7H), 7.77-8.00 (m, 4H), 8.49 (s, 1H), 8.55 (s, 1H); MS (FAB) m/z 547 (M ++ 1); C 30H 34N 4O 6HCl1.4H 2The analytical calculation value of O: C, 59.24; H, 6.26; N, 9.21; Cl, 5.83.Measured value: C, 59.42; H, 6.42; N, 9.04; Cl, 6.11.Embodiment 1454-[anti-form-1-[3-methoxyl group-4-[N '-(2-aminomethyl phenyl) urea groups] phenyl acetyl]-the 4-methylamino-(2S)-pyrrolidinyl] methoxybenzoic acid Under room temperature, to 4-(anti-form-1-tert-butoxycarbonyl-4-trifluoroacetamido-(2S)-pyrrolidinyl) methoxyl methyl benzoate (520mg, 1.17mmol) the agitating solution of DMF (10.0ml) in add potassium carbonate (321mg, 2.33mmol) and methyl iodide (330mg, 2.33mmol).In 50 ℃ this reactant mixture was stirred 18 hours.Water is added in this mixture, use ethyl acetate extraction.Wash organic layer with water, then through dried over sodium sulfate and vacuum concentration.Residue is through purification by silica gel column chromatography, with ethyl acetate-normal hexane (1: 2,, obtain 4-[anti-form-1-tert-butoxycarbonyl-4-(N-methyl-trifluoroacetamido)-(2S)-pyrrolidinyl v/v) as eluant] methoxyl methyl benzoate (390mg, 73%), is colorless oil. 1H-NMR (CDCl 3) δ 1.46 (s, 9H), 2.12-2.40 (m, 2H), 2.96 and 3.05 (they respectively are s, are total to 3H), 3.28-3.70 (m, 2H), 3.88 (s, 3H), 3.95-4.42 (m, 3H), 5.10-5.40 (m, 1H), 6.89-6.91 (m, 2H), 7.96-8.00 (m, 2H).
In 0 ℃, to 4-[anti-form-1-tert-butoxycarbonyl-4-(N-methyl-trifluoroacetamido)-(2S)-pyrrolidinyl] (390mg adds TFA (5.0ml) to methoxyl methyl benzoate in the agitating solution of dichloromethane 0.85mmol) (8.0ml).Under room temperature, stirred this reactant mixture 0.5 hour.This mixture of vacuum concentration adds saturated sodium bicarbonate in this residue, uses dichloromethane extraction.With salt water washing extract, through dried over sodium sulfate and vacuum concentration.Crude product is used for subsequent reaction without being further purified.In 0 ℃, to crude product, 3-methoxyl group-4-[N '-(2-aminomethyl phenyl) urea groups] phenylacetic acid (279mg, 0.89mmol), HOBt (143mg, 1.1mmol) and triethylamine (246ml, 1.77mmol) in the agitating solution of THF (8.0ml) and MeCN (8.0ml), add EDCHCl (255mg, 1.3mmol).Under room temperature, the mixture that obtains was stirred vacuum concentration 16 hours.Water is added in the residue, use ethyl acetate extraction.With saturated sodium bicarbonate, 2M citric acid and saturated sodium bicarbonate washing organic layer, then through dried over sodium sulfate and vacuum concentration.Residue is through purification by silica gel column chromatography; with ethyl acetate-normal hexane (4: 1; v/v) as eluant; obtain 4-[anti-form-1-[3-methoxyl group-4-[N '-(2-aminomethyl phenyl) urea groups] phenyl acetyl]-4-(N-methyl-trifluoroacetamido)-(2S)-pyrrolidinyl] methoxyl methyl benzoate (480mg; 82%), is colorless oil. 1H-NMR (CDCl 3) δ 2.18-2.35 (m, 2H), 2.31 (s, 3H), 2.87 and 2.97 (respectively is s, be total to 3H), 3.45-3.46 (m, 3H), 3.47 (s, 3H), 3.49 (s, 2H), 3.88 (s, 3H), 4.30-4.70 (m, 2H), and 5.20-5.40 (m, 1H), 6.38-6.43 (m, 1H), 6.67-6.86 (m, 4H), 7.09-7.24 (m, 4H), 7.51-7.54 (m, 1H), 7.93-8.08 (m, 3H).
To 4-[anti-form-1-[3-methoxyl group-4-[N '-(2-aminomethyl phenyl) urea groups] phenyl acetyl]-4-(N-methyl-N-TFA amino)-(2S)-pyrrolidinyl] methoxyl methyl benzoate (240mg; 0.37mmol) THF (5.0ml) and the agitating solution of methanol (3.0ml) in add the 1N sodium hydroxide (1.27ml, 1.27mmol).Stirred this mixture 18 hours in 60 ℃.This mixture of vacuum concentration adds water wherein, with 1N HCl neutralization.Collect the solid that produces, wash with water, vacuum drying obtains 153 (140mg, 70%), is white crystalline solid.MW 546.61 mp162-164 ℃ IR (KBr) 3338,1604,1535,1255,1033,755 cm -1 1H-NMR (DMSO-d 6) δ 1.85-1.95 (m, 1H), 2.10-2.20 (m, 1H), 2.24 (s, 3H), 2.34 and 2.39 (they respectively are s, are total to 3H), 3.41-3.71 (m, 3H), 3.58 (s, 2H), 3.80 (s, 3H), 4.05-4.20 (m, 2H), 4.36-4.60 (m, 1H), 6.73-7.16 (m, 7H), 7.77-8.01 (m, 4H), 8.45 (s, 1H), 8.53 (s, 1H); MS (FAB) m/z 547 (M ++ 1); C 30H 34N 4O 62.5H 2The analytical calculation value of O: C, 60.90; H, 6.64; N, 9.47.Measured value: C, 61.01; H, 6.50; N, 9.31.Embodiment 1464-[anti-form-1-[3-methoxyl group-4-[N '-(2-aminomethyl phenyl) urea groups] phenyl acetyl]-the 4-methylamino-(2S)-pyrrolidinyl] methoxyl methyl benzoate
Figure A0080951003881
Under room temperature; to 4-[anti-form-1-[3-methoxyl group-4-[N '-(2-aminomethyl phenyl) urea groups] phenyl acetyl]-4-(N-methyl trifluoro acetylamino)-(2S)-pyrrolidinyl] methoxyl methyl benzoate (240mg; 0.36mmol) THF (5.0ml) and the agitating solution of methanol (5.0ml) in add entry (2.0ml) and potassium carbonate (138mg, 1.0mmol).After stirring 18 hours under the room temperature, water is added in this mixture, use dichloromethane extraction.Wash extract with water, through dried over sodium sulfate and vacuum concentration.Residue is through purification by silica gel column chromatography, and (5/95-20/80 is v/v) as eluant with methanol-dichloromethane.This product is dissolved in the ethanol (5.0ml), to wherein add 1N HCl (in ethanol) (0.71ml, 0.71mmol).This mixture of vacuum concentration obtains 154 (180mg, 85%), is amorphous solid.MW 560.64 IR (KBr) 3311,2692,2453,1712,1604,1533 cm -1 1H-NMR (DMSO-d 6) δ 2.24 (s, 3H), 2.15-2.30 (m, 2H), 2.60 (br s, 3H), 3.60-4.20 (m, 5H), 3.78-3.81 (m, 8H), 4.47-4.70 (m, 1H), 6.71-7.16 (m, 7H), 7.77-8.00 (m, 4H), 8.48 (s, 1H), 8.55 (s, 1H), 9.21 (br s, 2H); MS (FAB) m/z 561 (M ++ 1); C 31H 36N 4O 6HCl1.4H 2The analytical calculation value of O: C, 59.83; H, 6.45; N, 9.00; Cl, 5.70.Measured value: C, 60.08; H, 6.51; N, 8.68; Cl, 5.99.Embodiment 1474-[is trans-4-dimethylamino-1-[3-methoxyl group-4-[N '-(2-aminomethyl phenyl) urea groups] and phenyl acetyl]-(2S)-and pyrrolidinyl] methoxybenzoic acid
In 0 ℃, to anti-form-1-tert-butoxycarbonyl-(2S)-hydroxymethyl-4-hydroxyl pyrrolidine (2.17g, 10.0mmol) and imidazoles (2.04g, add in the agitating solution of DMF 30.0mmol) (50ml) TBDPS-Cl (3.03g, 11.0mmol).In stirring this reactant mixture under the room temperature after 18 hours, water is added wherein, use ethyl acetate extraction.Wash extract with water, then through dried over sodium sulfate and vacuum concentration.Residue is through purification by silica gel column chromatography, with normal hexane-ethyl acetate (3: 2, v/v) as eluant, obtain the methyl-4-hydroxyl pyrrolidine (1.5g of anti-form-1-tert-butoxycarbonyl-(2S)-(tert-butyl diphenyl siloxy), 33%), is white crystalline solid. 1H-NMR (CDCl 3) δ 1.03 (s, 9H), 1.25 and 1.32 (respectively be s, 9H), 1.90-2.10 (m, 1H), 2.30-2.40 (m, 1H), 3.40-3.80 (m, 3H), 3.95-4.15 (m, 2H), 4.45-4.55 (m, 1H),, 7.37-7.39 (m, 6H), 7.63-7.64 (m, 4H).
Under room temperature, to the methyl-4-hydroxyl pyrrolidine (910mg of anti-form-1-tert-butoxycarbonyl-(2S)-(tert-butyl diphenyl siloxy), 2.0mmol) and triphenyl phasphine (628mg, add in the agitating solution of THF 2.4mmol) (20ml) carbon tetrabromide (993mg, 3.0mmol).Under room temperature, this mixture was stirred 0.5 hour.Normal hexane (40ml) is added wherein.Remove by filter the solid of generation, vacuum drying.Residue is through purification by silica gel column chromatography, with normal hexane to normal hexane-ethyl acetate (3: 2, v/v) as eluant, obtain the crassitude (1.0g of cis-4-bromo-1-tert-butoxycarbonyl-(2S)-(tert-butyl diphenyl siloxy), quantitatively), be faint yellow oily thing. 1H-NMR (CDCl 3) δ 1.06 (s, 9H), 1.31 and 1.45 (respectively be s, 9H), 2.63 (m, 2H), 3.49 (m, 1H), 3.89-4.14 (m, 5H), 7.35-7.42 (m, 6H), 7.64-7.66 (m, 4H).
Under room temperature, (480mg adds NaN in the agitating solution of DMF 0.93mmol) (5ml) to the crassitude of cis-4-bromo-1-tert-butoxycarbonyl-(2S)-(tert-butyl diphenyl siloxy) 3(241mg, 3.70mmol).In 70 ℃ this reactant mixture was stirred 3 days.Water is added wherein, use ethyl acetate extraction.Wash extract with water, then through dried over sodium sulfate and vacuum concentration.Crude product can be used for subsequent reaction without being further purified.Under room temperature and atmospheric pressure, with ethanol (10ml) solution of crude product through 10%Pd-C hydrogenation 4 hours.Remove by filter catalyst, vacuum concentrated filtrate obtains the crassitude (400mg, 95%) of trans-4-amino-1-tert-butoxycarbonyl-(2S)-(tert-butyl diphenyl siloxy), is colorless oil. 1H-NMR (CDCl 3) δ 1.06 (s, 9H), 1.32 and 1.45 (they respectively are s, are total to 9H), 2.20-2.35 (m, 1H), 3.05-3.18 (m, 1H), 3.55-4.05 (m, 6H), 7.35-7.41 (m, 6H), 7.61-7.69 (m, 4H).
In 0 ℃, to the crassitude (400mg of trans-4-amino-1-tert-butoxycarbonyl-(2S)-(tert-butyl diphenyl siloxy), 0.88mmol), (120ml 2.0mmol) and in the agitating solution of the methanol (10ml) of 37%HCHO aqueous solution (500ml) adds NaBH to acetic acid 3CN (111mg, 1.76mmol).Under room temperature, stirred reaction mixture 18 hours.Behind the vacuum concentration, add entry, use dichloromethane extraction.Through dried over sodium sulfate extract and vacuum concentration.Residue is through purification by silica gel column chromatography, with methanol-dichloromethane (3: 97, v/v) as eluant, obtain the methyl-4-dimethylamino pyrrolidine (330mg of anti-form-1-tert-butoxycarbonyl-(2S)-(tert-butyl diphenyl siloxy), 78%), is faint yellow oily thing. 1H-NMR (CDCl 3) δ 1.06 (s, 9H), 1.33 and 1.45 (they respectively are s, are total to 9H), 1.80-2.25 (m, 2H), 2.23 (br s, 6H), 2.95-4.05 (m, 6H), 7.36-7.39 (m, 6H), 7.63-7.65 (m, 4H).
In 0 ℃, to the methyl-4-dimethylamino pyrrolidine (330mg of anti-form-1-tert-butoxycarbonyl-(2S)-(tert-butyl diphenyl siloxy), 0.68mmol) the agitating solution of THF (5ml) in add TBAF (1.0M is solution in THF, 1.0ml, 1.0mmol).Under room temperature, stirred this reactant mixture 2 hours.This mixture of vacuum concentration.Residue is through purification by silica gel column chromatography, with methanol-dichloromethane (3: 97-20: 80, v/v) as eluant, obtain that anti-form-1-tert-butoxycarbonyl-4-dimethylamino-(2S)-(180mg quantitatively), is faint yellow oily thing to hydroxymethyl pyrrolidine. 1H-NMR(CDCl 3)δ1.47(s，9H)，2.23(s，6H)，1.65-1.75(m，2H)，2.75-4.10(m，4H)，3.61(d，J＝5.6Hz，2H)。
In 0 ℃, to anti-form-1-tert-butoxycarbonyl-4-dimethylamino-(2S)-hydroxymethyl pyrrolidine (180mg, 0.73mmol), 4-methyl hydroxybenzoate (114mg, 0.75mmol) and triphenyl phasphine (296mg, 1.13mmol) the agitating solution of THF (10ml) in add DIAD (227mg, 1.13mmol).This reactant mixture was stirred this mixture of vacuum concentration 18 hours in 70 ℃.Residue is through purification by silica gel column chromatography, with normal hexane-ethyl acetate (1: 2, v/v) to methanol-dichloromethane (5: 95, v/v) as eluant, obtain 4-[anti-form-1-tert-butoxycarbonyl-4-dimethylamino-(2S)-pyrrolidinyl] methoxyl methyl benzoate (180mg, 68%), is faint yellow oily thing. 1H-NMR(CDCl 3)δ1.46(s，9H)，1.80-1.95(m，1H)，2.20-2.23(m，1H)，2.24(s，6H)，2.90-2.95(m，1H)，3.10-3.30(m，1H)，3.50-3.65(m，1H)，3.88(s，3H)，3.95-4.35(m，3H)，6.93-6.95(m，2H)，7.96-7.98(m，2H)。
In 0 ℃, to 4-(anti-form-1-tert-butoxycarbonyl-4-dimethylamino-(2S)-pyrrolidinyl) methoxyl methyl benzoate (200mg, 0.53mmol) the agitating solution of dichloromethane (6ml) in add TFA (3ml), this reactant mixture was stirred under room temperature 0.5 hour.This mixture of vacuum concentration.Saturated sodium bicarbonate is added in this residue, use dichloromethane extraction.With salt water washing extract, through dried over sodium sulfate and vacuum concentration.Crude product is used for subsequent reaction without being further purified.In 0 ℃, to crude product, 3-methoxyl group-4-[N '-(2-aminomethyl phenyl) urea groups] phenylacetic acid (166mg, 0.53mmol), HOBt (71mg, 0.53mmol) and triethylamine (140ml, 1.10mmol) in the agitating solution of THF (5ml) and MeCN (5ml), add EDCHCl (152mg, 0.79mmol).Under room temperature, reactant mixture was stirred 16 hours vacuum concentration.Water is added in the residue, use ethyl acetate extraction.Wash extract with saturated sodium bicarbonate, then through dried over sodium sulfate and vacuum concentration.Residue is through purification by silica gel column chromatography; with ethyl acetate to methylene chloride-methanol (8: 92; v/v) as eluant; obtain 4-[trans-4-dimethylamino-1-[3-methoxyl group-4-[N '-(2-aminomethyl phenyl) urea groups] phenyl acetyl]-(2S)-pyrrolidinyl] methoxyl methyl benzoate (260mg; 86%), is colorless oil. 1H-NMR(CDCl 3)δ1.95-2.15(m，3H)，2.23(s，6H)，2.31(s，3H)，3.30-3.34(m，1H)，3.57(s，2H)，3.61(s，3H)，3.70-3.75(m，1H)，4.11-4.15(m，2H)，4.45-4.50(m，1H)，6.34(s，1H)，6.72-6.88(m，4H)，7.08-7.24(m，4H)，7.51-7.53(m，1H)，7.92-8.07(m，3H)。
To 4-[trans-4-dimethylamino-1-[3-methoxyl group-4-[N '-(2-aminomethyl phenyl) urea groups] phenyl acetyl]-the 2-pyrrolidinyl] methoxyl methyl benzoate (260mg; 0.45mmol) THF (4.0ml) and the agitating solution of methanol (2.0ml) in add the 1N sodium hydroxide (0.90ml, 0.90mmol).Stirred this mixture 24 hours in 70 ℃.This mixture of vacuum concentration adds water wherein, with 1N HCl neutralization.Collect the solid that produces, wash with water, vacuum drying obtains 155 (200mg, 79%), is white crystalline solid.MW 560.64 mp 145-150 ℃; IR (KBr) 3355,2948,1698,1604,1,533 1454, and 1417,1255,1226,1166,1035,755 cm -1 1H-NMR (DMSO-d 6) δ 1.82-1.98 (m, 1H), 2.08-2.11 (m, 1H), 2.20 (s, 6H), 2.25 (s, 3H), 3.40-3.60 (m, 3H), 3.64 (s, 2H), 3.82 (s, 3H), 4.01-4.16 (m, 2H), 4.36 (m, 1H), 6.74-7.15 (m, 7H), 7.77-8.02 (m, 4H), 8.44 (s, 1H), 8.54 (s, 1H); MS (FAB) m/z 561 (M ++ 1); C 31H 36N 4O 61.2H 2The analytical calculation value of O: C, 63.95; H, 6.65; N, 9.62.Measured value: C, 63.82; H, 6.72; N, 9.44.Embodiment 1484-[is trans-4-dimethylamino-1-[3-methoxyl group-4-[N '-(2-aminomethyl phenyl) urea groups] and phenyl acetyl]-(2S)-and pyrrolidinyl] the methoxyl methyl benzoate hydrochlorate In 0 ℃; to trans-4-[4-dimethylamino-1-[3-methoxyl group-4-[N '-(2-aminomethyl phenyl) urea groups] phenyl acetyl]-(2S)-and pyrrolidinyl] (80mg adds TMSCHN in the agitating solution of toluene 0.14mmol) (4.0ml) and methanol (1.0ml) to methoxybenzoic acid 2(2.0M in hexane, 100ml, 0.20mmol).Under room temperature, stirred this reactant mixture 1.5 hours.This mixture of vacuum concentration.Residue is through purification by silica gel column chromatography, with methanol-dichloromethane (5: 95, v/v) as eluant.Product is dissolved in the ethanol (5.0ml), and (in ethanol, 244 μ l 0.244mmol) add wherein with 1N HCl.This mixture of vacuum concentration obtains 156 (72mg, 88%), is amorphous solid.MW 574.67 IR (KBr) 3345,2950,2586,1712,1604,1511,1454,1284, and 1255,1170,1114,1029,850,771 cm -1 1H-NMR (DMSO-d 6) δ 2.25 (s, 3H), 2.35-2.37 (m, 2H), 2.77-2.81 (m, 6H), 3.62-3.71 (m, 2H), 3.79-3.81 (m, 8H), 3.99-4.16 (m, 3H), 4.50-4.70 (m, 1H), 6.74-7.16 (m, 7H), 7.77-8.01 (m, 4H), 8.48 (s, 1H), 8.55 (s, 1H); C 32H 38N 4O 61.0HCl1.2H 2The analytical calculation value of O: C, 60.74; H, 6.59; N, 8.85.Measured value: C, 61.03; H, 6.78; N, 8.33.Embodiment 1494-[cis-4-dimethylamino-1-[3-methoxyl group-4-[N '-(2-aminomethyl phenyl) urea groups] phenyl acetyl]-(2S)-and pyrrolidinyl] methoxybenzoic acid
In 0 ℃, to the methyl-4-hydroxyl pyrrolidine (1.82mg of cis-1-tert-butoxycarbonyl-(2S)-(tert-butyl diphenyl siloxy), 4.0mmol), phthalimide (647mg, 4.4mmol) and triphenyl phasphine (1.26g, 4.8mmol) the agitating solution of THF (20ml) in add DIAD (889mg, 4.4mmol).This reactant mixture was stirred 18 hours this mixture of vacuum concentration under room temperature.Residue is through purification by silica gel column chromatography, with normal hexane-ethyl acetate (5/1,, obtain the methyl-4-pyrrolidinyl of N-[cis-1-tert-butoxycarbonyl-(2S)-(tert-butyl diphenyl siloxy) v/v) as eluant] phthalimide (1.6g, 69%), is amorphous solid. 1H-NMR (CDCl 3) δ 1.07 (s, 9H), 1.30 and 1.44 (they respectively are s, are total to 9H), 2.27-2.37 (m, 1H), 2.94-2.96 (m, 1H), 3.81-4.09 (m, 5H), 4.72 (m, 1H), 7.37-7.38 (m, 6H), 7.67-7.74 (m, 6H), 7.84-7.86 (m, 2H).
Under room temperature, to the methyl-4-pyrrolidinyl of N-[cis-1-tert-butoxycarbonyl-(2S)-(tert-butyl diphenyl siloxy)] (1.60g adds NH in the agitating solution of ethanol 2.74mmol) (8ml) to phthalimide 2NH 2H 2O (206mg, 4.11mmol).In 70 ℃, this reactant mixture was stirred 1 hour.This mixture of vacuum concentration.Remove by filter the solid of generation, wash with chloroform.Vacuum concentrated filtrate.Remove by filter the solid of generation, wash with chloroform.Vacuum concentrated filtrate.Leach the solid of generation, wash with chloroform.Vacuum concentrated filtrate, (1.3g quantitatively), is faint yellow oily thing to obtain the crassitude of cis-4-amino-1-tert-butoxycarbonyl-(2S)-(tert-butyl diphenyl siloxy).Crude product is used for subsequent reaction without being further purified. 1H-NMR (CDCl 3) δ 1.06 (s, 9H), 1.30 and 1.45 (they respectively are s, are total to 9H), 1.59 (m, 1H), 1.85 (m, 1H), 2.94 (m, 1H), 3.44 (m, 1H), 3.78-4.07 (m, 4H), 7.36-7.41 (m, 6H), 7.51-7.65 (m, 4H).
In 0 ℃, to the crassitude (1.24g of cis-4-amino-1-tert-butoxycarbonyl-(2S)-(tert-butyl diphenyl siloxy), 2.74mmol), (374 μ l 5.48mmol) and in the agitating solution of the methanol (20ml) of 37%HCHO aqueous solution (1.0ml) add NaBH to acetic acid 3CN (345mg, 5.48mmol).Stirred reaction mixture is 18 hours under room temperature.Behind the vacuum concentration, add entry, use dichloromethane extraction.Through dried over sodium sulfate extract and vacuum concentration.Residue is through purification by silica gel column chromatography, with methanol-dichloromethane (3/97, v/v) as eluant, obtain the methyl-4-dimethylamino pyrrolidine (1.1g of cis-1-tert-butoxycarbonyl-(2S)-(tert-butyl diphenyl siloxy), 83%), is faint yellow oily thing. 1H-NMR (CDCl 3) δ 1.05 (s, 9H), 1.29 and 1.45 (they respectively are s, are total to 9H), 1.95-2.04 (m, 1H), 2.20-2.26 (m, 1H), 2.27 (s, 6H), 2.54 (m, 1H), 3.00-3.02 (m, 1H), 3.62-4.03 (m, 4H), 7.34-7.41 (m, 6H), 7.63-7.65 (m, 4H).
In 0 ℃, to cis-1-tert-butoxycarbonyl-2-(tert-butyl diphenyl siloxy) methyl-4-dimethylamino pyrrolidine (1.1g, 2.27mmol) the agitating solution of THF (10ml) in add TBAF (solution of 1.0M in THF, 4.5ml, 4.5mmol).Under room temperature, stirred this reactant mixture 2 hours.This mixture of vacuum concentration.Residue is through purification by silica gel column chromatography, with methanol-dichloromethane (3/97-20: 80, v/v) as eluant, obtain that cis-1-tert-butoxycarbonyl-4-dimethylamino-(2S)-(580mg quantitatively), is faint yellow oily thing to hydroxymethyl pyrrolidine. 1H-NMR(CDCl 3)δ1.47(s，9H)，1.25-1.96(m，2H)，2.25(S，6H)，2.53-2.58(m，1H)，3.17-4.02(m，5H)。
In 0 ℃, to cis-1-tert-butoxycarbonyl-4-dimethylamino-(2S)-hydroxymethyl pyrrolidine (555mg, 2.27mmol), 4-methyl hydroxybenzoate (380mg, 2.5mmol) and triphenyl phasphine (1.07g, 4.09mmol) the agitating solution of THF (10ml) in add DIAD (826mg, 4.09mmol).This reactant mixture was stirred this mixture of vacuum concentration 18 hours in 70 ℃.Residue is through purification by silica gel column chromatography, with normal hexane-ethyl acetate (1/2, v/v) methanol-dichloromethane (5/95, v/v) as eluant, obtain 4-(cis-1-tert-butoxycarbonyl-4-dimethylamino-(2S)-pyrrolidinyl) methoxyl methyl benzoate (260mg, 30%), is faint yellow oily thing. 1H-NMR(CDCl 3)δ1.45(s，9H)，1.70-1.90(m，1H)，2.26(s，6H)，2.33(m，1H)，2.57(m，1H)，3.06(m，1H)，3.85-4.23(m，4H)，3.88(s，3H)，6.93(m，2H)，7.95(m，2H)。
In 0 ℃, to 4-(cis-1-tert-butoxycarbonyl-4-dimethylamino-(2S)-pyrrolidinyl) methoxyl methyl benzoate (208mg, 0.55mmol) the agitating solution of dichloromethane (6ml) in add TFA (3ml), this reactant mixture was stirred under room temperature 0.5 hour.This mixture of vacuum concentration.Saturated sodium bicarbonate is added in this residue, use dichloromethane extraction.With salt water washing extract, through dried over sodium sulfate and vacuum concentration.Crude product is used for subsequent reaction without being further purified.In 0 ℃, to crude product, 3-methoxyl group-4-[N '-(2-aminomethyl phenyl) urea groups] phenylacetic acid (173mg, 0.55mmol), HOBt (74mg, 0.55mmol) and triethylamine (153 μ l, 1.1mmol) in the agitating solution of THF (6ml) and MeCN (6ml), add EDCHCl (160mg, 0.83mmol).Under room temperature, reactant mixture was stirred 16 hours vacuum concentration.Water is added in the residue, use ethyl acetate extraction.Wash extract with saturated sodium bicarbonate, then through dried over sodium sulfate and vacuum concentration.Residue is through purification by silica gel column chromatography; with ethyl acetate to methylene chloride-methanol (5/95; v/v) as eluant; obtain 4-[cis-4-dimethylamino-1-[3-methoxyl group-4-[N '-(2-aminomethyl phenyl) urea groups] phenyl acetyl]-(2S)-and pyrrolidinyl] methoxyl methyl benzoate (270mg; 47%), is colorless oil. 1H-NMR(CDCl 3)δ1.95-2.04(m，1H)，2.25(s，6H)，2.32(s，3H)，2.61(m，1H)，3.21(m，1H)，3.56-3.58(m，5H)，3.80-3.83(m，1H)，3.88(s，3H)，4.18-4.20(m，1H)，4.41-4.45(m，2H)，6.36(s，1H)，6.68-6.85(m，4H)，7.08-7.25(m，4H)，7.52-7.55(m，1H)，7.91-8.07(m，3H)。
To 4-[cis-4-monomethyl amino-1-[3-methoxyl group-4-[N '-(2-aminomethyl phenyl) urea groups] phenyl acetyl]-(2S)-and pyrrolidinyl] methoxyl methyl benzoate (270mg; 0.47mmol) THF (4.0ml) and the agitating solution of methanol (2.0ml) in add the 1N sodium hydroxide (1.0ml, 1.0mmol).Stirred this mixture 18 hours in 70 ℃.This mixture of vacuum concentration adds water wherein, with 1N HCl neutralization.Collect the solid that produces, wash with water, vacuum drying obtains 157 (170mg, 65%), is white crystalline solid.MW 560.64 mp 147-150 ℃; IR (KBr) 3353,2952,1700,1604,1,533 1454, and 1415,1255,1166,1035,755cm -1 1H-NMR (DMSO-d 6) δ: 1.83-1.84 (m, 1H), 2.08-2.10 (m, 1H), 2.21 (br s, 6H), 2.24 (s, 3H), 3.00 (m, 2H), 3.60 (s, 2H), 3.78 (s, 3H), 3.85-4.29 (m, 4H), 6.71-7.16 (m, 7H), 7.77-8.01 (m, 4H), 8.46 (s, 1H), 8.54 (s, 1H); MS (FAB) m/z 561 (M+H) +C 31H 36N 4O 62H 2The analytical calculation value of O: C, 62.40; H, 6.76; N, 9.39.Measured value: C, 62.51; H, 6.60; N, 9.36.Embodiment 1504-[cis-4-dimethylamino-1-[3-methoxyl group-4-[N '-(2-aminomethyl phenyl) urea groups] phenyl acetyl]-(2S)-and pyrrolidinyl] the methoxyl methyl benzoate hydrochlorate
In 0 ℃; to 4-[cis-4-dimethylamino-1-[3-methoxyl group-4-[N '-(2-aminomethyl phenyl) urea groups] phenyl acetyl]-(2S)-and pyrrolidinyl] (80mg adds TMSCHN in the agitating solution of toluene 0.14mmol) (4.0ml) and methanol (1.0ml) to methoxybenzoic acid 2(2.0M is in hexane) (100 μ l, 0.20mmol).Under room temperature, stirred this reactant mixture 1.5 hours.This mixture of vacuum concentration.Residue is through purification by silica gel column chromatography, with methanol-dichloromethane (5/95, v/v) as eluant.Product is dissolved in the ethanol (5.0ml), and (244 μ l 0.244mmol) add wherein with 1N HCl (in ethanol).This mixture of vacuum concentration obtains 158 (75mg, 79%), is amorphous solid.MW 574.67 IR (KBr) 3345,2950,2456,1712,1646,1604,1511,1454,1434,1415,1284,1257,1168,1114,1031,771cm -1 1H-NMR (DMSO-d 6) δ 2.10-2.20 (m, 2H), 2.25 (s, 3H), 2.83 (m, 6H), 3.60-3.62 (m, 2H), 3.76-3.81 (m, 8H), 4.20-4.33 (m, 4H), 6.71-7.17 (m, 6H), 7.77-7.98 (m, 5H), 8.47 (s, 1H), 8.55 (s, 1H); MS (FAB) m/z 574 (M+H) +C 32H 38N 4O 61.0HCl1.3H 2The analytical calculation value of O: C, 60.57; H, 6.61; N, 8.83.Measured value: C, 60.80; H, 6.82; N, 8.44.Embodiment 1514-[anti-form-1-[4-[N '-(2-chlorophenyl) urea groups]-3-methoxyphenyl acetyl group]-the 4-dimethylamino-(2S)-pyrrolidinyl] methoxybenzoic acid
In 0 ℃, (430mg adds TFA (5.0ml) to methoxyl methyl benzoate in the agitating solution of dichloromethane 1.1mmol) (10.0ml) to 4-(anti-form-1-tert-butoxycarbonyl-4-dimethylamino-(2S)-pyrrolidinyl).This reactant mixture was stirred under room temperature 0.5 hour.This mixture of vacuum concentration.Saturated sodium bicarbonate is added in this residue, use dichloromethane extraction.With salt water washing extract, through dried over sodium sulfate and vacuum concentration.Crude product is used for subsequent reaction without being further purified.In 0 ℃, to crude product, 4-[N '-(2-chlorophenyl) urea groups]-3-methoxybenzene guanidine-acetic acid (368mg, 1.1mmol), HOBt (162mg, 1.2mmol) and triethylamine (417ml, 3.0mmol) in the agitating solution of THF (10.0ml) and MeCN (10.0ml), add EDCHCl (288mg, 1.1mmol).Under room temperature, reactant mixture was stirred 16 hours vacuum concentration.Water is added in the residue, use ethyl acetate extraction.With saturated sodium bicarbonate, 2M citric acid and saturated sodium bicarbonate washing extract, then through dried over sodium sulfate and vacuum concentration.Residue is through purification by silica gel column chromatography; with normal hexane-ethyl acetate (1: 1; v/v) as eluant; obtain 4-[anti-form-1-[4-[N '-(2-chlorophenyl) urea groups]-3-methoxyphenyl acetyl group]-the 4-dimethylamino-(2S)-pyrrolidinyl] methoxybenzoic acid ester (530mg; 78%), is colorless oil. 1H-NMR(CDCL 3)δ1.94-1.99(m，1H)，2.48(s，9H)，3.06-3.12(m，1H)，3.33-3.38(m，1H)，3.60(s，2H)，3.68(s，3H)，3.69-3.80(m，1H)，3.88(s，3H)，4.13-4.20(m，2H)，4.56(m，1H)，6.76-7.00(m，5H)，7.22-7.34(m，3H)，7.92-8.00(m，3H)，8.17-8.19(m，1H)。For HCl salt: IR (KBr) 3324,2950,2454,1710,1604,1511,1284 cm -1 1H-NMR (DMSO-d 6) δ 2.30-2.40 (m, 2H), 2.77-2.80 (m, 6H), 3.60-3.75 (m, 2H), 3.75-3.85 (m, 8H), and 4.00-4.22 (m, 3H), 4.50-4.75 (m, 1H), 6.75-7.43 (m, 7H), 7.89-8.09 (m, 4H), 8.87 (s, 1H), 8.91 (s, 1H); MS (FAB) m/z 595 (M+H) +C 31H 36N 4O 6Cl1.0HCl1.0H 2The analytical calculation value of O: C, 57.23; H, 6.04; N, 8.61; Cl, 10.90.Measured value: C, 57.43; H, 6.08; N, 8.38; Cl, 10.73.
To 4-[anti-form-1-[4-[N '-(2-chlorophenyl) urea groups]-3-methoxyphenyl acetyl group]-the 4-dimethylamino-(2S)-pyrrolidinyl] methoxyl methyl benzoate (190mg; 0.32mmol) THF (3.0ml) and the agitating solution of methanol (2.0ml) in add the 1N sodium hydroxide (0.64ml, 0.64mmol).Stirred this mixture 24 hours in 70 ℃.This mixture of vacuum concentration adds water wherein, with 1N HCl neutralization.Collect the solid that produces, wash with water, vacuum drying obtains 159 (150mg, 83%), is white crystalline solid.MW 581.06 mp 159-161 ℃; IR (KBr) 3318,2938,1604,1531,1438,1340 cm -1 1H-NMR (DMSO-d 6) δ 2.10-2.40 (m, 8H), 2.50-2.70 (m, 2H), 3.85-3.90 (m, 5H), 4.02-4.18 (m, 3H), 4.30-4.60 (m, 1H), 6.75-7.43 (m, 7H), 7.86-8.09 (m, 4H), 8.86 (s, 1H), 8.91 (s, 1H); MS (FAB) m/z 581 (M+1) +C 30H 22N 4O 6Cl2H 2The analytical calculation value of O: C, 59.79; H, 5.92; N, 9.30.Measured value: C, 59.69; H, 5.93; N, 9.09.Embodiment 1524-[cis-1-[4-[N '-(2-chlorophenyl) urea groups]-3-methoxyphenyl acetyl group]-the 4-dimethylamino-(2S)-pyrrolidinyl] methoxybenzoic acid
In 0 ℃, (1.2g adds TFA (5.0ml) to methoxyl methyl benzoate in the agitating solution of dichloromethane 3.2mmol) (10.0ml) to 4-(cis-1-tert-butoxycarbonyl-4-dimethylamino-(2S)-pyrrolidinyl).This reactant mixture was stirred under room temperature 0.5 hour.This mixture of vacuum concentration.Saturated sodium bicarbonate is added in this residue, use dichloromethane extraction.With salt water washing extract, through dried over sodium sulfate and vacuum concentration.Crude product is used for subsequent reaction without being further purified.In 0 ℃, to crude product (278mg, 1.0mmol), 4-[N '-(2-chlorophenyl) urea groups]-3-methoxybenzene guanidine-acetic acid (335mg, 1.0mmol), HOBt (135mg, 1.0mmol) and triethylamine (417ml, 3.0mmol) in the agitating solution of THF (4.0ml) and MeCN (4.0ml), add EDCHCl (288mg, 1.5mmol).Under room temperature, reactant mixture was stirred 16 hours vacuum concentration.Water is added in the residue, use ethyl acetate extraction.With saturated sodium bicarbonate, 2M citric acid and saturated sodium bicarbonate washing extract, then through dried over sodium sulfate and vacuum concentration.Residue is through purification by silica gel column chromatography; with normal hexane-ethyl acetate (1: 1; v/v) as eluant; obtain 4-[cis-1-[4-[N '-(2-chlorophenyl) urea groups]-3-methoxyphenyl acetyl group]-the 4-dimethylamino-(2S)-pyrrolidinyl] methoxyl methyl benzoate (500mg; 84%), is colorless oil. 1H-NMR(CDCl 3)δ1.98-2.50(m，1H)，2.26(s，3H)，2.25-2.40(m，1H)，2.58-2.65(m，1H)，3.20-3.30(m，1H)，3.60(s，2H)，3.64(s，3H)，3.80-3.90(m，1H)，3.88(s，3H)，4.18-4.20(m，1H)，4.42-4.46(m，2H)，6.72-7.00(m，4H)，7.20-7.35(m，5H)，7.91-7.94(m，3H)，8.18-8.21(m，1H)。
To 4-[cis-1-[4-[N '-(2-chlorophenyl) urea groups]-3-methoxyphenyl acetyl group]-the 4-dimethylamino-(2S)-pyrrolidinyl] methoxyl methyl benzoate (250mg; 0.42mmol) THF (5.0ml) and the agitating solution of methanol (3.0ml) in add the 1N sodium hydroxide (1.0ml, 1.0mmol).Stirred this mixture 18 hours in 70 ℃.This mixture of vacuum concentration adds water wherein, with 1N HCl neutralization.Collect the solid that produces, wash with water, vacuum drying obtains 160 (170mg, 70%), is white crystalline solid.MW 581.06 mp 165-167 ℃; IR (KBr) 3328,1604,1531,1164,1033 cm -1 1H-NMR (DMSO-d 6) δ 1.80-1.90 (m, 1H), 2.20-2.50 (m, 7H), 3.60-3.70 (m, 2H), 3.77-3.81 (m, 5H), 4.00-4.30 (m, 4H), 6.72-7.44 (m, 7H), 7.86-8.10 (m, 4H), 8.88-8.92 (m, 2H); MS (FAB) m/z 581 (M ++ 1); C 30H 33N 4O 6Cl1.1H 2The analytical calculation value of O: C, 59.87; H, 6.06; N, 9.31.Measured value: C, 59.65; H, 5.76; N, 9.09.Embodiment 1534-[cis-1-[4-[N '-(2-chlorophenyl) urea groups]-3-methoxyphenyl acetyl group]-4-(2-naphthalene sulfonamido)-(2S)-pyrrolidinyl] methoxybenzoic acid
In 0 ℃, to 4-(cis-1-tert-butoxycarbonyl-4-amino-(2S)-pyrrolidinyl) methoxyl methyl benzoate (200mg, 0.57mmol) and TEA (317ml, and adding (2-naphthyl) sulfonic acid chloride in the agitating solution of chloroform 2.3mmol) (10.0ml) (155mg, 0.68mmol).This reactant mixture was stirred under room temperature 18 hours.This reactant mixture of vacuum concentration.Residue is through purification by silica gel column chromatography, with normal hexane-ethyl acetate (2: 1, v/v) as eluant, obtain 4-(cis-1-tert-butoxycarbonyl-4-(2-naphthyl sulfonamido)-(2S)-pyrrolidinyl) methoxyl methyl benzoate (240mg, 78%), is faint yellow oily thing. 1H-NMR(CDCl 3)δ1.25-1.45(br?s，9H)，1.70-1.80(m，1H)，2.20-2.40(m，1H)，3.20-3.50(m，2H)，3.90(s，3H)，3.85-4.15(m，3H)，4.55-4.65(m，1H)，6.90-7.10(m，2H)，7.58-8.04(m，8H)，8.43(s，1H)。
(240mg adds TFA (5.0ml) to methoxyl methyl benzoate in the agitating solution of dichloromethane 0.44mmol) (5.0ml) to 4-(cis-1-tert-butoxycarbonyl-4-(2-naphthyl sulfonamido)-(2S)-pyrrolidinyl).Under room temperature, stirred this reactant mixture 0.5 hour.This mixture of vacuum concentration adds saturated sodium bicarbonate in this residue, uses dichloromethane extraction.With salt water washing extract, through dried over sodium sulfate and vacuum concentration.Crude product is used for subsequent reaction without being further purified.In 0 ℃, to crude product, 4-[N '-(2-chlorophenyl) urea groups]-3-methoxybenzene guanidine-acetic acid (147mg, 0.44mmol), HOBt (59mg, 0.44mmol) and triethylamine (275ml, 1.9mmol) in the agitating solution of THF (6.0ml) and MeCN (6.0ml), add EDCHCl (127mg, 0.66mmol).Under room temperature, reactant mixture was stirred 16 hours vacuum concentration.Water is added in the residue, use ethyl acetate extraction.With saturated sodium bicarbonate, 2M citric acid and saturated sodium bicarbonate washing extract, then through dried over sodium sulfate and vacuum concentration.Residue is through purification by silica gel column chromatography; with normal hexane-ethyl acetate (1: 3; v/v) as eluant; obtain 4-[cis-1-[4-[N '-(2-chlorophenyl) urea groups]-3-methoxyphenyl acetyl group]-4-(2-naphthalene sulfonamido)-(2S)-pyrrolidinyl] methoxyl methyl benzoate (200mg; 65%), is colorless oil. 1H-NMR(CDCl 3)δ1.75-1.80(m，1H)，2.25-2.40(m，1H)，3.43(s，2H)，3.40-3.50(m，1H)，3.60(s，3H)，3.65-3.75(m，1H)，3.90(s，3H)，3.85-3.92(m，1H)，3.95-4.00(m，1H)，4.30-4.40(m，1H)，4.65-4.75(m，1H)，6.26(d，J＝9.3Hz，1H)，6.50(d，J＝8.3Hz，1H)，6.23(s，1H)，6.86(d，J＝8.8Hz，2H)，6.75-7.01(m，1H)，7.23-7.36(m，3H)，7.61-7.96(m，9H)，8.20(d，3＝8.1Hz，1H)，8.43(s，1H)。
To 4-[cis-1-[4-[N '-(2-chlorophenyl) urea groups]-3-methoxyphenyl acetyl group]-4-(2-naphthalene sulfonamido)-(2S)-pyrrolidinyl] methoxyl methyl benzoate (200mg; 0.26mmol) THF (6.0ml) and the agitating solution of methanol (3.0ml) in add the 1N sodium hydroxide (0.5ml, 0.5mmol).Stirred this mixture 18 hours in 70 ℃.This mixture of vacuum concentration adds water wherein, with 1N HCl neutralization.Collect the solid that produces, wash with water, vacuum drying, (210mg quantitatively), is white crystalline solid to obtain 161.MW 743.22 mp 135-142 ℃; IR (KBr) 3332,1685,1604,1531,1421,1159 cm -1 1H-NMR (DMSO-d6) δ 1.75-1.85 (m, 1H), 2.05-2.15 (m, 1H), 3.05-3.15 (m, 1H), 3.47 (s, 2H), 3.60-3.80 (m, 2H), 3.73 (s, 3H), 4.05-4.20 (m, 3H), 6.51 (d, J=8.5Hz, 1H), 6.74-7.04 (m, 5H), 7.27-7.31 (m, 1H), and 7.43-7.45 (m, 2H), 7.66-8.17 (m, 9H), 8.46 (s, 1H), 8.91 (d, J=9.5Hz, 1H); MS (FAB) m/z 743 (M ++ 1); C 38H 35N 4O 8ClS0.5H 2The analytical calculation value of O: C, 60.67; H, 4.82; N, 7.45; Cl, 4.26.Measured value: C, 60.77; H, 4.84; N, 7.21; Cl, 4.90.Embodiment 1544-[1-[4-[N '-(2-bromo phenyl) urea groups]-3-methoxyphenyl acetyl group]-4-(2-sulfonamido)-(2S)-pyrrolidinyl] methoxybenzoic acid
In 0 ℃, to 4-(cis-1-tert-butoxycarbonyl-4-amino-(2S)-pyrrolidinyl) methoxyl methyl benzoate (180mg, 0.51mmol) and TEA (283ml, and adding (2-) sulfonic acid chloride in the agitating solution of chloroform 2.0mmol) (10.0ml) (122mg, 0.56mmol).This reactant mixture was stirred under room temperature 18 hours.This reactant mixture of vacuum concentration.Residue is through purification by silica gel column chromatography, with normal hexane-ethyl acetate (3: 1, v/v) as eluant, obtain 4-(cis-1-tert-butoxycarbonyl-4-(2-sulfonamido)-(2S)-pyrrolidinyl) methoxyl methyl benzoate (170mg, 62%), is faint yellow oily thing. 1H-NMR(CDCl 3)δ1.40(s，9H)，1.85-1.95(m，1H)，2.29(s，3H)，2.35-2.45(m，1H)，2.62(s，6H)，3.80-4.15(m，3H)，3.89(s，3H)，3.50-3.65(m，1H)，6.94(s，2H)，6.94-7.00(m，2H)，7.99(d，J＝8.8Hz，2H)。
Under 0 ℃, (170mg adds TFA (5.0ml) in the agitating solution of dichloromethane 0.32mmol) (5.0ml) to (cis-1-tert-butoxycarbonyl-4-(2-sulfonamido)-(2S)-pyrrolidinyl) methoxyl methyl benzoate.Under room temperature, stirred this reactant mixture 0.5 hour.This mixture of vacuum concentration adds saturated sodium bicarbonate in this residue, uses dichloromethane extraction.With salt water washing extract, through dried over sodium sulfate and vacuum concentration.Crude product is used for subsequent reaction without being further purified.In 0 ℃, to crude product, 4-[N '-(2-bromo phenyl) urea groups]-3-methoxybenzene guanidine-acetic acid (121mg, 0.32mmol), HOBt (43mg, 0.32mmol) and triethylamine (139ml, 1.0mmol) add in the agitating solution in THF (5.0ml) and MeCN (5.0ml) EDCHCl (91mg, 0.48mmol).Under room temperature, reactant mixture was stirred 16 hours vacuum concentration.Water is added in the residue, use ethyl acetate extraction.With saturated sodium bicarbonate, 2M citric acid and saturated sodium bicarbonate washing extract, then through dried over sodium sulfate and vacuum concentration.Residue is through purification by silica gel column chromatography; with normal hexane-ethyl acetate (1: 4; v/v) as eluant; obtain 4-[1-[4-[N '-(2-bromo phenyl) urea groups]-3-methoxyphenyl acetyl group]-4-(2-sulfonamido)-(2S)-pyrrolidinyl] methoxyl methyl benzoate (210mg; 83%), is colorless oil. 1H-NMR(CDCl 3)δ1.85-1.90(m，1H)，1.95-2.05(m，1H)，2.32(s，3H)，2.60(s，6H)，3.40-3.50(m，3H)，3.60-3.70(m，2H)，3.68(s，3H)，3.89(s，3H)，3.96-3.99(m，1H)，3.35-3.45(m，1H)，3.70-3.75(m，1H)，6.00(d，J＝9.5Hz，1H)，6.57-7.08(m，9H)，7.29-7.34(m，1H)，7.51-7.53(m，1H)，7.92-7.96(m，3H)，8.15(d，J＝6.8Hz，1H)。
To 4-[1-[3-methoxyl group-4-[N '-(2-bromo phenyl) urea groups] phenyl acetyl]-4-(2-sulfonamido)-(2S)-pyrrolidinyl] methoxyl methyl benzoate (210mg; 0.26mmol) THF (5.0ml) and the agitating solution of methanol (3.0ml) in add the 1N sodium hydroxide (0.47ml, 0.47mmol).Stirred this mixture 24 hours in 70 ℃.This mixture of vacuum concentration adds water wherein, with 1N HCl neutralization.Collect the solid that produces, wash with water, vacuum drying obtains 162 (180mg, 87%), is white crystalline solid.MW 779.70 mp 130-132 ℃; IR (KBr) 3332,1689,1604,1529,1155cm -1 1H-NMR (DMSO-d 6) δ 1.70-1.85 (m, 1H), 2.02-2.12 (m, 1H), 2.25 (s, 3H), 2.52 (s, 6H), 3.05-3.12 (m, 1H), 3.48 (s, 2H), 3.60-3.70 (m, 2H), 3.77 (s, 3H), 3.90-4.20 (m, 3H), 6.59 (d, J=8.3Hz, 1H), 6.77-7.80 (m, 1H), 6.95-7.01 (m, 4H), 7.31-7.35 (m, 1H), 7.60 (d, J=8.1Hz, 1H), 7.86-7.97 (m, 5H), 8.72-8.76 (m, 1H), 8.89-8.93 (m, 1H); MS (FAB) m/z 779 (M +), 781 (M ++ 2); C 37H 39N 4O 8SBr0.5H 2The analytical calculation value of O: C, 56.35; H, 5.11; N, 7.10; Br, 10.13.Measured value: C, 56.39; H, 5.07; N, 6.89; Br, 10.25.Embodiment 1554-[1-[4-[N '-(2-bromo phenyl) urea groups]-3-methoxyphenyl acetyl group]-4-dansyl amino-(2S)-pyrrolidinyl] methoxybenzoic acid
Figure A0080951004041
In 0 ℃, to 4-(cis-1-tert-butoxycarbonyl-4-amino-(2S)-pyrrolidinyl) methoxyl methyl benzoate (180mg, 0.51mmol) and TEA (283ml, add in the agitating solution of chloroform 2.0mmol) (10.0ml) dansyl Cl (155mg, 0.68mmol).This reactant mixture was stirred under room temperature 18 hours.This reactant mixture of vacuum concentration.Residue is through purification by silica gel column chromatography, with normal hexane-ethyl acetate (2: 1, v/v) as eluant, obtain 4-(cis-1-tert-butoxycarbonyl-4-dansyl amino-(2S)-pyrrolidinyl) methoxyl methyl benzoate (200mg, 73%), is faint yellow oily thing. 1H-NMR(CDCl 3)δ1.34(s，9H)，1.50-1.60(m，1H)，2.15-2.25(m，1H)，2.87(s，6H)，3.15-3.22(m，1H)，3.35-3.45(m，1H)，3.48-3.52(m，1H)，3.80-4.10(m，3H)，3.91(s，3H)，7.01-7.25(m，4H)，7.50-7.53(m，1H)，8.03(d，J＝8.7Hz，2H)，8.16(d，J＝8.5Hz，1H)，8.28(d，J＝7.1Hz，1H)，8.53(d，J＝8.5Hz，1H)。
In 0 ℃, (200mg adds TFA (5.0ml) to methoxyl methyl benzoate in the agitating solution of dichloromethane 0.34mmol) (5.0ml) to 4-(cis-1-tert-butoxycarbonyl-4-dansyl amino-(2S)-pyrrolidinyl).Under room temperature, stirred this reactant mixture 0.5 hour.This mixture of vacuum concentration adds saturated sodium bicarbonate in this residue, uses dichloromethane extraction.With salt water washing extract, through dried over sodium sulfate and vacuum concentration.Crude product is used for subsequent reaction without being further purified.In 0 ℃, to crude product, 4-[N '-(2-bromo phenyl) urea groups]-3-methoxybenzene guanidine-acetic acid (129mg, 0.34mmol), HOBt (46mg, 0.34mmol) and triethylamine (142ml, 1.0mmol) in the agitating solution of THF (5.0ml) and MeCN (5.0ml), add EDCHCl (98mg, 0.51mmol).Under room temperature, reactant mixture was stirred 16 hours vacuum concentration.Water is added in the residue, use ethyl acetate extraction.With saturated sodium bicarbonate, 2M citric acid and saturated sodium bicarbonate washing extract, then through dried over sodium sulfate and vacuum concentration.Residue is through purification by silica gel column chromatography; with normal hexane-ethyl acetate (1: 4; v/v) as eluant; obtain 4-[1-[4-[N '-(2-bromo phenyl) urea groups]-3-methoxyphenyl acetyl group]-4-dansyl amino-(2S)-pyrrolidinyl] methoxyl methyl benzoate (250mg; 88%), is colorless oil. 1H-NMR(CDCl 3)δ1.55-1.65(m，1H)，2.15-2.25(m，1H)，2.87(s，6H)，3.20-3.35(m，3H)，3.50-3.55(m，1H)，3.67(s，3H)，3.78-3.81(m，1H)，3.88-3.93(m，1H)，3.91(s，3H)，4.28-4.31(m，1H)，4.65-4.70(m，1H)，6.35(d，J＝9.5Hz，1H)，6.54(d，J＝8.5Hz，1H)，6.63(s，1H)，6.90-7.13(m，6H)，7.22-7.31(m，2H)，7.50-7.56(m，2H)，7.88(d，J＝8.0Hz，1H)，7.99(d，J＝9.0Hz，1H)，8.13-8.16(m，2H)，8.27(d，J＝7.6Hz，1H)，8.56(d，J＝8.3Hz，1H)。
To 4-[1-[4-[N '-(2-bromo phenyl) urea groups]-3-methoxyphenyl acetyl group]-4-dansyl amino-2S-pyrrolidinyl] methoxyl methyl benzoate (250mg; 0.29mmol) THF (5.0ml) and the agitating solution of methanol (3.0ml) in add the 1N sodium hydroxide (0.52ml, 0.52mmol).Stirred this mixture 24 hours in 70 ℃.This mixture of vacuum concentration adds water wherein, with 1N HCl neutralization.Collect the solid that produces, wash with water, vacuum drying obtains 163 (230mg, 94%), is the green crystal solid.138-141 ℃ of MW 830.74mp; IR (KBr) 3340,2940,1604,1527,1421,1162,1145cm -1 1H-NMR (DMSO-d 6) δ 1.70-1.80 (m, 1H), 1.98-2.06 (m, 1H), 2.81 (s, 6H), 3.00-3.10 (m, 1H), 3.39-3.40 (m, 2H), 3.50-3.80 (m, 3H), 3.76 (s, 3H), 3.90-4.15 (m, 2H), 6.52 (d, J=9.0Hz, 1H), 6.73-7.00 (m, 4H), 7.22-7.35 (m, 2H), 7.55-7.64 (m, 3H), 7.83-8.48 (m, 8H), and 8.71-8.76 (m, 1H), 8.88-8.92 (m, 1H); MS (FAB) m/z 830 (M +), 832 (M ++ 2); C 40H 40N 5O 8BrS0.7H 2The analytical calculation value of O: C, 56.97; H .95; N, 8.30; Br, 9.47.Measured value: C, 57.06; H, 4.86; N, 7.98; Br, 9.66.Embodiment 1564-[4-methylsulfinyl amino-1-[4-[N '-(2-bromo phenyl) urea groups]-3-methoxyphenyl acetyl group]-(2S)-and pyrrolidinyl] methoxybenzoic acid
In 0 ℃, to 4-(cis-4-amino-1-tert-butoxycarbonyl-(2S)-pyrrolidinyl) methoxyl methyl benzoate (180mg, 0.51mmol) and TEA (283ml, add in the agitating solution of chloroform 2.0mmol) (10.0ml) mesyl chloride (88mg, 0.77mmol).This reactant mixture was stirred under room temperature 18 hours.This reactant mixture of vacuum concentration.Residue is through purification by silica gel column chromatography, with normal hexane-ethyl acetate (1: 1, v/v) as eluant, obtain 4-(cis-1-tert-butoxycarbonyl-4-methylsulfinyl amino-(2S)-pyrrolidinyl] methoxyl methyl benzoate (150mg, 69%), is faint yellow oily thing. 1H-NMR(CDCl 3)δ1.45(s，9H)，1.98-2.08(m，1H)，2.52-2.65(m，1H)，2.99(s，3H)，3.40-3.50(m，1H)，3.55-3.80(m，1H)，3.89(s，1H)，4.00-4.70(m，4H)，6.98-7.00(m，2H)，8.00(d，J＝8.8Hz，2H)。
(150mg adds TFA (5.0ml) to methoxyl methyl benzoate in the agitating solution of dichloromethane 0.43mmol) (5.0ml) to 4-(cis-1-tert-butoxycarbonyl-4-methylsulfinyl amino-(2S)-pyrrolidinyl).Under room temperature, stirred this reactant mixture 0.5 hour.This mixture of vacuum concentration adds saturated sodium bicarbonate in this residue, uses dichloromethane extraction.With salt water washing extract, through dried over sodium sulfate and vacuum concentration.Crude product is used for subsequent reaction without being further purified.In 0 ℃, to crude product, 4-[N '-(2-bromo phenyl) urea groups]-3-methoxybenzene guanidine-acetic acid (163mg, 0.42mmol), HOBt (58mg, 0.43mmol) and triethylamine (179ml, 1.3mmol) in the agitating solution of THF (5.0ml) and MeCN (5.0ml), add EDCHCl (144mg, 0.75mmol).Under room temperature, reactant mixture was stirred 16 hours vacuum concentration.Water is added in the residue, use ethyl acetate extraction.With saturated sodium bicarbonate, 2M citric acid and saturated sodium bicarbonate washing extract, then through dried over sodium sulfate and vacuum concentration.Residue is through purification by silica gel column chromatography; with normal hexane-ethyl acetate (1: 4; v/v) to ethanol-ethyl acetate (10%; v/v) as eluant; obtain 4-[4-methylsulfinyl amino-1-[4-[N '-(2-bromo phenyl) urea groups]-3-methoxyphenyl acetyl group]-(2S)-and pyrrolidinyl] methoxyl methyl benzoate (210mg; 73%), is colorless oil. 1H-NMR(CDCl 3)δ1.95-2.00(m，1H)，2.55-2.65(m，1H)，2.17(s，3H)，3.55-3.70(m，1H)，3.60(S，2H)，3.67(S，3H)，3.85-3.90(m，1H)，3.89(s，3H)，3.95-4.18(m，2H)，4.45-4.55(m，1H)，4.70-4.80(m，1H)，5.87(d，J＝9.3Hz，1H)，6.73-6.95(m，5H)，7.09(s，2H)，7.28-7.33(m，1H)，7.51(d，J＝8.0Hz，1H)，7.93-7.97(m，3H)，8.13(d，J＝8.3Hz，1H)。
To 4-[4-methylsulfinyl amino-1-[4-[N '-(2-bromo phenyl) urea groups]-3-methoxyphenyl acetyl group]-(2S)-and pyrrolidinyl] methoxyl methyl benzoate (210mg; 0.3mmol) THF (5.0ml) and the agitating solution of methanol (3.0ml) in add the 1N sodium hydroxide (0.8ml, 0.8mmol).Stirred this mixture 24 hours in 70 ℃.This mixture of vacuum concentration adds water wherein, with 1N HCl neutralization.Collect the solid that produces, wash with water, vacuum drying obtains 164 (170mg, 83%), is white crystalline solid.MW 675.55 mp 125-128 ℃ IR (KBr) 3353,1689,1604,1529,1419,1155cm -1 1H-NMR (DMSO-d 6) δ 1.88-2.00 (m, 1H), 2.35-245 (m, 1H), 2.96 (m, 3H), 3.15-3.23 (m, 1H), 3.60 (s, 2H), 3.50-3.70 (m, 1H), 3.78 (s, 3H), 3.80-3.90 (m, 1H), 3.95-4.05 (m, 1H), 4.10-4.30 (m, 2H), 6.71-7.03 (m, 4H), 7.32 (m, 1H), 7.45 (d, J=6.8Hz, 1H), 7.60 (d, J=7.8Hz, 1H), 7.87-7.95 (m, 4H), 8.74 (s, 1H), 8.92 (s, 1H); MS (FAB) m/z 675 (M +), 677 (M ++ 2); C 29H 31N 4O 8BrS0.6H 2The analytical calculation value of O: C, 50.75; H, 4.73; N, 8.16.Measured value: C, 51.04; H, 4.62; N, 7.79.Embodiment 1574-[1-[3-methoxyl group-4-[N '-(2-aminomethyl phenyl) urea groups] phenyl acetyl]-(2S)-and octahydro indyl methoxyl group] benzoic acid
Figure A0080951004071
In 0 ℃, (3.00g adds 1N sodium hydroxide (45ml) in the agitating solution of dioxane 17.7mmol) (20ml), stir this solution in 0 ℃ to octahydro indole-(2S)-formic acid.In 0 ℃, in this mixture, be added in (Boc) in the dioxane (25ml) 2(4.26g 19.5mmol), stirred this reactant mixture 1 day O under room temperature.With this mixture of 1N HCl acidify, use ethyl acetate extraction.With salt water washing extract,, obtain that 1-(tert-butoxycarbonyl) octahydro indole-(2S)-(4.78g q.y.), is colorless solid to formic acid through dried over sodium sulfate and evaporation.Mp 130-132 ℃; 1H-NMR (CDCl 3) δ 1.10-1.46 (a plurality of s and m, 14H altogether), and 1.65-1.76 (m, 3H), 1.90-2.18 (m, 2H), 2.26-2.35 (m, 1H), 3.77-3.86 (m, 1H), 4.22-4.34 (m, 1H); MS (ESI) m/z 270 (M ++ 1).
To 1-(tert-butoxycarbonyl) octahydro indole-(2S)-(1.00g adds BH in cold (0 ℃) solution of the stirring of THF 3.71mmol) (10ml) to formic acid 3DMS (530ml.5.59mmol) stirs this reactant mixture and to spend the night under room temperature.By adding entry quencher mixture, use ethyl acetate extraction.With salt water washing extract, through dried over sodium sulfate and evaporation.Residue is through purification by silica gel column chromatography, with chloroform-methanol (50: 1, v/v) as eluant, obtain 1-(tert-butoxycarbonyl) octahydro indole-(2S)-methanol (940mg, 99%), be colorless oil. 1H-NMR(CDCl 3)δ1.05-1.30(m，4H)，1.47(s，9H)，1.49-1.74(m，4H)，1.82-1.93(m，3H)，2.19-2.26(m，1H)，3.56-3.61(m，1H)，3.70-3.75(m，2H)，3.94-3.96(m，1H)；MS(FAB)m/z?256(M ++1)。
To 4-methyl hydroxybenzoate (560mg, 3.68mmol), 1-(tert-butoxycarbonyl) octahydro indole-(2S)-methanol (940mg, 3.68mmol) and triphenyl phasphine (1.16g, 4.42mmol) cold (0 ℃) solution of stirring of THF (20ml) in add DIAD (870ml, 4.42mmol), with this reaction mixture refluxed heating 8 hours.After being cooled to room temperature, evaporate this mixture.Residue is through purification by silica gel column chromatography, with normal hexane-ethyl acetate (5: 1,, obtain 4-[1-(tert-butoxycarbonyl)-(2S)-octahydro indyl methoxyl group v/v) as eluant] essence of Niobe (1.16g, 81%), be colorless oil. 1H-NMR (CDCl 3) δ 1.14-1.47 (a plurality of s and m, 13H altogether), 1.60-2.13 (a plurality of m, 6H altogether), 2.22-2.28 (m, 1H), (3.75-3.91 a plurality of s and m, 4H altogether), and 4.06-4.18 (m, 2H), 4.37 (m, 1H), and 6.94-6.96 (m, 2H), 7.96-7.98 (m, 2H); MS (FAB) m/z 390 (M ++ 1).
To 4-[1-(tert-butoxycarbonyl)-(2S)-octahydro indyl methoxyl group] (1.16g adds TFA (10ml) to essence of Niobe in the agitating solution of dichloromethane 2.98mmol) (10ml), under room temperature, reactant mixture was stirred 2 hours.This mixture of vacuum concentration makes it be alkalescence by adding saturated sodium bicarbonate, uses chloroform extraction.With salt water washing extract, dry and evaporation obtains 4-[(2S through potassium carbonate)-octahydro indyl methoxyl group] (860mg q.y.), is brown oil to essence of Niobe. 1H-NMR (CDCl 3) δ 1.23-1.78 (a plurality of m, 10H altogether), 2.00-2.09 (m, 2H), 3.14-3.18 (m, 1H), 3.55-3.62 (m, 1H), 3.88 (s, 3H), 3.96-4.06 (m, 2H), 6.92 (d, J=9.1Hz, 2H), 7.97 (d, J=9.1Hz, 2H); MS (FAB) m/z 290 (M ++ 1).
Under room temperature, with 3-methoxyl group-4-[N '-(2-aminomethyl phenyl) urea groups] phenylacetic acid (298mg, 0.95mmol), 4-[(2S)-octahydro indyl methoxyl group] essence of Niobe (274mg, 0.95mmol), EDCHCl (218mg, 1.14mmol), HOBt (154mg, 1.14mmol), (160ml, 1.15mmol) mixture in THF (7ml) stirs and to spend the night triethylamine.This mixture of dilute with water is used ethyl acetate extraction.With salt water washing extract, through dried over sodium sulfate and evaporation.Residue is through purification by silica gel column chromatography; with chloroform-methanol (100: 1-50: 1; v/v) as eluant; obtain 4-[1-[3-methoxyl group-4-[N '-(2-aminomethyl phenyl) urea groups] phenyl acetyl]-(2S)-and octahydro indyl methoxyl group] essence of Niobe (532mg; 96%), is the white foam thing. 1H-NMR (CDCl 3) δ 1.13-2.05 (a plurality of m, 9H altogether), 2.14-2.24 (m, 2H), 2.26 (s, 3H), 3.60 (s, 2H), 3.62 (s, 3H), 3.80-3.85 (m, 1H), 3.88 (s, 3H), 4.27-4.37 (m, 3H), 6.62 (s, 1H), 6.74-6.76 (m, 2H), 6.91 (d, J=8.8Hz, 2H), 7.09-7.13 (m, 1H), 7.20-7.24 (m, 3H), 7.55 (d, J=7.8Hz, 1H), 7.94 (d, J=8.8Hz, 2H), 8.04 (d, J=7.8Hz, 1H); MS (FAB) m/z 586 (M ++ 1).
To 4-[1-[3-methoxyl group-4-[N '-(2-aminomethyl phenyl) urea groups] phenyl acetyl]-(2S)-and octahydro indyl methoxyl group] essence of Niobe (532mg; 0.91mmol) add 0.5N sodium hydroxide (5ml) in the agitating solution in THF (5ml), with this reaction mixture refluxed heating 3 hours.After being cooled to room temperature, in this mixture impouring ice-1N HCl, collect the precipitation that produces.Make crude product solid recrystallization in methanol-chloroform-IPE, obtain 165 (278mg, 54%), be white crystalline powder.MW 571.66 mp 130-134 ℃; 1H-NMR (DMSO-d 6) δ 1.16-2.10 (a plurality of m, 9H altogether), 2.15-2.30 (a plurality of s and m, 4H altogether), 3.55-3.79 (m, 3H), 3.81 (s, 3H), 3.90-3.95 (m, 1H), 4.17-4.23 (m, 2H), 4.34-4.36 (m, 1H), 6.72-6.74 (m, 1H), 6.87-6.88 (m, 1H), 6.91-6.95 (m, 1H), 7.03 (d, J=8.8Hz, 2H), 7.10-7.16 (m, 2H), and 7.78-7.80 (m, 1H), 7.87 (d, J=8.8Hz, 2H), 7.98-8.00 (m, 1H), 8.45 (s, 1H), 8.54 (s, 1H), 12.61 (br s, 1H); MS (FAB) m/z 572 (M ++ 1); C 33H 37N 3O 61/4H 2The analytical calculation value of O: C, 68.79; H, 6.56; N, 7.29.Measured value: C, 68.70; H, 6.82; N, 6.97.Embodiment 1584-[1-[4-N '-(2-chlorophenyl) urea groups]-3-methoxyphenyl acetyl group]-(2S)-and octahydro indyl methoxyl group] benzoic acid
Under room temperature, with 4-[N '-(2-chlorophenyl) urea groups]-3-methoxybenzene guanidine-acetic acid (307mg, 0.92mmol), 4-[(2S)-octahydro indyl methoxyl group] essence of Niobe (265mg, 0.92mmol), EDCHCl (211mg, 1.10mmol), HOBt (148mg, 1.10mmol) and triethylamine (153ml, 1.10mmol) mixture in THF (7ml) stirs and to spend the night.This mixture of dilute with water is used ethyl acetate extraction.With salt water washing extract, through dried over sodium sulfate and evaporation.Residue is through purification by silica gel column chromatography; with chloroform-methanol (100: 1-50: 1; v/v) as eluant; obtain 4-[1-[4-[N '-(2-chlorophenyl) urea groups]-3-methoxyphenyl acetyl group]-(2S)-and octahydro indyl methoxyl group] essence of Niobe (550mg; 99%), is the white foam thing. 1H-NMR (CDCl 3) δ 1.15-2.02 (a plurality of m, 9H altogether), and 2.17-2.33 (m, 2H), 3.58 (s, 3H), 3.62 (s, 2H), (3.84-3.90 a plurality of s and m, 4H altogether), and 4.06-4.40 (m, 3H), 6.71-6.74 (m, 2H), 6.88-7.00 (m, 3H), 7.21-7.30 (m, 2H), 7.62 (s, 2H), 7.91-7.95 (m, 3H), 8.17-8.20 (m, 1H); MS (FAB) m/z 606 (M ++ 1).
To 4-[1-[4-[N '-(2-chlorophenyl) urea groups]-3-methoxyphenyl acetyl group]-(2S)-and octahydro indyl methoxyl group] essence of Niobe (550mg; 0.91mmol) add 0.5N sodium hydroxide (5ml) in the agitating solution in THF (5ml), with this reaction mixture refluxed heating 5 hours.After being cooled to room temperature, in this mixture impouring ice-1N HCl, collect the precipitation that produces.Make crude product solid recrystallization in methanol-chloroform-IPE, obtain 166 (286mg, 53%), be white crystalline powder.MW 388.29 mp 133-136 ℃; 1H-NMR (DMSO-d 6) δ 1.16-2.10 (a plurality of m, 10H altogether), and 2.24-2.27 (m, 1H), 3.55-3.75 (m, 2H), 3.80 (s, 3H), 3.90-3.96 (m, 1H), 4.17-4.23 (m, 2H), 4.34-4.36 (m, 1H), 6.73-6.75 (m, 1H), 6.88 (d, J=1.5Hz, 1H), and 6.99-7.05 (m, 3H), 7.25-7.30 (m, 1H), 7.43 (dd, J=1.5,8.1Hz, 1H), and 7.87-7.89 (m, 2H), 7.95 (d, J=8.1Hz, 1H), 8.08 (dd, J=1.5,8.3Hz, 1H), 8.88 (s, 1H), 8.92 (s, 1H), 12.61 (br s, 1H); MS (FAB) m/z 592 (M ++ 1); C 32H 34ClN 3O 61/4H 2The analytical calculation value of O: C, 64.42; H, 5.83; N, 7.04; Cl, 5.94.Measured value: C, 64.55; H, 6.09; N, 6.64; Cl, 5.93.Embodiment 1594-[1-[4-[N '-(2-bromo phenyl) urea groups]-3-methoxyphenyl acetyl group]-(2S)-and octahydro indyl methoxyl group] benzoic acid
Under room temperature, with 4-[N '-(2-bromo phenyl) urea groups]-3-methoxybenzene guanidine-acetic acid (457mg, 1.21mmol), 4-[(2S)-octahydro indyl methoxyl group] essence of Niobe (320mg, 1.21mmol), EDCHCl (277mg, 1.44mmol), HOBt (196mg, 1.45mmol) and triethylamine (200ml, 1.43mmol) mixture in THF (7ml) stirs and to spend the night.This mixture of dilute with water is used ethyl acetate extraction.With salt water washing extract, through dried over sodium sulfate and evaporation.Residue is through purification by silica gel column chromatography; with chloroform-methanol (100: 1,, obtain 4-[1-[4-[N '-(2-bromo phenyl) urea groups v/v) as eluant]-3-methoxyphenyl acetyl group]-(2S)-octahydro indyl methoxyl group] essence of Niobe (423mg; 54%), is the white foam thing. 1H-NMR (CDCl 3) δ 1.15-1.89 (a plurality of m, 8H altogether), and 1.96-2.02 (m, 1H), 2.16-2.32 (m, 2H), 3.63 (s, 2H), 3.65 (s, 3H), 3.82-3.86 (m, 1H), 3.88 (s, 3H), 4.30-4.39 (m, 3H), and 6.75-6.77 (m, 2H), 6.88-6.93 (m, 3H), 7.24-7.31 (m, 1H), 7.37-7.50 (s, 3H), 7.91-7.99 (m, 3H), 8.12-8.15 (m, 1H); MS (FAB) m/z 650 (M ++ 1).
To 4-[1-[4-[N '-(2-bromo phenyl) urea groups]-3-methoxyphenyl acetyl group]-(2S)-and octahydro indyl methoxyl group] essence of Niobe (420mg; 0.65mmol) add 0.5N sodium hydroxide (5ml) in the agitating solution in THF (5ml), with this reaction mixture refluxed heating 5 hours.After being cooled to room temperature, in this mixture impouring ice-1N HCl, collect the precipitation that produces.Make crude product solid recrystallization in methanol-chloroform-IPE, obtain 167 (197mg, 48%), be white crystalline powder.MW 636.53 mp 118-123 ℃; 1H-NMR (DMSO-d 6) δ 1.16-2.27 (a plurality of m, 10H altogether), and 3.56-3.75 (m, 3H), 3.81 (s, 3H), and 3.90-3.96 (m, 1H), 4.17-4.23 (m, 2H), 4.34-4.36 (m, 1H), 6.73-6.75 (m, 1H), 6.88-7.05 (m, 4H), and 7.30-7.34 (m, 1H), 7.59-7.61 (m, 1H), 7.87-7.96 (m, 4H), 8.73 (s, 1H), 8.91 (s, 1H), 12.64 (br s, 1H); MS (FAB) m/z 636 (M ++ 1); C 32H 34BrN 3O 61/4H 2The analytical calculation value of O: C, 59.96; H, 5.42; N, 6.55; Br, 12.46.Measured value: C, 60.12; H, 5.86; N, 6.09; Br, 12.47.Embodiment 1604-[3-[3-methoxyl group-4-[N '-(2-aminomethyl phenyl) urea groups] phenyl] acetyl group-4-thiazolidinyl] methoxybenzoic acid
Figure A0080951004121
(5.0g adds in the agitating solution of DMF 37.6mmol) (50.0ml) (Boc) to Thiazolidine-4-formic acid 2O (9.8g, 45.1mmol) and TEA (8.0ml).Under room temperature, stirred this reactant mixture 18 hours.Water is added in this mixture, use ethyl acetate extraction.Wash organic layer with water, through dried over sodium sulfate and vacuum concentration.Residue is through purification by silica gel column chromatography, with ethyl acetate-normal hexane (1: 3, v/v) as eluant, obtain 3-tert-butoxycarbonyl Thiazolidine-4-formic acid (6.5g, 74%), be white crystalline solid. 1H-NMR(CDCl 3)δ1.49(br?s，9H)，3.20-3.30(m，2H)，4.09-4.87(m，3H)。
In 0 ℃, (2.3g adds BH in the agitating solution of THF 10.0mmol) (30ml) to 3-tert-butoxycarbonyl Thiazolidine-4-formic acid 3THF (solution of 1.0M in THF, 20.0ml, 20.0mmol).After stirring 1.0 hours under the room temperature, with this reaction mixture refluxed heating 1.0 hours.After the cooling, this mixture of vacuum concentration.In 0 ℃ water is added wherein, use ethyl acetate extraction.Wash extract with water, through dried over sodium sulfate and vacuum concentration, (2.0g quantitatively), is colorless oil to obtain 3-tert-butoxycarbonyl-5-hydroxymethylthiazole alkane. 1H-NMR(CDCl 3)δ1.48(s，9H)，2.80-2.85(m，1H)，3.13-3.17(m，1H)，3.20-3.30(m，1H)，3.64-3.70(m，2H)，4.34(br?s，1H)，4.60(br?s，1H)。
In 0 ℃, to 3-tert-butoxycarbonyl-5-hydroxymethylthiazole alkane (1.9g, 8.7mmol), the 4-methyl hydroxybenzoate (1.3g, 8.7mmol) and triphenyl phasphine (3.2g adds DIAD (2.2g.10.4mmol) in the agitating solution of THF 12.2mmol) (10ml).Under room temperature, this reactant mixture was stirred 18 hours.This mixture of vacuum concentration.Residue is through purification by silica gel column chromatography, with ethyl acetate-normal hexane (1: 9, v/v) as eluant, obtain 4-(3-tert-butoxycarbonyl-4-thiazolidinyl) methoxyl methyl benzoate (1.6g, 52%), be faint yellow oily thing. 1H-NMR(CDCl 3)δ1.49(s，9H)，3.11-3.19(m，2H)，3.88(s，3H)，4.04-4.31(m，3H)，4.61(m，2H)，6.96(d，J＝8.8Hz，2H)，7.98(d，J＝8.8Hz，2H)。
In 0 ℃, (440mg adds TFA (3ml) in the agitating solution of dichloromethane 1.25mmol) (6ml) to 4-(3-tert-butoxycarbonyl-4-thiazolidinyl) methoxyl methyl benzoate.Under room temperature, reactant mixture was stirred 0.5 hour.This mixture of vacuum concentration adds saturated sodium bicarbonate in the residue, uses dichloromethane extraction.With salt water washing extract, through dried over sodium sulfate and vacuum concentration.Crude product is used for subsequent reaction without being further purified.In 0 ℃, to crude product (0.6mmol), 3-methoxyl group-4-[N '-(2-aminomethyl phenyl) urea groups] phenylacetic acid (188mg, 0.6mmol), HOBt (81mg, 0.6mmol) and triethylamine (280ml, 2.0mmol) THF (5ml) and the agitating solution of MeCN (5ml) in add EDCHCl (173mg, 0.9mmol).Under room temperature, reactant mixture was stirred 16 hours vacuum concentration.Water is added in the residue, use ethyl acetate extraction.With saturated sodium bicarbonate, 2M citric acid and saturated sodium bicarbonate washing extract, then through dried over sodium sulfate and vacuum concentration.Residue is through purification by silica gel column chromatography; with normal hexane-ethyl acetate (1: 3,, obtain 4-[3-[3-methoxyl group-4-[N '-(2-aminomethyl phenyl) urea groups v/v) as eluant] phenyl] acetyl group-4-thiazolidinyl] methoxyl methyl benzoate (340mg; quantitatively), be amorphous solid. 1H-NMR(CDCl 3)δ2.31(s，3H)，3.15-3.16(m，2H)，3.67-3.69(m，5H)，3.88(s，3H)，4.09-4.14(m，2H)，4.22-4.90(m，3H)，6.30(m，1H)，6.74-6.96(m，4H)，7.11-7.25(m，4H)，7.49-7.51(m，1H)，7.95-8.12(m，3H)。
To 4-[3-[3-methoxyl group-4-[N '-(2-aminomethyl phenyl) urea groups] phenyl] acetyl group-4-thiazolidinyl] methoxyl methyl benzoate (340mg; 0.62mmol) THF (5.0ml) and the agitating solution of ethanol (3.0ml) in add the 1N sodium hydroxide (0.62ml, 0.62mmol).Stirred this mixture 18 hours in 70 ℃.This mixture of vacuum concentration adds water wherein, with 1N HCl neutralization.Collect the solid that produces, wash with water, vacuum drying obtains 168 (290mg, 88%), is white crystalline solid.MW 535.62 mp 125-128 ℃; IR (KBr) 3357,2937,1604,1533,1419,1253,1166,1033,773cm -1 1H-NMR (DMSO-d 6) δ 2.25 (s, 3H), 3.05-3.20 (m, 2H), 3.71,3.83 and 3.85 (they respectively are s, are total to 5H), 4.03-4.15 (m, 3H), 4.52-4.76 (m, 2H), 6.15-6.17 (m, 7H), 7.78-8.30 (m, 4H), 8.30 (m, 1H), 8.56 (m, 1H); MS (FAB) m/z 536 (M ++ 1); C 28H 29N 3O 6S0.5H 2The analytical calculation value of O: C, 61.75; H, 5.55; N, 7.72.Measured value: C, 61.72; H, 5.55; N, 7.49.Embodiment 1614-[3-[4-[N '-(2-chlorophenyl) urea groups]-3-methoxyphenyl acetyl group]-the 4-thiazolidinyl] methoxybenzoic acid
Figure A0080951004141
In 0 ℃, (600mg adds TFA (6.0ml) in the agitating solution of dichloromethane 1.7mmol) (6.0ml) to 4-(3-tert-butoxycarbonyl-4-thiazolidinyl) methoxyl methyl benzoate.Under room temperature, reactant mixture was stirred 0.5 hour.This mixture of vacuum concentration adds saturated sodium bicarbonate in the residue, uses dichloromethane extraction.With salt water washing extract, through dried over sodium sulfate and vacuum concentration.Crude product is used for subsequent reaction without being further purified.In 0 ℃, to crude product, 4-[N '-(2-chlorophenyl) urea groups]-3-methoxybenzene guanidine-acetic acid (570mg, 1.7mmol), HOBt (230mg, 1.7mmol) and triethylamine (709ml, 5.1mmol) THF (10.0ml) and the agitating solution of MeCN (10.0ml) in add EDCHCl (490mg, 2.55mmol).Under room temperature, reactant mixture was stirred 16 hours vacuum concentration.Water is added in the residue, use ethyl acetate extraction.With saturated sodium bicarbonate, 2M citric acid and saturated sodium bicarbonate washing extract, then through dried over sodium sulfate and vacuum concentration.Residue is through purification by silica gel column chromatography; with normal hexane-ethyl acetate (1: 1,, obtain 4-[3-[4-[N '-(2-chlorophenyl) urea groups v/v) as eluant]-3-methoxyphenyl acetyl group]-the 4-thiazolidinyl] methoxyl methyl benzoate (900mg; 93%), is colorless oil. 1H-NMR(CDCl 3)δ3.15-3.18(m，2H)，3.70(s，2H)，3.78(s，3H)，3.86(s，3H)，4.09-4.93(m，5H)，6.80-7.01(m，5H)，7.19-7.35(m，4H)，7.94-8.18(m，4H)。
To 4-[3-[4-[N '-(2-chlorophenyl) urea groups]-3-methoxyphenyl acetyl group]-the 4-thiazolidinyl] methoxyl methyl benzoate (900mg; 1.6mmol) THF (8.0ml) and the agitating solution of methanol (4.0ml) in add the 1N sodium hydroxide (3.1ml, 3.1mmol).Stirred this mixture 24 hours in 70 ℃.This mixture of vacuum concentration adds water wherein, with 1N HCl neutralization.Collect the solid that produces, wash with water, vacuum drying obtains 169 (780mg, 89%), is white crystalline solid.MW 556.03 mp 126-129 ℃; IR (KBr) 3343,2937,1604,1531,1421,1245,1166,1035,752cm -1 1H-NMR (DMSO-d 6) δ 3.06-3.24 (m, 2H), 3.72-3.85 (m, 5H), 4.02-4.27 (m, 3H), 4.53-4.76 (m, 2H), 6.74-7.44 (m, 7H), 7.87-8.30 (m, 4H), 8.89-8.95 (m, 2H); MS (FAB) m/z 556 (M ++ 1); C 27H 27N 3O 6ClS0.7H 2The analytical calculation value of O: C, 56.93; H, 5.03; N, 7.38; Cl, 6.22.Measured value: C, 56.89; H, 4.84; N, 7.42; Cl, 6.35.Embodiment 1624-[3-[4-[N '-(2-bromo phenyl) urea groups]-3-methoxyphenyl acetyl group]-the 4-thiazolidinyl] methoxybenzoic acid
Figure A0080951004151
In 0 ℃, (560mg adds TFA (5.0ml) in the agitating solution of dichloromethane 1.6mmol) (5.0ml) to 4-(3-tert-butoxycarbonyl-4-thiazolidinyl) methoxyl methyl benzoate.Under room temperature, reactant mixture was stirred 0.5 hour.This mixture of vacuum concentration adds saturated sodium bicarbonate in the residue, uses dichloromethane extraction.With salt water washing extract, through dried over sodium sulfate and vacuum concentration.Crude product is used for subsequent reaction without being further purified.In 0 ℃, to crude product, 4-[N '-(2-bromo phenyl) urea groups]-3-methoxybenzene guanidine-acetic acid (599mg, 1.6mmol), HOBt (213mg, 1.6mmol) and triethylamine (659ml, 4.7mmol) THF (10.0ml) and the agitating solution of MeCN (10.0ml) in add EDCHCl (455mg, 2.4mmol).Under room temperature, reactant mixture was stirred 16 hours vacuum concentration.Water is added in the residue, use ethyl acetate extraction.With saturated sodium bicarbonate, 2M citric acid and saturated sodium bicarbonate washing extract, then through dried over sodium sulfate and vacuum concentration.Residue is through purification by silica gel column chromatography; with normal hexane-ethyl acetate (2: 3,, obtain 4-[3-[4-[N '-(2-bromo phenyl) urea groups v/v) as eluant]-3-methoxyphenyl acetyl group]-the 4-thiazolidinyl] methoxyl methyl benzoate (870mg; 89%), is colorless oil. 1H-NMR(CDCl 3)δ3.00-3.20(m，3H)，3.70(s，2H)，3.81(s，3H)，3.88(s，3H)，4.09-4.23(m，1H)，4.42(d，J＝8.5Hz，1H)，4.59(d，J＝8.5Hz，1H)，4.70-4.92(m，1H)，6.81-7.53(m，9H)，7.95-8.15(m，4H)。
To 4-[3-[4-[N '-(2-bromo phenyl) urea groups]-3-methoxyphenyl acetyl group]-the 4-thiazolidinyl] methoxyl methyl benzoate (870mg; 1.4mmol) THF (8.0ml) and the agitating solution of methanol (8.0ml) in add the 1N sodium hydroxide (2.8ml, 2.8mmol).Stirred this mixture 18 hours in 70 ℃.This mixture of vacuum concentration adds water wherein, with 1N HCl neutralization.Collect the solid that produces, wash with water, vacuum drying obtains 170 (740mg, 87%), is white crystalline solid.MW 600.48 mp 125-133 ℃; IR (KBr) 3332,2935,1604,1527,1421,1245,1166,1027,750cm -1 1H-NMR (DMSO-d 6) δ 3.01-3.25 (m, 2H), 3.72-3.85 (m, 5H), 4.02-4.30 (m, 2H), 4.54 (d, J=8.8Hz, 1H), and 4.74-4.87 (m, 2H), 6.76-7.07 (m, 5H), and 7.30-7.34 (m, 1H), 7.59 (d, J=8.1Hz, 1H), 7.86-7.98 (m, 4H), 8.74 (s, 1H), 8.92-8.94 (s, 1H); MS (FAB) m/z 600 (M ++ 1); C 27H 26N 3O 6BrS0.3H 2The analytical calculation value of O: C, 53.52; H, 4.43; N, 6.94.Measured value: C, 53.54; H, 4.45; N, 6.80.Embodiment 163 cis-4-[[1-[3-methoxyl group-4-[N '-(2-aminomethyl phenyl) urea groups] phenyl acetyl]-the 2-pyrrolidinyl] methylamino] naphthenic acid
In 0 ℃, to 2S-pyrrolidine carbinol (2.0g, 20.0mmol), 3-methoxyl group-4-[N '-(2-aminomethyl phenyl) urea groups] phenylacetic acid (6.28g, 20.0mmol), HOBt (71mg, 0.53mmol) and triethylamine (5.5ml, 40.0mmol) THF (50.0ml) and the agitating solution of MeCN (40.0ml) in add EDCHCl (5.7g, 30.0mmol).Under room temperature, reactant mixture was stirred 16 hours vacuum concentration.Water is added in the residue, use ethyl acetate extraction.With saturated sodium bicarbonate, 2M citric acid and saturated sodium bicarbonate washing organic layer, then through dried over sodium sulfate and vacuum concentration.Residue is through purification by silica gel column chromatography; with ethyl acetate to methanol-dichloromethane (1: 9,, obtain 1-[3-methoxyl group-4-[N '-(2-aminomethyl phenyl) urea groups v/v) as eluant] phenyl acetyl]-2S-pyrrolidine carbinol (7.0g; 89%), is white crystalline solid. 1H-NMR(CDCl 3)δ1.54-1.58(m，1H)，1.80-2.04(m，3H)，2.27(s，3H)，3.42-3.46(m，1H)，3.54-3.65(m，2H)，3.62(s，2H)，3.69(s，3H)，4.21-4.23(m，1H)，5.04(m，1H)，6.68-6.79(m，3H)，7.09-7.31(m，4H)，7.52(d，J＝7.8Hz，1H)，8.07(d，J＝8.0Hz，1H)。
In-78 ℃, to oxalyl chloride (0.3ml, add in the agitating solution of dichloromethane 3.3mmol) (30.0ml) DMSO (6.6ml, 0.51mmol).After 5 minutes, in this mixture, be added in 1-[3-methoxyl group-4-[N '-(2-aminomethyl phenyl) urea groups in the dichloromethane (5.0ml)] phenyl acetyl]-the 2-pyrrolidine carbinol (1.2g, 3.0mmol).In-78 ℃, mixture was stirred 30 minutes, the adding triethylamine (2.1ml, 15.0mmol).In-78 ℃ this mixture was stirred 30 minutes, under room temperature, stirred 30 minutes.Water is added in the mixture, use dichloromethane extraction.Wash extract with water, then through dried over sodium sulfate and vacuum concentration.Crude product is used for subsequent reaction without being further purified.In 0 ℃, (769mg 3.3mmol) and in the agitating solution of the DCE (10ml) of acetic acid (0.32ml) adds NaBH (OAc) to crude product, cis-4-aminocyclohexane benzyl formate 3(1.1g, 5.4mmol).Stirred reaction mixture is 18 hours under room temperature.This mixture of vacuum concentration.Saturated sodium bicarbonate is added in the residue, use dichloromethane extraction.With salt water washing extract, through dried over sodium sulfate and vacuum concentration.Residue is through purification by silica gel column chromatography; with methanol-dichloromethane (1: 9; v/v) as eluant; obtain cis-4-[[1-[3-methoxyl group-4-[N '-(2-aminomethyl phenyl) urea groups] phenyl acetyl]-the 2-pyrrolidinyl] methylamino] naphthenic acid benzyl ester (1.5g; 83%), is amorphous solid. 1H-NMR (CDCl 3) δ 1.40-1.65 (m, 6H), 1.80-1.98 (m, 6H), 2.26 (s, 3H), and 2.45-2.65 (m, 3H), 2.81-2.86 (m, 1H), 3.44-3.46 (m, 2H), 3.56 (s, 2H), 3.67 (s, 3H), and 3.90-4.15 (m, 1H), 5.09 and 5.11 (they respectively are s, are total to 2H), 6.74-6.83 (m, 3H), 7.07-7.20 (m, 4H), 7.31-7.35 (m, 5H), and 7.53-7.55 (m, 1H), 8.02-8.06 (m, 1H).
To cis-4-[[1-[3-methoxyl group-4-[N '-(2-aminomethyl phenyl) urea groups] phenyl acetyl]-the 2-pyrrolidinyl] methylamino] naphthenic acid benzyl ester (1.5g; 2.45mmol) THF (10.0ml) and the agitating solution of methanol (5.0ml) in add the 1N sodium hydroxide (3.68ml, 3.68mmol).Stirred this mixture 18 hours in 70 ℃.This mixture of vacuum concentration adds water wherein, with 1N HCl neutralization.This mixture of vacuum concentration.Residue is through purification by silica gel column chromatography; with methanol-dichloromethane (1: 5; v/v) as eluant; obtain cis-4-[[1-[3-methoxyl group-4-[N '-(2-aminomethyl phenyl) urea groups] phenyl acetyl]-the 2-pyrrolidinyl] methylamino] naphthenic acid 171 (940mg; 73%), is amorphous solid.MW 522.64 IR (KBr) 3283,2945,2860,1534,1453,1415cm -1 1H-NMR (DMSO-d 6) δ: 1.38-2.00 (m, 12H), 2.45 (s, 3H), 2.30-3.95 (m, 4H), 3.22-3.75 (m, 2H), 3.58 (s, 2H), 3.86 (s, 3H), 4.12 (m, 1H), 6.73-7.16 (m, 5H), 7.77-7.79 (m, 1H), 7.98-8.02 (m, 1H), 8.51-8.52 (m, 1H), 8.57-8.59 (m, 1H); MS (FAB) m/z 523 (M ++ 1); C 29H 38N 4O 50.5NaCl2.2H 2The analytical calculation value of O: C, 58.89; H, 7.23; N, 9.47.Measured value: C, 59.21; H, 7.11; N, 9.11.Embodiment 164 cis-4-[[1-[3-methoxyl group 4-[N '-(2-aminomethyl phenyl) urea groups] phenyl acetyl]-the 2-pyrrolidinyl] methylamino] the naphthenic acid methyl ester hydrochloride
Figure A0080951004191
In 0 ℃, with SOCl 2Add in the methanol.Stir after 5 minutes, add cis-4-[[1-[3-methoxyl group-4-[N '-(2-aminomethyl phenyl) urea groups] phenyl acetyl]-the 2-pyrrolidinyl] methylamino] naphthenic acid (200mg, 0.38mmol).Under room temperature, mixture was stirred 5 hours.This mixture of vacuum concentration.Sodium bicarbonate aqueous solution is added in the residue, use dichloromethane extraction.With salt water washing extract, then through dried over sodium sulfate and vacuum concentration.Residue is through purification by silica gel column chromatography, with methanol-dichloromethane (5: 95-18: 92, v/v) as eluant.Product is dissolved in the ethanol (5.0ml), and (1.0ml 1.0mmol) adds wherein with 1N HCl (in ethanol).This mixture of vacuum concentration obtains 172 (160mg, 74%), is amorphous solid.MW 536.66 IR (KBr) 3247,2950,2875,1731,1671,1612,1533,1454,1205cm -1 1H-NMR (DMSO-d 6) δ 1.45-2.10 (m, 12H), 2.25 (s, 3H), 2.60-2.70 (m, 1H), 2.90-3.20 (m, 3H), 2.50-2.55 (m, 2H), 3.63 (m, 5H), 3.86 (s, 3H), 4.15-4.30 (m, 1H), 6.74-7.16 (m, 5H), 7.76-7.78 (m, 1H), 8.00-8.09 (m, 1H), 8.54-8.70 (m, 2H); MS (FAB) m/z 537 (M ++ 1); C 30H 40N 4O 51.0HCl1.0H 2The analytical calculation value of O: C, 60.95; H, 7.33; N, 9.48; Cl, 6.00.Measured value: C, 60.87; H, 7.47; N, 8.97; Cl, 5.90.Embodiment 1654-[N-[1-[3-methoxyl group-4-[N '-(2-aminomethyl phenyl) urea groups] phenyl acetyl]-the 2-pyrrolidinyl] methyl]-the N-methylamino] naphthenic acid
Figure A0080951004192
In 0 ℃; to cis-4-[[1-[3-methoxyl group-4-[N '-(2-aminomethyl phenyl) urea groups] phenyl acetyl]-the 2-pyrrolidinyl] methylamino] naphthenic acid methyl ester (300mg; 0.55mmol), (66mg adds NaBH in the agitating solution of methanol 1.1mmol) (10.0ml) for HCHO (300ml) and acetic acid 3CN (70mg, 1.1mmol).Stirred reaction mixture is 18 hours under room temperature.Behind the vacuum concentration, add entry, use dichloromethane extraction.Wash extract with water, then through dried over sodium sulfate and vacuum concentration.Residue is through the TLC purification; with methanol-dichloromethane (3: 97; v/v) as eluant; obtain 4-[N-[1-[3-methoxyl group-4-[N '-(2-aminomethyl phenyl) urea groups] phenyl acetyl]-2-pyrrolidinyl methyl]-the N-methylamino] naphthenic acid methyl ester (160mg; 52%), is amorphous solid. 1H-NMR(CDCl 3)δ1.30-2.20(m，12H)，2.27-2.37(m，6H)，2.50-2.60(m，1H)，3.30-3.80(m，5H)，3.55(s，2H)，3.66-3.73(m，6H)，4.10-4.20(m，1H)，6.60-7.55(m，7H)，8.03(m，1H)，8.15(m，1H)。
To 4-[N-[1-[3-methoxyl group-4-[N '-(2-aminomethyl phenyl) urea groups] phenyl acetyl]-2-pyrrolidinyl methyl]-the N-methylamino] naphthenic acid methyl ester (100mg; 0.1 adding 1N sodium hydroxide in the agitating solution of THF 8mmol) (5.0ml) and methanol (2.5ml) (0.36ml, 0.36mmol).Stirred this mixture 18 hours in 60 ℃.This mixture of vacuum concentration adds water wherein, with 1N HCl neutralization.This mixture of vacuum concentration.Residue is through the TLC purification, with methanol-dichloromethane (1/4, v/v) as eluant, obtain 173 (10mg, 10%), be amorphous solid.MW 536.66 IR (KBr) 3440,2954,1697,1533,1454cm -1 1H-NMR (DMSO-d 6) δ: 1.20-2.30 (m, 13H), 2.24 (s, 3H), 2.35-4.00 (m, 13H), 6.50-8.10 (m, 8H), 8.50 (m, 1H); MS (FAB) m/z 537 (M ++ 1); C 30H 40N 4O 52.0NaCl0.8H 2The analytical calculation value of O: C, 53.94; H, 6.28; N, 8.39.Measured value: C, 54.08; H, 6.52; N, 8.04.Embodiment 1664-[[1-[3-methoxyl group-4-[N '-(2-aminomethyl phenyl) urea groups] phenyl acetyl]-(2S)-and pyrrolidinyl] methoxyl group] naphthenic acid
Figure A0080951004201
Under room temperature, with 4-(1-tert-butoxycarbonyl-(2S)-pyrrolidinyl) methoxyl methyl benzoate (1.0g, 2.9mmol) and 5% be carried on the hydrogenation 36 hours under 5 atmospheric pressure of the ethanol (10.0ml) of the Rh (500mg) on the aluminium oxide and the mixture in the acetic acid (1.0ml).Remove by filter catalyst, vacuum concentrated filtrate.Residue is through purification by silica gel column chromatography, with normal hexane-ethyl acetate (6: 1, v/v) as eluant, obtain cis-4-[(1-tert-butoxycarbonyl-(2S)-pyrrolidinyl) methoxyl group] naphthenic acid methyl ester (900mg, 89%), be faint yellow oily thing. 1H-NMR(CDCl 3)δ1.46(s，9H)，1.46-2.00(m，12H)，2.34(m，1H)，3.20-3.55(m，5H)，3.67(s，3H)，3.84-3.92(m，1H)。
In 0 ℃, to cis-4-[(1-tert-butoxycarbonyl-(2S)-pyrrolidinyl) methoxyl group] (900mg adds TFA (5.0ml) to the naphthenic acid methyl ester in the agitating solution of dichloromethane 2.6mmol) (5.0ml).Under room temperature, stirred this reactant mixture 0.5 hour.This mixture of vacuum concentration adds saturated sodium bicarbonate in this residue, uses dichloromethane extraction.With salt water washing extract, through dried over sodium sulfate and vacuum concentration.Crude product is used for subsequent reaction without being further purified.To crude product (200mg, 0.83mmol), 3-methoxyl group-4-[N '-(2-aminomethyl phenyl) urea groups] phenylacetic acid (260mg, 0.83mmol), HOBt (135mg, 1.0mmol) and triethylamine (344 μ l, and 1.35-2.10 (m, 12H), 2.15-2.38 (m, 1H), 2.29 (m, 3H), and 3.20-3.55 (m, 5H), 3.58 (s, 2H), 3.66 (s, 3H), 3.73 (s, 3H), 4.20-4.25 (m, 1H), 6.26-6.30 (m, 1H), and 6.78-6.81 (m, 2H), 7.06-7.23 (m, 3H), 7.51-7.52 (m, 1H), and 8.01-8.03 (m, 1H).
To 4-[[1-[3-methoxyl group-4-[N '-(2-aminomethyl phenyl) urea groups] phenyl acetyl]-(2S)-and pyrrolidinyl] methoxyl group] naphthenic acid methyl ester (460mg; 0.86mmol) THF (10.0ml) and the agitating solution of ethanol (5.0ml) in add 1N sodium hydroxide (1.4ml, 1.4 mmol).Stirred this mixture 18 hours in 60 ℃.This mixture of vacuum concentration adds water wherein, with 1N HCl neutralization.Collect the solid that produces, wash with water, vacuum drying obtains 174 (370mg, 83%), is white crystalline solid.MW 523.63 mp 110-113 ℃; IR (KBr) 3345,2937,1612,1533,1454cm -1 1H-NMR (DMSO-d 6) δ 1.00-2.00 (m, 12H), 2.24 (s, 3H), 2.20-2.30 (m, 1H), 3.20-3.80 (m, 5H), 3.55 (s, 2H), 3.85 (s, 3H), 4.00-4.18 (m, 1H), 6.71-7.16 (m, 5H), 7.78 (d, J=8.0Hz, 1H), 7.90 (d, J=8.3Hz, 1H), 8.45 (s, 1H), 8.54 (s, 1H); MS (FAB) m/z 524 (M ++ 1); C 29H 37N 3O 60.2H 2The analytical calculation value of O: C, 66.07; H, 7.15; N, 7.97.Measured value: C, 66.02; H, 7.14; N, 7.87.Embodiment 1674-[[1-[4-[N '-(2-chlorophenyl) urea groups]-3-methoxyphenyl acetyl group]-(2S)-and pyrrolidinyl] methoxyl group] naphthenic acid
Figure A0080951004221
In 0 ℃, to 4-[(1-tert-butoxycarbonyl-2-pyrrolidinyl) methoxyl group] (900mg adds TFA (5.0ml) to the naphthenic acid methyl ester in the agitating solution of dichloromethane 2.6mmol) (5.0ml).Under room temperature, stirred this reactant mixture 0.5 hour.This mixture of vacuum concentration adds saturated sodium bicarbonate in this residue, uses dichloromethane extraction.With salt water washing extract, through dried over sodium sulfate and vacuum concentration.Crude product is used for subsequent reaction without being further purified.In 0 ℃, to crude product (200mg, 0.83mmol), 4-[N '-(2-chlorophenyl) urea groups]-3-methoxybenzene guanidine-acetic acid (277mg, 0.83mmol), HOBt (135mg, 1.0mmol) and triethylamine (344ml, 2.48mmol) THF (10.0ml) and the agitating solution of MeCN (10.0ml) in add EDCHCl (238mg, 1.24mmol).Under room temperature, reactant mixture was stirred 16 hours vacuum concentration.Water is added in the residue, use ethyl acetate extraction.With saturated sodium bicarbonate, 2M citric acid and saturated sodium bicarbonate washing extract, then through dried over sodium sulfate and vacuum concentration.Residue is through purification by silica gel column chromatography; with normal hexane-ethyl acetate (1: 6; v/v) as eluant; obtain 4-[[1-[4-[N '-(2-chlorophenyl) urea groups]-3-methoxyphenyl acetyl group]-(2S)-and pyrrolidinyl] methoxyl group] naphthenic acid methyl ester (450mg; 97%), is colorless oil. 1H-NMR(CDCl 3)δ1.35-2.15(m，12H)，2.25-2.40(m，1H)，3.40-3.70(m，5H)，3.61(s，2H)，3.66(s，3H)，3.81(s，3H)，4.20-4.30(m，1H)，6.81-6.99(m，3H)，7.17-7.34(m，3H)，7.92-7.94(m，2H)，8.17-8.19(m，2H)。
To 4-[[1-[4-[N '-(2-chlorophenyl) urea groups]-3-methoxyphenyl acetyl group]-(2S)-and pyrrolidinyl] methoxyl group] naphthenic acid methyl ester (450mg; 0.86mmol) THF (10.0ml) and the agitating solution of methanol (5.0ml) in add the 1N sodium hydroxide (1.4ml, 1.4mmol).Stirred this mixture 24 hours in 70 ℃.This mixture of vacuum concentration adds water wherein, with 1N HCl neutralization.Collect the solid that produces, wash with water, vacuum drying obtains 175 (370mg, 84%), is white crystalline solid.MW 544.04 mp 111-115 ℃; IR (KBr) 3330,2938,1704,1594,1533,1438,1199cm -1 1H-NMR (DMSO-d 6) δ 1.00-2.00 (m, 12H), 2.20-2.30 (m, 1H), 3.20-3.80 (m, 5H), 3.55 (s, 2H), 3.85 (s, 3H), 4.00-4.20 (m, 1H), 6.73-6.75 (m, 1H), 6.87 (s, 1H), 6.99-7.03 (m, 1H), 7.25-7.29 (m, 1H), 7.42 (d, J=7.1Hz, 1H), 7.95 (d, J=8.3Hz, 1H), 8.08 (d, J=8.0Hz, 1H), 8.78 (s, 1H), 8.92 (s, 1H); MS (FAB) m/z 544 (M ++ 1); C 28H 34N 3O 6Cl0.2H 2The analytical calculation value of O: C, 61.41; H, 6.33; N, 7.67; Cl, 6.47.Measured value: C, 61.37; H, 6.32; N, 7.56; Cl, 6.55.Embodiment 1684-[1-[4-[N '-(2-bromo phenyl) urea groups]-3-methoxyphenyl acetyl group]-(2S)-and pyrrolidinyl] methoxyl group] naphthenic acid
Figure A0080951004231
In 0 ℃, to 4-[(1-tert-butoxycarbonyl-2-pyrrolidinyl) methoxyl group] (900mg adds TFA (5.0ml) to the naphthenic acid methyl ester in the agitating solution of dichloromethane 2.6mmol) (5.0ml).Under room temperature, stirred this reactant mixture 0.5 hour.This mixture of vacuum concentration adds saturated sodium bicarbonate in this residue, uses dichloromethane extraction.With salt water washing extract, through dried over sodium sulfate and vacuum concentration.Crude product is used for subsequent reaction without being further purified.In 0 ℃, to crude product (200mg, 0.83mmol), 4-[N '-(2-bromo phenyl) urea groups]-3-methoxybenzene guanidine-acetic acid (314mg, 0.83mmol), HOBt (135mg, 1.0mmol) and triethylamine (344ml, 2.48mmol) THF (10.0ml) and the agitating solution of MeCN (10.0ml) in add EDCHCl (238mg, 1.24mmol).Under room temperature, reactant mixture was stirred 16 hours vacuum concentration.Water is added in the residue, use ethyl acetate extraction.With saturated sodium bicarbonate, 2M citric acid and saturated sodium bicarbonate washing extract, then through dried over sodium sulfate and vacuum concentration.Residue is through purification by silica gel column chromatography; with normal hexane-ethyl acetate (1: 6; v/v) as eluant; obtain [4-[1-[4-[N '-(2-bromo phenyl) urea groups]-3-methoxyphenyl acetyl group]-(2S)-and pyrrolidinyl] methoxyl group] naphthenic acid methyl ester (450mg; 90%), is colorless oil. 1H-NMR(CDCl 3)δ1.30-2.10(m，12H)，2.35-2.40(m，1H)，3.25-3.70(m，5H)，3.84(s，3H)，4.10-4.25(m，1H)，6.81-7.06(m，4H)，7.25-7.32(m，2H)，7.50-7.52(m，1H)，7.90-7.92(m，1H)，8.13-8.15(m，1H)。
To 4-[1-[4-[N '-(2-bromo phenyl) urea groups]-3-methoxyphenyl acetyl group]-(2S)-and pyrrolidinyl] methoxyl group] naphthenic acid methyl ester (450mg; 0.74mmol) THF (10.0ml) and the agitating solution of methanol (5.0ml) in add the 1N sodium hydroxide (1.2ml, 1.2mmol).Stirred this mixture 24 hours in 70 ℃.This mixture of vacuum concentration adds water wherein, with 1N HCl neutralization.Collect the solid that produces, wash with water, vacuum drying obtains 176 (340mg, 77%), is white crystalline solid.MW 588.49 mp 108-111 ℃; IR (KBr) 3328,2938,1702,1594,1529,1434cm -1 1H-NMR (DMSO-d 6) δ 1.00-2.00 (m, 12H), 2.15-2.25 (m, 1H), 3.40-3.75 (m, 5H), 3.48 (s, 2H), 3.85 (s, 3H), 4.04-4.15 (m, 1H), 6.70-6.72 (m, 1H), 6.87 (s, 1H), and 6.94-6.98 (m, 1H), 7.29-7.33 (m, 1H), 7.59 (d, J=8.1Hz, 1H), 7.93-7.95 (m, 2H), 8.73-8.74 (m, 1H), 8.91-8.92 (m, 1H); MS (FAB) m/z 589 (M ++ 1); C 28H 34N 3O 6Br0.2H 2The analytical calculation value of O: C, 56.80; H, 5.86; N, 7.10; Br, 13.49.Measured value: C, 56.66; H, 5.83; N, 6.97; Br, 13.66.Embodiment 1694-[[1-[4-[N '-(2-aminomethyl phenyl) urea groups] phenyl acetyl]-(2S)-and pyrrolidinyl] methoxyl group] naphthenic acid
In 0 ℃, to 4-[(1-tert-butoxycarbonyl-2-pyrrolidinyl) methoxyl group] (450mg adds TFA (5.0ml) to the naphthenic acid methyl ester in the agitating solution of dichloromethane 1.3mmol) (5.0ml).Under room temperature, stirred this reactant mixture 0.5 hour.This mixture of vacuum concentration adds saturated sodium bicarbonate in this residue, uses dichloromethane extraction.With salt water washing extract, through dried over sodium sulfate and vacuum concentration.Crude product is used for subsequent reaction without being further purified.In 0 ℃, to crude product, 4-[N '-(2-aminomethyl phenyl) urea groups] phenylacetic acid (375mg, 1.3mmol), HOBt (178mg, 1.3mmol) and triethylamine (550ml, 3.9mmol) THF (6.0ml) and the agitating solution of MeCN (6.0ml) in add EDCHCl (380mg, 1.9mmol).Under room temperature, reactant mixture was stirred 16 hours vacuum concentration.Water is added in the residue, use ethyl acetate extraction.With saturated sodium bicarbonate, 2M citric acid and saturated sodium bicarbonate washing extract, then through dried over sodium sulfate and vacuum concentration.Residue is through purification by silica gel column chromatography; with normal hexane-ethyl acetate (1: 6,, obtain 4-[[1-[4-[N '-(2-aminomethyl phenyl) urea groups v/v) as eluant] phenyl acetyl]-(2S)-pyrrolidinyl] methoxyl group] naphthenic acid methyl ester (520mg; 78%), is colorless oil. 1H-NMR(CDCl 3)δ1.30-2.40(m，13H)，2.21(s，3H)，3.30-3.80(m，7H)，3.65(s，3H)，4.10-4.30(m，1H)，6.90-7.20(m，8H)，7.40-7.70(m，2H)。
To 4-[[1-[4-[N '-(2-aminomethyl phenyl) urea groups] phenyl acetyl]-(2S)-and pyrrolidinyl] methoxyl group] naphthenic acid methyl ester (520mg; 1.0mmol) THF (10.0ml) and the agitating solution of methanol (5.0ml) in add the 1N sodium hydroxide (1.5ml, 1.5mmol).Stirred this mixture 24 hours in 70 ℃.This mixture of vacuum concentration adds water wherein, with 1N HCl neutralization.Collect the solid that produces, wash with water, vacuum drying obtains 177 (450mg, 91%), is white crystalline solid.MW 493.60 mp 107-111 ℃; IR (KBr) 3353,2938,1704,1540,1454,1240cm -1 1H-NMR (DMSO-d 6) δ 1.20-2.00 (m, 12H), 2.23 (s, 3H), 2.22-2.24 (m, 1H), 3.20-3.80 (m, 7H), 4.00-4.18 (m, 1H), 6.90-6.94 (m, 1H), 7.10-7.16 (m, 5H), 7.36-7.38 (m, 2H), 7.82-7.87 (m, 2H), 8.89 (s, 1H), 12.0 (br s, 1H); MS (FAB) m/z 494 (M ++ 1); C 28H 35N 3O 50.2H 2The analytical calculation value of O: C, 67.64; H, 7.18; N, 8.45.Measured value: C, 67.66; H, 7.19; N, 8.24.Embodiment 170 cis-4-[1-[4-[N '-(2-chlorophenyl) urea groups]-3-methoxyphenyl acetyl group]-(2S)-and octahydro indyl methoxyl group] naphthenic acid
In 0 ℃, to [(the 1-tert-butoxycarbonyl-(2S)-the octahydro indyl] (1.0g adds BH in the agitating solution of THF 3.7mmol) (10.0ml) to formic acid 3(1.0M is in THF, 8.0ml) for THF.After stirring 1.0 hours under the room temperature, with this reaction mixture refluxed heating 1.5 hours.After the cooling, this mixture of vacuum concentration.In 0 ℃ water is added wherein, use ethyl acetate extraction.Wash extract with water, through dried over sodium sulfate and vacuum concentration, obtain [the 1-tert-butoxycarbonyl-(2S)-the octahydro indyl] (947mg quantitatively), is colorless oil to methanol.
In 0 ℃, to [the 1-tert-butoxycarbonyl-(2S)-the octahydro indyl] methanol (947mg, 3.7mmol), the 4-methyl hydroxybenzoate (565mg, 3.7mmol) and triphenyl phasphine (1.2g adds DIAD (984mg.4.5mmol) in the agitating solution of THF 4.5mmol) (10ml).Under room temperature, this reactant mixture was stirred 18 hours.This mixture of vacuum concentration.Residue is through purification by silica gel column chromatography, with normal hexane-ethyl acetate (9/1,, obtain 4-[1-tert-butoxycarbonyl-(2S)-octahydro indyl methoxyl group v/v) as eluant] essence of Niobe (700mg, 50%), be faint yellow oily thing. 1H-NMR(CDCl 3)δ1.10-2.25(m，11H)，1.45(s，9H)，3.88(s，3H)，3.70-4.20(m，3H)，4.36(br?s，1H)，6.94(d，J＝8.8Hz，2H)，7.96(d，J＝8.5Hz，2H)。
Under room temperature, with 4-[1-tert-butoxycarbonyl-(2S)-octahydro indyl methoxyl group] and essence of Niobe (700mg, 1.8mmol) and the hydrogenation 48 hours under 5 atmospheric pressure of 5% mixture that is carried on the ethanol (10.0ml) of the Rh (400mg) on the aluminium oxide and acetic acid (1.0ml).Remove by filter catalyst, vacuum concentrated filtrate.Residue is through purification by silica gel column chromatography, with normal hexane-ethyl acetate (7: 1,, obtain cis-4-[1-tert-butoxycarbonyl-(2S)-octahydro indyl methoxyl group v/v) as eluant] naphthenic acid methyl ester (600mg, 85%), be faint yellow oily thing. 1H-NMR(CDCl 3)δ1.10-2.35(m，20H)，1.44(s，9H)，3.45-3.90(m，5H)，3.80(s，3H)。
In 0 ℃, to cis-4-[1-tert-butoxycarbonyl-(2S)-octahydro indyl methoxyl group] (600mg adds TFA (6.0ml) to the naphthenic acid methyl ester in the agitating solution of dichloromethane 1.5mmol) (6.0ml).Under room temperature, stirred this reactant mixture 0.5 hour.This mixture of vacuum concentration adds saturated sodium bicarbonate in this residue, uses dichloromethane extraction.With salt water washing extract, through dried over sodium sulfate and vacuum concentration.Crude product is used for subsequent reaction without being further purified.In 0 ℃, to crude product (221mg, 0.75mmol), 4-[N '-(2-chlorophenyl) urea groups]-3-methoxybenzene guanidine-acetic acid (250mg, 0.75mmol), HOBt (101mg, 0.75mmol) and triethylamine (3 12ml, 2.3mmol) THF (10.0ml) and the agitating solution of MeCN (10.0ml) in add EDCHCl (216mg, 1.1mmol).Under room temperature, reactant mixture was stirred 16 hours vacuum concentration.Water is added in the residue, use ethyl acetate extraction.With saturated sodium bicarbonate, 2M citric acid and saturated sodium bicarbonate washing extract, then through dried over sodium sulfate and vacuum concentration.Residue is through purification by silica gel column chromatography; with normal hexane-ethyl acetate (1: 3; v/v) as eluant; obtain cis-4-[1-[4-[N '-(2-chlorophenyl) urea groups]-3-methoxyphenyl acetyl group]-(2S)-and octahydro indyl methoxyl group] naphthenic acid methyl ester (430mg; 94%), is colorless oil. 1H-NMR(CDCl 3)δ1.10-2.40(m，20H)，3.45(br?s，1H)，3.62(s，2H)，3.66(s，3H)，3.73(s，3H)，3.60-3.85(m，2H)，4.09-4.14(m，2H)，6.75-6.98(m，3H)，7.22-2.46(m，4H)，7.92(d，J＝8.0Hz，1H)，8.18(d，J＝8.3Hz，1H)。
To cis-4-[1-[4-[N '-(2-chlorophenyl) urea groups]-3-methoxyphenyl acetyl group]-(2S)-and octahydro indyl methoxyl group] naphthenic acid methyl ester (430mg; 0.7mmol) THF (10.0ml) and the agitating solution of methanol (5.0ml) in add the 1N sodium hydroxide (1.4ml, 1.4mmol).Stirred this mixture 24 hours in 70 ℃.This mixture of vacuum concentration adds water wherein, with 1N HCl neutralization.Collect the solid that produces, wash with water, vacuum drying obtains 178 (360mg, 86%), is white crystalline solid.MW 598.13 mp 120-121 ℃ IR (KBr) 3338,2933,2859,1614,1533,1438cm -1 1H-NMR (CDCl 3) δ 1.05-2.40 (m, 20H), 3.38-3.50 (m, 2H), 3.63 and 3.65 (respectively is s, be total to 2H), 3.71 and 3.75 (they respectively are s, are total to 3H), 3.70-3.80 (m, 1H), 3.93-3.97 (m, 1H), 4.15 (br s, 1H), 6.75-6.77 (m, 2H), and 6.93-6.97 (m, 1H), 7.20-7.32 (m, 3H), 7.60-7.63 (m, 1H), 7.85 (d, J=8.3Hz, 1H), 8.16 (d, J=8.3Hz, 1H); MS (FAB) m/z 598 (M ++ 1); C 32H 40N 3O 6C10.5H 2The analytical calculation value of O: C, 63.30; H, 6.81; N, 6.92; Cl, 5.84.Measured value: C, 63.68; H, 6.81; N, 6.81; Cl, 5.98.Embodiment 171 cis-4-[1-[4-[N '-(2-bromo phenyl) urea groups]-3-methoxyphenyl acetyl group]-(2S)-and octahydro indyl methoxyl group] naphthenic acid
In 0 ℃, to cis-4-[1-tert-butoxycarbonyl-(2S)-octahydro indyl methoxyl group] (600mg adds TFA (6.0ml) to the naphthenic acid methyl ester in the agitating solution of dichloromethane 1.5mmol) (6.0ml).Under room temperature, stirred this reactant mixture 0.5 hour.This mixture of vacuum concentration adds saturated sodium bicarbonate in this residue, uses dichloromethane extraction.With salt water washing extract, through dried over sodium sulfate and vacuum concentration.Crude product is used for subsequent reaction without being further purified.In 0 ℃, to crude product (221mg, 0.75mmol), 4-[N '-(2-bromo phenyl) urea groups]-3-methoxybenzene guanidine-acetic acid (284mg, 0.75mmol), HOBt (101mg, 0.75mmol) and triethylamine (312ml, 2.3mmol) THF (10.0ml) and the agitating solution of MeCN (10.0ml) in add EDCHCl (216mg, 1.1mmol).Under room temperature, reactant mixture was stirred 16 hours vacuum concentration.Water is added in the residue, use ethyl acetate extraction.With saturated sodium bicarbonate, 2M citric acid and saturated sodium bicarbonate washing extract, then through dried over sodium sulfate and vacuum concentration.Residue is through purification by silica gel column chromatography; with normal hexane-ethyl acetate (1: 3; v/v) as eluant; obtain cis-4-[1-[4-[N '-(2-bromo phenyl) urea groups]-3-methoxyphenyl acetyl group]-(2S)-and octahydro indyl methoxyl group] naphthenic acid methyl ester (480mg; 96%), is colorless oil. 1H-MR(CDCl 3)δ1.10-2.40(m，20H)，3.45(br?s，1H)，3.61(s，2H)，3.66(s，3H)，3.76(s，3H)，3.60-3.80(m，2H)，4.11-4.14(m，2H)，6.76-6.92(m，3H)，7.25-7.32(m，3H)，7.49(d，J＝7.1Hz，1H)，7.90(d，J＝8.0Hz，1H)，8.13(d，J＝8.0Hz，1H)。
To cis-4-[1-[4-[N '-(2-bromo phenyl) urea groups]-3-methoxyphenyl acetyl group]-(2S)-and octahydro indyl methoxyl group] naphthenic acid methyl ester (480mg; 0.73mmol) THF (10.0ml) and the agitating solution of methanol (5.0ml) in add 1N sodium hydroxide (1.5ml, 1.5 mmol).Stirred this mixture 24 hours in 70 ℃.This mixture of vacuum concentration adds water wherein, with 1N HCl neutralization.Collect the solid that produces, wash with water, vacuum drying obtains 179 (400mg, 85%), is white crystalline solid.MW 642.58 mp 115-120 ℃; IR (KBr) 3332,2933,2859,1704,1592,1529,1434cm -1 1H-NMR (CDCl 3) δ 1.10-2.40 (m, 20H), 3.40-3.50 (m, 2H), 3.61 with 3.63 (they respectively are s, are total to 2H), 3.75 and 3.78 (they respectively are s, are total to 3H), 3.70-3.80 (m, 1H), 3.90-3.93 (m, 1H), 4.15 (br s, m), 6.76-6.92 (m, 3H), 7.26-7.30 (m, 1H), 7.43-7.52 (m, 3H), 7.84-7.86 (m, 1H), 8.10-8.12 (m, 1H); MS (FAB) m/z 643 (M ++ 1); C 32H 40N 3O 6Br0.4H 2The analytical calculation value of O: C, 59.15; H, 6.33; N, 6.49; Br, 12.30.Measured value: C, 59.26; H, 6.33; N, 6.36; Br, 12.37.Embodiment 1724-[[1-[3-methoxyl group-4-[N '-(2-aminomethyl phenyl) urea groups] phenyl acetyl]-the 2-pyrrolidinyl] carbonylamino] naphthenic acid
In 0 ℃, to 4-aminocyclohexane benzyl formate (900mg, 3.9mmol), boc-proline (830mg, 3.9mmol), HOBt (521mg, 3.9mmol) and triethylamine (1.6ml, add in the agitating solution of dichloromethane 11.6mmol) (30.0ml) EDCHCl (1.1g, 5.8mmol).Under room temperature, reactant mixture was stirred 16 hours vacuum concentration.Water is added in the residue, use ethyl acetate extraction.With saturated sodium bicarbonate, 2M citric acid and saturated sodium bicarbonate washing extract, then through dried over sodium sulfate and vacuum concentration.Residue is through purification by silica gel column chromatography, with normal hexane-ethyl acetate (6: 1, v/v) as eluant, obtain cis-4-[(1-tert-butoxycarbonyl-2-pyrrolidinyl) carbonylamino] naphthenic acid benzyl ester (600mg, 36%), be amorphous solid. 1H-NMR(CDCl 3)δ1.44(s，9H)，1.50-1.90(m，12H)，2.20-2.26(m，1H)，3.25-3.50(m，2H)，3.80-3.90(m，1H)，4.10-4.25(m，1H)，5.12(s，2H)，7.35-7.36(m，5H)。
In 0 ℃, to cis-4-[(1-tert-butoxycarbonyl-2-pyrrolidinyl) carbonylamino] (600mg adds TFA (3.0ml) to naphthenic acid benzyl ester in the agitating solution of dichloromethane 1.4mmol) (6.0ml).Under room temperature, stirred this reactant mixture 0.5 hour.This mixture of vacuum concentration adds saturated sodium bicarbonate in this residue, uses dichloromethane extraction.With salt water washing extract, through dried over sodium sulfate and vacuum concentration.Crude product is used for subsequent reaction without being further purified.In 0 ℃, to crude product (300mg, 0.7mmol), 3-methoxyl group-4-[N '-(2-aminomethyl phenyl) urea groups] phenylacetic acid (220mg, 0.7mmol), HOBt (94mg, 0.7mmol) and triethylamine (291ml, 2.1mmol) THF (10.0ml) and the agitating solution of MeCN (10.0ml) in add EDCHCl (201mg, 1.1mmol).Under room temperature, reactant mixture was stirred 16 hours vacuum concentration.Water is added in the residue, use ethyl acetate extraction.With saturated sodium bicarbonate, 2M citric acid and saturated sodium bicarbonate washing extract, then through dried over sodium sulfate and vacuum concentration.Collect the solid that produces,, obtain 4-[[1-[3-methoxyl group-4-[N '-(2-aminomethyl phenyl) urea groups with the ethyl acetate washing] phenyl acetyl]-the 2-pyrrolidinyl] carbonylamino] naphthenic acid benzyl ester (380mg, 87%), be white crystalline solid. 1H-NMR(CDCl 3)δ1.40-2.15(m，12H)，2.28(m，3H)，2.30-2.50(m，2H)，3.40-3.55(m，2H)，3.61(s，2H)，3.71(s，3H)，3.82(m，1H)，4.53(d，J＝6.3Hz，1H)，5.10(s，2H)，6.42(s，1H)，6.77-6.79(m，2H)，7.04-7.34(m，9H)，7.50-7.52(m，1H)，8.05-8.07(m，1H)。
To 4-[[1-[3-methoxyl group-4-[N '-(2-aminomethyl phenyl) urea groups] phenyl acetyl]-the 2-pyrrolidinyl] carbonylamino] naphthenic acid benzyl ester (380mg; 0.6mmol) THF (10.0ml) and the agitating solution of ethanol (5.0ml) in add the 1N sodium hydroxide (0.9ml, 0.9mmol).Stirred this mixture 18 hours in 50 ℃.This mixture of vacuum concentration adds water wherein, with 1N HCl neutralization.Collect the solid that produces, wash with water, vacuum drying obtains 180 (230mg, 71%), is white crystalline solid.MW 636.62 mp 136-142 ℃; IR (KBr) 3345,2940,1650,1625,1535,1454cm -1 1H-NMR (DMSO-d 6) δ 1.40-2.00 (m, 12H), 2.24 (s, 3H), 2.30-2.40 (m, 1H), 3.45-3.80 (m, 5H), and 3.86-3.87 (m, 3H), 4.30-4.43 (m, 1H), 6.65-7.30 (m, 5H), 7.70-7.80 (m, 1H), 7.98-8.09 (m, 1H), 8.46-8.57 (m, 1H); MS (FAB) m/z 537 (M ++ 1); C 29H 36N 4O 60.5H 2The analytical calculation value of O: C, 63.84; H, 6.83; N, 10.27.Measured value: C, 64.18; H, 6.91; N, 9.85.Embodiment 1734-[[1-[4-[N '-(2-chlorophenyl) urea groups]-3-methoxyphenyl acetyl group]-the 2-pyrrolidinyl] carbonylamino] naphthenic acid
Figure A0080951004311
In 0 ℃, to cis-4-[(1-tert-butoxycarbonyl-2-pyrrolidinyl) carbonylamino] (600mg adds TFA (3.0ml) to naphthenic acid benzyl ester in the agitating solution of dichloromethane 1.4mmol) (6.0ml).Under room temperature, stirred this reactant mixture 0.5 hour.This mixture of vacuum concentration adds saturated sodium bicarbonate in this residue, uses dichloromethane extraction.With salt water washing extract, through dried over sodium sulfate and vacuum concentration.Crude product is used for subsequent reaction without being further purified.In 0 ℃, to crude product (300mg, 0.7mmol), 4-[N '-(2-chlorophenyl) urea groups]-3-methoxybenzene guanidine-acetic acid (237mg, 0.7mmol), HOBt (94mg, 0.7mmol) and triethylamine (291ml, 2.1mmol) THF (10.0ml) and the agitating solution of MeCN (10.0ml) in add EDCHCl (201mg, 1.1mmol).Under room temperature, reactant mixture was stirred 16 hours vacuum concentration.Water is added in the residue, use ethyl acetate extraction.With saturated sodium bicarbonate, 2M citric acid and saturated sodium bicarbonate washing extract, then through dried over sodium sulfate and vacuum concentration.Collect the solid that produces,, obtain 4-[[1-[4-[N '-(2-chlorophenyl) urea groups with the ethyl acetate washing]-3-methoxyphenyl acetyl group]-the 2-pyrrolidinyl] carbonylamino] naphthenic acid benzyl ester (310mg, 68%), be white crystalline solid. 1H-NMR(CDCL 3)δ1.40-2.15(m，12H)，2.30-2.60(m，2H)，3.42-3.55(m，2H)，3.64(s，2H)，3.84(s，3H)，4.55(d，J＝6.1Hz，1H)，5.12(s，2H)，6.81-7.35(m，12H)，7.96-7.98(m，1H)，8.17-8.19(m，1H)。
To 4-[[1-[4-[N '-(2-chlorophenyl) urea groups]-3-methoxyphenyl acetyl group]-the 2-pyrrolidinyl] carbonylamino] naphthenic acid benzyl ester (310mg; 0.86mmol) THF (10.0ml) and the agitating solution of methanol (5.0ml) in add the 1N sodium hydroxide (0.7ml, 0.7mmol).Stirred this mixture 18 hours in 50 ℃.This mixture of vacuum concentration adds water wherein, with 1N HCl neutralization.Collect the solid that produces, wash with water, vacuum drying obtains 181 (260mg, 98%), is white crystalline solid.MW 557.03 mp 135-140 ℃; IR (KBr) 3328,2938,1594,1533,1438,1203cm -1 1H-NMR (DMSO-d 6) δ 1.40-2.20 (m, 12H), 2.30-2.40 (m, 1H), 3.40-3.80 (m, 5H), 3.65-3.85 (m, 3H), 4.30-4.43 (m, 1H), 6.66-7.31 (m, 5H), 7.42-8.10 (m, 2H), 8.89-8.94 (m, 2H); MS (FAB) m/z 557 (M ++ 1); C 28H 33N 4O 6Cl0.3H 2The analytical calculation value of O: C, 59.79; H, 6.02; N, 9.96; Cl, 6.30.Measured value: C, 59.86; H, 6.10; N, 9.60; Cl, 6.34.Embodiment 1744-[1-[3-methoxyl group-4-[N '-(2-aminomethyl phenyl) urea groups] phenyl acetyl]-(3S)-and pyrrolidinyl oxygen base] benzoic acid
To 4-methyl hydroxybenzoate (1.84g, 12.1mmol), (R)-1-benzyl-3-pyrrolidinol (2.00ml, 12.1mmol) and triphenyl phasphine (3.81g, 14.5mmol) cold (0 ℃) solution of stirring of THF (25ml) in add DIAD (2.86ml, 14.5mmol), with this reaction mixture refluxed heating 10 hours.After being cooled to room temperature, evaporate this mixture.Residue is through purification by silica gel column chromatography, with normal hexane-ethyl acetate (3: 1,, obtain 4-[1-benzyl-(3S)-pyrrolidinyl oxygen base v/v) as eluant] essence of Niobe (3.66g, 97%), be faint yellow oily thing. 1H-NMR(CDCl 3)δ1.95-2.02(m，1H)，2.28-2.37(m，1H)，2.57-2.63(m，1H)，2.72-2.81(m，2H)，2.96-3.01(m，1H)，3.63-3.71(m，2H)，3.87(s，3H)，4.84-4.89(m，1H)，6.84(d，J＝8.8Hz，2H)，7.23-7.34(m，5H)，7.95(d，J＝8.8Hz，2H)。
With 4-[1-benzyl-(3S)-pyrrolidinyl oxygen base] (3.66g, the solution of methanol 11.8mmol) (25ml) is through Pd (OH) for essence of Niobe 2/ C (0.73g, 20wt%) spend the night by hydrogenation.Filter this reactant mixture to remove catalyst, evaporate this solution, obtain 4-[(3S)-pyrrolidinyl oxygen base] (2.60g q.y.), is faint yellow oily thing to essence of Niobe. 1H-NMR(CDCl 3)δ1.94-2.01(m，2H)，2.09-2.18(m，1H)，2.91-2.97(m，1H)，3.04-3.09(m，1H)，3.16-3.23(m，2H)，3.88(s，3H)，4.88-4.91(m，1H)，6.86(m，2H)，7.96-7.98(m，2H)；MS(ESI)m/z?222(M ++1)。
Under room temperature, with 3-methoxyl group-4-[N '-(2-aminomethyl phenyl) urea groups] phenylacetic acid (449mg, 1.43mmol), 4-[(3S)-pyrrolidinyl oxygen base] essence of Niobe (316mg, 1.43mmol), EDCHCl (330mg, 1.72mmol), HOBt (193mg, 1.43mmol) and triethylamine (240ml, 1.72mmol) mixture in THF (5ml) stirs and to spend the night.This reactant mixture of dilute with water is used ethyl acetate extraction.With salt water washing extract, through dried over sodium sulfate and evaporation.Residue is through purification by silica gel column chromatography; with chloroform-methanol (100: 1-50: 1,, obtain 4-[1-[3-methoxyl group-4-[N '-(2-aminomethyl phenyl) urea groups v/v) as eluant] phenyl acetyl]-(3S)-pyrrolidinyl oxygen base] essence of Niobe (735mg; 99%), is faint yellow oily thing. 1H-NMR (CDCl 3) δ 2.03-2.31 (a plurality of s and m, 5H altogether), 3.57-3.78 (a plurality of m, 9H altogether), 3.88 (s, 3H), 4.95-4.99 (m, 1H), 6.73-7.00 (m, 5H), 7.06-7.10 (m, 1H), and 7.18-7.22 (m, 2H), 7.42-7.46 (m, 1H), 7.57-7.62 (m, 1H), and 7.95-8.08 (m, 3H); MS (ESI) m/z 518 (M ++ 1).
To 4-[1-[3-methoxyl group-4-[N '-(2-aminomethyl phenyl) urea groups] phenyl acetyl]-(3S)-and pyrrolidinyl oxygen base] essence of Niobe (627mg; 1.21mmol) the agitating solution of THF (5ml) in add 0.5N sodium hydroxide (5ml), with this reaction mixture refluxed heating 3 hours.After being cooled to room temperature, in this mixture impouring ice-1N HCl, the precipitation that produces is collected in decompression.Make crude product solid recrystallization in methanol-chloroform-IPE, obtain 182 (235mg, 39%), be white crystalline powder.MW 503.55 mp 131-135 ℃; 1H-NMR (DMSO-d 6) δ 2.06-2.27 (a plurality of m, 5H altogether), and 3.57-3.64 (m, 4H), 3.71-3.88 (a plurality of s and m, be total to 5H), 5.11 and 5.20 (they respectively are m, are total to 1H), 6.73-6.77 (m, 1H), 6.88-6.95 (m, 2H), 7.02-7.05 (m, 2H), 7.11-7.17 (m, 2H), and 7.79-7.81 (m, 1H), 7.88-7.90 (m, 2H), 7.98-8.03 (m, 1H), 8.45-8.47 (m, 1H), 8.55-8.57 (m, 1H), 12.66 (br s, 1H); MS (ESI) m/z504 (M ++ 1); C 28H 29N 3O 63/4H 2The analytical calculation value of O: C, 65.04; H, 5.95; N, 8.13.Measured value: C, 65.11; H, 5.99; N, 7.66.Embodiment 1754-[1-[4-[N '-(2-chlorophenyl) urea groups]-3-methoxyphenyl acetyl group]-(3S)-and pyrrolidinyl oxygen base] benzoic acid
Under room temperature, with 4-[N '-(2-chlorophenyl) urea groups]-3-methoxybenzene guanidine-acetic acid (410mg, 1.22mmol), 4-[(3S)-pyrrolidinyl oxygen base] essence of Niobe (270mg, 1.22mmol), EDCHCl (280mg, 1.46mmol), HOBt (200mg, 1.48mmol) and triethylamine (205ml, 1.47mmol) mixture in THF (8ml) stirs and to spend the night.This reactant mixture of dilute with water is used ethyl acetate extraction.With salt water washing extract, through dried over sodium sulfate and evaporation.Residue is through purification by silica gel column chromatography; with chloroform-methanol (100: 1-60: 1,, obtain 4-[1-[4-[N '-(2-chlorophenyl) urea groups v/v) as eluant]-3-methoxyphenyl acetyl group]-(3S)-pyrrolidinyl oxygen base] essence of Niobe (652mg; 99%), is white solid.Mp 200-203 ℃; 1H-NMR (CDCl 3) δ 2.06-2.32 (m, 2H), 3.60-3.82 (a plurality of m, 9H altogether), 3.88 (s, 3H), 4.97-5.01 (m, 1H), 6.76-6.86 (m, 4H), 6.95-6.99 (m, 1H), 7.23-7.47 (m, 4H), 7.91-7.99 (m, 3H), 8.19-8.21 (m, 1H); MS (ESI) m/z 537 (M +).
To 4-[1-[4-[N '-(2-chlorophenyl) urea groups]-3-methoxyphenyl acetyl group]-(3S)-and pyrrolidinyl oxygen base] essence of Niobe (650mg; 1.21mmol) the agitating solution of THF (5ml) in add 0.5N sodium hydroxide (5ml), with this reaction mixture refluxed heating 5 hours.After being cooled to room temperature, in this mixture impouring ice-1N HCl, the precipitation that produces is collected in decompression.Make crude product solid recrystallization in methanol-chloroform-IPE, obtain 183 (443mg, 70%), be light yellow crystalline powder.MW 523.97 mp 190-193 ℃; 1H-NMR (DMSO-d 6) δ 2.06-2.27 (m, 2H), 3.56-3.62 (m, 4H), 3.71-3.88 (a plurality of s and m, be total to 5H), 5.11 and 5.20 (they respectively are m, are total to 1H), 6.74-6.78 (m, 1H), 6.89-6.91 (m, 1H), 7.00-7.05 (m, 3H), 7.26-7.30 (m, 1H), 7.43-7.45 (m, 1H), 7.88-7.98 (m, 3H), 8.08-8.10 (m, 1H), 8.89-8.95 (m, 2H), 12.67 (br s, 1H); MS (ESI) m/z 524 (M ++ 1); C 27H 26ClN 3O 61/4H 2The analytical calculation value of O: C, 61.36; H, 5.05; N, 7.95; Cl, 6.71.Measured value: C, 61.69; H, 5.45; N, 7.29; Cl, 6.91.Embodiment 1764-[1-[4-[N '-(2-bromo phenyl) urea groups]-3-methoxyphenyl acetyl group]-(3S)-and pyrrolidinyl oxygen base] benzoic acid
Under room temperature, with 4-[N '-(2-bromo phenyl) urea groups]-3-methoxybenzene guanidine-acetic acid (540mg, 1.42mmol), 4-[(3S)-pyrrolidinyl oxygen base] essence of Niobe (315mg, 1.42mmol), EDCHCl (328mg, 1.71mmol), HOBt (230mg, 1.70mmol) and triethylamine (240ml, 1.72mmol) mixture in THF (8ml) stirs and to spend the night.This reactant mixture of dilute with water is used ethyl acetate extraction.With salt water washing extract, through dried over sodium sulfate and evaporation.Residue is through purification by silica gel column chromatography; with chloroform-methanol (1 00: 1-50: 1,, obtain 4-[1-[4-[N '-(2-bromo phenyl) urea groups v/v) as eluant]-3-methoxyphenyl acetyl group]-(3S)-pyrrolidinyl oxygen base] essence of Niobe (620mg; 74%), is the white foam thing. 1H-NMR (CDCl 3) δ 2.06-2.33 (m, 2H), 3.60-3.82 (a plurality of m, 9H altogether), 3.89 (s, 3H), 4.97-5.01 (m, 1H), 6.77-7.00 (m, 5H), 7.27-7.40 (m, 3H), 7.49-7.51 (m, 1H), 7.91-7.99 (m, 3H), 8.13-8.17 (m, 1H); MS (ESI) m/z 583 (M ++ 1).
To 4-[1-[4-[N '-(2-bromo phenyl) urea groups]-3-methoxyphenyl acetyl group]-(3S)-and pyrrolidinyl oxygen base] essence of Niobe (620mg; 1.06mmol) the agitating solution of THF (5ml) in add 0.5N sodium hydroxide (5ml), with this reaction mixture refluxed heating 2.5 hours.After being cooled to room temperature, in this mixture impouring ice-1N HCl, the precipitation that produces is collected in decompression.Make crude product solid recrystallization in methanol-chloroform-IPE, obtain 184 (421mg, 70%), be white crystalline powder.MW 568.42 mp 173-175 ℃; 1H-NMR (DMSO-d 6) δ 2.06-2.28 (m, 2H), 3.56-3.65 (m, 4H), (3.71-3.88 a plurality of s and m, 5H altogether), 5.12 and 5.20 (they respectively are m, are total to 1H), 6.74-6.78 (m, 1H), 6.89-7.05 (m, 4H), 7.31-7.34 (m, 1H), 7.59-7.61 (m, 1H), 7.88-7.98 (m, 4H), 8.73-8.74 (m, 1H), 8.91-8.93 (m, 1H), 12.67 (br s, 1H); MS (ESI) m/z 569 (M ++ 1); C 27H 26BrN 3O 6The analytical calculation value: C, 57.05; H, 4.61; N, 7.39; Br, 14.06.Measured value: C, 57.57; H, 5.12; N, 6.81; Br, 13.96.Embodiment 1774-[1-[3-methoxyl group-4-[N '-(2-aminomethyl phenyl) urea groups] phenyl acetyl]-(3R)-and pyrrolidinyl oxygen base] benzoic acid
To 4-methyl hydroxybenzoate (1.78g, 11.7mmol), 1-benzyl-(3S)-pyrrolidinol (2.07g, 11.7mmol) and triphenyl phasphine (3.68g, 14.0mmol) cold (0 ℃) solution of stirring of THF (25ml) in add DIAD (2.76ml.14.0mmol), with this reaction mixture refluxed heating 10 hours.After being cooled to room temperature, evaporate this mixture.Residue is through purification by silica gel column chromatography, with normal hexane-ethyl acetate (3: 1,, obtain 4-[1-benzyl-(3R)-pyrrolidinyl oxygen base v/v) as eluant] essence of Niobe (3.56g, 98%), be faint yellow oily thing. 1H-NMR(CDCl 3)δ1.95-2.03(m，1H)，2.28-2.37(m，1H)，2.57-2.63(m，1H)，2.73-2.81(m，2H)，2.97-3.01(m，1H)，3.63-3.72(m，2H)，3.87(s，3H)，4.85-4.90(m，1H)，6.83-6.85(m，2H)，7.27-7.34(m，5H)，7.94-7.97(m，2H)；MS(ESI)m/z312(M ++1)。
With 4-[1-benzyl-(3R)-pyrrolidinyl oxygen base] (3.56g, the solution of methanol 11.4mmol) (25ml) is through Pd (OH) for essence of Niobe 2/ C (0.72g, 20wt%) spend the night by hydrogenation.Filter this reactant mixture to remove catalyst, evaporate this solution, obtain 4-[(3R)-pyrrolidinyl oxygen base] (2.53g q.y.), is faint yellow oily thing to essence of Niobe. 1H-NMR(CDCl 3)δ1.94-2.18(m，3H)，2.91-2.97(m，1H)，3.04-3.09(m，1H)，3.16-3.22(m，2H)，3.88(s，3H)，4.88-4.91(m，1H)，6.86-6.89(m，2H)，7.97-7.99(m，2H)；MS(ESI)m/z263[M ++1+41，(+MeCN)]。
Under room temperature, with 3-methoxyl group-4-[N '-(2-aminomethyl phenyl) urea groups] phenylacetic acid (460mg, 1.46mmol), 4-[(3R)-pyrrolidinyl oxygen base] essence of Niobe (324mg, 1.46mmol), EDCHCl (337mg, 1.76mmol), HOBt (237mg, 1.75mmol) and triethylamine (245ml, 1.76mmol) mixture in THF (10ml) stirs and to spend the night.This reactant mixture of dilute with water is used ethyl acetate extraction.With salt water washing extract, through dried over sodium sulfate and evaporation.Residue is through purification by silica gel column chromatography; with chloroform-methanol (100: 1-50: 1,, obtain 4-[1-[3-methoxyl group-4-[N '-(2-aminomethyl phenyl) urea groups v/v) as eluant] phenyl acetyl]-(3R)-pyrrolidinyl oxygen base] essence of Niobe (583mg; 77%), is colorless oil. 1H-NMR (CDCl 3) δ 2.06-2.23 (m, 2H), 2.30 (s, 3H), 3.58-3.89 (a plurality of s and m, 12H altogether), 4.95-4.99 (m, 1H), 6.75-7.00 (m, 4H), 7.13-7.30 (m, 5H), 7.52-7.57 (m, 1H), 7.96-8.03 (m, 3H); MS (ESI) m/z 518 (M ++ 1).
To 4-[1-[3-methoxyl group-4-[N '-(2-aminomethyl phenyl) urea groups] phenyl acetyl]-(3R)-and pyrrolidinyl oxygen base] essence of Niobe (583mg; 1.13mmol) the agitating solution of THF (5ml) in add 0.5N sodium hydroxide (5ml), with this reaction mixture refluxed heating 3 hours.After being cooled to room temperature, in this mixture impouring ice-1N HCl, the precipitation that produces is collected in decompression.Make crude product solid recrystallization in methanol-chloroform-ether, obtain 185 (297mg, 52%), be white crystalline powder.MW 503.55 mp 158-162 ℃; 1H-NMR (DMSO-d 6) δ 2.08-2.31 (a plurality of s and m, 5H altogether), 3.54-3.89 (a plurality of m, 9H altogether), 5.11 with 5.20 (they respectively are m, are total to 1H), 6.72-7.17 (a plurality of m, 6H altogether), 7.78-7.80 (m, 1H), 7.87-7.90 (m, 2H), and 7.98-8.02 (m, 2H), 8.46-8.47 (m, 1H), 8.55-8.57 (m, 1H), 12.66 (br s, 1H); MS (ESI) m/z 504 (M ++ 1); C 28H 29N 3O 61/4H 2The analytical calculation value of O: C, 66.19; H, 5.85; N, 8.27.Measured value: C, 66.12; H, 5.77; N, 8.21.Embodiment 1784-[1-[4-[N '-(2-chlorophenyl) urea groups]-3-methoxyphenyl acetyl group]-(3R)-and pyrrolidinyl oxygen base] benzoic acid
Figure A0080951004371
Under room temperature, with 4-[N '-(2-chlorophenyl) urea groups]-3-methoxybenzene guanidine-acetic acid (498mg, 1.49mmol), 4-[(3R)-pyrrolidinyl oxygen base] essence of Niobe (329mg, 1.49mmol), EDCHCl (342mg, 1.78mmol), HOBt (241mg, 1.78mmol) and triethylamine (250ml, 1.79mmol) mixture in THF (10ml) stirs and to spend the night.This reactant mixture of dilute with water is used ethyl acetate extraction.With salt water washing extract, through dried over sodium sulfate and evaporation.Residue is through purification by silica gel column chromatography; with chloroform-methanol (100: 1-50: 1,, obtain 4-[1-[4-[N '-(2-chlorophenyl) urea groups v/v) as eluant]-3-methoxyphenyl acetyl group]-(3R)-pyrrolidinyl oxygen base] essence of Niobe (561mg; 70%), is the white foam thing. 1H-NMR (CDCl 3) δ 2.05-2.34 (m, 2H), 3.59-4.07 (a plurality of s and m, 12H altogether), 4.97-5.02 (m, 1H), 6.75-6.86 (m, 4H), 6.94-7.00 (m, 1H), 7.22-7.33 (m, 2H), 7.59-7.66 (m, 2H), 7.92-7.99 (m, 3H), 8.19-8.22 (m, 1H); MS (ESI) m/z 538 (M ++ 1).
To 4-[1-[4-[N '-(2-chlorophenyl) urea groups]-3-methoxyphenyl acetyl group]-(3R)-and pyrrolidinyl oxygen base] essence of Niobe (561mg; 1.04mmol) the agitating solution of THF (5ml) in add 0.5N sodium hydroxide (5ml), with this reaction mixture refluxed heating 5 hours.After being cooled to room temperature, in this mixture impouring ice-1N HCl, the precipitation that produces is collected in decompression.Make crude product solid recrystallization in methanol-chloroform-ether, obtain 186 (361mg, 66%), be white crystalline powder.MW 523.97 mp 193-194 ℃; 1H-NMR (DMSO-d 6) δ 2.06-2.28 (m, 2H), 3.58-3.62 (m, 4H), 3.71-3.76 (m, 1H), 3.83-3.89 (a plurality of s and m, 4H altogether), 5.12 and 5.20 (respectively are m, are total to 1H), 6.74-6.78 (m, 1H), 6.90-6.91 (m, 1H), 7.01-7.05 (m, 3H), and 7.27-7.30 (m, 1H), 7.43-7.45 (m, 1H), 7.88-7.98 (m, 3H), 8.08-8.10 (m, 1H), 8.89-8.96 (m, 2H), 12.67 (br s, 1H); MS (ESI) m/z524 (M ++ 1); C 27H 26ClN 3O 61/4H 2The analytical calculation value of O: C, 61.36; H, 5.05; N, 7.95; Cl, 6.71.Measured value: C, 61.49; H, 5.11; N, 7.72; Cl, 7.08.Embodiment 1794-[1-[4-[N '-(2-bromo phenyl) urea groups]-3-methoxyphenyl acetyl group]-(3R)-and pyrrolidinyl oxygen base] benzoic acid
Under room temperature, with 4-[N '-(2-bromo phenyl) urea groups]-3-methoxybenzene guanidine-acetic acid (460mg, 1.21mmol), 4-[(3R)-pyrrolidinyl oxygen base] essence of Niobe (269mg, 1.21mmol), EDCHCl (280mg, 1.46mmol), HOBt (197mg, 1.46mmol) and triethylamine (205ml, 1.47mmol) mixture in THF (10ml) stirs and to spend the night.This reactant mixture of dilute with water is used ethyl acetate extraction.With salt water washing extract, through dried over sodium sulfate and evaporation.Residue is through purification by silica gel column chromatography; with chloroform-methanol (100: 1-60: 1,, obtain 4-[1-[4-[N '-(2-bromo phenyl) urea groups v/v) as eluant]-3-methoxyphenyl acetyl group]-(3R)-pyrrolidinyl oxygen base] essence of Niobe (555mg; 78%), is the white foam thing. 1H-NMR (CDCL 3) δ 2.05-2.33 (m, 2H), 3.60-4.07 (a plurality of s and m, 12H altogether), 4.96-5.01 (m, 1H), 6.76-6.93 (m, 5H), 7.28-7.30 (m, 1H), 7.48-7.58 (m, 3H), 7.92-7.99 (m, 3H), 8.12-8.16 (m, 1H); MS (ESI) m/z 582 (M +).
To 4-[1-[4-[N '-(2-bromo phenyl) urea groups]-3-methoxyphenyl acetyl group]-(3R)-and pyrrolidinyl oxygen base] essence of Niobe (555mg; 0.95mmol) the agitating solution of THF (5ml) in add 0.5N sodium hydroxide (5ml), with this reaction mixture refluxed heating 5 hours.After being cooled to room temperature, in this mixture impouring ice-1N HCl, the precipitation that produces is collected in decompression.Make crude product solid recrystallization in methanol-chloroform-ether, obtain 187 (330mg, 61%), be white crystalline powder.MW 568.42 mp 175-177 ℃; 1H-NMR (DMSO-d 6) δ 1.99-2.27 (m, 2H), 3.56-3.63 (m, 4H), 3.71-3.76 (m, 1H), and 3.83-3.89 (a plurality of s and m, 4H), 5.12 and 5.20 (they respectively are m, are total to 1H), 6.74-6.78 (m, 1H), 6.89-7.05 (m, 4H), 7.30-7.35 (m, 1H), 7.59-7.61 (m, 1H), 7.88-7.98 (m, 4H), 8.74-8.76 (m, 1H), 8.92-8.95 (m, 1H), 12.68 (br s, 1H); MS (ESI) m/z 569 (M ++ 1); C 27H 26BrN 3O 6The analytical calculation value: C, 57.05; H, 4.61; N, 7.39; Br, 14.06.Measured value: C, 56.91; H, 4.66; N, 7.20; Br, 14.59.Embodiment 180
Figure A0080951004401
1g Tentagel PHB resin (load 0.29mmol/g) is dissolved among the 25ml DMF, and adding Fmoc-(4-carboxymethyl)-piperidines (318mg, 0.87mmol).This resin of jolting 5 minutes, add then DIC (220mg, 0.27ml, 1.74mmol) and DMAP (106mg, 0.87mmol) and this resin of jolting 24 hours.
Discharge this resin, with DMF (3x), methanol (3x) and dichloromethane (3x) washing, dry under vacuum then.Resin provides negative bromobenzene basket (bromophenylblue) test.Make resin be dissolved in the DMF solution of 20% piperidines and jolting 4 hours.Discharge this resin, with DMF (3x), methanol (3x) and dichloromethane (3x) washing, dry under vacuum then.Resin provides positive bromobenzene basket test.This resin is dissolved among the 25ml DMF, and adding Fmoc-L-morpholine-2-formic acid (307mg, 0.87mmol).This resin of jolting 5 minutes, add then PyBroP (406mg, 0.87mmol) and DIEA (123mg, 0.15ml, 0.87mmol) and jolting resin 24 hours.
Discharge this resin, with DMF (3x), methanol (3x) and dichloromethane (3x) washing, dry under vacuum then.Resin provides negative bromobenzene basket test.Make resin be dissolved in the DMF solution of 20% piperidines and jolting 4 hours.Discharge this resin, with DMF (3x), methanol (3x) and dichloromethane (3x) washing, dry under vacuum then.Resin provides positive bromobenzene basket test.This resin is dissolved among the 25ml DMF, add 4-neighbour-tolyl urea groups phenylacetic acid (247mg, 0.87mmol) and this resin of jolting 5 minutes.Add PyBroP (406mg, 0.87mmol) and DIEA (123mg, 0.15ml, 0.87mmol) and jolting resin 24 hours.
Discharge this resin, with DMF (3x), methanol (3x) and dichloromethane (3x) washing, dry under vacuum then.Resin provides negative bromobenzene basket test.Make resin be dissolved in the dichloromethane solution of 90%TFA and jolting 4 hours.Discharge this resin, collect eluent.Resin is dissolved in the fresh dichloromethane and jolting 30 minutes.Discharge this resin, collect eluent, and merge with first.Solvent removed in vacuo makes residue recrystallization from ethyl acetate-hexane, obtains 85mg 188.Embodiment 181
1g Tentagel PHB resin (load 0.29mmol/g) is dissolved among the 25ml DMF, and adding Fmoc-(4-carboxymethyl)-piperidines (318mg, 0.87mmol).This resin of jolting 5 minutes, add then DIC (220mg, 0.27ml, 1.74mmol) and DMAP (106mg, 0.87mmol) and this resin of jolting 24 hours.
Discharge this resin, with DMF (3x), methanol (3x) and dichloromethane (3x) washing, dry under vacuum then.Resin provides negative bromobenzene basket test.Make resin be dissolved in the DMF solution of 20% piperidines and jolting 4 hours.Discharge this resin, with DMF (3x), methanol (3x) and dichloromethane (3x) washing, dry under vacuum then.Resin provides positive bromobenzene basket test.This resin is dissolved among the 25ml DMF, and adding Fmoc-L-4-phenyl proline (307mg, 0.87mmol).This resin of jolting 5 minutes, add then PyBroP (406mg, 0.87mmol) and DIEA (123mg, 0.15ml, 0.87mmol) and jolting resin 24 hours.
Discharge this resin, with DMF (3x), methanol (3x) and dichloromethane (3x) washing, dry under vacuum then.Resin provides negative bromobenzene basket test.Make resin be dissolved in the DMF solution of 20% piperidines and jolting 4 hours.Discharge this resin, with DMF (3x), methanol (3x) and dichloromethane (3x) washing, dry under vacuum then.Resin provides positive bromobenzene basket test.This resin is dissolved among the 25ml DMF, add 4-neighbour-tolyl urea groups phenylacetic acid (247mg, 0.87mmol) and this resin of jolting 5 minutes.Add PyBroP (406mg, 0.87mmol) and DIEA (123mg, 0.15ml, 0.87mmol) and jolting resin 24 hours.
Discharge this resin, with DMF (3x), methanol (3x) and dichloromethane (3x) washing, dry under vacuum then.Resin provides negative bromobenzene basket test.Make resin be dissolved in the dichloromethane solution of 90%TFA and jolting 4 hours.Discharge this resin, collect eluent.Resin is dissolved in the fresh dichloromethane and jolting 30 minutes.Discharge this resin, collect eluent, and merge with first.Solvent removed in vacuo makes residue recrystallization from ethyl acetate-hexane, obtains 82mg 189.Embodiment 182
1g Tentagel PHB resin (load 0.29mmol/g) is dissolved among the 25ml DMF, and adding Fmoc-4-carboxymethyl-piperazine (318mg, 0.87mmol).This resin of jolting 5 minutes, add then DIC (220mg, 0.27ml, 1.74mmol) and DMAP (106mg, 0.87mmol) and this resin of jolting 24 hours.
Discharge this resin, with DMF (3x), methanol (3x) and dichloromethane (3x) washing, dry under vacuum then.Resin provides negative bromobenzene basket test.Make resin be dissolved in the DMF solution of 20% piperidines and jolting 4 hours.Discharge this resin, with DMF (3x), methanol (3x) and dichloromethane (3x) washing, dry under vacuum then.Resin provides positive bromobenzene basket test.This resin is dissolved among the 25ml DMF, and adding Fmoc-L-proline (294mg, 0.87mmol).This resin of jolting 5 minutes, add then PyBroP (406mg, 0.87mmol) and DIEA (123mg, 0.15ml, 0.87mmol) and jolting resin 24 hours.
Discharge this resin, with DMF (3x), methanol (3x) and dichloromethane (3x) washing, dry under vacuum then.Resin provides negative bromobenzene basket test.Make resin be dissolved in the DMF solution of 20% piperidines and jolting 4 hours.Discharge this resin, with DMF (3x), methanol (3x) and dichloromethane (3x) washing, dry under vacuum then.Resin provides positive bromobenzene basket test.This resin is dissolved among the 25ml DMF, add 4-neighbour-tolyl urea groups phenylacetic acid (247mg, 0.87mmol) and this resin of jolting 5 minutes.Add PyBroP (406mg, 0.87mmol) and DIEA (123mg, 0.15ml, 0.87mmol) and jolting resin 24 hours.
Discharge this resin, with DMF (3x), methanol (3x) and dichloromethane (3x) washing, dry under vacuum then.Resin provides negative bromobenzene basket test.Make resin be dissolved in the dichloromethane solution of 90%TFA and jolting 4 hours.Discharge this resin, collect eluent.Resin is dissolved in the fresh dichloromethane and jolting 30 minutes.Discharge this resin, collect eluent, and merge with first.Solvent removed in vacuo makes residue recrystallization from ethyl acetate-hexane, obtains 78mg 190.Embodiment 183
Figure A0080951004431
1g Tentagel PHB resin (load 0.29mmol/g) is dissolved among the 25ml DMF, and the adding different piperidine carboxylic acid of Fmoc-(isonipecotic acid) (306mg, 0.87mmol).This resin of jolting 5 minutes, add then DIC (220mg, 0.27ml, 1.74mmol) and DMAP (106mg, 0.87mmol) and this resin of jolting 24 hours.
Discharge this resin, with DMF (3x), methanol (3x) and dichloromethane (3x) washing, dry under vacuum then.Resin provides negative bromobenzene basket test.Make resin be dissolved in the DMF solution of 20% piperidines and jolting 4 hours.Discharge this resin, with DMF (3x), methanol (3x) and dichloromethane (3x) washing, dry under vacuum then.Resin provides positive bromobenzene basket test.This resin is dissolved among the 25ml DMF, and adding Fmoc-L-proline (294mg, 0.87mmol).This resin of jolting 5 minutes, add then PyBroP (406mg, 0.87mmol) and DIEA (123mg, 0.15ml, 0.87mmol) and jolting resin 24 hours.
Discharge this resin, with DMF (3x), methanol (3x) and dichloromethane (3x) washing, dry under vacuum then.Resin provides negative bromobenzene basket test.Make resin be dissolved in the DMF solution of 20% piperidines and jolting 4 hours.Discharge this resin, with DMF (3x), methanol (3x) and dichloromethane (3x) washing, dry under vacuum then.Resin provides positive bromobenzene basket test.This resin is dissolved among the 25ml DMF, add 4-neighbour-tolyl urea groups phenylacetic acid (247mg, 0.87mmol) and this resin of jolting 5 minutes.Add PyBroP (406mg, 0.87mmol) and DIEA (123mg, 0.15ml, 0.87mmol) and jolting resin 24 hours.
Discharge this resin, with DMF (3x), methanol (3x) and dichloromethane (3x) washing, dry under vacuum then.Resin provides negative bromobenzene basket test.Make resin be dissolved in the dichloromethane solution of 90%TFA and jolting 4 hours.Discharge this resin, collect eluent.Resin is dissolved in the fresh dichloromethane and jolting 30 minutes.Discharge this resin, collect eluent, and merge with first.Solvent removed in vacuo makes residue recrystallization from ethyl acetate-hexane, obtains 73mg 191.Embodiment 184
Figure A0080951004441
1g Tentagel PHB resin (load 0.29mmol/g) is dissolved among the 25ml DMF, and adding Fmoc-(4-carboxymethyl)-piperidines (318mg, 0.87mmol).This resin of jolting 5 minutes, add then DIC (220mg, 0.27ml, 1.74mmol) and DMAP (106mg, 0.87mmol) and this resin of jolting 24 hours.
Discharge this resin, with DMF (3x), methanol (3x) and dichloromethane (3x) washing, dry under vacuum then.Resin provides negative bromobenzene basket test.Make resin be dissolved in the DMF solution of 20% piperidines and jolting 4 hours.Discharge this resin, with DMF (3x), methanol (3x) and dichloromethane (3x) washing, dry under vacuum then.Resin provides positive bromobenzene basket test.This resin is dissolved among the 25ml DMF, and adding Fmoc-L-proline (294mg, 0.87mmol).This resin of jolting 5 minutes, add then PyBroP (406mg, 0.87mmol) and DIEA (123mg, 0.15ml, 0.87mmol) and jolting resin 24 hours.
Discharge this resin, with DMF (3x), methanol (3x) and dichloromethane (3x) washing, dry under vacuum then.Resin provides negative bromobenzene basket test.Make resin be dissolved in the DMF solution of 20% piperidines and jolting 4 hours.Discharge this resin, with DMF (3x), methanol (3x) and dichloromethane (3x) washing, dry under vacuum then.Resin provides positive bromobenzene basket test.This resin is dissolved among the 25ml DMF, add Fmoc-3-amino-2-OXo-1-pyrrolidine acetas (331mg, 0.87mmol) and this resin of jolting 5 minutes.Add PyBroP (406mg, 0.87mmol) and DIEA (123mg, 0.15ml, 0.87mmol) and jolting resin 24 hours.
Discharge this resin, with DMF (3x), methanol (3x) and dichloromethane (3x) washing, dry under vacuum then.Resin provides negative bromobenzene basket test.Make resin be dissolved in the DMF solution of 20% piperidines and jolting 4 hours.Discharge this resin, with DMF (3x), methanol (3x) and dichloromethane (3x) washing, dry under vacuum then.Resin provides positive bromobenzene basket test.This resin is dissolved in the dichloromethane, and adding Carbimide. neighbour-toluene ester (193mg, 1.45mmol, 0.18ml).This resin of jolting 24 hours.
Discharge this resin, with DMF (3x), methanol (3x) and dichloromethane (3x) washing, dry under vacuum then.Resin provides negative bromobenzene basket test.Make resin be dissolved in the dichloromethane solution of 90%TFA and jolting 4 hours.Discharge this resin, collect eluent.Resin is dissolved in the fresh dichloromethane and jolting 30 minutes.Discharge this resin, collect eluent, and merge with first.Solvent removed in vacuo makes residue recrystallization from ethyl acetate-hexane, obtains 69mg 192.Embodiment 185
Will (10.0g 67.1mmol) packs at the bottom of the garden of a 250ml in the flask at the neighbour of the isothiocyanic acid in the 150ml dichloromethane-toluene ester.This solution is cooled to-78 ℃, and bubbling fed ammonia (excessive) 10 minutes.Form precipitation immediately and be found to be required product neighbour-tolylthiourea.Filter this reactant mixture, with the solid of cold dichloromethane thorough washing collection.The vacuum drying white solid obtains the required neighbour-tolylthiourea of 10.12g (yield 92%).
(9.1g, absolute methanol 62mmol) (100ml) solution makes neighbour-tolylthiourea, and (10.12g 61mmol) methylates by adding methyl iodide then.Under room temperature, stirred this reactant 6 hours, then vacuum concentration.Ethyl acetate extraction 3 times will also be used in the residue impouring aqueous ammonium chloride solution.Dry Organic substance and the vacuum concentration that merges obtains 8.7g (yield 84%) 2-methyl-2-sulfo--neighbour-tolyl pseudo-urea.Under room temperature, make pseudo-urea (8.7g, 5.1mmol) be dissolved in methanol (100ml) and piperidines (8.7g, 102mmol) in.This mixture stirred spend the night, vacuum concentration then obtains the product ester of 9.2g, is faint yellow solid.Make this solid saponification with Lithium hydrate, obtain required 9.0g end-product carboxylic acid 193.Embodiment 186
Make 4-aminophenyl methyl acetate (4.0g 25mmol) is dissolved in the dichloromethane (100ml), in this solution, add isothiocyanic acid neighbour-toluene ester (3.7g, 25mmol).With this reaction mixture refluxed heating 4 hours, be cooled to room temperature then.In this solution impouring 1N HCl, use ethyl acetate extraction then 3 times, through dried over mgso and vacuum concentration, obtain 5.2g (yield 67%) thiourea methyl ester.Make this ester saponification with Lithium hydrate, obtain the required 4-of 5.0g (neighbour-tolylthiourea base) phenylacetic acid 194.Embodiment 187
Figure A0080951004462
Under room temperature; with 4-[1-[3-methoxyl group-4-[N '-(2-aminomethyl phenyl) urea groups] phenyl acetyl]-(2S; 3R; 4R)-3; 4-isopropylidene dioxy base-2-pyrrolidinyl carbonyl]-1-piperazinyl ethyl acetate (1.27g; 1.99mmol) solution stirring in saturated HCl (gas)-methanol (20ml) 2 hours, evaporate methanol.Residue is dissolved in the saturated sodium bicarbonate solution, and the usefulness chloroform-methanol (4: 1, v/v) extract.With salt water washing extract, through dried over mgso and be concentrated into dried.Residue is through chromatography; with chloroform-methanol (5: 1; v/v) as eluant; obtain 4-[1-[3-methoxyl group-4-[N '-(2-aminomethyl phenyl) urea groups] phenyl acetyl]-(2S; 3R, 4R)-3,4-dihydroxy-2-pyrrolidinyl carbonyl]-1-piperazinyl ethyl acetate (990mg; 83%), is yellow amorphous solid.IR(KBr)3338，2937，2830，1743，1625，1600，1532，1454cm -1； 1H-NMR(CDCl 3)δ1.25(t，J＝7.1Hz，3H)，2.20(s，3H)，2.46-2.56(m，4H)，3.15(s，2H)，3.40-3.72(m，9H)，3.62(s，3H)，4.01-4.08(m，2H)，4.16(q，J＝7.1Hz，2H)，4.22(m，1H)，4.72(d，J＝2.9Hz，1H)，6.68(d，J＝7.6Hz，1H)，6.72(s，1H)，7.06(d，J＝7.6Hz，1H)，7.15-7.18(m，3H)，7.52-7.56(m，2H)，7.90(d，J＝8.3Hz，1H)；MS(FAB)m/z?598(M ++1)。
To 4-[1-[3-methoxyl group-4-[N '-(2-aminomethyl phenyl) urea groups] phenyl acetyl]-(2S; 3R; 4R)-3; 4-dihydroxy-2-pyrrolidinyl carbonyl]-1-piperazinyl ethyl acetate (870mg; 1.46mmol) the solution of THF (15ml) in add the 0.25N sodium hydroxide (7.00ml, 1.75mmol).After stirring 3.5 hours under the room temperature, concentrate this reactant mixture.The dilute with water residue neutralizes with 1N HCl in 0 ℃.Concentrate this mixture, spent ion exchange resin (HP-20, Mitsubishi Chemical) purification obtains 195 (645mg, 78%), is colourless amorphous solid.IR (KBr) 3330,2937,1627,1535,1454cm -1 1H-NMR (DMSO-d 6) δ 2.25 (s, 3H), 2.38 (m, 1H), 2.42-2.58 (m, 2H), 2.64 (m, 1H), 3.01 (s, 2H), 3.13-3.71 (m, 8H), 3.88 (s, 3H), 3.89 (m, 1H), 4.05 (m, 1H), 4.58 (d, J=3.2Hz, 1H), 6.76 (dd, J=8.3,1.5Hz, 1H), 6.91-6.95 (m, 2H), 7.10-7.16 (m, 2H), 7.79 (d, J=8.3Hz, 1H), 8.00 (d, J=8.3Hz, 1H), 8.49 (s, 1H), 8.57 (s, 1H); MS (FAB) m/z 570 (M ++ 1); C 28H 35N 5O 82.75H 2The analytical calculation value of O: C, 54.32; H, 6.59; N, 11.31.Measured value: C, 54.07; H, 6.11; N, 11.00.Embodiment 188
To 2-amino-4-thiazolyl acetic acid (4g, 1: 1 dichloromethane 25mmol): add in the suspension of acetone (100ml) Carbimide. neighbour-toluene ester (3.5g, 26mmol).This mixture heated to refluxing 8 hours, is formed yellow mercury oxide this moment.Filter this precipitate, with 1: 1 a large amount of dichloromethane: this solid of washing with acetone.Make this solid recrystallization with hot methanol, vacuum drying obtains the required 2-of 4.8g (yield 66%) (neighbour-tolyl urea groups)-4-thiazolyl acetic acid 196.Embodiment 189
In flask at the bottom of the garden, (3.00g 16.13mmol) is dissolved in dioxane-water (1: 1), and adding solid sodium carbonate is 8-9 until pH to make 3-bromo benzylamine.Add Boc 2(3.87g 17.74mmol), stirred this reactant 12 hours to O under room temperature.In this reactant mixture impouring 1NHCl, use ethyl acetate extraction water layer 3 times.The organic layer that merges of water, salt water washing then, it is after anhydrous magnesium sulfate drying.Filter this solution, removal of solvent under reduced pressure.Product is through flash chromatography (4: 1 hexane-ethyl acetate) purification.Obtain 4.39g 197.
Figure A0080951004482
Under argon atmospher, (2.00g 6.99mmol) is dissolved among the anhydrous THF benzylamine that Boc-is protected.Make reactant be cooled to-78 ℃.With added in 10 minutes two (trimethyl silyl) lithamides (13.98ml, 13.98mmol).Stirred these reactants 1 hour in-78 ℃, add fast then iodomethane (1.98g, 13.98mmol, 0.87ml).Making this reactant slowly be warmed to room temperature and stir spends the night.In this reactant impouring 1N HCl, use ethyl acetate extraction water layer 3 times.Filter this solution, removal of solvent under reduced pressure.Product separates through flash chromatography (7: 1 hexane-ethyl acetate).Obtain 1.68g 198.
Figure A0080951004483
In a pressurization test tube, and the 3-benzyl bromide methylamine of the Boc-that packs into protection (1.68g, 5.60mmol).Then with DMF, sodium acetate (0.51g, 6.16mmol), P (neighbour-tolyl) 3 (0.51g, 6.16mmol) and Pd (OAc) 2(0.25g 1.12mmol) adds in this test tube.With argon purge test tube 10 minutes, add then acrylic acid methyl ester. (0.53g, 0.53mmol, 0.55ml).Seal this test tube, in 135 ℃ of heating 24 hours.Make reactant be cooled to 0 ℃.Slowly open test tube.In this solution impouring 1N HCl, use ethyl acetate extraction water layer 3 times.The organic layer that merges of water, salt water washing then, it is after dried over mgso.Filter this solution, removal of solvent under reduced pressure.Product separates through flash chromatography (6: 1 hexane-ethyl acetate).Obtain 1.60g 199.
Figure A0080951004491
In a pressurization test tube, and the 3-benzyl bromide methylamine of the Boc-that packs into protection (1.00g, 3.33mmol).Then with DMF, sodium acetate (0.30g, 3.36mmol), P (neighbour-tolyl) 3 (0.20,0.66mmol) and Pd (OAc) 2(0.15g 0.66mmol) adds in this test tube.With argon purge test tube 10 minutes, add then methylmethacrylate (.37g, 3.66mmol, 0.39ml).Seal this test tube, in 135 ℃ of heating 24 hours.Make reactant be cooled to 0 ℃.Slowly open test tube.In this solution impouring 1N HCl, use ethyl acetate extraction water layer 3 times.The organic layer that merges of water, salt water washing then, it is after dried over mgso.Filter this solution, removal of solvent under reduced pressure.Product separates through flash chromatography (6: 1 hexane-ethyl acetate).Obtain 1.01g 200.
Figure A0080951004492
With α, (1.60g 5.49mmol) places Paar container and be dissolved in ethyl acetate to β-undersaturated ester.Add Pd/C (0.3g), use H 2This container is forced into 50psi.Stirred this container 12 hours.With argon purge Paar container, remove catalyst by diatomite filtration.Removal of solvent under reduced pressure.Obtain 1.60g 201.
Figure A0080951004493
With Alpha-Methyl, (1.01g 3.16mmol) places Paar container and be dissolved in ethyl acetate to β-undersaturated ester.Add Pd/C, use H 2This container is forced into 50psi.Stirred this container 12 hours.With argon purge Paar container, remove catalyst by diatomite filtration.Removal of solvent under reduced pressure.Obtain 996.41mg 202.
(304mg 1.04mmol) is dissolved in the dichloromethane, adds excessive trifluoroacetic acid to make the Boc ester.Stirred this reactant then 2 hours.Remove and desolvate, residue is dissolved in the ethyl acetate, wash with saturated sodium bicarbonate solution.Water, salt water washing organic layer are then through dried over sodium sulfate.Filter this solution, removal of solvent under reduced pressure.Make residue be dissolved in dichloromethane-DMF and HOBt (154.30mg, 1.14mmol) in, add 4-[N '-(neighbour-tolyl urea)-phenylacetic acid (324.11mg, 1.14mmol) and EDCI (218.53mg, 1.14mmol).Stirred this reactant 24 hours.In this solution impouring 1N HCl, use ethyl acetate extraction water layer 3 times.The organic layer that merges of water, salt water washing then, it is after dried over mgso.Filter this solution, removal of solvent under reduced pressure.Product separates through flash chromatography (ethyl acetate).Obtain 380.32mg 203.
Figure A0080951004501
(380.32mg 0.80mmol) is dissolved in the alcohol-water (4: 1), adds sodium hydroxide to make described ester.With this reactant be heated to 50 ℃ 2 hours.TLC (ethyl acetate) shows does not have raw material to exist.Make reactant be cooled to room temperature.In this solution impouring 1N HCl, use ethyl acetate extraction water layer 3 times.The organic layer that merges of water, salt water washing then, it is after dried over mgso.Filter this solution, removal of solvent under reduced pressure.Residue is recrystallization from ethyl acetate-hexane.Obtain 319.40mg 204.Embodiment 190
Make the Boc ester (209.60,0.65mmol) be dissolved in the dichloromethane, add excessive trifluoroacetic acid.Stirred this reactant then 2 hours.Remove and desolvate, residue is dissolved in the ethyl acetate, wash with saturated sodium bicarbonate solution.Water, salt water washing organic layer are then through dried over sodium sulfate.Filter this solution, removal of solvent under reduced pressure.Make residue be dissolved in dichloromethane-DMF and HOBt (97.45mg, 0.72mmol) in, add 4-[N '-(neighbour-tolyl urea)-phenylacetic acid (204.70mg, 0.72mmol) and EDCI (138.03mg, 0.72mmol).Stirred this reactant 24 hours.In this solution impouring 1N HCl, use ethyl acetate extraction water layer 3 times.The organic layer that merges of water, salt water washing then, it is after dried over mgso.Filter this solution, removal of solvent under reduced pressure.Product separates through flash chromatography (ethyl acetate).Obtain 237.8mg 205.
Figure A0080951004511
(237.8mg 0.49mmol) is dissolved in the alcohol-water (4: 1), adds sodium hydroxide to make described ester.With this reactant be heated to 50 ℃ 2 hours.TLC (ethyl acetate) shows does not have raw material to exist.Make reactant be cooled to room temperature.In this solution impouring 1N HCl, use ethyl acetate extraction water layer 3 times.The organic layer that merges of water, salt water washing then, it is after dried over mgso.Filter this solution, removal of solvent under reduced pressure.Residue is recrystallization from ethyl acetate-hexane.Obtain 207.8mg 206.Embodiment 191
Make 1,2,3, (12.20g 91.60mmol) in the vitriolization (40ml), is cooled to-10 ℃ to the 4-tetrahydroisoquinoline.Concentrated nitric acid (9.0ml) is slowly added in this solution, keep interior Wen Yu-10 ℃ simultaneously.Add finish after, this reactant is left standstill and with slowly being warmed to room temperature in 12 hours.This reactant mixture is slowly joined in the ice, with the ammonium chloride aqueous solution that alkalizes.With chloroform extraction water layer 4 times.Wash the organic layer of merging with water, it is after dried over sodium sulfate.Filter this solution, removal of solvent under reduced pressure.The brown oil that obtains is dissolved in the ethanol, adds concentrated hydrochloric acid.Filter and collect white solid and the vacuum drying that obtains.Obtain 8.0g 207.
Figure A0080951004513
Make 6-nitro-1,2,3, (1.00g 5.61mmol) is dissolved in the ethanol 4-tetrahydroisoquinoline.Add then the monobromo-acetic acid methyl ester (0.86g, 5.61mmol, 0.53ml) and triethylamine (1.17g, 11.59mmol, 1.62ml), with this mixture heated to refluxing 5 hours.Make this solution be cooled to room temperature, this solution of vacuum concentration.This solution is added in the entry, use ethyl acetate extraction water layer 3 times.Organic layer through dried over sodium sulfate merges filters and removal of solvent under reduced pressure.Product separates through flash chromatography (3: 1 hexane-ethyl acetate).Obtain 702mg 208.
(702mg 2.81mmol) places Paar container and be dissolved in ethanol with top ester.Add Pd/C (100mg), use H 2This container is forced into 50psi.Stirred this container 24 hours.With argon purge Paar container, remove catalyst by diatomite filtration.Removal of solvent under reduced pressure.1H-NMR shows only required product.Obtain 587mg 209.
Under argon atmospher, (587.0mg 2.66mmol) is dissolved in anhydrous methylene chloride and the pyridine to make aniline.Make this reactant be cooled to 0 ℃.With adding 3-methoxyl group-4-(N '-phenyl urea groups) phenyl chloroacetic chloride (837.70mg, dichloromethane solution 2.66mmol) in 5 minutes.Making this reactant be warmed to room temperature and stir then spends the night.With among this reactant mixture impouring 1N HCl, use ethyl acetate extraction water layer 3 times again.The organic layer of using saturated sodium bicarbonate, water, salt water washing to merge then is again through dried over mgso.Filter this solution, removal of solvent under reduced pressure.Product separates through flash chromatography (ethyl acetate).Obtain 618.36mg 210.
Figure A0080951004523
With top methyl ester (618.36mg 1.20mmol) is dissolved in THF-water, add Lithium hydrate (558.07mg, 13.30mmol).Under room temperature, stirred this reactant mixture 24 hours.In this reactant impouring 1N HCl, use ethyl acetate extraction water layer 3 times.The organic layer that merges of water, salt water washing then is again through dried over mgso.Filter this solution, removal of solvent under reduced pressure.Product is through recrystallization (hexane-ethyl acetate) purification.Obtain 600mg 211.Embodiment 192
Under argon atmospher, (1.00g 5.12mmol) is dissolved among the anhydrous THF to make 3-nitro-phenylpropionic acid.This reactant is cooled to 0 ℃, with 10 minutes adding BH 3-THF (1.0M, 15.37mmol, 15.37ml).Stirred this reactant 1 hour, the slow quencher of water then in 0 ℃.Make this solution slowly be warmed to room temperature, among the impouring 1N HCl, use ethyl acetate extraction water layer 3 times then.The organic layer of using saturated sodium bicarbonate, water, salt water washing to merge then is again through dried over mgso.Filter this solution, removal of solvent under reduced pressure.Product separates through flash chromatography (1: 1 hexane-ethyl acetate).Obtain 909.0mg 212.
Figure A0080951004532
(909.0mg 5.02mmol) is dissolved in the anhydrous methylene chloride to make 3-nitro-phenyl propanol.In flask at the bottom of second garden, under argon atmospher, with (COCl) 2(700.65mg, 5.52mmol 0.48ml) add in the anhydrous methylene chloride.Then with (COCl) 2--dichloromethane solution is cooled to-60 ℃, slowly add DMSO (862.56mg, 11.4mmol, 0.78ml).Stir these reactants 5 minutes in-60 ℃, added described alcoholic solution with 5 minutes by conduit then.Stirred these reactant mixtures 1 hour in-60 ℃, (2.54g, 25.10mmol 3.50ml), make this reactant slowly be warmed to room temperature to add triethylamine then.In this reactant impouring 1N HCl, use ethyl acetate extraction water layer 3 times.The organic layer of using saturated sodium bicarbonate, water, salt water washing to merge then is again through dried over mgso.Filter this solution, removal of solvent under reduced pressure.1H-NMR shows that no raw material exists.This aldehyde 213 need not to be further purified and the former state use.
Figure A0080951004533
In flask at the bottom of the garden, under argon atmospher, (132.48mg 5.52mmol) forms slurry in anhydrous THF to make sodium hydride.By syringe slowly add the 2-phosphono propanoic acid triethyl that is dissolved among the anhydrous THF (1.31g, 5.52mmol, 1.18ml).Under room temperature, stirred this reactant mixture 30 minutes.With 10 minutes, the aldehyde above will being dissolved among the anhydrous THF under argon atmospher joined in the above-mentioned phosphonate ester solution by syringe.This reactant mixture was stirred 12 hours.In this reactant impouring 1N HCl, use ethyl acetate extraction water layer 3 times.The organic layer of using saturated sodium bicarbonate, water, salt water washing to merge then is again through dried over mgso.Filter this solution, removal of solvent under reduced pressure.Product separates through flash chromatography (1: 1 ethyl acetate-hexane).Obtain 992.0mg214.
With above α, (992.0mg 3.77mmol) places Paar container and be dissolved in ethanol to β-undersaturated ester.With this container of argon purge, add Pd/C (200.0mg).Under 50psi, replace argon with H2.Jolting Paar container is 12 hours then.With argon hydrogen is blown away from this container, removed catalyst by diatomite filtration.Removal of solvent under reduced pressure.1H-NMR shows only required product.Obtain 851.2mg 215.
Figure A0080951004542
Under argon atmospher, (850.0mg 3.61mmol) is dissolved in anhydrous methylene chloride and the pyridine to make top aniline.Make this reactant be cooled to 0 ℃.With adding 3-methoxyl group-4-(N '-phenyl urea groups) phenyl chloroacetic chloride (1.14g, dichloromethane solution 3.61mmol) in 5 minutes.Making this reactant be warmed to room temperature and stir then spends the night.With among this reactant mixture impouring 1N HCl, use ethyl acetate extraction water layer 3 times again.The organic layer of using saturated sodium bicarbonate, water, salt water washing to merge then is again through dried over mgso.Filter this solution, removal of solvent under reduced pressure.Product separates through flash chromatography (ethyl acetate).Obtain 576.0mg 216.
Figure A0080951004543
(576.0mg mmol) is dissolved in the alcohol-water, adds sodium hydroxide with top ethyl ester.With this reactant mixture be heated to 50 ℃ 2 hours.Make this reactant be cooled to room temperature, among the impouring 1N HCl, use ethyl acetate extraction water layer 3 times again.The organic layer that merges of water, salt water washing then is again through dried over mgso.Filter this solution, removal of solvent under reduced pressure.Product is through the Sep-Pak column purification.Obtain 534mg 217.Embodiment 193
Figure A0080951004551
With 1g Wang resin (tentagel S-PHB, 0.3mmol load) be suspended in 3-(Fmoc-amino) phenylpropionic acid (361.31mg, 0.90mmol), DMAP (109.95mg, 0.90mmol), HOBt (243.63mg, 0.90mmol) and DIC (227.16mg, 1.80mmol, 0.28ml) in the solution in the mixture of DMF and dichloromethane.This mixture of jolting 20 hours is also discharged.With DMF, methanol, washed with dichloromethane resin 218 and drying under reduced pressure.
Resin upward (500mg, add in 0.15mmol) piperidines-DMF (50%v/v, 4ml), this mixture of jolting 4 hours.With DMF, methanol, washed with dichloromethane resin.Adding TMOF and isobutylaldehyde (isobutrylaldehyde) in this resin (108.17mg, 1.50mmol, 0.14ml).This mixture of jolting 4 hours.Discharge resin, add the TMOF and the isobutylaldehyde of new preparation.This mixture of jolting is 12 hours then.Discharge resin and make it to be dissolved in methanol-1% acetic acid adding NaCNBH 3(150.0mg, 2.39mmol).This resin of jolting 6 hours.Discharge resin, with methanol, methanol-triethylamine, methanol, DMF, washed with dichloromethane.This resin is dissolved among the DMF, add 3-methoxyl group-4-(N '-phenyl urea groups) phenylacetic acid (141.45mg, 0.45mmol), PyBrop (209.78mg, 0.45mmol) and DIEA (58.16mg, 0.45mmol, 0.08ml).The jolting resin is 24 hours then, discharges again.With DMF, methanol, this resin of washed with dichloromethane.The dichloromethane solution of adding TFA in this resin (30%v/v, 3ml), this mixture of jolting 5 hours.Filter this mixture, vacuum concentrated filtrate.Through Sep-Pak column purification residue.Except that after desolvating, ether is joined in this residue, collect solid, obtain 15mg 219, be crystalline solid.Embodiment 194
Tentagel PHB resin (1.0g, load 0.29mmol/g) is dissolved among the 25ml DMF, and the adding 6-bromonexanoic acid (169mg, 0.87mmol).This resin of jolting 5 minutes, add then DIC (220mg, 0.27ml, 1.74mmol) and DMAP (35mg, 0.29mmol) and this resin of jolting 14 hours.Discharge this resin, with DMF (3x), methanol (3x) and dichloromethane (3x) washing, dry under vacuum then.
In this resin, be added in 2 among the 15ml DMF, 2-dimethyl-1,3-dioxolanes-4-methylamine (227mg, 1.74mmol) and lithium iodide (232mg, 1.74mmol).The jolting resin is 14 hours under room temperature.Discharge resin, with DMF (3x), methanol (3x) and dichloromethane (3x) washing, dry under vacuum then.Resin provides positive bromobenzene basket test.
This resin is dissolved among the 25ml DMF, add 4-neighbour-tolyl urea groups-3-methoxybenzene guanidine-acetic acid (247mg, 0.87mmol) and this resin of jolting 5 minutes.Add PyBroP (406mg, 0.87mmol) and DIEA (123mg, 0.15ml, 0.87mmol) and jolting resin 14 hours.Discharge this resin, with DMF (3x), methanol (3x) and dichloromethane (3x) washing, dry under vacuum then.Resin provides negative bromobenzene basket test.
Make resin be dissolved in the dichloromethane solution of 90%TFA and jolting 4 hours.Discharge this resin, collect eluent.Resin is dissolved in the fresh dichloromethane and jolting 30 minutes.Discharge this resin, collect eluent, and merge with first.Solvent removed in vacuo makes residue recrystallization from ethyl acetate-hexane, obtains 56mg 220.Embodiment 195
Figure A0080951004562
1g Tentagel PHB resin (load 0.29mmol/g) is dissolved among the 25ml DMF, and adding Fmoc-7-aminoheptylic acid (319mg, 0.87mmol).This resin of jolting 5 minutes, add then DIC (220mg, 0.27ml, 1.74mmol) and DMAP (106mg, 0.87mmol) and this resin of jolting 24 hours.Discharge this resin, with DMF (3x), methanol (3x) and dichloromethane (3x) washing, dry under vacuum then.Resin provides negative bromobenzene basket test.Make resin be dissolved in the DMF solution of 20% piperidines and jolting 4 hours.Discharge this resin, with DMF (3x), methanol (3x) and dichloromethane (3x) washing, dry under vacuum then.Resin provides positive bromobenzene basket test.
This resin is dissolved among the 25ml DMF, add 4-neighbour-tolyl urea groups-3-methoxybenzene guanidine-acetic acid (247mg, 0.87mmol) and this resin of jolting 5 minutes.Add PyBrOP (406mg, 0.87mmol) and DIEA (123mg, 0.15ml, 0.87mmol) and jolting resin 14 hours.Discharge this resin, with DMF (3x), methanol (3x) and dichloromethane (3x) washing, dry under vacuum then.Resin provides negative bromobenzene basket test.
Make resin be dissolved in the dichloromethane solution of 90%TFA and jolting 4 hours.Discharge this resin, collect eluent.Resin is dissolved in the fresh dichloromethane and jolting 30 minutes.Discharge this resin, collect eluent, and merge with first.Solvent removed in vacuo makes residue recrystallization from ethyl acetate-hexane, obtains 66mg 221.Embodiment 196
In-78 ℃, with 30 fens clockwise oxalyl chlorides (3.8g, drip in dichloromethane 30mmol) (100ml) solution DMSO (2.4g, 31mmol).With drip in 15 fens this solution of clockwise N-Boc-dried meat ammonia alcohol (5.0g, 25mmol) in.In-78 ℃, this reactant was stirred 3 hours, by adding cold 1N HCl quencher, use ethyl acetate extraction 3 times then, dry and vacuum concentration obtains crude product dried meat ammonium aldehyde, and it through chromatography (25% ethyl acetate/hexane), is obtained the required product of 3.8g.
Preparation (the inferior phosphoranyl (triphenylphosphoranylidene) of triphen) methyl butyrate (6.9g, THF 19mmol) (100ml) solution.(the 2.0M solution of 10ml 20mmol), stirred 1 hour then to add LiHMDS in-78 ℃.(3.8g 19mmol), made this mixture be warmed to room temperature with 4 hours to the dried meat ammonium aldehyde of-inferior property above the adding.By adding this reactant of 1N HCl quencher, use ethyl acetate extraction 3 times, dry and vacuum concentration obtains crude product alkene, and it through chromatography (25% ethyl acetate/hexane), is obtained the required product of 2.9g.
By (2.9g 10mmol) places ethanol (20ml) and add the 10%Pd/C of catalytic amount, then under 40psi, makes olefin hydrogenation 4 hours in the Parr hydrogenator, and the alkane that obtains can need not purification and use with this alkene.Under room temperature, remove the Boc group by the TFA/ dichloromethane that added 1: 1.Stirred this reactant 2 hours, solvent removed in vacuo.Crude product amine 1.9g can need not to be further purified and use.
Unhindered amina (1.9g, dichloromethane 10mmol) (100ml) solution above the preparation.Under room temperature, in this solution, add EDCI (2.95g, 10mmol), DMAP (1.2g, 10mmol) and 4-neighbour-tolyl urea groups-3-methoxybenzene guanidine-acetic acid (3.15g, 10mmol).Stirred this reactant mixture 4 hours, and by adding 1N HCl quencher, used ethyl acetate extraction 3 times then, dry and vacuum concentration.The crude product amide obtains the required product of 1.95g through chromatography (5% ethanol/methylene).
(1.95g 4.2mmol) is dissolved in 1: 1 the THF-water, adds Lithium hydrate under room temperature to make this ester.Stirred reaction mixture is 3 hours then.In this solution impouring 1N HCl, use ethyl acetate extraction water layer 3 times.The organic layer that water, salt water washing merge, it is after anhydrous magnesium sulfate drying.Filter this solution, removal of solvent under reduced pressure.Grind this solid with cold ether then, obtain the required carboxylic acid of 1.65g 222.Embodiment 197
Figure A0080951004581
To 8-aminocaprylic acid methyl ester (2.0g, 1: 1 dioxane 12mmol): add in the solution of water (100ml) the Boc anhydride (2.8g, 13mmol) and potassium carbonate (10g).Under room temperature, stirred this solution 14 hours.With among this reactant impouring 1N HCl, use ethyl acetate extraction 3 times then, dry and vacuum concentration.The crude product carbamate obtains the required product of 2.7g through chromatography (50% ethyl acetate/hexane).
Place THF (75ml) by amine with the Boc protection; then add LiHMDS (the 2.0M solution of 25ml in-78 ℃; 50mmol); stirred this solution then 30 minutes; disposable adding methyl iodide (7.2g; 50mmol), make this reactant mixture be warmed to ambient temperature overnight amine-methylated with Boc protection.By adding this reactant of 1N HCl quencher, with ethyl acetate extraction 3 times and vacuum concentration.The methylated carbamate of crude product obtains the required dimethyl product of 1.9g through chromatography (50% ethyl acetate/hexane).
Under room temperature, remove the Boc group by the TFA/ dichloromethane that added 1: 1.Stirred this reactant 2 hours, solvent removed in vacuo.Crude product amine 900mg can need not to be further purified and use.
Unhindered amina (900mg, dichloromethane 4.5mmol) (100ml) solution above the preparation.Under room temperature, in this solution, add EDCI (1.33g, 4.5mmol), DMAP (567mg, 4.5mmol) and 4-neighbour-tolyl urea groups-3-methoxybenzene guanidine-acetic acid (1.45g, 4.6mmol).Stirred this reactant mixture 4 hours, and by adding 1N HCl quencher, used ethyl acetate extraction 3 times then, dry and vacuum concentration.The crude product amide obtains the required product of 1.2g through chromatography (5% ethanol/methylene).
(1.2g 2.4mmol) is dissolved in 1: 1 the THF-water, adds Lithium hydrate under room temperature to make this ester.Stirred reaction mixture is 3 hours then.In this solution impouring 1N HCl, use ethyl acetate extraction water layer 3 times.The organic layer that water, salt water washing merge, it is after anhydrous magnesium sulfate drying.Filter this solution, removal of solvent under reduced pressure.Grind this solid with cold ether then, obtain the required carboxylic acid of 1.01g 223.Embodiment 198
To 4-aminophenyl acetic acid (10g, 1: 1 dichloromethane 66mmol): add in the suspension of acetone (100ml) Carbimide. neighbour-toluene ester (8.8g, 66mmol).This mixture heated to refluxing 4 hours, is formed white precipitate this moment.Filtering-depositing is with 1: 1 a large amount of dichloromethane: this solid of washing with acetone.Make this solid recrystallization with hot methanol, vacuum drying obtains the required 4-of 14.1g (yield 75%) (neighbour-tolyl urea groups) phenylacetic acid 224.Embodiment 199
To 2-amino-4-thiazolyl acetic acid (4g, 1: 1 dichloromethane 25mmol): add in the suspension of acetone (100ml) Carbimide. neighbour-toluene ester (3.5g, 26mmol).This mixture heated to refluxing 8 hours, is formed yellow mercury oxide this moment.Filtering-depositing is with 1: 1 a large amount of dichloromethane: this solid of washing with acetone.Make this solid recrystallization with hot methanol, vacuum drying obtains the required 2-of 4.8g (yield 66%) (neighbour-tolyl urea groups)-4-thiazolyl acetic acid 225.Embodiment 200
(5.00g 25.25mmol) is dissolved among the DMF to make 3-bromo-4-hydroxy benzonitrile.Add benzyl bromide a-bromotoluene (4.75g, 27.78mmol, 3.30ml) and cesium carbonate (16.45g, 50.50mmol), with this reactant be heated to 50 ℃ 2 hours.This solution is cooled among room temperature and the impouring 1N HCl, uses ethyl acetate extraction water layer 3 times.The organic layer that water, salt water washing merge, it is after dried over mgso.Filter this solution, removal of solvent under reduced pressure.Product is through flash chromatography (hexane to 8: 1 hexane-ethyl acetate) separate.Obtain 8.90g 226.
Figure A0080951004603
Under argon atmospher, (1.50g 5.21mmol) is dissolved among the anhydrous THF, and this solution is cooled to 0 ℃, adds BH with 5 minutes by syringe to make 3-bromo-4-benzyloxy benzonitrile 3-THF (10.41ml, 10.41mm).This reactant mixture is warmed to room temperature, is heated to then and refluxed 12 hours.Make this solution be cooled to 0 ℃, slowly add methanol.When not observing more gas and emit, this solution is warmed to room temperature, add excessive 1N sodium hydroxide solution.Add Boc 2(1.25g 5.73mmol), stirred this reactant mixture 12 hours to O under room temperature.In this solution impouring 1N HCl, use ethyl acetate extraction water layer 3 times.The organic layer that merges of water, salt water washing then, it is after dried over mgso.Filter this solution, removal of solvent under reduced pressure.Product separates through flash chromatography (7: 1 ethyl acetate-hexanes).Obtain 1.80g 227.
Under argon atmospher, (1.80g 4.59mmol) is dissolved among the anhydrous THF benzylamine that Boc-is protected.Make reactant be cooled to-78 ℃.With added in 10 minutes two (trimethyl silyl) lithamides (13.77ml, 13.77mmol).Stirred these reactants 1 hour in-78 ℃, add fast then iodomethane (1.95ml, 13.77mmol, 0.86ml).Making this reactant slowly be warmed to room temperature and stir spends the night.In this reactant impouring 1N HCl, use ethyl acetate extraction water layer 3 times.Filter this solution, removal of solvent under reduced pressure.Product separates through flash chromatography (7: 1 hexane-ethyl acetate).Obtain 1.70g 228.
(1.70g 4.18mmol) places a pressurization test tube with 4-(N-methyl-Boc-amino methyl)-2-bromo benzyloxy phenol.Then with DMF, sodium acetate (0.38g, 4.60mmol), dppp (0.35g, 0.84mmol) and Pd (OAc) 2(0.19g 0.84mmol) adds in this test tube.With argon purge test tube 10 minutes, add then acrylic acid methyl ester. (0.40g, 4.60mmol, 0.41ml).Seal this test tube, in 135 ℃ of heating 24 hours.Make reactant be cooled to 0 ℃, slowly open test tube.In this solution impouring 1N HCl, use ethyl acetate extraction water layer 3 times.The organic layer that merges of water, salt water washing then, it is after dried over mgso.Filter this solution, removal of solvent under reduced pressure.Product separates through flash chromatography (6: 1 hexane-ethyl acetate).Obtain 1.12g 229.
(307.40mg 0.75mmol) is dissolved in the dichloromethane, adds excessive TFA with undersaturated ester.Under room temperature, stirred this reactant 4 hours.Removal of solvent under reduced pressure makes residue dry under fine vacuum.Remove and desolvate, residue is dissolved in the ethyl acetate, with the saturated sodium bicarbonate solution washing, water, salt water washing organic layer are then through dried over sodium sulfate.Filter this solution, removal of solvent under reduced pressure.Make residue be dissolved in dichloromethane-DMF and HOBt (110.99mg, 0.82mmol) in, add 3-methoxyl group-4-(N '-phenyl urea groups) phenylacetic acid (258.31mg, 0.82mmol) and EDCI (157.20mg, 0.82mmol).Stirred this reactant 24 hours.In this solution impouring 1N HCl, use ethyl acetate extraction water layer 3 times.The organic layer that water, salt water washing merge, it is after dried over mgso.Filter this solution, removal of solvent under reduced pressure.Product separates through flash chromatography (ethyl acetate).Obtain 296.30mg 230.
Figure A0080951004622
(296.30mg 0.49mmol) is dissolved in the ethyl acetate, adds Pd/C (75mg) under argon atmospher, replaces argon and stirs 24 hours with 1 atmospheric pressure hydrogen to make beta-unsaturated esters.Remove hydrogen, replace argon.Remove catalyst by diatomite filtration, wash Celite pad 3 times with ethyl acetate.Removal of solvent under reduced pressure.H 1-NMR shows only required product.Do not need to be further purified.Obtain 233.00mg 231.
Figure A0080951004623
Make this ester (233.00mg 0.45mmol) is dissolved in the THF-water (4: 1), add Lithium hydrate (94.41mg, 2.25mmol).Under room temperature, stirred reaction mixture 24 hours.In this solution impouring 1N HCl, use ethyl acetate extraction water layer 3 times.The organic layer that water, salt water washing merge, it is after dried over mgso.Filter this solution, removal of solvent under reduced pressure.Wash this product with ether-hexane (1: 1), dry under fine vacuum.Obtain 211.58mg 232.Embodiment 201
Figure A0080951004631
Make carboxylic acid (65.00mg 0.13mmol) is dissolved in the benzene, add right-toluenesulfonic acid (10.00mg, 0.06mmol).Add dean stark trap, this solution is heated to refluxed 24 hours.Reactant is cooled in room temperature and the impouring saturated sodium bicarbonate.Separate organic layer, use ethyl acetate extraction water layer 3 times.The organic layer that water, salt water washing merge is through dried over mgso.Filter this solution, removal of solvent under reduced pressure.Product separates through flash chromatography (hexane-ethyl acetate was to ethyl acetate in 4: 1).Obtain 29.00mg 233.Embodiment 202
Figure A0080951004632
(5.15g 25.49mmol) is dissolved in the ethanol, adds sulphuric acid (1ml), this solution is heated to refluxed 24 hours to make 5-bromo nicotinic acid.Make this solution be cooled to room temperature and concentrated.Then this solution is added in the saturated sodium bicarbonate, use ether extraction water layer 3 times.Organic layer through dried over sodium sulfate merges filters and removal of solvent under reduced pressure.Product has enough purity to be used for next step.Obtain 5.42g 234.
Make 5-bromo ethyl nicotinate (5.40g 23.47mmol) is dissolved in 95% ethanol, under room temperature, slowly add sodium borohydride (8.31g, 225.69mmol).After the adding, under room temperature, stirred this solution 24 hours.Water is slowly joined in this solution, stirred this mixture then 4 hours.Ethanol is removed in decompression, uses dichloromethane extraction water layer 3 times.Organic layer through dried over sodium sulfate merges filters and removal of solvent under reduced pressure.Product separates through flash chromatography (2: 1 ethyl acetate-hexanes).Obtain 2.12g 235.
Figure A0080951004641
(2.12g 11.28mmol) is dissolved in the ether, and HCl gas bubbling was fed this solution 10 minutes to make benzyl alcohol.Under room temperature, stirred this solution 1 hour, then solid collected by filtration.Wash this solid with ether, dry then.Hydrochlorate is added SOCl 2In, with this mixture heated to refluxing 1.5 hours.Make this solution be cooled to room temperature, add ether to be settled out product.Filter and collect this solid, with ether washing and dry under vacuum.Obtain 2.42g 236.
Figure A0080951004642
Under room temperature, (2.42g 9.96mmol) added CH with benzyl chloride with 1 hour 3NH 2In (75.9ml, 2.5M is in ethanol).Under room temperature, stirred this reactant 48 hours.Concentrated this solution also joins in the saturated sodium bicarbonate.With ethyl acetate extraction water layer 3 times.Organic layer through dried over sodium sulfate merges filters and removal of solvent under reduced pressure.Obtain 1.19g 237.
Make 3-bromo-5-(N-methyl-amino methyl)-pyridine (1.19g 5.01mmol) is dissolved among the DMF, add triethylamine (0.90g, 1.24ml, 8.89mmol).Add Boc 2O (1.55g, 7.10mmol).Under room temperature, stirred this reactant mixture 48 hours.In this solution impouring 1N HCl, use ethyl acetate extraction water layer 3 times.The organic layer that water, salt water washing merge, it is after dried over mgso.Filter this solution, removal of solvent under reduced pressure.Product separates through flash chromatography (2% methanol-dichloromethane).Obtain 1.6g 238.
Figure A0080951004651
Make the a-Sodium methacrylate. (5.00g 46.27mmol) is dissolved among the DMF, under room temperature, add benzyl bromide a-bromotoluene (8.70g, 50.89mmol).Add then potassium carbonate (7.03g, 50.89mmol), with this solution be heated to 50 ℃ 24 hours.In this solution impouring 1N HCl, use ether extraction water layer 3 times.The organic layer that water, salt water washing merge, it is after dried over mgso.Filter this solution, decompression careful removing is down desolvated.Product separates through flash chromatography (2% ether-pentane).Obtain 6.93g 239.
Figure A0080951004652
(700.00mg 2.33mmol) places a pressurization test tube with 3-bromo-5-(N-methyl-Boc-amino methyl)-pyridine.With DMF, triethylamine (260.05mg, 2.57mmol, 0.36ml), dPpp (193.85mg, 0.47mmol) and Pd (OAc) 2(105.52mg 0.47mmol) adds in this test tube.With argon purge test tube 10 minutes, add then benzyl methacrylate (452.86mg, 2.57ml).Seal this test tube, be heated to 135 ℃ 24 hours.Make reactant be cooled to 0 ℃, slowly open test tube.In this solution impouring 1N HCl, use ethyl acetate extraction water layer 3 times.The organic layer that merges of water, salt water washing then, it is after dried over mgso.Filter this solution, removal of solvent under reduced pressure.Product separates through flash chromatography (6: 1 hexane-ethyl acetate).Obtain 785.23mg 240.
(392.61mg 0.99mmol) is dissolved in the dichloromethane, adds excessive TFA with undersaturated ester.Under room temperature, stirred this reactant 4 hours.Removal of solvent under reduced pressure makes residue dry under fine vacuum.Remove and desolvate, residue is dissolved in the ethyl acetate, with the saturated sodium bicarbonate solution washing, water, salt water washing organic layer are then through dried over sodium sulfate.Filter this solution, removal of solvent under reduced pressure.Make residue be dissolved in dichloromethane-DMF and HOBt (147.53mg, 1.09mmol) in, add 3-methoxyl group-4-(N '-phenyl urea groups) phenylacetic acid (342.64mg, 1.09mmol) and EDCI (208.96mg, 1.09mmol).Stirred this reactant 24 hours.In this solution impouring 1N HCl, use ethyl acetate extraction water layer 3 times.The organic layer that water, salt water washing merge, it is after dried over mgso.Filter this solution, removal of solvent under reduced pressure.Product separates through flash chromatography (ethyl acetate).Obtain 363.79mg 241.
(363.00mg 0.61mmol) is dissolved in the methanol, adds Pd/C (100.00mg) under argon atmospher to make beta-unsaturated esters.Replace argon and stirred 24 hours with 1 atmospheric pressure hydrogen.Remove hydrogen, replace argon.Remove catalyst by diatomite filtration, wash Celite pad 3 times with ethyl acetate.Removal of solvent under reduced pressure.H 1-NMR shows only required product.Wash this solid with ether, dry under fine vacuum then.Obtain 254.79mg 242.Embodiment 203
(9.41g 71.76mmol) is dissolved in the ethanol and is cooled to 0 ℃ to make the 3-cyanobenzaldehyde.Repeatedly add on a small quantity sodium borohydride (2.71g, 71.76mmol).Stirred this solution 30 minutes in 0 ℃, be warmed to room temperature then and stirred 1 hour.In the slow impouring 1N of this reactant HCl, use ethyl acetate extraction water layer 3 times.The organic layer that water, salt water washing merge, it is after dried over mgso.Filter this solution, removal of solvent under reduced pressure.Residue is dissolved among the DMF, and the adding imidazoles (2.08g, 30.50mmol).(4.61g, 16.78mmol 4.36ml), stirred this reactant 12 hours under room temperature to add TBDPSCl then.In this solution impouring 1N HCl, use ethyl acetate extraction water layer 3 times.The organic layer that water, salt water washing merge, it is after dried over mgso.Filter this solution, removal of solvent under reduced pressure.Product is through flash chromatography (7: 1 hexane-ethyl acetate to 4: 1 hexane-ethyl acetate) separate.Obtain 16.23g 243.
(8.50g 34.36mmol) is dissolved in the ethyl acetate 3-cyano group benzyl alcohol that silicyl is protected, and adds Boc 2O (8.25g, 37.79mmol).Add Pd/C (1.0g), with 50psi pressurized with hydrogen Parr container.This container of jolting 24 hours, reuse argon purge hydrogen removes by filter catalyst by Celite pad, washs kieselguhr 3 times with ethyl acetate.Removal of solvent under reduced pressure, product separates through flash chromatography (10: 1 hexane-ethyl acetate).Obtain 11.10g 244.
Under argon atmospher, (5.00g 14.22mmol) is dissolved among the anhydrous THF benzyl alcohol that O-silicyl-N-Boc-is protected.Make reactant be cooled to-78 ℃.With added in 10 minutes two (trimethyl silyl) lithamides (42.67ml, 42.67mmol).Stirred these reactants 1 hour in-78 ℃, add fast then iodomethane (6.06g, 42.67mmol, 2.66ml).Making this reactant slowly be warmed to room temperature and stir spends the night.In this reactant impouring 1N HCl, use ethyl acetate extraction water layer 3 times.Filter this solution, removal of solvent under reduced pressure.Product separates through flash chromatography (2% ethyl acetate-hexane).Obtain 4.7g 245.
Under room temperature, (4.7g 9.60mmol) is dissolved among THF and the TBAF (14.39ml, 1.0M is in THF) to make the benzyl alcohol of O-silicyl-Boc-N-methyl protection.Stirred this solution 4 hours.TLC shows does not have raw material to exist.In this reactant impouring 1N HCl, use ethyl acetate extraction water layer 3 times.Filter this solution, removal of solvent under reduced pressure.Product is through flash chromatography (4: 1 hexane-ethyl acetate to 1: 1 hexane-ethyl acetate) separate.Obtain 2.39g 246.
Under argon atmospher, (1.00g 3.98mmol) is dissolved in the anhydrous methylene chloride benzyl alcohol that N-methyl Boc is protected.(1.46g 5.57mmol), is cooled to 0 ℃ with this solution to add triphenyl phasphine.With added in 10 minutes the carbon tetrabromide be dissolved in the anhydrous methylene chloride (1.85g, 5.57mmol).Stirred this solution 1 hour in 0 ℃, then removal of solvent under reduced pressure.Residue is dissolved in the ether, removes by filter the solid of generation, collect filtrate, removal of solvent under reduced pressure.Product separates through flash chromatography (2% ether-pentane).Obtain 1.15g 247.
Figure A0080951004682
Under-78 ℃, argon, (3.23ml 3.23mmol) adds among the anhydrous DME with LHMDS.(300mg, 2.94mmol 0.33ml) will add among the LHMDS, stir these solution 1 hour in-78 ℃ with being dissolved in methyl butyrate among the anhydrous DME in 15 minutes.(1.02g 3.23mmol) adds in this enolate solution, makes this solution slowly be warmed to-20 ℃ and stirred 4 hours again with the benzyl bromide a-bromotoluene that will be dissolved in the 3-N-methyl-N-Boc protection among the anhydrous DME in 15 minutes.In this reactant impouring lN HCl, use ethyl acetate extraction water layer 3 times.Filter this solution, removal of solvent under reduced pressure.Through flash chromatography (3% ethyl acetate-hexane) separated product.Obtain 414mg 248.
(121.60mg 0.36mmol) is dissolved in the dichloromethane, adds excessive trifluoroacetic acid to make the Boc ester.Stirred this reactant then 2 hours.Remove and desolvate, residue is dissolved in the ethyl acetate, wash with saturated sodium bicarbonate solution.Water, salt water washing organic layer are then through dried over sodium sulfate.Filter this solution, removal of solvent under reduced pressure.Make residue be dissolved in dichloromethane-DMF and HOBt (54.10mg, 0.40mmol) in, add 3-methoxyl group-4-(N '-phenyl urea groups) phenylacetic acid (125.74mg, 0.40mmol) and EDCI (77.0mg, 0.40mmol).Stirred this reactant 24 hours.In this solution impouring 1N HCl, use ethyl acetate extraction water layer 3 times.The organic layer that water, salt water washing merge, it is after dried over mgso.Filter this solution, removal of solvent under reduced pressure.Product separates through flash chromatography (ethyl acetate).Obtain 165.20mg 249.
Figure A0080951004691
Make described ester (165.20,0.31mmol) be dissolved in the alcohol-water (4: 1), add sodium hydroxide.With this reactant be heated to 50 ℃ 2 hours.TLC (ethyl acetate) shows does not have raw material to exist.Make reactant be cooled to room temperature.In this solution impouring 1N HCl, use ethyl acetate extraction water layer 3 times.The organic layer that merges of water, salt water washing then, it is after dried over mgso.Filter this solution, removal of solvent under reduced pressure.Residue is recrystallization from ethyl acetate-hexane.Obtain 120.00mg 250.Embodiment 204
Under-78 ℃, argon, (250mg, 2.90mmol 223.20ml) were added among the LHMDS (2.90ml, 1.0M is in hexane) among the THF with butyrolactone with 10 minutes.Stirred this solution 1 hour in-78 ℃.(991.24mg 2.90mmol) adds in the enolate solution, makes this solution slowly be warmed to room temperature again and stirs 12 hours with the benzyl bromide a-bromotoluene that will be dissolved in the 3-N-methyl-N-Boc protection among the anhydrous DME in 15 minutes.In this reactant impouring 1N HCl, use ethyl acetate extraction water layer 3 times.Filter this solution, removal of solvent under reduced pressure.Through flash chromatography (4: 1 hexane-ethyl acetate to 1: 1 ethyl acetate-hexane) separated product.Obtain 501.18mg 251.
Figure A0080951004693
(250.00mg 0.78mmol) is dissolved in the dichloromethane, adds excessive trifluoroacetic acid to make the Boc ester.Stirred this reactant then 2 hours.Remove and desolvate, residue is dissolved in the ethyl acetate, wash with saturated sodium bicarbonate solution.Water, salt water washing organic layer are then through dried over sodium sulfate.Filter this solution, removal of solvent under reduced pressure.Make residue be dissolved in dichloromethane-DMF and HOBt (116.40mg, 0.86mmol) in, add 3-methoxyl group-4-(N '-phenyl urea groups) phenylacetic acid (270.33mg, 0.86mmol) and EDCI (165.06mg, 0.86mmol).Stirred this reactant 24 hours.In this solution impouring 1N HCl, use ethyl acetate extraction water layer 3 times.The organic layer that water, salt water washing merge, it is after dried over mgso.Filter this solution, removal of solvent under reduced pressure.Product separates through flash chromatography (ethyl acetate).Obtain 119.00mg 252.Embodiment 2053-methoxyl group-4-[2-[3-methoxyl group-4-[N '-(2-aminomethyl phenyl) urea groups] phenyl acetyl]-the N-methyl amino ethoxy] benzoic acid
With 15 fens clockwise N-methylethanolamine (3.10g that stir, cold (0 ℃), 41.27mmol), triethylamine (11.80ml, DMF-water 84.66mmol) (3: 1, v/v, 40ml) drip 30% benzyl chloroformate (25.40g, toluene solution 49.13mmol) in the solution.The mixture that stirring obtains under room temperature 1 day.With this mixture of ethyl acetate extraction.With saturated sodium bicarbonate, salt water washing extract, through dried over sodium sulfate and evaporation.Residue is through purification by silica gel column chromatography, with normal hexane-ethyl acetate (3: 1, v/v), use chloroform then as eluant, obtain 4.67g (54%) N-methyl-N-(benzyloxycarbonyl) ethanolamine, be colorless oil. 1H-NMR(CDCl 3)d1.82(bs，1H)，3.00(s，3H)，3.46(bs，2H)，3.77(bs，2H)，5.13(s，2H)，7.29-7.36(m，5H)。
To vanillic acid ethyl ester (2.01g, 10.25mmol), N-methyl-N-(benzyloxycarbonyl) ethanolamine (2.11g, 10.08mmol), triphenyl phasphine (3.26g, 12.43mmol) the agitating solution of THF in add DIAD (2.65ml.13.46mmol), with this reactant mixture heated overnight under refluxing.Evaporate this mixture, residue is through silica gel short column chromatography, with normal hexane-ethyl acetate (5: 1,, obtain 3-methoxyl group-4-[2-methyl-2-(benzyloxycarbonyl) amino ethoxy v/v) as eluant] ethyl benzoate, be crude product.
(5.20g adds acetic acid (5ml) in ethanol 13.42mmol) (50ml) solution, makes this solution through 5%Pd/C hydrogenation 4 hours to crude product.Filter this mixture to remove catalyst, evaporated filtrate.Dilute residue with chloroform, with saturated sodium bicarbonate, salt water washing, through dried over sodium sulfate and evaporation.Residue is through silica gel column chromatography, with chloroform-methanol (10: 1, v/v) as eluant, obtain 510mg (2 steps, 20%) 3-methoxyl group-4-(2-methyl amino ethoxy) ethyl benzoate, be yellow oil. 1H-NMR(CDCl 3)d?1.39(t，3H，J＝7.3Hz)，1.82(bs，1H)，2.52(s，3H)，3.04(t，2H，J＝5.3Hz)，3.91(s，3H)，4.18(t，2H，J＝5.3Hz)，4.36(q，2H，J＝7.3Hz)，6.90(d，1H，J＝8.3Hz)，7.55(d，1H，J＝2.0Hz)，7.65(dd，1H，J＝2.0，8.3Hz)。
To 3-methoxyl group-4-(2-methyl amino ethoxy) ethyl benzoate (510mg, 2.01mmol) the agitating solution of DMF (13ml) in add 3-methoxyl group-4-[N '-(2-aminomethyl phenyl) urea groups] pentafluorophenyl esters (900mg of phenylacetic acid, 1.87mmol) and triethylamine (0.420ml, 3.01mmol), the mixture that obtains was stirred 2 days.Dilute this mixture with ethyl acetate, with 1N HCl, saturated sodium bicarbonate, salt water washing and through dried over sodium sulfate.After the evaporation; residue is through purification by silica gel column chromatography; with chloroform-methanol (50: 1; v/v) eluting; obtain 880mg (85%) 3-methoxyl group-4-[2-[3-methoxyl group-4-[N '-(2-aminomethyl phenyl) urea groups] phenyl acetyl]-the N-methyl amino ethoxy] ethyl benzoate, be colourless amorphous solid. 1H-NMR (CDCl 3) d 1.37-1.41 (m, 3H), 2.28 (s, 3H), 3.03 and 3.18 (s, 3H), 3.56 (s, 2H), 3.65 (s, 2H), 3.75-3.87 (m, 6H), 4.06-4.24 (2H, m), 4.33-4.39 (m, 2H), 6.68-8.08 (a plurality of m, 12H).
To 3-methoxyl group-4-[2-[3-methoxyl group-4-[N '-(2-aminomethyl phenyl) urea groups] phenyl acetyl]-the N-methyl amino ethoxy] ethyl benzoate (880mg; 1.601mmol) the solution of THF (15ml) in add 0.25N sodium hydroxide (15ml), with this reaction mixture refluxed heated overnight.In this mixture impouring 1N HCl (100ml), collect solid.Make crude product solid recrystallization from methanol-chloroform, obtain 253, be white powder.IR (KBr) 1700cm -1 1H-NMR (DMSO-d 6) δ 2.25 (s, 3H), 2.50 (s, 2H), 2.91 and 3.12 (s, 3H), 3.53-3.76 (m, 2H), 3.80 (s, 3H), 3.84 (s, 3H), 4.16-4.21 (m, 2H), 6.72-8.56 (a plurality of m, 12H), 12.68 (bs, 1H); MS (FAB) m/z 522 (M ++ 1); C 28H 31N 3O 71H 2The analytical calculation value of O: C, 62.33; H, 6.16; N, 6.63.Measured value: C, 62.17; H, 6.05; N, 7.57.Embodiment 2064-[[2-[3-methoxyl group-4-[N '-(2-aminomethyl phenyl) urea groups] phenyl acetyl] methylamino] ethyoxyl] isophthalic acid
Figure A0080951004721
Under room temperature, to N-methyl-N-benzyloxycarbonyl ethanolamine (1.05g, 5.02mmol), 4-hydroxyisophthalic acid dimethyl ester (1.05g, 5.00mmol), triphenyl phasphine (1.59g, 6.06mmol) the agitating solution of THF (20ml) in add DIAD (1.28ml, 6.50mmol).With the mixture heated overnight under refluxing that obtains.After being cooled to room temperature, evaporate this mixture.Residue is dissolved in the ethanol, adds 5%Pd/C (200mg).With the hydrogenation 2 hours that stirs the mixture under 1 atmospheric pressure that obtains.Filter this mixture to remove catalyst, evaporated filtrate.Residue is through purification by silica gel column chromatography, with chloroform-methanol (30: 1, v/v) as eluant, obtain 480mg (36%, two step) 4-(2-methyl amino ethoxy) dimethyl isophthalate, be grease. 1H-NMR(CDCl 3)d?1.68(s，1H)，2.53(s，3H)，3.01-3.04(m，2H)，3.89(s，3H)，3.90(s，3H)，4.21-4.23(m，2H)，7.00(d，1H，J＝8.8Hz)，8.14(dd，1H，J＝2.4，8.8Hz)，8.50(d，1H，J＝2.4Hz)；MS(FAB)m/z?268(M ++1)。
To 4-(2-methyl amino ethoxy) dimethyl isophthalate (410mg, 1.53mmol) the agitating solution of DMF (13ml) in add 3-methoxyl group-4-[N '-(2-aminomethyl phenyl) urea groups] pentafluorophenyl esters (700mg of phenylacetic acid, 1.46mmol) and triethylamine (340 μ l, 2.44mmol), the mixture stirring that obtains is spent the night.Dilute this mixture with ethyl acetate, with 1N HCl, saturated sodium bicarbonate and salt water washing.Through this solution of dried over sodium sulfate, evaporation obtains 780mg (95%) 4-[2-[3-methoxyl group-4-[N '-(2-aminomethyl phenyl) urea groups] phenyl acetyl] methyl amino ethoxy] dimethyl isophthalate, be crystalline powder. 1H-NMR (CDCl 3) d 2.29 (s, 3H), 3.24 (s, 3H), 3.59 (s, 3H), 3.67-3.68 (m, 2H), 3.84 (s, 3H), 3.91 (s, 3H), 3.81-3.86 (m, 2H), 4.25-4.28 (m, 2H), 651-8.48 (a plurality of m, 12H); MS (FAB) m/z 564 (M ++ 1).
To 4-[2-[3-methoxyl group-4-[N '-(2-aminomethyl phenyl) urea groups] phenyl acetyl] methyl amino ethoxy) (780mg adds 0.25N sodium hydroxide (30ml) to dimethyl isophthalate in the solution of THF 1.384mmol) (30ml).Then the mixture reflux that obtains is spent the night.In this mixture impouring ice-1N HCl (200ml), collect solid.Make crude product solid recrystallization from methanol-chloroform, obtain 420mg (57%) 4-[[2-[3-methoxyl group-4-[N '-(2-aminomethyl phenyl) urea groups] phenyl acetyl] methylamino] ethyoxyl] isophthalic acid 254, be white crystalline powder.Mp139-141 ℃; IR (KBr) 1700cm -1 1H-NMR (DMSO-d 6) δ 2.94 (s, 3H), 3.18 (s, 3H), 3.62-3.86 (m, 8H altogether), 4.24-4.28 (m, 2H), 6.74-8.58 (a plurality of m, 12H altogether), 12.91 (bs, 1H); MS (FAB) m/z 536 (M ++ 1); C 28H 29N 3O 82.5HCl the analytical calculation value: C, 53.66; H, 5.07; N, 6.70.Measured value: C, 53.80; H, 4.64; N, 6.70.Embodiment 2073-methoxyl group-4-[2-[3-methoxyl group-4-[N '-(2-aminomethyl phenyl) urea groups] phenyl acetyl] amino ethoxy] benzoic acid
Figure A0080951004731
Under room temperature, to the 2-ethanolamine (5.16g, 84.48mmol), triethylamine (23.50ml, dioxane-water 168.60ml) (1/1,160ml) drip (Boc) in the solution 2O (23.40ml, 101.86mmol).Stirred reaction mixture is 2 days under room temperature.Dilute the mixture that obtains with chloroform.With 0.5N HCl, saturated sodium bicarbonate and salt water washing.Through the isolating organic layer of dried over sodium sulfate, evaporation obtains 11.86g (87%) N-Boc-2-ethanolamine, is grease. 1H-NMR(CDCl 3)d?1.45(s，9H)，3.29-3.31(m，2H)，3.71-3.72(m，2H)。
To vanillic acid ethyl ester (1.46g, 7.44mmol), N-Boc ethanolamine (1.19g, 7.38mmol), triphenyl phasphine (2.53g, 9.65mmol) the agitating solution of THF (30ml) in add DIAD (1.90ml.9.65mmol), then with the mixture heated overnight under refluxing that obtains.Evaporate this mixture and obtain the crude product jelly.Crude product is dissolved among dichloromethane (20ml) and the TFA (20ml).The mixture that stirring obtains under room temperature 2.5 hours.This mixture of vacuum concentration makes residue be alkalescence with saturated sodium bicarbonate, uses chloroform extraction.With salt water washing extract, through dried over sodium sulfate and evaporation, obtain 1.61g (90%, two step) 3-methoxyl group-4-(2-amino ethoxy) ethyl benzoate, be yellow oil. 1H-NMR(CDCl 3)d?1.39(t，3H，J＝7.3Hz)，3.14-3.17(m，2H)，3.92(s，3H)，4.09-4.11(m，2H)，4.36(q，2H，J＝7.3Hz)，6.89(d，1H，J＝8.3Hz)，7.56(d，1H，J＝2.0Hz)，7.66(dd，1H，J＝2.0，8.3Hz)。
To 3-methoxyl group-4-(2-amino ethoxy) ethyl benzoate (250mg, 1.04mmol) and 3-methoxyl group-4-[N '-(2-aminomethyl phenyl) urea groups] pentafluorophenyl esters (500mg of phenylacetic acid, 1.04mmol) agitating solution in add triethylamine (210 μ l, 3.01mmol), the mixture that obtains was stirred 2 days.0.25N sodium hydroxide (20ml) and THF (20ml) are added in this mixture, down the mixture heated that obtains is spent the night in refluxing.After the cooling, evaporate this mixture, make the residue acidify by adding 1N HCl.With this mixture of dichloromethane extraction, with salt water washing extract.Through dried over sodium sulfate and evaporation; the crude product solid recrystallization from chloroform that obtains; obtain 110mg (20%, two step) 3-methoxyl group-4-[2-[3-methoxyl group-4-[N '-(2-aminomethyl phenyl) urea groups] phenyl acetyl] amino ethoxy] benzoic acid 255, be white crystalline powder.Mp 180-181 ℃; IR (KBr) 1687cm -1 1H-NMR (DMSO-d 6) d 2.24 (s, 3H), 3.37 (s, 2H), 3.38 (s, 2H), 3.41-3.50 (m, 2H), 3.81 (s, 3H), 3.83 (s, 3H), 4.06-4.08 (m, 2H), 6.76-8.55 (a plurality of m, 12H altogether); MS (FAB) m/z 508 (M ++ 10); C 27H 29N 3O 71/2H 2The analytical calculation value of O: C, 62.78; H, 5.85; N, 8.13.Measured value: C, 62.46; H, 5.69; N, 8.03.Embodiment 2083-methoxyl group-4-[2-[3-methoxyl group-4-[N '-(2-aminomethyl phenyl) urea groups] phenyl acetyl] the ethylamino ethyoxyl] benzoic acid
To 3-methoxyl group-4-(2-amino ethoxy) ethyl benzoate (1.93g, 8.07mmol) and triethylamine (2.00ml adds TFAA (1.35ml.9.56mmol) in the solution of 14.35mmol) cold (0 ℃), under room temperature the mixture that obtains is stirred and spends the night.Mixture with the ether dilution obtains washs in proper order with saturated sodium bicarbonate, 1N HCl, water and saline.Through dried over sodium sulfate extract and evaporation, obtain 1.22g (45%) 3-methoxyl group-4-(2-N-trifluoroacetamido ethyoxyl) ethyl benzoate, be grease. 1H-NMR(CDCl 3)d?1.39(t，3H，J＝7.3Hz)，3.77-3.81(m，2H)，3.92(s，3H)，4.18-4.20(m，2H)，4.37(q，2H，J＝7.3Hz)，6.92(d，1H，J＝8.7Hz)，7.59(d，1H，J＝2.0Hz)，7.67(dd，1H，J＝2.0，8.7Hz)；MS(FAB)m/z?335(M +)，290(M +-OEt)。
Under room temperature, to 3-methoxyl group-4-(2-N-trifluoroacetamido ethyoxyl) ethyl benzoate (1.20g, add in the agitating solution of DMF 3.58mmol) (15ml) potassium carbonate (0.98g, 7.09mmol) and EtI (0.43ml, 5.38mmol).In 60 ℃ the mixture that obtains was stirred 2 days.Dilute this mixture with ethyl acetate, wash successively with 1N HCl, saline, through dried over sodium sulfate.Evaporating solvent, residue be through purification by silica gel column chromatography, with normal hexane-ethyl acetate (2: 1, v/v) as eluant, obtain 990mg (76%) 3-methoxyl group-4-[2-(N-ethyl-N-TFA amino) ethoxy benzonitrile acetoacetic ester, be the yellow crystal solid. 1H-NMR(CDCl 3)d?1.28-1.31(m，3H)，1.37-1.40(m，3H)，3.64-3.69(m，2H)，3.81-3.84(m，2H)，3.92(s，3H)，4.27-4.30(m，2H)，4.34-4.39(m，2H)，6.89(d，1H，J＝8.3Hz)，7.55(d，1H，J＝2.0Hz)，7.66(dd，1H，J＝2.0，8.3Hz)；MS(FAB)m/z?364(M ++1)。
To 3-methoxyl group-4-[2-(N-ethyl-N-TFA amino) ethoxy benzonitrile acetoacetic ester (990mg, 2.73mmol) the THF-methanol-water (2: 1: 1, v/v adds potassium carbonate (560mg in agitating solution 20ml), 4.05mmol), the mixture stirring that obtains is spent the night.The mixture that dilute with water obtains is used ethyl acetate extraction.Wash extract in proper order with saturated sodium bicarbonate, saline.Through dried over sodium sulfate and evaporation, obtain 800mg (q.y.) 3-methoxyl group-4-(2-ethylamino ethyoxyl) ethyl benzoate, be grease. 1H-NMR(CDCl 3)d?1.15(t，3H，J＝7.3Hz)，1.39(t，3H，J＝7.3Hz)，1.76(bs，1H)，2.74(q，2H，J＝7.3Hz)，3.08(t，2H，J＝5.4Hz)，3.91(s，3H)，4.18(t，2H，J＝5.4Hz)，4.36(q，2H，J＝7.3Hz)，6.90(d，1H，J＝8.3Hz)，7.55(d，1H，J＝2.0Hz)，7.66(dd，1H，J＝2.0，8.3Hz)；MS(FAB)m/z?268(M ++1)。
To 3-methoxyl group-4-(2-ethylamino ethyoxyl) ethyl benzoate (290mg, 1.08mmol) and 3-methoxyl group-4-[N '-(2-aminomethyl phenyl) urea groups] pentafluorophenyl esters (502mg of phenylacetic acid, 1.05mmol) the agitating solution of DMF (7ml) in add triethylamine (250 μ l, 1.79mmol), the mixture stirring that obtains is spent the night.Dilute this mixture with ethyl acetate, with 0.5N HCl, salt water washing.Through dried over sodium sulfate.Evaporating solvent; residue is through purification by silica gel column chromatography; with chloroform-methanol (40: 1; v/v) as eluant; obtain 550mg (93%) 3-methoxyl group-4-[2-[3-methoxyl group-4-[N '-(2-aminomethyl phenyl) urea groups] phenyl acetyl] the ethylamino ethyoxyl] ethyl benzoate, be amorphous solid. 1H-NMR (CDCl 3) d 1.11-1.18 (m, 3H), 1.37-1.41 (m, 3H), 2.30 (s, 3H), 3.47-3.53 (m, 2H), 3.61-3.75 (m, 7H), 3.84 (s, 3H), 4.03-4.27 (m, 2H), 4.33-4.39 (m, 2H), 6.34-8.07 (a plurality of m, 12H altogether).
To 3-methoxyl group-4-[2-[3-methoxyl group-4-[N '-(2-aminomethyl phenyl) urea groups] phenyl acetyl] the ethylamino ethyoxyl] (550mg adds 0.25N sodium hydroxide (15ml) to ethyl benzoate in the solution of THF 0.98mmol) (15ml).Then with the mixture reflux that obtains 2 days.In this mixture impouring 1N HCl, collect solid.Make crude product solid recrystallization from ethanol-chloroform, obtain 182mg (35%) 3-methoxyl group-4-[2-[3-methoxyl group-4-[N '-(2-aminomethyl phenyl) urea groups] phenyl acetyl] the ethylamino ethyoxyl] benzoic acid 256, be white crystalline powder.Mp 115-118 ℃; IR (KBr) 1707cm -1 1H-NMR (DMSO-d 6) δ 1.02-1.12 (m, 3H), 2.25 (s, 3H), 2.50 (s, 2H), 3.35-3.89 (m, 10H), 4.11-4.16 (m, 2H), 6.71-8.56 (a plurality of m, 12H altogether), 12.56 (br s, 1H); MS (FAB) m/z 536 (M ++ 1); C 29H 33N 3O 73/4H 2The analytical calculation value of O: C, 63.43; H, 6.33; N, 7.65.Measured value: C, 63.34; H, 6.28; N, 7.28.Embodiment 2093-nitro-4-[2-[3-methoxyl group-4-[N '-(2-aminomethyl phenyl) urea groups] phenyl acetyl] amino ethoxy] benzoic acid
Under room temperature, (5.18g, (4: 1, v/v added TMSCHN in agitating solution 140ml) to benzene-methanol 28.29mmol) to 4-hydroxyl-3-nitrobenzoic acid 2(14.10ml, 28.20mmol, the hexane solution of 2M) stirs the mixture overnight that obtains.Evaporate this mixture, residue as eluant, obtains 4.18g (75%) 3-nitro-4-methyl hydroxybenzoate with chloroform through purification by silica gel column chromatography, is the yellow crystal solid. 1H-NMR(CDCl 3)d?3.95(s，3H)，7.22(d，1H，J＝8.8Hz)，8.24(dd，1H，J＝2.0，8.8Hz)，8.83(d，1H，J＝2.0Hz)，10.89(s，1H)。
To 3-nitro-4-methyl hydroxybenzoate (1.98g, 10.04mmol), N-Boc ethanolamine (1.63g, 10.11mmol) and triphenyl phasphine (3.43g, 13.08mmol) the agitating solution of THF (40ml) in add DIAD (2.57ml, 13.05mmol), then with reactant mixture heated overnight under refluxing.The mixture that evaporation obtains obtains a jelly.Residual crude product jelly is dissolved among dichloromethane (30ml) and the TFA (30ml), under room temperature, stirred the mixture 1 hour.This mixture of vacuum concentration makes residue be alkalescence with saturated sodium bicarbonate.With this mixture of chloroform extraction, use the salt water washing, through dried over sodium sulfate.Vacuum evaporating solvent obtains the oily residue, with it through purification by silica gel column chromatography, use chloroform, use then chloroform-methanol (20: 1, v/v) as eluant, obtain 930mg (27%, two step) 3-nitro-4-(2-amino ethoxy) essence of Niobe, be jelly. 1H-NMR (CDCl 3) d 3.16-3.19 (m, 1H), 3.53-3.57 (m, 1H), 3.90 and 3.94 (s, 3H), 3.95-3.98 (m, 1H), 4.21-4.24 (m, 1H), 6.89-6.91 and 7.11-7.13 (m, 1H), 8.03-8.19 and 8.21 (m, 1H), 8.52 and 8.86 (m, 1H).
To 3-methoxyl group-4-[N '-(2-aminomethyl phenyl) urea groups] pentafluorophenyl esters (1.86g of phenylacetic acid, 3.87mmol) and 3-nitro-4-(2-amino ethoxy) essence of Niobe (0.93g, 3.87mmol) the agitating solution of DMF (27ml) in add triethylamine (0.90ml, 6.46mmol), the mixture stirring that obtains is spent the night.In this mixture impouring 0.5N HCl, collect the solid that produces.Make the crude product solid be dissolved in the THF-0.25N sodium hydroxide (1: 1,20ml) in, the mixture heated that obtains is spent the night to refluxing.With this mixture of ethyl acetate extraction.Use the salt water washing.Through dried over sodium sulfate and evaporation.Make crude product solid recrystallization from chloroform-ethanol, obtain 60mg (3%, two step) 3-nitro-4-[2-[3-methoxyl group-4-[N '-(2-aminomethyl phenyl) urea groups] phenyl acetyl] amino ethoxy] benzoic acid 257, be the yellow crystal solid.Mp 112-115 ℃; 1H-NMR (DMSO-d 6) δ 2.24 (s, 3H), 3.37-3.66 (m, 7H), 3.84 (s, 3H), 4.27-4.30 (m, 1H), 6.74-8.56 (a plurality of m, 12H altogether); MS (FAB) m/z 523 (M ++ 1); C 26H 26N 4O 83/2H 2The analytical calculation value of O: C, 56.83; H, 5.32; N, 10.20.Measured value: C56.66; H, 4.90; N, 9.33.Embodiment 2103-methoxyl group-4-[2-[3-methoxyl group-4-[N '-(2-fluoro phenyl) urea groups] phenyl acetyl] the ethylamino ethyoxyl] benzoic acid
Figure A0080951004781
To 3-methoxyl group-4-[N '-(2-fluoro phenyl) urea groups] pentafluorophenyl esters (135mg of phenylacetic acid, 0.28mmol) and 3-methoxyl group-4-(2-ethylamino ethyoxyl) ethyl benzoate (78mg, 0.29mmol) agitating solution in add triethylamine (0.1ml, 0.72mmol), the mixture stirring that obtains is spent the night.Dilute this mixture with ethyl acetate, wash in proper order, through dried over sodium sulfate and evaporation with 0.5N HCl, saline.Residue is through purification by silica gel column chromatography; with chloroform-methanol (50: 1; v/v), obtain 160mg (q.y.) 3-methoxyl group-4-[2-[3-methoxyl group-4-[N '-(2-fluoro phenyl) urea groups as eluant] phenyl acetyl] the ethylamino ethyoxyl] ethyl benzoate, be grease. 1H-NMR (CDCl 3) d 1.13-1.23 (m, 3H), 1.37-1.40 (m, 3H), 2.90-3.89 (m, 12H), 4.09-4.28 (m, 2H), 4.33-4.39 (m, 2H), 6.70-8.21 (a plurality of m, 12H altogether).
To 3-methoxyl group-4-[2-[3-methoxyl group-4-[N '-(2-fluoro phenyl) urea groups] phenyl acetyl] the ethylamino ethyoxyl] ethyl benzoate (160mg; 0.28mmol) the agitating solution of THF (5m1) in add 0.25N sodium hydroxide (5ml), the mixture reflux that obtains is spent the night.In this mixture impouring 1N HCl, collect solid.Make crude product solid recrystallization from ethanol-chloroform-normal hexane, obtain 70mg (46%) 3-methoxyl group-4-[2-[3-methoxyl group-4-[N '-(2-fluoro phenyl) urea groups] phenyl acetyl] the ethylamino ethyoxyl] benzoic acid 258, be the yellow crystal powder.Mp105-110 ℃; IR (KBr) 1687cm -1 1H-NMR (DMSO-d 6) δ 1.00-1.10 (m, 3H), 2.48 (s, 2H), 3.35-3.81 (m, 10H), 4.13-4.14 (m, 2H), 6.70-9.15 (a plurality of m, 12H); MS (FAB) m/z 540 (M ++ 1); C 28H 30FN 3O 71/2H 2The analytical calculation value of O: C, 61.31; H, 5.82; N, 7.47.Measured value: C, 61.05; H, 5.82; N, 7.47.Embodiment 2114-[4-[3-methoxyl group-4-[N '-(2-aminomethyl phenyl) urea groups] phenyl] acetyl group]-1-piperazinyl phenyl formic acid
Figure A0080951004791
With the 1-piperazinecarboxylic acid tert-butyl ester (1.00g, 5.37mmol), 4-fluorinated acid ethyl ester (903mg, 5.37mmol) and potassium carbonate (1.11g, DMF 8.06mmol) (10ml) stirs the mixture in 120 ℃ of heated overnight.After the cooling, (300ml) dilutes this mixture with ethyl acetate, and then (2 * 200ml) washings are through dried over mgso and evaporation with saline.Residue is through silica gel column chromatography, with chloroform-ethyl acetate (20: 1 to 4: 1,, obtain 257mg (14%) 4-[4-(tert-butoxycarbonyl)-1-piperazinyl v/v) as eluant] ethyl benzoate, be faint yellow amorphous solid.IR (KBr) 1701,1612cm -1 1H-NMR (CDCl 3) d 1.37 (3H, t, J=7.3Hz), 1.49 (9H, s), 3.30 (4H, t, J=5.4Hz), 3.58 (4H, t, J=5.4Hz), 4.33 (2H, q, J=7.3Hz), 6.87 (2H, d, J=8.8Hz), 7.94 (2H, dt, J=8.8,2.4Hz); MS (FAB) m/z 335 (M ++ 1); C 18H 26N 2O 4The analytical calculation value: C, 64.54; H, 7.84; N, 8.38.Measured value: C, 64.39; H, 7.89; N, 8.38.
To 4-[4-(tert-butoxycarbonyl)-1-piperazinyl] (240mg adds TFA (5ml) to ethyl benzoate in the agitating solution of dichloromethane 0.718mmol) (5ml), the mixture that stirring obtains 3 hours.This mixture of vacuum concentration makes residue be alkalescence by adding saturated sodium bicarbonate, and then (2 * 100ml) extract with chloroform.Through dry extract that merges of sodium carbonate and evaporation, obtain 168mg 4-(1-piperazinyl) ethyl benzoate (100%), be yellow oil. 1H-NMR(CDCl 3)d?1.37(3H，t，J＝7.3Hz)，3.03(4H，t，J＝4.9Hz)，3.29(4H，t，J＝4.9Hz)，4.33(2H，q，J＝7.3Hz)，6.87(2H，dt，J＝8.8，2.4Hz)，7.91-7.94(2H，m)。
To 4-(1-piperazinyl) ethyl benzoate (170mg, 0.730mmol) and 3-methoxyl group-4-[N '-(2-aminomethyl phenyl) urea groups] phenylacetic acid (229mg, 0.730mmol) the agitating solution of DMF (10ml) in add EDCHCl (210mg, 1.10mmol), DMAP (catalytic amount) and HOBt (catalytic amount), mixture stirred spends the night.In this mixture impouring water (100ml), the collected at suction solid.Residue is recrystallization from chloroform-normal hexane, obtains 290mg (75%) 4-[4-[3-methoxyl group-4-[N '-(2-aminomethyl phenyl) urea groups] phenyl] acetyl group]-1-piperazinyl phenyl Ethyl formate, be the colourless crystallization powder.Mp 208-210 ℃; IR (KBr) 1711,1695cm -1 1H-NMR (CDCl 3) d 1.37 (3H, t, J=7.3Hz), 2.29 (3H, s), 3.14 (2H, t, J=4.9Hz), 3.28 (2H, t, J=4.9Hz), 3.62 (2H, t, J=4.9Hz), 3.71 (3H, s), 3.72 (2H, s), 3.79 (2H, t, J=4.9Hz), 4.33 (2H, q, J=7.3Hz), 6.38 (1H, s), 6.78-6.99 (4H, m), 7.13-7.24 (4H, m), 7.50 (1H, d, J=7.8Hz), 7.92 (2H, d, J=8.8Hz), 8.12 (1H, d, J=7.8Hz); MS (FAB) m/z 531 (M ++ 1); C 30H 34N 4O 50.5H 2The analytical calculation value of O: C, 66.77; H, 6.54; N, 10.38.Measured value: C, 66.89; H, 6.39; N, 10.45.
To 4-[4-[3-methoxyl group-4-[N '-(2-aminomethyl phenyl) urea groups] phenyl] acetyl group]-1-piperazinyl phenyl Ethyl formate (290mg; 0.547mmol) methanol-THF (2: 1; v/v; add 0.25N sodium hydroxide (5ml in agitating solution 15ml); 1.25mmol), with this mixture reflux 3 hours.In this mixture impouring ice-1N HCl (100ml), the collected at suction solid.Residue is recrystallization from chloroform-methanol, obtains 190mg (69%) 4-[4-[3-methoxyl group-4-[N '-(2-aminomethyl phenyl) urea groups] phenyl] acetyl group]-1-piperazinyl phenyl formic acid 259, be the yellow crystal powder.Mp 240-245 ℃; 1H-NMR (DMSO) d 2.24 (3H, s), 3.17-3.50 (8H, m), 3.72 (2H, s), 3.86 (3H, s), 6.77 (1H, d, J=8.3Hz), 6.90 (1H, s), 6.91-6.96 (3H, m), 7.11-7.17 (2H, m), 7.76-7.80 (3H, m), 8.03 (1H, d, J=8.3Hz), 8.47 (1H, s), 8.58 (1H, s), 12.30 (1H, s); C 28H 30N 4O 5H 2The analytical calculation value of O: C, 64.60; H, 6.20; N, 10.76.Measured value: C, 64.64; H, 5.85; N, 10.51.Embodiment 212 (R)-3-methoxyl group-4-[2-[3-methoxyl group-4-[N '-(2-aminomethyl phenyl) urea groups] the phenyl acetylamino]-the 1-propoxyl group] benzoic acid
Figure A0080951004801
To (R)-2-amino-1-propanol (3.01g, 0.04mmol) and the DMF-water of triethylamine (6.70ml, 0.05 mmol) (1: 1, v/v) add (Boc) in cold (0 ℃) solution of (40ml) 2(10.0ml 0.04mmol), stirred the mixture that obtains 2 days O under room temperature.Dilute this mixture with ethyl acetate, water, salt water washing, through dried over sodium sulfate and the evaporation, obtain 6.91g (98%) (R)-2-N-tert-butoxycarbonyl amino-1-propanol, be colorless oil. 1H-NMR(CDCl 3)δ1.15(d，3H，J＝6.8Hz)，1.45(s，9H)，3.48-3.53(m，1H)，3.62-3.66(m，1H)，3.76-3.77(m，1H)；MS(FAB)m/z?176(M ++1)，120(M +- tBu)。
To vanillic acid ethyl ester (7.74g, 0.04mmol), (R)-2-N-tert-butoxycarbonyl amino-1-propanol (6.91g, 0.04mmol) and triphenyl phasphine (13.44g, 0.05mmol) the agitating solution of THF (70ml) in add azo-2-carboxylic acid's diisopropyl ester (DIAD) (10.0ml, 0.05mmol), the mixture reflux that obtains is spent the night.After being cooled to room temperature, evaporating solvent.This mixture is dissolved among dichloromethane (50ml) and the TFA (30ml), under room temperature, stirred this solution 1 hour.Behind the vacuum concentration, in residue impouring saturated sodium bicarbonate, use chloroform extraction.With salt water washing extract, through dried over sodium sulfate and evaporation.Residue is through purification by silica gel column chromatography, with the chloroform solution of 5% methanol as eluant, obtain 7.93g (two steps 79%) (R)-3-methoxyl group-4-(2-amino-1-propoxyl group) ethyl benzoate, be yellow oil. 1H-NMR(CDCl 3)δ1.90(d，3H，J＝6.8Hz)，1.39(t，3H，J＝7.3Hz)，1.72(bs，2H)，3.42-3.47(m，1H)，3.74-3.89(m，1H)，3.91(s，3H)，3.96-4.00(m，1H)，4.35(q，2H，J＝7.3Hz)，6.88(d，1H，J＝8.3Hz)，7.55(d，1H，J＝2.0Hz)，7.65(dd，1H，J＝2.0，8.3Hz)；MS(FAB)m/z?254(M ++1)。
To 3-methoxyl group-4-[N '-(2-aminomethyl phenyl) urea groups] pentafluorophenyl esters (459mg of phenylacetic acid, 0.96mmol) and (R)-3-methoxyl group-4-(the amino propoxyl group of 2-) ethyl benzoate (242mg, 0.96mmol) the agitating solution of DMF (5ml) in add triethylamine (200 μ l, 1.43mmol), the mixture that obtains was stirred 2 hours.Dilute this mixture with ethyl acetate, with 0.5N HCl, salt water washing, through dried over sodium sulfate and evaporation.Residue is through purification by silica gel column chromatography; with chloroform-methanol (50: 1; v/v) as eluant, obtain 360mg (69%) (R)-3-methoxyl group-4-[2-[3-methoxyl group-4-[N '-(2-aminomethyl phenyl) urea groups] phenyl acetyl amino]-the 1-propoxyl group] ethyl benzoate, be the colourless crystallization solid. 1H-NMR (CDCl 3) d 1.23-1.28 (m, 3H), 1.38-1.41 (t, 3H, J=7.3Hz), 2.32 (s, 3H), 3.50-4.13 (m, 11H altogether), 4.36 (q, 2H, J=7.3Hz), 6.65-8.13 (a plurality of m, 12H altogether).
To (R)-3-methoxyl group-4-[2-[3-methoxyl group-4-[N '-(2-aminomethyl phenyl) urea groups] phenyl acetyl amino]-the 1-propoxyl group] ethyl benzoate (360mg; 0.66mmol) at THF-methanol (20ml; 9: 1; v/v) add 0.25N sodium hydroxide (10ml) in the agitating solution, the mixture reflux that obtains is spent the night.In this mixture impouring ice-1N HCl, the collecting precipitation thing.Make crude product solid recrystallization from chloroform-normal hexane, obtain 172mg (50%) 260, be white crystalline powder.Mp168-169 ℃; IR (KBr) 1687cm -1 1H-NMR (DMSO-d 6) δ 1.18 (d, 3H, J=6.8Hz), 2.24 (s, 3H), 2.50-2.51 (m, 2H), 3.80 (s, 3H), 3.84 (s, 3H), 3.87-4.06 (m, 2H), 4.07-4.14 (m, 1H), 6.76-8.57 (a plurality of m, 12H altogether), 12.66 (bs, 1H); MS (FAB) m/z 522 (M ++ 1); C 28H 31N 30 73/4H 2The analytical calculation value of O: C, 62.85; H, 6.12; N, 7.85.Measured value: C, 62.77; H, 5.95; N, 7.79.Embodiment 2134-[[2-[3-methoxyl group-4-[N '-(2-aminomethyl phenyl) urea groups] phenyl acetyl] allyl amino] ethyoxyl] benzoic acid
To 4-(2-N-trifluoroacetyl group amino ethoxy)-3-methoxybenzoic acid ethyl ester (3.5g; 10.4mmol) and potassium carbonate (2.3g; 16.4mmol) the stirring the mixture of DMF (20ml) in add allyl bromide, bromoallylene (14.2ml 16.5mmol), stir the mixture that obtains 45 minutes in 65 ℃.After the cooling, water is added this mixture, uses ethyl acetate extraction, with salt water washing extract, through dried over sodium sulfate and vacuum evaporation, make residue be dissolved in the THF-methanol-water (1: 1: 1, v/v/v) in (30ml), add potassium carbonate (2.3g, 16.4mmol).The mixture that stirring obtains under room temperature 16 hours.Dilute this mixture with ethyl acetate, use the salt water washing, through dried over sodium sulfate and evaporation.Residue is through silica gel column chromatography, with chloroform-methanol (95: 5 to 95: 5, v/v) as eluant, obtain 2.9g (100%) 4-(2-allyl amino ethyoxyl)-3-methoxybenzoic acid ethyl ester, be faint yellow oily thing. 1H-NMR(CDCl 3，400MHz)δ1.39(t，3H，J＝7.3Hz)，3.07(t，2H，J＝5.3Hz)，3.34(d，2H，J＝5.9Hz)，3.91(s，3H)，4.18(t，2H，J＝5.4Hz)，4.35(dd，2H，J＝7.3Hz，14.1Hz)，5.12(d，1H，J＝10.3Hz)，5.22(dd，1H，J＝1.5Hz，17.1Hz)，5.92(m，2H)，6.90(d，1H，J＝8.3Hz)，7.55(d，1H，J＝1.5Hz)，7.65(dd，1H，J＝2.0Hz，8.3Hz)；MS(FAB)m/z?278，280(M+H) +。
Under room temperature, to 4-(2-allyl amino ethyoxyl)-3-methoxybenzoic acid ethyl ester (578mg, 2.1mmol), 3-methoxyl group-4-[N '-(2-aminomethyl phenyl) urea groups] phenylacetic acid (650mg, 2.1mmol), HOBt (420mg, 3.11mmol) and the stirring the mixture of the DMF (4ml) of DMAP (catalytic amount) in add EDC (596mg, 3.11mmol).With the mixture that obtains restir 18 hours under room temperature.In this mixture impouring 1N HCl, use ethyl acetate extraction.With salt water washing extract; through dried over sodium sulfate and vacuum evaporation; residue is through silica gel column chromatography; with chloroform-ethyl acetate (95: 5 to 1: 1; v/v) as eluant; obtain 1g (84%) 3-methoxyl group-4-[[2-[3-methoxyl group-4-[N '-(2-aminomethyl phenyl) urea groups] phenyl acetyl] allyl amino] ethyoxyl] benzoic acid, be faint yellow gluey thing. 1H-NMR (CDCl 3, 400MHz) δ 1.39 (t, 3H, J=7.3Hz), 2.29 (s, 3H), 3.58 with 3.63 (s, 3H altogether), 3.70-3.77 (m, 2H), 3.83 and 3.87 (s, 3H), 4.05-4.13 (m, 2H), 4.25 (m, 1H), 4.36 (q, 1H, J=7.0Hz), 5.04-5.22 (m, 2H), 5.73 (m, 1H), 6.32 and 6.47 (s, 1H), 6.69-6.85 (m, 2H), 7.12 (m, 2H), 7.23 (m, 2H), 7.50-7.65 (m, 2H), 8.05 (d, 1H, J=7.8Hz); MS (FAB) m/z 576 (M+H) +
With 3-methoxyl group-4-[[2-[3-methoxyl group-4-[N '-(2-aminomethyl phenyl) urea groups] phenyl acetyl] allyl amino] ethyoxyl] benzoic acid (50mg; 0.09mmol) THF-MeOH (1: 1; v/v) (2ml) solution and 1N sodium hydroxide (0.135ml; 0.135mmol) mixture under room temperature, stirred 15 hours, stirred 3 hours in 50 ℃.In this mixture impouring ice-1N HCl.Collect solid, wash with water also air-dry.Make crude product solid recrystallization from chloroform-normal hexane, obtain 38mg (77%) 261, be white crystalline material.mp?125-130℃；IR(KBr)3319，2939，1687，1647，1601，1535，1456，1417，1269，1223，1034，760cm -1； 1H-NMR(DMSO-d 6，400MHz)δ2.29(s，3H)，3.68(s，2H)，3.75-3.85(m，8H)，4.05(br，1H)，4.19(m，3H)，5.10-5.25(m，2H)，5.65-5.90(m，1H)，6.75(m，1H)，6.85(s，1H)，6.92(m，1H)，7.02-7.20(m，3H)，7.48(d，1H，J＝10.2Hz)，7.56(m，1H)，7.79(d，1H，J＝6.8Hz)，8.01(m，1H)，8.46(s，1H)，8.56(d，1H，J＝4.4Hz)，12.7(br，1H)；MS(FAB)m/z?548(M+H) +。C 30H 33N 3O 70.5H 2The analytical calculation value of O: C, 64.74; H, 6.16; N, 7.55.Measured value: C, 64.72; H, 6.07; N, 7.55.Embodiment 2143-methoxyl group-4-[[2-[3-methoxyl group-4-[N '-(2-aminomethyl phenyl) urea groups] phenyl acetyl]-(2-morpholinyl) ethylamino] ethyoxyl] benzoic acid
To 3-methoxyl group-4-[[2-[3-methoxyl group-4-[N '-(2-aminomethyl phenyl) urea groups] phenyl acetyl] allyl amino] ethyoxyl] benzoic acid (950mg; 1.65mmol) THF: add N-methylmorpholine-N-oxide (579mg in the agitating solution of water (7ml); 4.95mmol) and Osmic acid. (aqueous solution of 0.2M) (0413ml, 0.08mmol).The mixture that obtains was stirred under room temperature 3 hours.Saturated sodium sulfite is added in this mixture, by this mixture of diatomite filtration.Use ethyl acetate extraction filtrate.With salt water washing extract, through dried over mgso and vacuum evaporation.Make residue be dissolved in methanol-THF-water (1: 1: 1, v/v) in (12ml), add sodium metaperiodate (318mg, 1.5mmol).The mixture that stirring obtains under room temperature 1 hour.With the ethyl acetate dilution, use the salt water washing, through dried over mgso.Vacuum evaporating solvent obtains 862mg (90%) 3-methoxyl group-4-[[2-[3-methoxyl group-4-[N '-(2-aminomethyl phenyl) urea groups] phenyl acetyl]-N-formoxyl methylamino] ethyoxyl] ethyl benzoate, be faint yellow gluey thing. 1H-NMR(CDCl 3，400MHz)δ1.39(t，3H，J＝7.3Hz)，2.29(s，3H)，3.31-3.95(m，11H)，4.10-4.42(m，5H)，6.51-6.82(m，3H)，7.10-7.25(m，3H)，7.50(m，2H)，7.60(m，1H)，8.10(m，1H)，9.50(m，1H)；MS(FAB)m/z?578(M+H) +。
Under room temperature; to 3-methoxyl group-4-[[2-[3-methoxyl group-4-[N '-(2-aminomethyl phenyl) urea groups] phenyl acetyl]-N-formoxyl methylamino] ethyoxyl] ethyl benzoate (265mg; 0.46mmol), morpholine (0.40ml; 4.59mmol) and acetic acid (0.263ml adds NaBH in the agitating solution of ethanol 4.6mmol) (3ml) 3CN (288mg, 4.6mmol).The mixture that obtains was stirred under room temperature 15 hours.Dilute this mixture with ethyl acetate, add saturated sodium bicarbonate in 0 ℃.The mixture that obtains in 0 ℃ of stirring 0.5 hour.With this mixture of ethyl acetate extraction.With salt water washing extract; through dried over mgso and vacuum evaporation; residue is through silica gel column chromatography; with chloroform-methanol (95: 5; v/v) as eluant; obtain 213mg (71%) 3-methoxyl group-4-[[2-[3-methoxyl group-4-[N '-(2-aminomethyl phenyl) urea groups] phenyl acetyl]-(2-morpholinyl) ethylamino] ethyoxyl] ethyl benzoate, be grease. 1H-NMR (CDCl 3, 400MHz) δ 1.28 (t, 3H, J=7.0Hz), 2.31 (s, 3H), 2.48 (brs, 4H), 2.52 (m, 2H), 3.60-3.91 (m, 16H), 4.11 and 4.28 (m, 2H altogether), 4.39 (q, 2H, J=7.0Hz), 6.70-6.85 (m, 4H), 7.15 (m, 2H), 7.50-7.63 (m, 3H), 8.08 (d, 1H, J=8.0Hz); MS (FAB) m/z 649 (M+H) +
With 3-methoxyl group-4-[[2-[3-methoxyl group-4-[N '-(2-aminomethyl phenyl) urea groups] phenyl acetyl]-(2-morpholinyl) ethylamino] ethyoxyl] (265mg, 0.46mmol) mixture in THF (4ml) and 1N sodium hydroxide (0.984ml) stirred 15 hours in 50 ℃ ethyl benzoate.By adding the pH regulator to 7.4 of 1N HCl, use chloroform with this mixture: methanol (9: 1, v/v) extract.With salt water washing extract, through dried over mgso and vacuum evaporation.Residue is crystallization from ether, obtains 160mg (78%) 262, is white crystalline material.mp?125-130℃；IR(KBr)，3346，2956，2937，1705，1622，1599，1537，1456，1417，1299，1114，1032，752cm -1； 1H-NMR(CD 3OD，400MHz)δ2.29(s，3H)，2.49-2.64(m，6H)，3.65-3.85(m，16H)，4.13(m，1H)，4.26(m，1H)，6.78-7.04(m，4H)，7.18(m，2H)，7.55-7.64(m，3H)，7.99(m，2H)；MS(FAB)m/z?621(M+H) +。C 33H 40N 4O 82.5H 2The analytical calculation value of O: C, 59.54; H, 6.81; N, 8.42.Measured value: C, 59.71; H, 6.35; N, 7.98.Embodiment 2153-methoxyl group-4-[[2-[3-methoxyl group-4-[N '-(2-aminomethyl phenyl) urea groups] phenyl acetyl]-[2-[4-methyl isophthalic acid-piperazinyl) ethylamino] ethyoxyl] benzoic acid
Under room temperature; to 3-methoxyl group-4-[[2-[3-methoxyl group-4-[N '-(2-aminomethyl phenyl) urea groups] phenyl acetyl]-N-formoxyl methylamino] ethyoxyl] ethyl benzoate (242mg; 0.42mmol), N methyl piperazine (0.465ml; 4.2mmol) and acetic acid (0.240ml, the middle NaBH of adding that stirs the mixture of ethanol 4.2mmol) (3ml) 3CN (263mg, 4.2mmol).The mixture that obtains was stirred under room temperature 15 hours.Dilute this mixture with ethyl acetate, add saturated sodium bicarbonate in 0 ℃.The mixture that obtains in 0 ℃ of stirring 0.5 hour.With this mixture of ethyl acetate extraction.With salt water washing extract; through dried over mgso and vacuum evaporation; residue is through silica gel column chromatography; with chloroform-methanol (95: 5; v/v) as eluant; obtain 195mg (70%) 3-methoxyl group-4-[[2-[3-methoxyl group-4-[N '-(2-aminomethyl phenyl) urea groups] phenyl acetyl]-[2-(4-methyl isophthalic acid-piperazinyl) ethylamino] ethyoxyl] ethyl benzoate, be grease. 1H-NMR(CDCl 3，400MHz)δ1.23(t，3H，J＝7.0Hz)，2.25(s，3H)，2.29(s，3H)，2.50(br?m，12H)，3.44-3.85(m，12H)，4.10(br，1H)，4.22(br，1H)，4.35(m，2H)，6.70-6.85(m，3H)，6.98(s，1H)，7.10(m，1H)，7.20(m，2H)，7.40(m，1H)，7.60-7.70(m，3H)，8.05(d，1H，J＝7.8Hz)；MS(FAB)m/z?662(M+H) +。
With 3-methoxyl group-4-[[2-[3-methoxyl group-4-[N '-(2-aminomethyl phenyl) urea groups] phenyl acetyl]-[2-(4-methyl isophthalic acid-piperazinyl) ethylamino] ethyoxyl] ethyl benzoate (195mg; 0.30mmol) at THF: (4: 1, v/v) mixture in (5ml) and the 1N sodium hydroxide (0.885ml) stirred 15 hours in 50 ℃ methanol.By adding the pH regulator to 7.4 of 1N HCl, use chloroform with this mixture: methanol (9: 1, v/v) extract.With salt water washing extract, through dried over mgso and vacuum evaporation.Residue is crystallization from ether, obtains 141mg (75%) 263, is white crystalline material.mp?155-160℃；IR(KBr)2937，1537，783cm -1； 1H-NMR(CD 3OD，400MHz)δ2.29(s，3H)，2.49-2.80(m，15H)，3.60-3.85(m，9H)，3.92(s，1H)，4.12(m，1H)，4.25(m，1H)，6.78-7.20(m，6H)，7.61(m，3H)，8.00(m，1H)；MS(FAB)m/z?632(M) +。C 34H 43N 5O 72.5H 2The analytical calculation value of O: C, 60.16; H, 7.13; N, 10.32.Measured value: C, 59.72; H, 6.86; N, 9.97.Embodiment 2163-methoxyl group-4-[[2-[3-methoxyl group-4-[N '-(2-aminomethyl phenyl) urea groups] phenyl acetyl]-[2-cyclopropyl amino] ethylamino] ethyoxyl] benzoic acid
Figure A0080951004871
Under room temperature; to 3-methoxyl group-4-[[2-[3-methoxyl group-4-[N '-(2-aminomethyl phenyl) urea groups] phenyl acetyl]-N-formoxyl methylamino] ethyoxyl] ethyl benzoate (267mg; 0.46mmol), cyclopropylamine (0.32ml; 4.6mmol) and acetic acid (0.264ml, the middle NaBH of adding that stirs the mixture of ethanol 4.6mmol) (3ml) 3CN (290mg, 4.6mmol).The mixture that obtains was stirred under room temperature 15 hours.Dilute this mixture with ethyl acetate, add saturated sodium bicarbonate in 0 ℃.The mixture that obtains in 0 ℃ of stirring 0.5 hour.With this mixture of ethyl acetate extraction.With salt water washing extract, through dried over mgso and vacuum evaporation.Residue is through silica gel column chromatography; with chloroform-methanol (95: 5; v/v) as eluant; obtain 1 56mg (55%) 3-methoxyl group-4-[[2-[3-methoxyl group-4-[N '-(2-aminomethyl phenyl) urea groups] phenyl acetyl]-[2-cyclopropyl amino] ethylamino] ethyoxyl] ethyl benzoate, be grease. 1H-NMR(CDCl 3，400MHz)δ0.35(m，4H)，1.22(br?s，3H)，2.10(m，1H)，2.20(s，3H)，2.42(br，2H)，2.90(br?s，2H)，3.60-3.80(m，10H)，4.10(br，1H)，4.22(br，1H)，4.33(br，2H)，6.72(m，3H)，7.05-7.30(m，4H)，7.55(m，4H)，8.06(br?s，1H)；MS(FAB)m/z?619(M+H) +。
With 3-methoxyl group-4-[[2-[3-methoxyl group-4-[N '-(2-aminomethyl phenyl) urea groups] phenyl acetyl]-N-[2-cyclopropyl amino] ethylamino] ethyoxyl] ethyl benzoate (195mg; 0.30mmol) at THF: (4: 1, v/v) mixture in (5ml) and the 1N sodium hydroxide (0.756ml) stirred 15 hours in 50 ℃ methanol.By adding the pH regulator to 7.4 of 1N HCl, use chloroform with this mixture: methanol (9: 1, v/v) extract.With salt water washing extract, through dried over mgso and vacuum evaporation.Residue is crystallization from ether, obtains 57mg (38%) 3-methoxyl group-4-[[2-[3-methoxyl group-4-[N '-(2-aminomethyl phenyl) urea groups] phenyl acetyl]-[2-cyclopropyl amino] ethylamino] ethyoxyl] benzoic acid 264, be white crystalline material.mp?135-140℃；IR(KBr)3324，2937，1535，1032，754cm -1； 1H-NMR(CD 3OD，400MHz)δ0.50-0.73(m，4H)，2.29(s，3H)，2.53(m，1H)，2.98(m，1H)，3.21(m，1H)，3.58-3.88(m，11H)，3.91(s，1H)，4.09(m，1H)，4.25(m，1H)，6.76-6.92(m，3H)，7.01(m，1H)，7.18(m，2H)，7.60(m，3H)，8.00(d，J＝8.3Hz，1H)；MS(FAB)m/z?591(M+H) +。C 32H 38N 4O 73.0H 2The analytical calculation value of O: C, 59.62; H, 6.88; N, 8.69.Measured value: C, 59.25; H, 6.29; N, 8.29.Embodiment 2174-[[2-[3-methoxyl group-4-[N '-(2-fluoro phenyl) urea groups] phenyl acetyl]-the N-methylamino] ethyoxyl] benzoic acid 3-chloro-4-[[2-[3-methoxyl group-4-[N '-(2-fluoro phenyl) urea groups] phenyl acetyl]-the N-methyl amino ethoxy] benzoic acid
To 2-(N-benzyloxycarbonyl-N-methyl) ethanolamine (3.01g, 14.4mmol), 3-chloro-4-methyl hydroxybenzoate (2.68g, 14.4mmol), triphenyl phasphine (5.65g, 21.5mmol) cold (0 ℃) solution of stirring of THF (30ml) in add azo-2-carboxylic acid's diisopropyl ester (DIAD) (4.25ml, 21.6mmol), the mixture reflux that obtains is spent the night.Evaporating solvent, residue as eluant, obtain 3.90g (72%) 3-chloro-4-[2-(N-benzyloxycarbonyl-N-methylamino) ethyoxyl with chloroform through purification by silica gel column chromatography] essence of Niobe, be faint yellow solid. 1H-NMR(CDCl 3)δ3.15(s，3H)，3.74-3.76(m，2H)，3.89(s，3H)，4.17-4.27(m，2H)，5.14(s，2H)，6.81-6.94(m，1H)，7.33-7.36(m，5H)，7.85-7.92(m，1H)，8.05(bs，1H)。
With 3-chloro-4-[2-(N-benzyloxycarbonyl-N-methylamino) ethyoxyl] (3.90g, (40ml, 1: 1, v/v) solution was through 5%Pd-C (1.95g, 50wt%) hydrogenation 2 hours under 3 atmospheric pressure for ethyl acetate-acetic acid 10.3mmol) for essence of Niobe.Filter this mixture, use the saturated sodium bicarbonate wash filtrate, with chloroform extraction alkalescence water layer, with salt water washing and evaporation, obtain 3-chloro-4-(N-methyl amino ethoxy) essence of Niobe and 4-[2-(N-methylamino) ethyoxyl] the unsegregated mixture (1.61g) of essence of Niobe purpose chemical compound, be faint yellow oily thing. 1H-NMR (CDCl 3) δ 2.52-2.52 and 2.53-2.54 (respectively be m, 3H), 2.98-3.00 and 3.03-3.05 (respectively being m, each 2H), 3.88 with 3.99 (respectively being s, each 3H), 4.11-4.14 and 4.18-4.20 (respectively are m, each 2H), 6.91-6.96 and 7.90-8.05 (a plurality of m, 7H altogether).
Under room temperature, with 3-methoxyl group-4-[N '-(2-fluoro phenyl) urea groups] phenylacetic acid (392mg), 3-chloro-4-[2-(N-methylamino) ethyoxyl] essence of Niobe and 4-[2-(N-methylamino) ethyoxyl] and essence of Niobe mixture (305mg), EDC (hydrochlorate) (354mg), HOBt (250mg) and the mixture of DMAP (250mg) in DMF (8ml) stirred 6 hours.Dilute this mixture with ethyl acetate, with 0.5HCl, salt water washing, through dried over sodium sulfate and evaporation.Make residue through purification by silica gel column chromatography; with the chloroformic solution of 1% methanol as eluant; obtain 3-chloro-4-[[2-[3-methoxyl group-4-[N '-(2-fluoro phenyl) urea groups] phenyl acetyl]-the N-methyl amino ethoxy] essence of Niobe and 4-[[2-[3-methoxyl group-4-[N '-(2-fluoro phenyl) urea groups] phenyl acetyl]-the N-methyl amino ethoxy] mixture (505mg) of essence of Niobe, be brown amorphous solid.
To 3-chloro-4-[[2-[3-methoxyl group-4-[N '-(2-fluoro phenyl) urea groups] phenyl acetyl]-the N-methyl amino ethoxy] essence of Niobe and 4-[[2-[3-methoxyl group-4-[N '-(2-fluoro phenyl) urea groups] phenyl acetyl]-the N-methyl amino ethoxy] THF of mixture (550mg) of essence of Niobe: methanol (20ml; 1: 1; v/v) add 0.5N sodium hydroxide (10ml) in the agitating solution in, the mixture that obtains was heated 6 hours under refluxing.In this mixture impouring ice-1N HCl, collect solid.The crude product solid is through preparation property TLC purification, obtains 265 (56mg is white amorphous solid) and 266 (88mg is brown amorphous solid) with the chloroformic solution of 10% methanol as eluant.Embodiment 2184-[[2-[3-methoxyl group-4-[N '-(2-aminomethyl phenyl) urea groups] phenyl acetyl] methylamino] ethyoxyl] benzoic acid
Figure A0080951004901
3-chloro-4-[[2-[3-methoxyl group-4-[N '-(2-aminomethyl phenyl) urea groups] phenyl acetyl] methylamino] ethyoxyl] benzoic acid
Figure A0080951004902
Under room temperature, to 4-[2-(N-methyl-2-amino) ethyoxyl]-3-chlorobenzene methyl formate (292mg, 1.2mmol), 3-methoxyl group-4-[N '-(2-aminomethyl phenyl) urea groups] phenylacetic acid (377mg, 1.2mmol), EDC (345mg, 1.8mmol), HOBt (243mg, 1.8mmol) and DMAP (29mg, 0.24mmol) mixture in DMF (2.7ml) stirred 6 hours.In this mixture impouring ice-1N HCl, use ethyl acetate extraction.With salt water washing extract, through dried over mgso and vacuum evaporation.Residue is through silica gel column chromatography; with chloroform-ethyl acetate (95: 5 to 0: 100; v/v) as eluant; obtain 3-chloro-4-[[2-[3-methoxyl group-4-[N '-(2-aminomethyl phenyl) urea groups] phenyl acetyl] methylamino] ethyoxyl] essence of Niobe and 4-[[2-[3-methoxyl group-4-[N '-(2-aminomethyl phenyl) urea groups] phenyl acetyl] methylamino] ethyoxyl] the unsegregated mixture (489mg) of essence of Niobe, be faint yellow oily thing.
With 3-chloro-4-[[2-[3-methoxyl group-4-[N '-(2-aminomethyl phenyl) urea groups] phenyl acetyl] methylamino] ethyoxyl] essence of Niobe and 4-[[2-[3-methoxyl group-4-[N '-(2-aminomethyl phenyl) urea groups] phenyl acetyl] methylamino] ethyoxyl] mixture (480mg of essence of Niobe; be mixture) at THF: methanol (4ml; 1: 1; v/v) in, stirred 15 hours in 50 ℃.In this mixture impouring ice-1N HCl.Collect solid, wash with water also air-dry.The crude product solid is used chloroform through preparation property TLC purification: and methanol (93: 7, v/v) as eluant, obtain 267 (180mg in two steps 31%, is crystalline material) and 268 (280mg in two steps 44%, is crystalline material).267:mp 145-150 ℃; 1H-NMR (DMSO-d 6, 400MHz) δ 2.31 (s, 3H), 3.05 and 3.19 (s, 3H), 3.35 and 3.38 (s, 3H), 3.72-3.85 (m, 7H), 4.09 and 4.23 (m, altogether 2H), 6.79-7.20 (m, 7H), 7.60 (m, 1H), 7.86-8.09 (m, 3H); MS (FAB) m/z 493 (M+H) +C 27H 29N 3O 61.75H 2The analytical calculation value of O: C, 62.00; H, 6.26; N, 8.03.Measured value: C, 62.16; H, 5.88; N, 7.82.268:mp 145-150 ℃; 1H-NMR (DMSO-d 6, 400MHz) δ 2.29 (s, 3H), 3.06 and 3.26 (s, 3H), 3.31 and 3.35 (s, 3H), 3.85-3.94 (m, 4H), 4.18 and 4.32 (m, altogether 2H), 6.75-6.85 (m, 2H), 6.99-7.20 (m, 4H), 7.59 (m, 1H), 7.90-8.02 (m, 3H); MS (FAB) m/z 526 (M+H) +C 27H 28ClN 3O 62.0H 2The analytical calculation value of O: C, 52.70; H, 5.74; N, 7.48.Measured value: C, 57.99; H, 5.53; N, 7.07.Embodiment 2194-[3-[3-methoxyl group-4-[N '-(2-aminomethyl phenyl) urea groups] phenyl acetyl]-the N-methylamino]-the 1-propyl group] benzoic acid
(1.22g, (4.0ml 4.0mmol), stirs the mixture that obtains 1 hour under uniform temp to add NaHMDS (1.0M is in THF) in THF 4.05mmol) (10ml) solution to the 4-(phosphonomethyl) benzoic acid triethyl of cold (78 ℃) that stir.Under this temperature, (700mg, THF 3.38mmol) (5ml) solution slowly adds in this solution, stirs down, makes this mixture be warmed to room temperature with 2 hours with 2-(N-benzyloxycarbonyl-N-methylamino) acetaldehyde.By adding this solution of saturated ammonium chloride (100ml) quencher, use ethyl acetate extraction.With saline (200ml) washing extract, through dried over mgso and evaporation.Residue is through silica gel column chromatography, with chloroform-ethyl acetate (20: 1, v/v) as eluant, obtain (E)-4-[3-(N-benzyloxycarbonyl-N-methylamino)-1-propenylbenzene Ethyl formate of 810mg (68%), be yellow oil. 1H-NMR (CDCl 3) δ 1.40 (t, J=7.3Hz, 3H), 2.95 (m, 2H), 4.09 (m, 2H), 4.35-4.40 (m, 2H), 5.17 (s, 2H), 6.26-6.64 (a plurality of m, 2H), 7.36 (m, 7H), 7.99 (d, J=8.3Hz, 2H).
With (E)-4-[3-(N-benzyloxycarbonyl-N-methylamino)-1-propenylbenzene Ethyl formate (810mg, ethanol-acetic acid 2.29mmol) (10: 1, v/v, agitating solution 22ml) was through 5%Pd-C (1g) hydrogenation 3 days.Filter this mixture, evaporated filtrate.Make residue be alkalescence with saturated sodium bicarbonate, use chloroform extraction.Through sodium carbonate dry extract and evaporation, obtain 438mg (86%) 4-(3-methylamino-1-propyl group) ethyl benzoate, be yellow oil. 1H-NMR(CDCl 3)δ1.39(t，J＝7.3Hz，3H)，1.82(m，2H)，2.43(s，3H)，2.61(t，J＝7.3Hz，2H)，2.72(t，J＝7.3Hz，2H)，3.33(br?s，1H)，4.36(q，J＝7.3Hz，2H)，7.25(d，J＝8.3Hz，2H)，7.96(d，J＝8.3Hz，2H)。
To 3-methoxyl group-4-[N '-(2-aminomethyl phenyl) urea groups] phenylacetic acid (456mg, 1.45mmol) and 4-(3-methylamino-1-propyl group) ethyl benzoate (220mg, 1.45mmol) agitating solution in be added in EDCHCl (417mg among the DMF (10ml), 2.16mmol), HOBt (catalytic amount) and DMAP (catalytic amount), the mixture that obtains stirred spends the night.(300ml) dilutes this mixture with ethyl acetate, uses the salt water washing, through dried over mgso and evaporation.Residue with chloroform-ethanol (10: 1) eluting, obtains 4-[3-[3-methoxyl group-4-[N '-(2-aminomethyl phenyl) urea groups of 503mg (71%) through silica gel column chromatography] phenyl acetyl]-the N-methylamino]-the 1-propyl group] ethyl benzoate, be yellow oil.MS(FAB)m/z?518(M+H) +。
To 4-[3-[3-methoxyl group-4-[N '-(2-aminomethyl phenyl) urea groups] phenyl acetyl]-the N-methylamino]-the 1-propyl group] ethyl benzoate (500mg; 0.966mmol) the agitating solution of THF (8ml) in add 0.25N sodium hydroxide (8ml), this mixture reflux is spent the night.Among the solution impouring ice-1N HCl (100ml) with gained, the collected at suction solid.Solid is dissolved in the chloroform (100ml), through dried over mgso.Except that after desolvating; residue is through silica gel column chromatography; with chloroform-methanol (10: 1 to 5: 1; v/v) eluting; obtain 4-[3-[3-methoxyl group-4-[N '-(2-aminomethyl phenyl) urea groups of 131mg (28%)] phenyl acetyl]-the N-methylamino]-the 1-propyl group] benzoic acid 269, be faint yellow amorphous solid. 1H-NMR (DMSO-d 6) δ 1.68-1.80 (m, 2H), 2.24 (s, 3H), 2.57 (m, 2H), 2.81 and 2.97 (they respectively are s, are total to 3H), 3.33 (m, 2H), 3.57-3.61 (m, 2H), 3.84 (s, 3H), 6.71 (dd, J=29.8,8.3Hz, 1H), 6.87 (d, J=11.2Hz, 1H), 6.93 (t, J=7.3Hz, 1H), 7.15 (m, 2H), 7.28 (m, 2H), 7.79 (d, J=8.3Hz, 1H), and 7.84-7.87 (m, 2H), 8.02 (d, J=8.3Hz, 1H), 8.49 (d, J=7.3Hz, 1H), 8.58 (d, J=4.9Hz, 1H); MS (FAB) m/z 490 (M ++ 1); C 28H 31N 3O 51/2H 2The analytical calculation value of O: C, 67.45; H, 6.47; N, 8.43.Measured value: C, 67.27; H, 6.51; N, 8.02.Embodiment 2204-[[2-[4-[N '-(2-aminomethyl phenyl) urea groups] phenyl acetyl]-the N-methylamino] ethyoxyl] benzoic acid
Figure A0080951004931
3-chloro-4-[[2-[4-[N '-(2-aminomethyl phenyl) urea groups] phenyl acetyl]-the N-methylamino] ethyoxyl] benzoic acid
Figure A0080951004932
Under room temperature, with 4-[N '-(2-aminomethyl phenyl) urea groups] pentafluorophenyl esters of phenylacetic acid (562mg, 1.29mmol), 3-chloro-4-[2-(N-methylamino) ethyoxyl] essence of Niobe and 4-[2-(N-methylamino) ethyoxyl] mixture (304mg) of essence of Niobe and the solution stirring of triethylamine (260ml) in DMF (8ml) 4 hours.Dilute this mixture with ethyl acetate, with 0.5N HCl, salt water washing, through dried over sodium sulfate and evaporation.Make residue through purification by silica gel column chromatography; with chloroform-methanol (50: 1; v/v) as eluant; obtain 3-chloro-4-[[2-[4-[N '-(2-aminomethyl phenyl) urea groups] phenyl acetyl]-the N-methylamino] ethyoxyl] essence of Niobe and 4-[[2-[4-[N '-(2-aminomethyl phenyl) urea groups] phenyl acetyl]-the N-methylamino] ethyoxyl] mixture (670mg) of essence of Niobe, be grease.
THF to this mixture (670mg): methanol (20ml, 1: 1, add 0.5N sodium hydroxide (10ml) in stirred suspension v/v), the mixture that obtains was heated 6 hours under refluxing.In this solution impouring ice-1N HCl, collect solid.The crude product solid is through preparation of lamina chromatography (TLC) purification, and obtaining 73mg with the chloroformic solution of 10% methanol as eluant is that 270 and 110mg of amorphous solid is 271 of white amorphous solid.270?MS(FAB)m/z?462(M ++1)。271?MS(FAB)m/z?496(M ++1)。Embodiment 221 (S)-4-[2-[3-methoxyl group-4-[N '-(2-aminomethyl phenyl) urea groups] phenyl acetyl amino]-the 1-propoxyl group] benzoic acid
Figure A0080951004941
To (S)-2-amino-1-propanol (2.08g, 27.7mmol) and triethylamine (4.63ml, (40ml 1: 1, adds in cold (0 ℃) solution v/v) (Boc) DMF-water 33.2mmol) 2O (6.36ml.27.7mmol) stirred the solution that obtains 2 days under room temperature.Water is added in this mixture, use ethyl acetate extraction.With salt water washing extract, through dried over sodium sulfate.Evaporating solvent obtain 4.24g (87%) (S)-2-(N-tert-butoxycarbonyl amino)-1-propanol, be colorless oil. 1H-NMR(CDCl 3)δ1.14(d，3H，J＝6.8Hz)，1.45(s，9H)，3.51-3.52(m，1H)，3.63-3.66(m，1H)，3.77(m，1H)，4.62(m，1H)。
To (S)-2-(N-tert-butoxycarbonyl amino)-1-propanol (1.02g, 5.82mmol), 4-methyl hydroxybenzoate (0.89g, 5.85mmol) and triphenyl phasphine (1.98g, 7.55mmol) cold (0 ℃) solution of THF (20ml) in add azo-2-carboxylic acid's diisopropyl ester (DIAD) (1.49ml.7.57mmol), the mixture reflux that obtains is spent the night.Evaporate this solution and residue is dissolved among dichloromethane (20ml) and the TFA (10ml).Under room temperature, stirred this mixture 1.5 hours.This solution of vacuum concentration is handled residue with saturated sodium bicarbonate.With this mixture of chloroform extraction, use the salt water washing, through dried over sodium sulfate and evaporation.Residue is through purification by silica gel column chromatography, with chloroform-methanol (50: 1, v/v) as eluant, obtain 480mg (two steps 39%) (S)-4-(2-amino-1-propoxyl group) essence of Niobe, be faint yellow oily thing. 1H-NMR(CDCl 3)δ1.19(d，3H，J＝6.4Hz)，3.35-3.39(m，1H)，3.72-3.76(m，1H)，3.89(s，3H)，3.90-3.94(m，1H)，6.92(d，2H，J＝8.8Hz)，7.99(d，2H，J＝8.8Hz)。
With 3-methoxyl group-4-[N '-(2-aminomethyl phenyl) urea groups] pentafluorophenyl esters (505mg of phenylacetic acid, 1.05mmol), (S)-4-(2-amino-1-propoxyl group) essence of Niobe (220mg, 1.05mmol) and triethylamine (0.220ml, 1.58mmol) mixture in DMF (8ml) stirred 3 hours under room temperature.Dilute this mixture with ethyl acetate, with 0.5N HCl, salt water washing, through dried over sodium sulfate.Remove desolvate after, make residue recrystallization from methanol-chloroform-normal hexane, obtain 290mg (55%) (S)-4-[2-[3-methoxyl group-4-[N '-(2-aminomethyl phenyl) urea groups] phenyl acetyl amino]-the 1-propoxyl group] essence of Niobe, be white crystalline powder. 1H-NMR (DMSO-d 6) δ 1.18 (d, 3H, J=6.8Hz), 2.24 (s, 3H), 3.36 (s, 2H), 3.80 (s, 3H), 3.82 (s, 3H), 3.93-4.03 (m, 2H), 4.09-4.14 (m, 1H), 6.75-8.57 (a plurality of m, 13H altogether).
To (S)-4-[2-[3-methoxyl group-4-[N '-(2-aminomethyl phenyl) urea groups] phenyl acetyl amino]-the 1-propoxyl group] essence of Niobe (290mg; 0.57mmol) at THF-methanol (20ml; 1: 1; v/v) add 0.5N sodium hydroxide (20ml) in the agitating solution, with this solution reflux 2 hours.In this mixture impouring ice-1N HCl, and the usefulness chloroform-methanol (10: 1, v/v) extract.With salt water washing extract, through dried over sodium sulfate and evaporation.Residue obtains 158mg (56%) 272 through recrystallization from methanol-chloroform-normal hexane, is white crystalline powder.Mp 198-201 ℃; 1H-NMR (DMSO-d 6) δ 1.18 (d, 3H, J=6.3Hz), 2.24 (s, 3H), 3.36 (s, 2H), 3.82 (s, 3H), 3.87-4.10 (m, 2H), 4.10-4.16 (m, 1H), and 6.75-8.78 (m, 1H), 6.92-7.02 (m, 4H), 7.11-7.18 (m, 2H), and 7.78-7.80 (m, 1H), 7.86-7.89 (m, 2H), 7.98-8.00 (m, 1H), 8.12-8.14 (m, 1H), 8.46 (s, 1H), 8.55 (s, 1H), 12.62 (bs, 1H); MS (FAB) m/z 492 (M ++ 1); C 27H 29N 3O 61/2H 2The analytical calculation value of O: C, 64.79; H, 6.04; N, 8.21.Measured value: C, 64.36; H, 5.85; N, 8.21.Embodiment 222 (S)-4-[2-[4-[N '-(2-aminomethyl phenyl) urea groups] phenyl acetyl amino]-the 1-propoxyl group] benzoic acid
With 4-[N '-(2-aminomethyl phenyl) urea groups] pentafluorophenyl esters (560mg of phenylacetic acid, 1.24mmol), (S)-4-(2-amino-1-propoxyl group) essence of Niobe (260mg, 1.24mmol), (0.260ml, 1.87mmol) mixture in DMF (8ml) stirred 3 hours under room temperature triethylamine.Dilute this mixture with ethyl acetate, with 0.5N HCl, this solution of salt water washing, through dried over sodium sulfate.Remove desolvate after, make residue recrystallization purifying from methanol-chloroform-normal hexane, obtain 210mg (36%) (S)-4-[2-[3-methoxyl group-4-[N '-(2-aminomethyl phenyl) urea groups] phenyl acetyl amino]-the 1-propoxyl group] benzoic acid, be white crystalline powder. 1H-NMR(DMSO-d 6)δ1.17(d，3H，J＝6.8Hz)，2.24(s，3H)，3.32(s，2H)，3.81(s，3H)，3.92-4.03(m，2H)，4.08-4.15(m，1H)，6.92-6.95(m，1H)，7.04-7.06(m，2H)，7.12-7.18(m，4H)，7.35-7.39(m，2H)，7.83-7.85(m，1H)，7.89-7.92(m，3H)，8.12-8.14(m，1H)，8.97(s，1H)。
To (S)-4-[2-[4-[N '-(2-aminomethyl phenyl) urea groups] phenyl acetyl amino]-the 1-propoxyl group] essence of Niobe (200mg; 0.42mmol) at THF-methanol (10ml; 1: 1, add 0.5N sodium hydroxide (10ml) in agitating solution v/v), with this mixture reflux 2 hours.In this mixture impouring ice-1N HCl, collect solid.The crude product solid obtains 68mg (34%) 273 through recrystallization from methanol-chloroform-normal hexane, is white crystalline powder.Mp 262-265 ℃; 1H-NMR (DMSO-d 6) δ 1.17 (d, 3H, J=6.8Hz), 2.24 (s, 3H), 3.32 (s, 2H), 3.91-4.02 (m, 2H), 4.09-4.15 (m, 1H), and 6.92-6.96 (m, 1H), 7.01-7.03 (m, 2H), 7.12-7.20 (m, 4H), 7.36-7.40 (m, 2H), 7.83-7.95 (m, 4H), 8.12-8.14 (m, 1H), 8.99 (s, 1H), 12.63 (bs, 1H); MS (FAB) m/z 462 (M ++ 1); C 26H 27N 3O 51/4H 2The analytical calculation value of O: C, 67.01; H, 5.95; N, 9.02.Measured value: C, 67.13; H, 5.90; N, 9.02.Embodiment 223 (S)-3-chloro-4-[2-[4-[N '-(2-aminomethyl phenyl) urea groups] phenyl acetyl amino]-the 1-propoxyl group] benzoic acid
Figure A0080951004961
To (S)-2-(N-tert-butoxycarbonyl amino)-1-propanol (1.05g, 5.99mmol), 3-chloro-4-methyl hydroxybenzoate (1.12g, 6.00mmol) and triphenyl phasphine (2.36g, 9.00mmol) cold (0 ℃) solution of THF (20ml) in add azo-2-carboxylic acid's diisopropyl ester (DIAD) (1.77ml, 8.99mmol), with the mixture reflux that obtains 2 days.Solvent evaporated is dissolved among dichloromethane (20ml) and the TFA (10ml) residue.The mixture that stirring obtains under room temperature 1.5 hours.This solution of vacuum concentration is dissolved in the chloroform residue.With this mixture of water extraction, make water layer be alkalescence by adding saturated sodium bicarbonate.Should the alkalescence water layer with chloroform extraction.With salt water washing extract, through dried over sodium sulfate and evaporation, obtain 660mg (two step, 32%) (S)-3-chloro-4-(2-amino-1-propoxyl group) essence of Niobe, be colorless oil. 1H-NMR(CDCl 3)δ1.21(d，3H，J＝6.4Hz)，3.41-3.48(m，1H)，3.77-3.81(m，1H)，3.89(s，3H)，3.98-4.01(m，1H)，6.91-6.94(m，1H)，7.90-7.93(m，1H)，8.05-8.06(m，1H)。
With 4-[N '-(2-aminomethyl phenyl) urea groups] pentafluorophenyl esters (508mg of phenylacetic acid, 1.13mmol), (S)-3-chloro-4-(2-amino-1-propoxyl group) essence of Niobe (275mg, 1.13mmol) and triethylamine (0.240ml, 1.72mmol) mixture in DMF (10ml) stirs under room temperature and spends the night.Dilute this mixture with ethyl acetate, with 0.5N HCl, saturated sodium bicarbonate, this solution of salt water washing, through dried over sodium sulfate and evaporation.Make residue recrystallization from methanol-chloroform-normal hexane, obtain 240mg (42%) (S)-3-chloro-4-[2-[4-[N '-(2-aminomethyl phenyl) urea groups] phenyl acetyl amino]-the 1-propoxyl group] essence of Niobe, be white crystalline powder. 1H-NMR(DMSO-d 6)δ1.21(d，3H，J＝6.4Hz)，2.25(s，3H)，3.33(s，2H)，3.82(s，3H)，4.04-4.14(m，3H)，6.90-6.94(m，1H)，7.11-7.16(m，4H)，7.29-7.38(m，3H)，7.83-7.94(m，3H)，8.13-8.17(m，2H)，9.34(s，1H)；MS(FAB)m/z?510(M +)。
To (S)-3-chloro-4-[2-[4-[N '-(2-aminomethyl phenyl) urea groups] phenyl acetyl amino]-the 1-propoxyl group] essence of Niobe (240mg; 0.47mmol) at THF-methanol (10ml; 1: 1; v/v) add 0.5N sodium hydroxide (10ml) in the agitating solution, the mixture reflux that obtains is spent the night.In this mixture impouring ice-1N HCl, collect solid.The crude product solid obtains 98mg (42%) 274 through recrystallization from methanol-chloroform-normal hexane, is white crystalline powder.Mp 228-231 ℃; 1H-NMR (DMSO-d 6) δ 1.20 (d, 3H, J=6.3Hz), 2.24 (s, 3H), 3.34 (s, 2H), 4.02-4.18 (m, 3H), 6.92-6.95 (m, 1H), 7.12-7.42 (a plurality of m, 7H altogether), 7.82-8.18 (a plurality of m, 5H altogether), 9.12 (s, 1H); MS (FAB) m/z 496 (M +), 497 (M ++ 1); C 26H 26ClN 3O 51/2H 2The analytical calculation value of O: C, 61.84; H, 5.39; Cl, 7.02; N, 8.32.Measured value: C, 61.76; H, 5.25; Cl, 7.09; N, 8.25.Embodiment 224 (S)-3-chloro-4-[2-[3-methoxyl group-4-[N '-(2-aminomethyl phenyl) urea groups] phenyl acetyl amino]-the 1-propoxyl group] benzoic acid
With 3-methoxyl group-4-[N '-(2-aminomethyl phenyl) urea groups] pentafluorophenyl esters (513mg of phenylacetic acid, 1.07mmol), (S)-3-chloro-4-(2-amino-1-propoxyl group) essence of Niobe (260mg, 1.07mmol) and triethylamine (220 μ l, 1.58mmol) mixture in DMF (10ml) stirs under room temperature and spends the night.Dilute this mixture with ethyl acetate, wash this solution, through dried over sodium sulfate and evaporation with saturated sodium bicarbonate.Make residue recrystallization from methanol-chloroform-ethyl acetate-normal hexane; obtain 400mg (69%) (S)-3-chloro-4-[2-[3-methoxyl group-4-[N '-(2-aminomethyl phenyl) urea groups] phenyl acetyl amino]-the 1-propoxyl group] essence of Niobe, be the light brown crystalline powder. 1H-NMR(DMSO-d 6)δ1.21(d，3H，J＝6.4Hz)，2.24(s，3H)，3.37(s，2H)，3.82(s，3H)，3.83(s，3H)，4.04-4.12(m，3H)，6.75-6.77(m，1H)，6.91-6.95(m，2H)，7.11-7.17(m，2H)，7.29-7.31(m，1H)，7.78-7.99(m，4H)，8.12-8.13(m，1H)，8.46(s，1H)，8.55(s，1H)。
To (S)-3-chloro-4-[2-[3-methoxyl group-4-[N '-(2-aminomethyl phenyl) urea groups] phenyl acetyl amino]-the 1-propoxyl group] essence of Niobe (400mg; 0.74mmol) at THF-methanol (20ml; 1: 1; v/v) add 0.5N sodium hydroxide (20ml) in the agitating solution, the mixture reflux that obtains is spent the night.In this mixture impouring ice-1N HCl, collect solid.The crude product solid obtains 200mg (51%) 275 through recrystallization from methanol-chloroform-ether, is the light brown crystalline powder.Mp 198-201 ℃; 1H-NMR (DMSO-d 6) δ 1.21 (d, 3H, J=6.8Hz), 2.24 (s, 3H), 3.37 (s, 2H), 3.84 (s, 3H), 4.00-4.15 (m, 3H), 6.76-7.28 (a plurality of m, 6H altogether), 7.77-8.14 (a plurality of m, 5H altogether), 8.46 (s, 1H), 8.56 (s, 1H); MS (FAB) m/z 526 (M +), 528 (M ++ 2); C 27H 28ClN 3O 61/4H 2The analytical calculation value of O: C, 61.13; H, 5.42; Cl, 6.68; N, 7.92.Measured value: C, 60.97; H, 5.48; Cl, 6.86; N, 7.89.Embodiment 2253-dimethylamino-4-[[2-[3-methoxyl group-4-[N '-(2-aminomethyl phenyl) urea groups] phenyl acetyl] methylamino] ethyoxyl] benzoic acid
To 2-(N-Boc-N-methylamino) ethanol (3g, 17mmol), 4-hydroxyl-3-nitrobenzoic acid methyl ester (3.38g, 17mmol) and triphenyl phasphine (5.4g, add azo-2-carboxylic acid's diisopropyl ester (DIAD) in cold (0 ℃) solution of the stirring of THF 21mmol) (20ml) (4ml.21mmol), with the mixture reflux that obtains 15 hours.Solvent evaporated.Residue is through purification by silica gel column chromatography, with chloroform-methanol (100: 0 to 4: 1,, obtain 4-[2-(N-methyl-2-amino) ethyoxyl of 2.5g (39%) v/v) as eluant]-3-nitrobenzoic acid methyl ester, be faint yellow oily thing. 1H-NMR(CDCl 3，400MHz)δ2.82(s，3H)，3.50(t，2H，J＝4.5Hz)，3.95(s，3H)，4.54(t，2H，J＝4.5Hz)，7.26(d，1H，J＝8.8Hz)，8.25(d，1H，J＝8.8Hz)，8.56(s，1H)；MS(FAB)m/z?255(M ++1)。
With 3-methoxyl group-4-[N '-(2-aminomethyl phenyl) urea groups] phenylacetic acid (992mg, 3.11mmol), 4-(N-methyl-2-amino ethoxy)-3-nitrobenzoic acid methyl ester (800mg, 3.11mmol) and 4-DMAP (77mg, 0.63mmol), HOBt (640mg, 4.7mmol) and EDC (904mg, 4.7mmol) mixture in DMF (20ml) stirs under room temperature and spends the night.Dilute this mixture with ethyl acetate, with 0.5N HCl, saturated sodium bicarbonate, this solution of salt water washing, through dried over sodium sulfate and evaporation.Residue is through the silica gel column chromatography purification; with chloroform-ethyl acetate (95: 5 to 0: 100; v/v) as eluant; obtain 587mg (34%) 3-nitro-4-[[2-[3-methoxyl group-4-[N '-(2-aminomethyl phenyl) urea groups] phenyl acetyl] methylamino] ethyoxyl] essence of Niobe, be faint yellow oily thing. 1H-NMR (CDCl 3, 400MHz) δ 2.31 (s, 3H), 3.05 (s, 1H), 3.23 (s, 2H), 3.71 (s, 2H), 3.83 (s, 3H), 3.85 (m, 3H), 3.94 (s, 3H), 4.19 and 4.39 (m, 2H altogether), 6.80 (m, 2H), 7.05 (m, 1H), 7.22 (m, 3H), 7.62 (d, 1H, J=8.2Hz), 8.02 (d, 1H, J=8.2Hz), 8.21 (dd, 1H, J=2.1Hz, 8.8Hz), 8.55 (d, 1H, J=2.1Hz); MS (FAB) m/z 551 (M ++ 1).
With 3-nitro-4-[[2-[3-methoxyl group-4-[N '-(2-aminomethyl phenyl) urea groups] phenyl acetyl] methylamino] ethyoxyl] essence of Niobe (587mg; 1.1mmol) and 5%Pd-C (600mg) THF-methanol-acetic acid (1: 1: 1; v/v, the 150ml) mixture in hydrogenation 18 hours under 45psi.Insoluble catalyst is removed in suction, and vacuum evaporation filtrate obtains 555mg (100%) 3-amino-4-[[2-[3-methoxyl group-4-[N '-(2-aminomethyl phenyl) urea groups] phenyl acetyl] methylamino] ethyoxyl] essence of Niobe, be faint yellow gluey thing. 1H-NMR (CDCl 3, 400MHz) δ 2.29 (s, 3H), 3.08 (m, 1H), 3.19 (s, 2H), 3.65 (s, 1H), 3.73-3.80 (m, 3H), 3.84 (s, 3H), 3.87 (s, 3H), 4.19 and 4.40 (m, 2H altogether), 6.70-6.82 (m, 2H), and 7.02-7.29 (m, 6H), 7.60 (d, 1H, J=7.8Hz), 7.92-7.99 (m, 3H); MS (FAB) m/z 521 (M ++ 1).
Under room temperature; to 3-amino-4-[[2-[3-methoxyl group-4-[N '-(2-aminomethyl phenyl) urea groups] phenyl acetyl] methylamino] ethyoxyl] essence of Niobe (555mg; 1.1mmol), (0.58ml adds NaBH in the agitating solution of MeCN 10mmol) (10ml) for formaldehyde (10ml) and acetic acid 3(0.67g 10mmol), stirred the mixture that obtains 15 hours CN under room temperature.Saturated sodium bicarbonate is added in this mixture, use chloroform extraction.With salt water washing extract, through dried over mgso and vacuum evaporation.Residue is through silica gel column chromatography; with toluene-acetone (7: 3 to 1: 1; v/v), obtain 123mg (21%) 3-dimethylamino-4-[[2-[3-methoxyl group-4-[N '-(2-aminomethyl phenyl) urea groups as eluant] phenyl acetyl] methylamino] ethyoxyl] essence of Niobe, be grease. 1H-NMR (CDCl 3, 400MHz) δ 2.30 (s, 3H), 2.70 (s, 3H), 2.75 (s, 3H), 3.05 (s, 1H), 3.18 (s, 2H), 3.61 (s, 3H), 3.70 (s, 1H), 3.80 (m, 3H), 3.86 (s, 3H), 4.07 and 4.22 (m, 2H altogether), 6.28 (m, 1H), 6.70-6.80 (m, 3H), 7.03 (m, 1H), 7.15-7.25 (m, 4H), 7.46-7.65 (m, 2H), 8.02 (m, 1H); MS (FAB) m/z 548 (M ++ 1).
With 3-dimethylamino-4-[[2-[3-methoxyl group-4-[N '-(2-aminomethyl phenyl) urea groups] phenyl acetyl] methylamino] ethyoxyl] essence of Niobe (123mg; 0.22mmol) in THF (15ml) and 1N sodium hydroxide (0.885ml, the heating 15 hours that stirs the mixture under refluxing in 0.885mmol).By adding the pH regulator to 5.0 of 1N HCl, use chloroform with this mixture: methanol (9: 1, v/v) extract.With salt water washing extract, through dried over mgso and vacuum evaporation, residue is crystallization from ether-normal hexane, obtains 118mg (100%) 276, is white crystalline material.Mp 125-130 ℃; IR (KBr) 3346,2940,1620,1597,1535,1456,1417,1227,1039,754cm -1 1H-NMR (CD 3OD, 400MHz) δ 2.29 (s, 3H), 2.70 (s, 3H), 2.79 (s, 2H), 3.05 (s, 1H), 3.22 (s, 2H), 3.75 (s, 3H), 3.85 (m, 4H), 4.15 and 4.28 (m, 2H), 6.78-7.05 (m, 4H), 7.18 (m, 2H), 7.55-7.70 (m, 3H), 7.98 (m, 1H); MS (FAB) m/z 535 (M ++ 1); C 29H 34N 4O 62.0H 2The analytical calculation value of O: C, 61.04; H, 6.71; N, 9.82.Measured value: C, 61.15; H, 6.43; N, 8.94.Embodiment 2263-dimethylamino-4-[[2-[3-methoxyl group-4-[N '-(2-fluoro phenyl) urea groups] phenyl acetyl] methylamino] ethyoxyl] benzoic acid
Figure A0080951005011
With 3-methoxyl group-4-[N '-(2-fluoro phenyl) urea groups] phenylacetic acid (1g, 3.11mmol), 4-[2-(N-methyl-2-amino) ethyoxyl]-3-nitrobenzoic acid methyl ester (800mg, 3.11mmol) and 4-DMAP (77mg, 0.63mmol), HOBt (640mg, 4.7mmol) and EDC (904mg, 4.7mmol) mixture in DMF (20ml) stirs under room temperature and spends the night.Dilute this mixture with ethyl acetate, with 0.5N HCl, saturated sodium bicarbonate, this solution of salt water washing, through dried over sodium sulfate and evaporation.Residue is through the silica gel column chromatography purification; with chloroform-ethyl acetate (95: 5 to 0: 100; v/v) as eluant; obtain 420mg (19%) 3-nitro-4-[[2-[3-fluoro-4-[N '-(2-fluoro phenyl) urea groups] phenyl acetyl] methylamino] ethyoxyl] essence of Niobe, be faint yellow oily thing. 1H-NMR (CDCl 3, 400MHz) δ 3.04 (s, 1H), 3.24 (s, 2H), 3.72 (s, 1H), 3.85 (s, 3H), 3.90 (m, 3H), 3.93 (s, 3H), 4.16 and 4.39 (2m, 3H), 6.80 (m, 2H), 6.99 (m, 1H), 7.05 (m, 2H), 7.22 (d, 1H, J=8.8Hz), 7.51 (s, 2H), 8.00 (m, 1H), 8.08 (m, 1H), 8.21 (d, 1H, J=8.6Hz), 8.51 (s, 1H); MS (FAB) m/z555 (M ++ 1).
With 3-nitro-4-[[2-[3-methoxyl group-4-[N '-(2-fluoro phenyl) urea groups] phenyl acetyl] methylamino] ethyoxyl] essence of Niobe (420mg; 0.76mmo1) and 5%Pd-C (1g) THF-methanol-acetic acid (1: 1: 1; v/v, the 150ml) mixture in hydrogenation 18 hours under 45psi.Insoluble catalyst is removed in suction, and vacuum evaporation filtrate obtains 397mg (100%) 3-amino-4-[[2-[3-methoxyl group-4-[N '-(2-fluoro phenyl) urea groups] phenyl acetyl] methylamino] ethyoxyl] essence of Niobe, be faint yellow gluey thing. 1H-NMR (CDCl 3, 400MHz) δ 3.05 and 3.13 (s, 3H altogether), 3.66 (s, 3H), 3.70 (s, 2H), 3.65-3.90 (m, 4H), 3.86 (s, 3H), 4.10 and 4.23 (m, 2H), 6.70-6.83 (m, 3H), 6.98-7.15 (m, 6H), 7.25-7.43 (m, 2H), 7.99 (m, 1H), 8.13 (m, 1H); MS (FAB) m/z 525 (M ++ 1).
Under room temperature; to 3-amino-4-[[2-[3-methoxyl group-4-[N '-(2-fluoro phenyl) urea groups] phenyl acetyl] methylamino] ethyoxyl] essence of Niobe (397mg; 0.76mmol), (0.43ml adds NaBH in the agitating solution of MeCN 7.6mmol) (10ml) for formaldehyde (10ml) and acetic acid 3(0.48g 7.6mmol), stirred the mixture that obtains 15 hours CN under room temperature.Saturated sodium bicarbonate is added in this mixture, use chloroform extraction.With salt water washing extract; through dried over mgso and vacuum evaporation; residue is through silica gel column chromatography; with toluene-acetone (7: 3 to 1: 1; v/v) as eluant; obtain 123mg (21%) 3-dimethylamino-4-[[2-[3-methoxyl group-4-[N '-(2-aminomethyl phenyl) urea groups] phenyl acetyl] methylamino] ethyoxyl] essence of Niobe, be grease. 1H-NMR (CDCl 3, 400MHz) δ 2.74 (s, 3H), 2.77 (s, 3H), 3.08 (s, 1H), 3.22 (s, 2H), 3.52 (s, 3H), 3.61 (s, 1H), 3.83 (m, 3H), 3.88 (s, 3H), 4.12 and 4.23 (m, be total to 2H), 6.68 (s, 1H), 6.78 (m, 2H), 6.98 (m, 2H), 7.10 (m, 1H), 7.55-7.68 (m, 4H), 7.99 (m, 1H), 8.16 (t, 1H, J=8.3Hz); MS (FAB) m/z 553 (M ++ 1).
With 3-dimethylamino-4-[[2-[3-methoxyl group-4-[N '-(2-fluoro phenyl) urea groups] phenyl acetyl] methylamino] ethyoxyl] essence of Niobe (61mg; 0.11mmol) in THF (15ml) and 1N sodium hydroxide (0.22ml, the heating 15 hours that stirs the mixture under refluxing in 0.22mmol).By adding the pH regulator to 5.0 of 1N HCl, use chloroform with this mixture: methanol (9: 1, v/v) extract.With salt water washing extract, and usefulness methanol-acetone (93: 7, v/v) as eluant, obtain 37mg (63%) 277, be white crystalline material.Mp 120-125 ℃; 1H-NMR (CD 3OD, 400MHz) δ 2.60 (s, 4H), 2.78 (s, 2H), 3.06 (s, 1H), 3.22 (s, 2H), 3.75 (s, 3H), 3.85-3.92 (m, 4H), 4.17 and 4.29 (m, 2H altogether), 6.80-7.12 (m, 6H), 7.61-7.70 (m, 2H), 8.00 (m, 1H), 8.08 (m, 1H); MS (FAB) 539 (M ++ 1); C 28H 31FN 4O 62.75H 2The analytical calculation value of O: C, 57.18; H, 6.26; N, 9.53.Measured value: C, 57.20; H, 5.62; N, 9.06.Embodiment 2273-dimethylamino-4-[[2-[4-[N '-(2-aminomethyl phenyl) urea groups] phenyl acetyl] methylamino] ethyoxyl] benzoic acid
Figure A0080951005031
Will [4-[N '-(2-aminomethyl phenyl) urea groups] phenyl] acetic acid pentafluorophenyl esters (1.42g, 3.15mmol), 4-[2-(N-methyl-2-amino) ethyoxyl]-3-nitrobenzoic acid methyl ester (800mg, 3.15mmol) and triethylamine (0.66ml, 4.73mmol) mixture in DMF (8ml) stirred 15 hours in 50 ℃.In this mixture impouring ice-1N HCl, use chloroform extraction.With salt water washing extract, through dried over sodium sulfate and evaporation.Residue is through the silica gel column chromatography purification; with chloroform-ethyl acetate (95: 5 to 0: 100; v/v), obtain 1.04g (63%) 3-nitro-4-[[2-[4-[N '-(2-aminomethyl phenyl) urea groups as eluant] phenyl acetyl] methylamino] ethyoxyl] essence of Niobe, be faint yellow oily thing. 1H-NMR (CDCl 3, 400MHz) δ 2.30 (s, 3H), 2.90,3.02 and 3.05 (s, altogether 1H), 3.22 (s, 2H), 3.71 (s, 1H), 3.85 (3H), 3.93 (s, 3H), 4.19 and 4.39 (m, 2H), 7.02 (m, 1H), 7.19 (m, 4H), 7.35 (d, 1H, J=8.3Hz), 7.40 (d, 1H, J=8.0Hz), 7.55 (s, 2H), 7.70 (m, 1H), 8.22 (d, 1H, J=6.7Hz), 8.51 (s, 1H); MS (FAB) m/z 521 (M ++ 1).
With 3-nitro-4-[[2-[4-[N '-(2-aminomethyl phenyl) urea groups] phenyl acetyl] methylamino] ethyoxyl] essence of Niobe (1.04g; 2mmol) and 5%Pd-C (1.2g) THF-methanol-acetic acid (1: 1: 1; v/v, the 150ml) mixture in hydrogenation 18 hours under 45psi.Insoluble catalyst is removed in suction, and vacuum evaporation filtrate obtains 3-amino-4-[[2-[4-[N '-(2-aminomethyl phenyl) urea groups] phenyl acetyl] methylamino] ethyoxyl] essence of Niobe, be faint yellow gluey thing.
Under room temperature, to 3-amino-4-[[2-[4-[N '-(2-aminomethyl phenyl) urea groups] phenyl acetyl] methylamino] ethyoxyl] (1.14ml adds NaBH in the agitating solution of MeCN 20mmol) (5ml) for benzoate, formaldehyde (5ml) and acetic acid 3(1.26g 20mmol), stirred the mixture that obtains 15 hours CN under room temperature.Saturated sodium bicarbonate is added in this mixture, use chloroform extraction.With salt water washing extract, through dried over mgso and vacuum evaporation.Residue is through silica gel column chromatography; with toluene-acetone (7: 3 to 1: 1, v/v) as eluant, obtain 85mg (two the step; 8%) 3-dimethylamino-4-[[2-[4-N '-(2-aminomethyl phenyl) urea groups] phenyl acetyl] methylamino] ethyoxyl] essence of Niobe, be grease. 1H-NMR(CDCl 3，400MHz)δ2.12(s，3H)，2.73(s，3H)，2.75(s，3H)，3.05(s，1H)，3.20(s，2H)，3.60(s，1H)，3.80(m，3H)，3.88(s，3H)，4.17(m，2H)，6.95-7.28(m，8H)，7.55-7.75(m，3H)；MS(FAB)m/z?518(M ++1)。
With 3-dimethylamino-4-[[2-[4-[N '-(2-aminomethyl phenyl) urea groups] phenyl acetyl] methylamino] ethyoxyl] essence of Niobe (ap315201) (85mg; 0.16mmol) in THF (15ml) and 1N sodium hydroxide (0.32ml, the heating 15 hours that stirs the mixture under refluxing in 0.32mmol).By adding the pH regulator to 5.0 of 1N HCl, use chloroform with this mixture: methanol (9: 1, v/v) extract.With salt water washing extract, through dried over mgso and vacuum evaporation, residue is crystallization from ether, obtains 53mg (65%) 278, is white crystalline material.Mp 110-115 ℃; 1H-NMR (CD 3OD, 400MHz) δ 2.29 (s, 3H), 2.75 (s, 3H), 2.76 (s, 3H), 3.02 (s, 1H), 3.20 (s, 2H), 3.72 (s, 1H), 3.85 (m, 3H), 4.18 and 4.28 (m, 2H altogether), 6.95-7.03 (m, 2H), 7.18 (m, 4H), 7.34 (d, 1H, J=8.3Hz), 7.38 (d, 1H, J=8.8Hz), 7.62 (d, 1H, J=8.3Hz), 7.66 (s, 1H), 7.80 (m, 1H); MS (FAB) m/z 505 (M ++ 1).Embodiment 2283-isopropyl amino-4-[[2-[3-methoxyl group-4-[N '-(2-fluoro phenyl) urea groups] phenyl acetyl] methylamino] ethyoxyl] benzoic acid
Figure A0080951005041
To 3-amino-4-[[2-[3-methoxyl group-4-[N '-(2-fluoro phenyl) urea groups] phenyl acetyl] methylamino] ethyoxyl] essence of Niobe (100mg; 0.19mmol) acetone/acetic acid/DMF (13ml; 6: 6: 1, add NaBH in cold (0 ℃) solution of stirring v/v/v) 3CN (300mg) stirs the mixture that obtains 60 hours under room temperature.In this mixture impouring saturated sodium bicarbonate, the collected at suction solid.Precipitation is dissolved in the chloroform (20ml), with this solution of salt water washing, through dried over mgso and reduction vaporization.Residue is through the silica gel plate chromatography; with toluene-acetone (2: 1; v/v), obtain 108mg (100%) 3-isopropyl amino-4-[[2-[3-methoxyl group-4-[N '-(2-fluoro phenyl) urea groups as eluant] phenyl acetyl] methylamino] ethyoxyl] essence of Niobe, be colorless oil. 1H-NMR (CDCl 3, 400MHz) δ 1.20 (m, 1H), 2.88 (s, 6H), 3.02 with 3.12 (s, 3H altogether), 3.53 (s, 2H), 3.60-3.80 (m, 7H), 3.85 (s, 3H), 4.10 and 4.20 (m, 2H), 6.65-6.75 (m, 2H), 6.90-7.08 (m, 2H), 7.22-7.35 (m, 2H), 8.02 (s, 2H), 8.10 (m, 2H), 8.21 (br, 1H), 8.33 (br, 1H); MS (FAB) m/z 566 (M ++ 1).
With 3-isopropyl amino-4-[[2-[3-methoxyl group-4-[N '-(2-fluoro phenyl) urea groups] phenyl acetyl] methylamino] ethyoxyl] essence of Niobe (116mg, 0.2mmol), the heating 8 hours that stirs the mixture under refluxing of 0.25N sodium hydroxide (6ml) and THF (6ml).In this mixture impouring water (200ml), through 1N HCl acidify, collected at suction solid.Make solid recrystallization from chloroform-normal hexane-Di Iso Propyl Ether, obtain 56mg (50%) 279, be the colourless crystallization powder.Mp195-200 ℃; 1H-NMR (CD 3OD, 400MHz) δ 1.15-1.20 (m, 6H), 3.01 (s, 1H), 3.12 (s, 2H), 3.48-3.60 (m, 1H), 3.68 (s, 3H), 3.75 (s, 2H), 3.82 (s, 1H), 3.86 (m, 3H), 4.15-4.23 (m, 2H), 6.80 (m, 3H), 4.15-4.23 (m, 2H), 6.80 (m, 3H), 6.98 (m, 1H), 7.10 (m, 2H), 7.20 and 7.25 (s, 1H altogether), 7.32 (m, 1H), 7.98 (m, 1H), 8.05 (m, 1H); MS (FAB) m/z 552 (M+H) +C 29H 33FN 4O 61.0H 2The analytical calculation value of O: C, 61.04; H, 6.18; N, 9.82.Measured value: C, 61.36; H, 6.25; N, 9.45.Embodiment 2294-[[1-[4-[N '-(2-fluoro phenyl) urea groups]-3-methoxyphenyl acetyl group]-the 2-methylamino]-2-methyl-2-propoxyl group] benzoic acid
Figure A0080951005061
In 5-10 ℃, to 2-amino-2-methyl-1-propanol (8.4g, 93.89mmol) and triethylamine (11.4g, DMF-water 0.113mol) (1: 1, v/v, add in agitating solution 100ml) Bis(tert-butoxycarbonyl)oxide (25g, 0.115mol).The solution that obtains was stirred under room temperature 2 hours.Water (100ml) dilutes this mixture, uses ethyl acetate extraction.With salt water washing extract, through dried over sodium sulfate and evaporation.Residue as eluant, obtains 12g (68%) 2-tert-butoxycarbonyl amino-2-methyl-1-propanol with dichloromethane through silica gel column chromatography, is syrup. 1H-NMR(CDCl 3)δ1.25(s，6H)，1.43(s，9H)，3.59(d，J＝8.3Hz，2H)，4.68(br，1H)。
Under room temperature, to 2-tert-butoxycarbonyl amino-2-methyl-1-propanol (5.7g, 30.11mmol) and Powdered sodium hydroxide (6.7g, add in ether 0.151mol) (200ml) agitating solution p-toluenesulfonyl chloride (6.9g, 36.14mmol).The mixture that is produced that stirs was heated 8 hours under refluxing.After the cooling, will ice-water (100ml) and add in this solution.With the isolating ether layer of salt water washing, through dried over sodium sulfate and evaporation.Add in the residue normal hexane and grinding.Collect solid obtain 8.5g (82.2%) right-toluenesulfonic acid 2-tert-butoxycarbonyl amino-2-methyl-1-propyl ester, be crystalline material. 1H-NMR(CDCl 3)δ1.26(s，6H)，1.38(s，9H)，2.37(s，3H)，4.05(s，2H)，4.49(br，1H)，7.34(d，J＝7.8Hz，2H)，7.78(d，J＝7.8Hz，2H)。
With right-toluenesulfonic acid 2-tert-butoxycarbonyl amino-2-methyl-1-propyl ester (8.2g, 23.88mmol) and powdered sodium hydroxide (6.7g, the agitating solution of ether 0.151mol) (200ml) heating 10 hours under refluxing.After the cooling, filter this mixture.Water, saline wash filtrate are through dried over sodium sulfate and evaporation.Residue is used normal hexane through silica gel column chromatography: and ethyl acetate (6: 1, v/v) as eluant, obtain 2.7g (66.2%) 1-tert-butoxycarbonyl-2-metering system imines, be grease. 1H-NMR(CDCl 3)δ1.29(s，6H)，1.47(s，9H)，2.05(s，2H)。
Under room temperature, to 1-tert-butoxycarbonyl-2-metering system imines (1.03g, 6.01mmol) and the 4-methyl hydroxybenzoate (800mg, add in the agitating solution of dichloromethane 6.26mmol) (10ml) boron trifluoride diethyl etherate (0.127ml, 1mmol).Under room temperature with the solution restir that obtains 3 hours.Water, this mixture of salt water washing are through dried over sodium sulfate and evaporation.Residue is used normal hexane through silica gel column chromatography: and ethyl acetate (6: 1, v/v) as eluant, obtain 550mg (33%) 4-(1-tert-butoxycarbonyl amino-2-methyl-2-propoxyl group) essence of Niobe, be jelly. 1H-NMR(CDCl 3)δ1.33(s，6H)，1.47(s，9H)，3.36(d，J＝6.3Hz，2H)，3.90(s，3H)，5.05(br，1H)，6.99(d，J＝8.8Hz，2H)，7.97(d，J＝8.8Hz，2H)。
With 4-(1-tert-butoxycarbonyl amino-2-methyl-2-propoxyl group) essence of Niobe (460mg, 1.42mmol) and methyl phenyl ethers anisole (0.155ml, 1.42mmol) mixture in dichloromethane (15ml) and TFA (3ml) stirred 3 hours under room temperature.Evaporate this mixture.Make residue be dissolved in dichloromethane (30ml), make by adding 0.5N sodium hydroxide to be alkalescence.The separate dichloromethane layer is through dried over sodium sulfate and evaporation.Residue as eluant, obtains 370mg (100%) 4-(1-amino-2-methyl-2-propoxyl group) essence of Niobe with dichloromethane through silica gel column chromatography, is jelly. 1H-NMR(CDCl 3)δ1.34(s，6H)，2.87(s，2H)，3.90(s，3H)，7.10(d，J＝8.8Hz，2H)，7.97(d，J＝8.8Hz，2H)。
In 0 ℃, to 4-(1-amino-2-methyl-2-propoxyl group) essence of Niobe (370mg, 1.66mmol) and triethylamine (0.35ml, add in the agitating solution of dichloromethane 2.49mmol) (15ml) trifluoroacetic anhydride (0.316ml, 2.24mmol).After stirring 1 hour under the identical temperature, water is added in this solution.The separate dichloromethane layer washes with water, through dried over sodium sulfate and evaporation.Residue as eluant, obtains 530mg (100%) 4-(1-trifluoroacetamido-2-methyl-2-propoxyl group) essence of Niobe with dichloromethane through silica gel (20ml) chromatography, is jelly.This chemical compound can need not to be further purified and be used for subsequent reaction.
Under room temperature, to 4-(1-trifluoroacetamido-2-methyl-2-propoxyl group) essence of Niobe (530mg, 1.66mmol) and potassium carbonate (345mg, 2.49mmol) the stirring the mixture of DMF (10ml) in add methyl iodide (0.14ml, 2.37mmol), the mixture that obtains was stirred under room temperature 18 hours.In this mixture impouring water, use ethyl acetate extraction.With salt water washing extract, through dried over sodium sulfate and evaporation.Residual jelly can need not to be further purified and be used for subsequent reaction.
Top crude product residue is dissolved in the methanol (10ml).(352mg 3.32mmol) adds in the solution of this stirring, stirs the mixture that obtains 5 hours under room temperature with water (5ml) and sodium carbonate.In this mixture impouring water, use chloroform extraction.Wash extract with water, through dried over sodium sulfate and evaporation.Residue as eluant, obtains 390mg (100%) 4-(1-methylamino-2-methyl-2-propoxyl group) essence of Niobe with chloroform through silica gel column chromatography, is jelly. 1H-NMR(CDCl 3)δ1.37(s，6H)，2.51(s，3H)，3.89(s，3H)，6.92(d，J＝8.8Hz，2H)，7.97(d，J＝8.8Hz，2H)。
Under room temperature, to 4-(1-methylamino-2-methyl-2-propoxyl group) essence of Niobe (20mg, 0.84mmol), 4-[N '-(2-fluoro phenyl) urea groups]-3-methoxybenzene guanidine-acetic acid (268mg, 0.84mmol), 4-DMAP (125mg, 1.0mmol) the stirring the mixture of DMF (5ml) in add EDC (220mg, 1.14mmol).With the mixture that obtains restir 10 hours under room temperature.In this mixture impouring water, use ethyl acetate extraction.With salt water washing extract; through dried over sodium sulfate and evaporation; residual jelly grinds with dichloromethane and ether; obtain 200mg (44.1%) 4-[[1-[4-[N '-(2-fluoro phenyl) urea groups]-3-methoxyphenyl acetyl group] methylamino]-2-methyl-2-propoxyl group] essence of Niobe, be crystalline material. 1H-NMR (CDCl 3) δ 1.36 and 1.31 (respectively be s, 6H), 3.24-3.88 (a plurality of s, 13H), 6.65-8.20 (a plurality of m, 14H).
With 4-[[1-[4-[N '-(2-fluoro phenyl) urea groups]-3-methoxyphenyl acetyl group] methylamino]-2-methyl-2-propoxyl group] (180mg, 0.335mmol) mixture in THF (3ml) and 0.25N sodium hydroxide (4ml) stirred 5 hours under room temperature essence of Niobe.In this mixture impouring ice-1N HCl (5ml).Collect solid.Wash with water also air-dry.Make crude product solid recrystallization from ethanol-chloroform-normal hexane, obtain 70mg (40%) 280, be the fine acicular material.Mp200-207 ℃; 1H-NMR (DMSO-d 6) δ 1.26 and 1.33 (respectively be s, 6H), 3.70-3.81 (a plurality of s, 7H), 3.83 (s, 3H), 6.75-8.20 (a plurality of m, 10H), 8.71 (s, 1H), 9.17 (br s, 1H), 12.72 (s, 1H).Embodiment 230 (S)-3-chloro-4-[2-[3-methoxyl group-4-[N '-(2-fluoro phenyl) urea groups] phenyl acetyl amino]-the 1-propoxyl group] benzoic acid
Figure A0080951005091
Under room temperature, with 3-methoxyl group-4-[N '-(2-fluoro phenyl) urea groups] phenylacetic acid (327mg, 1.03mmol), (S)-3-chloro-4-(2-amino-1-propoxyl group) essence of Niobe (250mg, 1.03mmol), EDC (hydrochlorate) (295mg, 1.54mmol), HOBt (208mg, 1.54mmol) and DMAP (25mg, 0.20mmol) mixture in DMF (8ml) stirs under room temperature and spends the night.Dilute this mixture with ethyl acetate, with 0.5N HCl, salt water washing, through dried over sodium sulfate and evaporation.Make residue recrystallization from chloroform-ethyl acetate, obtain 308mg (55%), be white crystalline powder. 1H-NMR(CDCl 3)δ1.29(d，3H，J＝6.8Hz)，3.51(s，2H)，3.84(s，3H)，3.88(s，3H)，4.01-4.08(m，2H)，4.38-4.39(m，1H)，6.25(d，1H，J＝8.3Hz)，6.78-6.83(m，2H)，6.90-6.95(m，2H)，7.02-7.11(m，2H)，7.89(dd，1H，J＝2.0，8.8Hz)，8.02(d，1H，J＝2.4Hz)，8.20(d，1H，J＝8.3Hz)，8.27-8.31(m，1H)，8.53(s，1H)，8.84(s，1H)。
To (S)-3-chloro-4-[2-[3-methoxyl group-4-[N '-(2-fluoro phenyl) urea groups] phenyl acetyl amino]-the 1-propoxyl group] essence of Niobe (308mg; 0.57mmol) at THF-methanol (10ml; 1: 1; v/v) add 0.5N sodium hydroxide (10ml) in the agitating solution, reactant mixture was heated 1 hour under refluxing.In this mixture impouring ice-1N HCl, collect solid.Make crude product solid recrystallization purifying from methanol-chloroform-normal hexane, obtain 196mg (65%) 281, be white crystalline powder.Mp 188-191 ℃; 1H-NMR (DMSO-d 6) δ 1.21 (d, 3H, J=6.4Hz), 3.38 (s, 2H), 3.83 (s, 3H), 4.02-4.18 (m, 3H), 6.78-7.29 (a plurality of m, 6H altogether), 7.85-8.00 (m, 3H), 8.12-8.19 (m, 2H), 8.70 (s, 1H), 9.17 (s, 1H), 12.98 (bs, 1H); MS (FAB) m/z 530 (M +), 531 (M ++ 1), 532 (M ++ 2); C 26H 25ClFN 3O 61/4H 2The analytical calculation value of O: C, 58.43; H, 4.81; Cl, 6.63; F, 3.55; N, 7.86.Measured value: C, 58.45; H, 4.83; Cl, 6.68; F, 3.38; N, 7.79.Embodiment 2314-[[2-[3-methoxyl group-4-[N '-(2-aminomethyl phenyl) urea groups] phenyl acetyl] methylamino] ethylamino] benzoic acid
(1.77g, (1.29g 8.5mmol), stirred the mixture under room temperature 1 hour to add the 4-Methyl anthranilate in the agitating solution of toluene 1.1mmol) (3ml) to N-benzyloxycarbonyl-N-methylamino acetaldehyde.The reduction vaporization reactant mixture is dissolved among the MeCN (15ml) residue.In this solution, add acetic acid (2.44ml, 43mmol) and NaBH 3(2.67g 43mmol), stirred the mixture that obtains 15 hours CN under room temperature.By adding this reactant of saturated sodium bicarbonate quencher.With this mixture of ethyl acetate extraction, use the salt water washing, through dried over mgso and evaporation.Residue is through silica gel column chromatography, with normal hexane-ethyl acetate (7: 3,, obtain 1.26g (43%) 4-[2-(N-benzyloxycarbonyl-N-methylamino)-ethylamino v/v) as eluant] essence of Niobe, be colorless oil. 1H-NMR (CDCl 3, 400MHz) δ 2.96 (s, 3H), 3.32 (br m, 2H), 3.50-3.60 (m, 2H), 3.82 (s, 3H), 5.15 (respectively be d, 2H, J=14.6Hz), 6.35 and 6.58 (m, altogether 2H), 7.36 (s, 5H), 7.76 and 7.84 (m, 2H altogether); MS (FAB) m/z 342 (M ++ 1).
To 4-[2-(N-benzyloxycarbonyl-N-methylamino) ethylamino] (1.26g adds 5wt Pd-C (700mg), this mixture of hydrogenation under room temperature (3 atm) 4 hours to essence of Niobe in the agitating solution of methanol 3.68mmol) (20ml).Filter this mixture, reduction vaporization filtrate obtains 600mg (78%) 4-[2-(N-methylamino) ethylamino] essence of Niobe, be colorless oil. 1H-NMR(CDCl 3，400MHz)δ2.46(s，3H)，2.88(t，2H，J＝5.5Hz)，3.27(br?s，2H)，3.85(s，3H)，6.57(d，2H，J＝8.8Hz)，7.85(d，2H，J＝8.8Hz)；MS(FAB)m/z?209(M ++1)。
To 4-[2-(N-methylamino) ethylamino] essence of Niobe (590mg, 2.83mmol), 3-methoxyl group-4-[N '-(2-aminomethyl phenyl) urea groups] phenylacetic acid (891mg, 2.83mmol) the agitating solution of DMF (14ml) in add EDC (815mg, 4.25mmol), HOBt (574mg, 4.25mmol) and 4-DMAP (519mg, 4.25mmol), the mixture that obtains stirred under room temperature spend the night.In this mixture impouring 1N HCl, the collected at suction solid.Make the crude product solid through silica gel column chromatography (middle pressure) purification; with chloroform-ethyl acetate (10: 0 to 7: 3; v/v) as eluant; obtain 4-[[2-[3-methoxyl group-4-[N '-(2-aminomethyl phenyl) urea groups of 1.25g (87%)] phenyl acetyl] methylamino] ethylamino] essence of Niobe, be faint yellow oily thing. 1H-NMR (CDCl 3, 400MHz) δ 3.18 (s, 2H), 3.05 and 3.32 (s, be total to 2H), and 3.72-3.85 (m, 9H), 4.12 and 4.23 (m, be total to 2H), 6.78 (m, 3H), 7.05 (t, 1H, J=7.5Hz), 7.20 (m, 2H), 7.43-7.50 (m, 3H), 7.62 (d, 1H, J=8.8Hz), 8.05 (d, 1H, J=8.8Hz); MS (FAB) m/z 505 (M ++ 1).
With 4-[[2-[3-methoxyl group-4-[N '-(2-aminomethyl phenyl) urea groups] phenyl acetyl] methylamino] ethylamino] essence of Niobe (300mg, 0.6mmol) reflux 8 hours of stirring the mixture in 0.25N sodium hydroxide (6ml) and THF (6ml).In this mixture impouring water, with 1N HCl acidify.The collected at suction solid.Make crude product solid recrystallization from normal hexane-Di Iso Propyl Ether, obtain 282 of 202mg (69%), be light yellow crystalline powder.Mp 115-120 ℃; IR (KBr) 3346,2935,1603,1531,1454,1417,1257,1174,1036,754cm -1 1H-NMR (CD 3OD, 400MHz) δ 2.28 (s, 3H), 2.98 (s, 1H), 3.10 (s, 2H), 3.35-3.42 (m, 2H), 3.53-3.65 (m, 3H), 3.70 (s, 1H), 3.80 and 3.82 (s, 3H), 6.60-6.85 (m, 4H), 7.02 (m, 1H), 7.18 (m, 2H), 7.58 (m, 1H), 7.82 (m, 2H), 7.96 (m, 1H); MS (FAB) m/z 491 (M ++ 1).Embodiment 232 (S)-3-chloro-4-[2-[N-methyl-N-[3-methoxyl group-4-[N '-(2-fluoro phenyl) urea groups] phenyl acetyl] amino]-the 1-propoxyl group] benzoic acid
Figure A0080951005111
Under room temperature, to (S)-3-chloro-4-(2-amino-1-propoxyl group) essence of Niobe (1.74g, 7.14mmol) cold (0 ℃) solution of dichloromethane (20ml) in add triethylamine (1.19ml, 8.54mmol) and trifluoroacetic anhydride (TFAA) (1.11ml, 7.86mmol), reactant mixture stirred under room temperature spend the night.Dilute this mixture with chloroform, with 0.5N HCl, salt water washing, through dried over sodium sulfate and evaporation.Make residue recrystallization from chloroform-normal hexane, obtain 1.59g (66%) (S)-3-chloro-4-(2-trifluoroacetamido-1-propoxyl group] benzoate, be white crystalline material.Mp 122-125 ℃; 1H-NMR (CDCl 3) δ 1.48 (d, 3H, J=6.8Hz), 3.91 (s, 3H), 4.10-4.19 (m, 2H), 4.47-4.52 (m, 1H), 6.68 (bs, 1H), 6.92-6.94 (m, 1H), 7.93-7.95 (m, 1H), 8.07-8.08 (m, 1H); MS (FAB) m/z 340 (M ++ 1); C 13H 13ClF 3NO 4The analytical calculation value: C, 45.96; H, 3.86; Cl, 10.44; F, 16.78; N, 4.12.Measured value: C, 45.88; H, 3.97; Cl, 10.24; F, 16.72; N, 4.18.
To (S)-3-chloro-4-(2-trifluoroacetamido-1-propoxyl group] essence of Niobe (800mg, 2.36mmol) in the agitating solution of DMF (5ml), add potassium carbonate (651mmol, 4.71mmol) and methyl iodide (0.22ml 3.53mmol), spends the night reactant mixture in 60 ℃ of stirrings.Dilute this mixture with ethyl acetate, with 0.5N HCl, salt water washing, through dried over sodium sulfate and evaporation.Residue is through purification by silica gel column chromatography, with the chloroformic solution of 5% ethyl acetate as eluant, obtain 850mg (100%) (S)-3-chloro-4-[2-(N-methyl-N-TFA amino)-1-propoxyl group] essence of Niobe, be colorless oil. 1H-NMR (CDCl 3) δ 1.43-1.46 (m, 3H), 3.03 and 3.21 (s, 3H), 3.90 (s, 3H), 4.04-4.22 (m, 2H), 4.81-4.87 (m, 1H), 6.91 (d, 1H, J=8.3Hz), 7.93 (dd, 1H, J=2.0,8.3Hz), 8.06 (d, 1H, J=2.0Hz).
Under room temperature, to (S)-3-chloro-4-[2-(N-methyl-N-TFA amino)-1-propoxyl group] essence of Niobe (880mg, methanol-water 2.49mmol) (10ml, 1: 1, add in agitating solution v/v) potassium carbonate (516mg, 3.73mmol).The mixture that obtains stirred under room temperature spend the night.Dilute this mixture with ethyl acetate, water, salt water washing, through dried over sodium sulfate and the evaporation, obtain 460mg (72%) (S)-3-chloro-4-(2-methylamino-1-propoxyl group) benzoate, be colorless oil. 1H-NMR(CDCl 3)δ1.20(d，3H，J＝6.4Hz)，2.51(s，3H)，3.07-3.12(m，1H)，3.89(s，3H)，3.92-4.04(m，2H)，6.93-6.95(m，1H)，7.90-7.93(m，1H)，8.05-8.06(m，1H)。
Under room temperature, with 3-methoxyl group-4-[N '-(2-fluoro phenyl) urea groups] phenylacetic acid (296mg, 0.93mmol), (S)-3-chloro-4-(2-methylamino-1-propoxyl group) essence of Niobe (240mg, 0.93mmol), EDC (hydrochlorate) (268mg, 1.40mmol), HOBt (189mg, 1.40mmol) and DMAP (23mg, 0.19mmol) mixture in DMF (8ml) stirred 1.5 hours under room temperature.Dilute this mixture with ethyl acetate, with 0.5N HCl, salt water washing, through dried over sodium sulfate and evaporation.Residue is through purification by silica gel column chromatography; with the chloroformic solution of 5% methanol as eluant; obtain 520mg (100%) (S)-3-chloro-4-[2-[N-methyl-N-[3-methoxyl group-4-[N '-(2-fluoro phenyl) urea groups] phenyl acetyl] amino]-the 1-propoxyl group] essence of Niobe, be red-brown amorphous solid.
(520mg adds 0.5N sodium hydroxide (10ml) in the agitating solution of THF 0.93mmol) (10ml), the mixture that obtains was heated 3 hours under refluxing to this product.In this mixture impouring ice-1N HCl, collect solid.Make crude product solid recrystallization from chloroform-ether, obtain 170mg (two steps, 37%) 283, be white crystalline powder.Mp 142-147 ℃; 1H-NMR (DMSO-d 6) δ 1.13-1.20 (m, 3H), 2.74 and 2.94 (s, 3H), 3.65 (s, 2H), 3.82 and 3.84 (s, 3H), 4.13-4.22 (m, 2H), 4.53 and 4.91-4.92 (m, 1H), (6.71-7.29 a plurality of m, 6H altogether), and 7.86-8.02 (m, 3H), 8.15-8.19 (m, 1H), 8.71 (s, 1H), 9.17 (s, 1H), 13.00 (bs, 1H); MS (FAB) m/z 544 (M +), 545 (M ++ 1); C 27H 27ClFN 3O 61/4H 2The analytical calculation value of O: C, 59.13; H, 5.05; Cl, 6.46; F, 3.46; N, 7.66.Measured value: C, 59.19; H, 4.99; Cl, 6.64; F, 3.23; N, 7.55.Embodiment 233 (S)-3-chloro-4-[2-[N-methyl-N-[3-methoxyl group-4-[N '-(2-aminomethyl phenyl) urea groups] phenyl acetyl] amino-1-propoxyl group] benzoic acid
Figure A0080951005131
Under room temperature, with 3-methoxyl group-4-[N '-(2-aminomethyl phenyl) urea groups] phenylacetic acid (261mg, 0.83mmol), (S)-3-chloro-4-(2-methylamino-1-propoxyl group) essence of Niobe (214mg, 0.83mmol), EDC (hydrochlorate) (239mg, 1.25mmol), HOBt (168mg, 1.24mmol) and DMAP (20mg, 0.16mmol) mixture in DMF (8ml) stirred 2 hours under room temperature.Dilute this mixture with ethyl acetate, with 0.5N HCl, salt water washing, through dried over sodium sulfate and evaporation.Residue is through purification by silica gel column chromatography; with chloroform-methanol (50: 1; v/v) as eluant; obtain 470mg (100%) (S)-3-chloro-4-[2-[N-methyl-N-[3-methoxyl group-4-[N '-(2-aminomethyl phenyl) urea groups] phenyl acetyl] amino]-the 1-propoxyl group] essence of Niobe, be faint yellow amorphous solid.
(470mg adds 0.5N sodium hydroxide (10ml) to product upward in the agitating solution of THF 0.95mmol) (10ml), reactant mixture was heated 3 hours under refluxing.In this mixture impouring ice-1N HCl, collect solid.Make crude product solid recrystallization from chloroform-ether, obtain 168mg (two steps, 33%) 284, be light yellow crystalline powder.Mp 125-130 ℃; 1H-NMR (DMSO-d 6) δ 1.13-1.19 (m, 3H), 2.24 (s, 3H), 2.74 and 2.94 (s, 3H), 3.65 (s, 2H), 3.83 and 3.85 (s, 3H), 4.14-4.22 (m, 2H), 4.91-4.93 (m, 1H), 6.70-6.74 (m, 1H), 6.84 (m, 1H), 6.92-6.95 (m, 1H), 7.11-7.17 (m, 2H), 7.25-7.29 (m, 1H), 7.78-7.80 (m, 1H), 7.85-7.93 (m, 2H), 7.99-8.02 (m, 1H), 8.46 (s, 1H), 8.56 (s, 1H), 12.99 (bs, 1H); MS (FAB) m/z 540 (M +), 541 (M ++ 1); C 28H 30ClN 3O 61/2H 2The analytical calculation value of O: C, 61.26; H, 5.69; N, 7.65.Measured value: C, 61.15; H, 5.58; N, 7.51.Embodiment 2343-isopropyl amino-4-[[2-[3-methoxyl group-4-[N '-(2-aminomethyl phenyl) urea groups] phenyl acetyl] methylamino] ethyoxyl] benzoic acid
To 3-amino-4-[[2-[3-methoxyl group-4-[N '-(2-aminomethyl phenyl) urea groups] phenyl acetyl] methylamino] ethyoxyl] essence of Niobe (300mg; 0.58mmol) acetone-acetic acid-DMF (13ml; 6: 6: 1, add NaBH in cold (0 ℃) solution of stirring v/v/v) 3CN (300mg), the mixture that stirring obtains under room temperature 60 hours.In this mixture impouring saturated sodium bicarbonate, use chloroform extraction.With salt water washing extract,, obtain 280mg (86%) 3-isopropyl amino-4-[[2-[3-methoxyl group-4-[N '-(2-aminomethyl phenyl) urea groups through dried over mgso and evaporation] phenyl acetyl] methylamino] ethyoxyl] essence of Niobe, be colorless oil. 1H-NMR (CDCl 3, 400MHz) δ 1.21 (m, 6H), 2.30 (s, 3H), 3.05 and 3.10 (s, be total to 2H), 3.59 (s, 3H), 3.62-3.81 (m, 6H), 3.89 (s, 3H), 4.10 and 4.21 (m, 2H), 6.65-6.80 (m, 4H), 7.10 (m, 1H), 7.20 (s, 3H), 7.32 (m, 2H), 7.54 (d, 1H, J=8.3Hz), 8.00 (s, 1H), 8.07 (d, 1H, J=8.3Hz); MS (FAB) m/z 563 (M ++ 1).
With 3-isopropyl amino-4-[[2-[3-methoxyl group-4-[N '-(2-aminomethyl phenyl) urea groups] phenyl acetyl] methylamino] ethyoxyl] essence of Niobe (280mg, 0.5mmol), the heating 8 hours that stirs the mixture under refluxing of 0.25N sodium hydroxide (6ml) and THF (6ml).In this mixture impouring water (200ml), through 1N HCl acidify, collected at suction solid.Make solid recrystallization from chloroform-normal hexane-Di Iso Propyl Ether, obtain 202mg (74%) 285, be light yellow crystalline powder.Mp 130-135 ℃; 1H-NMR (CD 3OD, 400MHz) (d is total to 6H, J=6.3Hz) to δ 1.18 and 1.22,2.29 with 2.32 (s, 3H altogether), 3.04 and 3.14 (s, 2H altogether), 3.60-3.90 (m, 9H), 4.16 and 4.25 (m, 2H altogether), 6.80 (m, 3H), 7.02 (m, 1H), 7.12-7.24 (m, 3H), 7.29-7.38 (m, 1H), 7.59 (m, 1H), 8.02 (m, 1H); MS (FAB) m/z 548 (M+1) +C 30H 36N 4O 6The analytical calculation value: C, 65.68; H, 6.61.Measured value: C, 65.80; H, 6.83.Embodiment 2354-[[2-[3-methoxyl group-4-[N '-(2-aminomethyl phenyl) urea groups] phenyl acetyl] methylamino] ethyl] the methylamino benzoic acid
To 4-[[2-[3-methoxyl group-4-[N '-(2-aminomethyl phenyl) urea groups] phenyl acetyl] methylamino] ethyl] Methyl anthranilate (300mg; 0.6mmol) MeCN-formaldehyde-acetic acid (11ml; 8: 2: 1, add NaBH in cold (0 ℃) solution of stirring v/v/v) 3CN (187mg, 2.40mmol), the mixture that stirring obtains under room temperature 18 hours.In this mixture impouring saturated sodium bicarbonate, use chloroform extraction.With salt water washing extract; through dried over mgso and evaporation; obtain 309mg (100%) 4-[[2-[3-methoxyl group-4-[N '-(2-aminomethyl phenyl) urea groups] phenyl acetyl]-2-N '-methylamino] ethyl] N-methylamino essence of Niobe, be colorless oil. 1H-NMR(CDCl 3，400MHz)δ2.30(m，3H)，2.98(m，7H)，3.46-3.72(m，9H)，3.82(m，3H)，6.32(m，1H)，6.55-6.75(m，4H)，7.05-7.50(m，2H)，7.82-8.02(m，4H)；MS(FAB)m/z?518(M ++1)。
With 4-[[2-[3-methoxyl group-4-[N '-(2-aminomethyl phenyl) urea groups] phenyl acetyl]-2-N '-methylamino] ethyl] the methylamino essence of Niobe (309mg, heated 8 hours under refluxing by stirring the mixture of 1N sodium hydroxide (2.4ml) 0.6mmol) and THF (10ml).In this mixture impouring water (200ml), by adding 1N HCl acidify.The collected at suction solid.Make crude product solid recrystallization from chloroform-normal hexane-Di Iso Propyl Ether, obtain 211mg (70%) 286, be light yellow crystalline powder.Mp 125-130 ℃; IR (KBr) 3338,2933,1601,1529,1182,1036,752cm -1 1H-NMR (CD 3OD, 400MHz) δ 2.28 and 2.29 (s, 3H), 2.95-3.02 (m, 6H), 3.60 (br, 6H), 3.80 (s, 1H), 3.86 (s, 2H), 6.60-6.82 (m, 4H), 7.01-7.18 (m, 3H), 7.58 (m, 1H), 7.82-7.99 (m, 3H); MS (FAB) m/z504 (M ++ 1).Embodiment 236 (S)-3-chloro-4-[2-[N-benzyl-N-[3-methoxyl group-4-[N '-(2-fluoro phenyl) urea groups] phenyl acetyl] amino]-the 1-propoxyl group] benzoic acid
To NaBH 3CN (985mg, 15.68mmol) the agitating solution of methanol (5ml) in add (S)-3-chloro-4-(2-amino-1-propoxyl group] essence of Niobe (382mg, 1.57mmol) and methanol (5ml) solution of benzaldehyde (0.19ml), under room temperature, stir the mixture overnight that obtains.By adding this mixture of entry quencher, use chloroform extraction.With salt water washing extract, through dried over sodium sulfate and evaporation, residue is through purification by silica gel column chromatography, with chloroform-methanol (30: 1, v/v) as eluant, obtain 336mg (64%) (S)-3-chloro-4-(2-N-benzylamino-1-propoxyl group) benzoate, be colorless oil. 1H-NMR (CDCl 3) δ 1.22 (d, 3H, J=6.4Hz), 3.21-3.25 (m, 1H), 3.84-4.03 (m, 7H altogether), 6.89-6.91 (m, 1H), 7.23-7.37 (m, 5H), 7.89-7.91 (m, 1H), 8.05 (m, 1H).
Under room temperature, with 3-methoxyl group-4-[N '-(2-fluoro phenyl) urea groups] phenylacetic acid (320mg, 1.01mmol), (S)-3-chloro-4-(2-N-benzylamino-1-propoxyl group) essence of Niobe (336mg, 1.01mmol), EDC (hydrochlorate) (289mg, 1.51mmol), HOBt (204mg, 1.51mmol) and DMAP (25mg, 0.20mmol) mixture in DMF (7ml) stirred 2 days under room temperature.Dilute this mixture with ethyl acetate, with 0.5N HCl, salt water washing, through dried over sodium sulfate and evaporation.Residue is through purification by silica gel column chromatography; with chloroform-methanol (50: 1; v/v) as eluant; obtain 607mg (95%) (S)-3-chloro-4-[2-[N-benzyl-N-[3-methoxyl group-4-[N '-(2-fluoro phenyl) urea groups] phenyl acetyl] amino]-the 1-propoxyl group] essence of Niobe, be faint yellow amorphous solid. 1H-NMR (CDCl 3) δ 1.20-1.27 (m, 3H), 3.62 (s, 2H), 3.73-4.16 (a plurality of m, 9H altogether), 4.71 (bs, 2H), 6.67-7.38 (a plurality of m, 12H altogether), 7.77-8.17 (a plurality of m, 5H altogether).
To (S)-3-chloro-4-[2-[N-benzyl-N-[3-methoxyl group-4-[N '-(2-fluoro phenyl) urea groups] phenyl acetyl] amino]-the 1-propoxyl group] essence of Niobe (607mg; 0.96mmol) the agitating solution of THE (6ml) in add 0.5N sodium hydroxide (6ml), with the mixture heated overnight under refluxing that obtains.In this mixture impouring ice-1N HCl, collect solid.Make crude product solid recrystallization, obtain 192mg (32%) 287, be white crystalline powder.Mp 125-130 ℃; 1H-NMR (DMSO-d 6) δ 1.07-1.23 (m, 3H), 3.76 (s, 2H), 3.85 (s, 3H), 3.90-4.26 (m, 3H), 4.56 (s, 2H), 6.66-7.38 (a plurality of m, 10H altogether), 7.82-8.20 (a plurality of m, 5H altogether), 8.70-8.74 (m, 1H), 9.18-9.20 (m, 1H), 13.02 (bs, 1H); MS (FAB) m/z 621 (M ++ 1); C 33H 31ClFN 3O 61/4H 2The analytical calculation value of O: C, 63.46; H, 5.08; Cl, 5.68; F, 3.04; N, 6.73.Measured value: C, 63.67; H, 5.16; Cl, 5.75; F, 2.95; N, 6.55.Embodiment 2373-(N-isopropyl-N-methylamino)-4-[[2-[3-methoxyl group-4-[N '-(2-aminomethyl phenyl) urea groups] phenyl acetyl] methylamino] ethyoxyl] benzoic acid
To 3-isopropyl amino-4-[[2-[3-methoxyl group-4-[N '-(2-aminomethyl phenyl) urea groups] phenyl acetyl] methylamino] ethyoxyl] essence of Niobe (324mg; 0.58mmol) acetonitrile-formaldehyde-acetic acid (11ml; 8: 2: 1, add NaBH in cold (0 ℃) solution of stirring v/v/v) 3CN (145mg, 2.30mmol), the mixture that stirring obtains under room temperature 18 hours.In this mixture impouring saturated sodium bicarbonate, the collected at suction solid.The crude product solid is dissolved in the chloroform (20ml); with this solution of salt water washing; through dried over mgso and evaporation; obtain 317mg (95%) 3-(N-isopropyl-N-methylamino)-4-[[2-[3-methoxyl group-4-[N '-(2-aminomethyl phenyl) urea groups] phenyl acetyl] methylamino] ethyoxyl] essence of Niobe, be colorless oil. 1H-NMR (CDCl 3) δ 1.02 (m, 6H), 2.29 (s, 3H), 2.63 (m, 3H), and 3.02-3.20 (m, 3H), 3.49-3.80 (m, 8H), 3.88 (s, 3H), 4.06 and 4.21 (m, be total to 2H), 6.59 (m, 1H), 6.76 (m, 2H), and 7.11-7.23 (m, 3H), 7.50-7.62 (m, 3H), 8.05 (d, 1H, J=8.3Hz); MS (FAB) m/z576 (M ++ 1).
With 3-(N-isopropyl-N-methylamino)-4-[[2-[3-methoxyl group-4-[N '-(2-aminomethyl phenyl) urea groups] phenyl acetyl] methylamino] ethyoxyl] (317mg 0.55mmol) heated 8 hours under stirring the mixture of 0.25N sodium hydroxide (8.2ml) and THF (8ml) refluxed essence of Niobe.In this mixture impouring water (200ml), by adding 1N HCl acidify, collected at suction solid.Make crude product solid recrystallization from chloroform-normal hexane-Di Iso Propyl Ether, obtain 288, be light yellow crystalline powder.Mp 130-135 ℃; IR (KBr) 2970,1537,1038,754cm -1 1H-NMR (CD 3OD) δ 1.12 (m, 6H), 2.29 (s, 3H), 2.76 (m, 1H), 2.89 (s, 2H), 3.02 (s, 1H), 3.21 (s, 2H), 3.72 (s, 2H), 3.80 (s, 3H), 3.84 (m, 3H), 4.13 with 4.40 (m, 2H altogether), 6.76 (d, 1H, J=7.8Hz), 6.86 (s, 1H), 7.00 (m, 2H), 7.18 (m, 3H), 7.57 (d, 1H, J=7.8Hz), 7.95 (m, 2H); MS (FAB) m/z 562 (M ++ 1); C 31H 38N 4O 62.0H 2The analytical calculation value of O: C, 62.19; H, 7.07; N, 9.36.Measured value: C, 62.54; H, 6.85; N, 8.90.Embodiment 2383-(piperidino)-4-[[2-[3-methoxyl group-4-[N '-(2-fluoro phenyl) urea groups] phenyl acetyl] methylamino] ethyoxyl] benzoic acid
Figure A0080951005191
In 0 ℃; to 3-amino-4-[[2-[3-methoxyl group-4-[N '-(2-fluoro phenyl) urea groups] phenyl acetyl] methylamino] ethyoxyl] essence of Niobe (300mg; 0.57mmol) the solution of stirring of THF (2ml) in add glutaraldehyde (50% aqueous solution) (0.13ml, the solution in methanol 0.69mmol) (1.4ml), THF (0.993ml) and the 3N sulphuric acid (0.952ml).Under room temperature, in this solution, add NaBH 3CN (150mg), DMF (5ml) and methanol (2ml) stir the mixture that obtains 15 hours.In this mixture impouring saturated sodium bicarbonate, use chloroform extraction.With salt water washing extract, through dried over mgso and reduction vaporization.Residue is through silica gel column chromatography; use toluene: acetone (7: 3; v/v), obtain 145mg (43%) 3-(piperidino)-4-[[2-[3-methoxyl group-4-[N '-(2-fluoro phenyl) urea groups as eluant] phenyl acetyl] methylamino] ethyoxyl] essence of Niobe, be colorless oil. 1H-NMR(CDCl 3，400MHz)δ1.58-1.72(m，6H)，2.80-2.92(m，4H)，3.26(s，2H)，3.58(s，2H)，3.69(s，2H)，3.80-3.89(m，7H)，4.22(m，2H)，6.72(s，1H)，6.78(m，2H)，6.95-7.18(m，2H)，7.32(s，1H)，7.60(s，1H)，7.63(d，1H，J＝7.8Hz)，7.98(d，1H，J＝7.8Hz)，8.18(m，1H)；MS(FAB)m/z?593(M ++1)。
With 3-(piperidino)-4-[[2-[3-methoxyl group-4-[N '-(2-fluoro phenyl) urea groups] phenyl acetyl] methylamino] ethyoxyl] essence of Niobe (290mg, 0.49mmol), stirring the mixture under refluxing the heating 4 hour of 1N sodium hydroxide (1.95ml) in methanol (5ml) and THF (10ml).In this mixture impouring water (200ml), usefulness 1N HCl acidify (pH=4.0), the usefulness chloroform-methanol (9: 1, v/v) extract.The extract that merges through dried over mgso also evaporates.Make residue crystallization from Di Iso Propyl Ether-hexane, obtain 201mg (71%) 289, be light yellow crystalline powder.Mp 125-130 ℃; IR (KBr) 3338,2935,1599,1537,1041,752cm -1 1H-NMR (CD 3OD) δ 1.52-1.73 (m, 6H), 2.88 (s, 3H), 2.98 (br, 1H), 3.09 (s, 1H), 3.23 (s, 2H), 3.66 (s, 2H), 3.75 (s, 1H), 3.82 (s, 4H), 4.13 and 4.29 (m, 2H altogether), 6.78 (m, 2H), 6.99 (m, 2H), 7.1 0 (m, 2H), 7.60-7.70 (m, 2H), 7.96-8.07 (m, 2H); MS (FAB) m/z 578 (M ++ 1); C 31H 35FN 4O 60.5H 2The analytical calculation value of O: C, 63.36; H, 6.17; N, 9.53.Measured value: C, 63.22; H, 6.15; N, 9.16.Embodiment 2393-amino-4-[[2-[3-methoxyl group-4-[N '-(2-aminomethyl phenyl) urea groups] phenyl acetyl] methylamino] ethyoxyl] benzoic acid
To 3-amino-4-[[2-[3-methoxyl group-4-[N '-(2-aminomethyl phenyl) urea groups] phenyl acetyl] methylamino] ethyoxyl] essence of Niobe (300mg; 0.58mmol) methanol-THF (2: 1; v/v; add 0.25N sodium hydroxide (9ml) in agitating solution 12ml), the mixture that obtains was heated 16 hours under refluxing.In this mixture impouring water (100ml), by adding 1N HCl acidify.The collected at suction solid.Make residue recrystallization from Di Iso Propyl Ether, obtain 243mg (83%) 290, be the yellow crystal powder.Mp 125-130 ℃; IR (KBr) 3346,2935,1533,1211,1034,756,637cm -1 1H-NMR (DMSO-d 6) δ 2.29 (s, 3H), 3.05 (s, 1H), 3.72-3.85 (m, 7H), 4.12 and 4.25 (m, 2H altogether), 6.76-6.89 (m, 3H), 7.00 (m, 1H), 7.18 (m, 2H), 7.39 (m, 2H), 7.60 (d, 1H, J=7.8Hz), 7.96 (m, 1H); C 27H 30N 4O 60.5H 2The analytical calculation value of O: C, 62.90; H, 6.06; N, 10.87.Measured value: C, 63.03; H, 6.14; N, 10.56.Embodiment 2403-(piperidino)-4-[[2-[3-methoxyl group-4-[N '-(2-aminomethyl phenyl) urea groups] phenyl acetyl] methylamino] ethyoxyl] benzoic acid
To 3-amino-4-[[2-[3-methoxyl group-4-[N '-(2-aminomethyl phenyl) urea groups] phenyl acetyl] methylamino] ethyoxyl] essence of Niobe (573mg; 1.1mmol) THF-methanol (2: 1; v/v; add glutaraldehyde (0.423ml in refrigerative agitating solution 6ml); 2.2mmol), 3N sulphuric acid (1.83ml, 5.5mmol) and NaBH 3CN (276mg, 4.4mmol).Under room temperature, the mixture that obtains was stirred 18 hours.In this mixture impouring saturated sodium bicarbonate (100ml), the collected at suction solid.The crude product solid is through purification by silica gel column chromatography; use toluene: acetone (10: 0-4: 1; v/v) eluting obtains 240mg (37%) 3-(piperidino)-4-[[2-[3-methoxyl group-4-[N '-(2-aminomethyl phenyl) urea groups] phenyl acetyl] methylamino] ethyoxyl] essence of Niobe, be jelly. 1H-NMR (CD 3OD) δ 1.52-1.72 (m, 6H), 2.30 (s, 3H), 2.36 (s, 2H), 2.89 (br s, 3H), 2.98 (m, 1H), 3.05 (s, 1H), 3.20 (s, 2H), 3.68 (s, 2H), 3.69 (m, 2H), 3.79 (m, 2H), 3.88 (s, 2H), 4.08 and 4.21 (m, 2H), 6.35 and 6.42 (s, 1H altogether), 6.70 (s, 1H), 6.70 (s, 1H), 6.79 (m, 2H), 7.09-7.24 (m, 4H), 7.50-7.65 (m, 3H), 8.05 (d, 1H, J=8.3Hz); MS (FAB) m/z 588 (M ++ 1).
With 3-(piperidino)-4-[[2-[3-methoxyl group-4-[N '-(2-aminomethyl phenyl) urea groups] phenyl acetyl] methylamino] ethyoxyl] essence of Niobe (240mg, 0.41mmol), stirring the mixture under refluxing the heating 11 hour of 1N sodium hydroxide (1.63ml) in methanol (6.5ml), water (5ml) and THF (6.5ml).In this mixture impouring water (200ml), be acidified to pH=4.0, the collected at suction solid by adding 1N HCl.Extract water layer with chloroform-methanol (9: 1, v/v, 30ml * 3).Precipitate is dissolved in the organic extract of merging.Through dried over mgso organic layer and evaporation.Make residue crystallization from Di Iso Propyl Ether, obtain 151mg (65%) 291, be light yellow crystalline powder.Mp 120-125 ℃; IR (KBr) 3354,2935,1535,1252,1217,1034,754,638cm -1 1H-NMR (CD 3OD) δ 1.55-1.72 (m, 6H), 2.30 (s, 3H), 2.89 (br, 2H), 2.98 (br, 1H), 3.09 (s, 1H), 3.22 (s, 2H), 3.69 (s, 3H), 3.74 (s, 1H), 3.83 (m, 4H), 4.12 and 4.28 (m, 2H altogether), 6.75 (m, 2H), 6.88-7.02 (m, 1H), 7.17 (m, 2H), 7.58-7.73 (m, 4H), 7.99 (d, 1H, J=8.3Hz); MS (FAB) m/z574 (M ++ 1); C 32H 38N 4O 60.5H 2The analytical calculation value of O: C, 65.85; H, 6.73; N, 9.60.Measured value: C, 65.94; H, 6.88; N, 9.03.Embodiment 2413-amino-4-[[2-[3-methoxyl group-4-[N '-(2-fluoro phenyl) urea groups] phenyl acetyl] methylamino] ethyoxyl] benzoic acid
With 3-amino-4-[[2-[3-methoxyl group-4-[N '-(2-fluoro phenyl) urea groups] phenyl acetyl] methylamino] ethyoxyl] essence of Niobe (250mg, 0.48mmol) heated 10 hours under refluxing by stirring the mixture in 1N sodium hydroxide (1.19ml), methanol (8ml), water (6ml) and THF (8ml).In this mixture impouring water (200ml), by adding 1N HCl acidify (pH=4.8), collected at suction solid.Make crude product solid recrystallization from Di Iso Propyl Ether, obtain 209mg (86%) 292, be light yellow crystalline powder.Mp 125-130 ℃; IR (KBr) 3325,2935,1537,1209,1032,752,449cm -1 1H-NMR (CD 3OD) δ 3.05 (s, 1H), 3.31 (s, 2H), 3.77 (m, 3H), 3.80 (s, 1H), 3.84 (m, 3H), 4.14-4.26 (m, 2H), 6.80-6.87 (m, 3H), 7.01 (m, 1H), 7.10 (m, 2H), 7.39 (m, 2H), 7.80 (m, 1H), 8.07 (m, 1H); MS (FAB) m/z 510 (M ++ 1); C 26H 27FN 4O 60.5H 2The analytical calculation value of O: C, 60.11; H, 5.43; F, 3.66; N, 10.78.Measured value: C, 60.29; H, 5.40; F, 3.60; N, 10.59.Embodiment 242 (S)-3-amino-4-[2-[3-methoxyl group-4-[N '-(2-aminomethyl phenyl) urea groups] phenyl acetyl amino]-the 1-propoxyl group] benzoic acid
Figure A0080951005231
Under room temperature, to (S)-2-(N-tert-butoxycarbonyl amino)-1-propanol (0.90g, 5.13mmol), 4-hydroxyl-3-nitrobenzoic acid methyl ester (1.01g, 5.12mmol) and triphenyl phasphine (1.75g, 6.67mmol) the agitating solution of THF (15ml) in add azo-2-carboxylic acid's diisopropyl ester (DIAD) (1.31ml, 6.65mmol), the mixture reflux that obtains is spent the night.Evaporate this solution, residue is dissolved among dichloromethane (20ml) and the TFA (10ml).Under room temperature, stirred this mixture 3 hours.This mixture of vacuum concentration is dissolved in the chloroform residue.With saturated sodium bicarbonate, salt water washing, through dried over sodium sulfate and evaporation.Residue is through purification by silica gel column chromatography, with the chloroformic solution of 5% methanol as eluant, obtain 313mg (24%) (S)-3-nitro-4-(2-amino-1-propoxyl group) essence of Niobe, be faint yellow oily thing. 1H-NMR(CDCl 3)δ1.22(d，3H，J＝6.8Hz)，3.43-3.48(m，1H)，3.69-3.87(m，2H)，3.89(s，3H)，6.93-6.95(m，1H)，7.99-8.02(m，1H)，8.18-8.21(m，1H)。
With 3-methoxyl group-4-[N '-(2-aminomethyl phenyl) urea groups] pentafluorophenyl esters (591mg of phenylacetic acid, 1.23mmol), (S)-3-nitro-4-(2-amino-1-propoxyl group) essence of Niobe (313mg, 1.23mmol) and triethylamine (257ml, 1.84mmol) mixture in DMF (5ml) stirred 2 days under room temperature.Dilute this mixture with ethyl acetate, with 0.5N HCl, salt water washing, through dried over sodium sulfate and evaporation.Residue is through purification by silica gel column chromatography; with the chloroformic solution of 5% methanol as eluant; obtain 310mg (46%) (S)-3-nitro-4-[2-[3-methoxyl group-4-[N '-(2-aminomethyl phenyl) urea groups] phenyl acetyl amino]-the 1-propoxyl group] essence of Niobe, be jelly. 1H-NMR(CDCl 3)δ1.28(d，3H，J＝6.8Hz)，2.31(s，3H)，3.49(s，2H)，3.71(s，3H)，3.92(s，3H)，4.14-4.16(m，2H)，4.38-4.41(m，1H)，5.88-5.90(m，1H)，6.49(s，1H)，6.70-6.71(m，1H)，6.77-6.79(m，1H)，7.05-7.30(m，4H)，7.55-7.57(m，1H)，8.02-8.06(m，2H)，8.16-8.19(m，1H)，8.51-8.52(m，1H)；MS(FAB)m/z?551(M ++1)。
With (S)-3-nitro-4-[2-[3-methoxyl group-4-[N '-(2-aminomethyl phenyl) urea groups] phenyl acetyl amino]-the 1-propoxyl group] essence of Niobe (310mg; 0.56mmol) methanol-THF (10ml; 1: 1, (50mg, 16wt%) spent the night by hydrogenation through 5%Pd-C for agitating solution v/v).Filter this mixture, to remove catalyst, evaporated filtrate obtains (S)-3-amino-4-[2-[3-methoxyl group-4-[N '-(2-aminomethyl phenyl) urea groups] phenyl acetyl amino]-the 1-propoxyl group] essence of Niobe (240mg), be jelly.
(10ml 1: 1, adds 0.5N sodium hydroxide (10ml), with the mixture reflux that obtains 2 hours to the THF-methanol of crude product upward (220mg) in agitating solution v/v).In this mixture impouring ice-water, make water layer be acid (pH=4.8) by adding 1N HCl.Collect solid, make crude product solid recrystallization from methanol-chloroform-normal hexane, obtain 116mg (two steps, 44%) 293, be light yellow crystalline powder.Mp 200-204 ℃; 1H-NMR (DMSO-d 6) δ 1.21 (d, 3H, J=6.8Hz), 2.24 (s, 3H), 3.37 (s, 2H), 3.80 (s, 3H), 3.83-3.99 (m, 2H), 4.10-4.19 (m, 1H), 4.99 (bs, 2H), (6.75-7.23 a plurality of m, 8H altogether), 7.79 (d, 1H, J=7.8Hz), 7.98 (d, 1H, J=7.8Hz), 8.17 (d, 1H, J=8.3Hz), 8.46 (s, 1H), 8.55 (s, 1H); MS (FAB) m/z 507 (M ++ 1); C 27H 30N 4O 61/2H 2The analytical calculation value of O: C, 62.90; H, 6.06; N, 10.87.Measured value: C, 62.85; H, 6.10; N, 10.51.Embodiment 243 (S)-4-[2-[3-methoxyl group-4-[N '-(2-bromo phenyl) urea groups] phenyl acetyl amino]-the 1-propoxyl group] benzoic acid
Figure A0080951005241
To (S)-2-(N-tert-butoxycarbonyl amino)-1-propanol (6.74g, 0.04mol), 4-hydroxy benzoic acid benzyl ester (8.78g, 0.04mol) and triphenyl phasphine (15.13g, 0.06mol) cold (0 ℃) solution of stirring of THF (100ml) in add azo-2-carboxylic acid's diisopropyl ester (DIAD) (11.4ml, 0.06mol), the mixture reflux that obtains is spent the night.Evaporate this solution, residue is dissolved among dichloromethane (30ml) and the TFA (30ml).Under room temperature, stirred resulting solution 30 minutes.This solution of vacuum evaporation.Residue is dissolved in the chloroform, with the saturated sodium bicarbonate washing, through dried over sodium sulfate and evaporation.Residue is through purification by silica gel column chromatography, with chloroform to chloroform: methanol (9: 1, v/v) as eluant, obtain 6.63g (two steps, 60%) (S)-4-(2-amino-1-propoxyl group) benzyl benzoate, be yellow oil. 1H-NMR(CDCl 3)δ1.18(d，J＝6.3Hz，3H)，3.35-3.39(m，1H)，3.71-3.75(m，1H)，3.90-3.93(m，1H)，5.34(s，2H)，6.90-6.93(m，2H)，7.32-7.46(m，5H)，8.01-8.04(m，2H)。
With 3-methoxyl group-4-[N '-(2-bromo phenyl) urea groups] phenylacetic acid (480mg, 1.27mmol), (S)-4-(2-amino-1-propoxyl group) benzyl benzoate (361mg, 1.27mmol), EDC (hydrochlorate) (364mg, 1.90mmol), HOBt (256mg, 1.89mmol) and 4-DMAP (31mg, 0.25mmol) mixture in DMF (8ml) stirs under room temperature and spends the night.Dilute this mixture with ethyl acetate, with 0.5N HCl, salt water washing, through dried over sodium sulfate and evaporation.Residue is through purification by silica gel column chromatography; with the chloroformic solution of chloroform to 5% methanol as eluant; obtain 476mg (58%) (S)-4-[2-[3-methoxyl group-4-[N '-(2-bromo phenyl) urea groups] phenyl acetyl amino]-the 1-propoxyl group] benzyl benzoate, be brown solid. 1H-NMR(CDCl 3)δ1.25-1.28(m，3H)，3.51(s，2H)，3.74(s，3H)，3.95-3.97(m，2H)，4.36-4.39(m，1H)，5.33(s，2H)，6.75-6.94(m，5H)，7.26-7.70(m，8H)，7.99-8.26(m，5H)。
To (S)-4-[2-[3-methoxyl group-4-[N '-(2-bromo phenyl) urea groups] phenyl acetyl amino]-the 1-propoxyl group] benzyl benzoate (476mg; 1.39mmol) the agitating solution of THF (10ml) in add 0.5N sodium hydroxide (10ml), with the heating 2 hours under refluxing of the mixture that obtains.In this mixture impouring ice-1N HCl, collect solid.Make crude product solid recrystallization from methanol-chloroform-normal hexane, obtain 240mg (59%) 294, be white crystalline powder.Mp 202-205 ℃; 1H-NMR (DMSO-d 6) δ 1.18 (d, J=6.8Hz, 3H), 3.37 (s, 2H), 3.82 (s, 3H), 3.92-4.00 (m, 2H), 4.03-4.15 (m, 1H), and 6.77-6.79 (m, 1H), 6.93-7.03 (m, 4H), 7.30-7.34 (m, 1H), 7.59-7.61 (m, 1H), 7.87-7.97 (m, 4H), 8.13-8.15 (m, 1H), 8.73 (s, 1H), 8.91 (s, 1H), 12.63 (bs, 1H); MS (FAB) m/z 557 (M ++ 1); C 26H 26BrN 3O 6The analytical calculation value: C, 56.12; H, 4.71; Br, 14.36; N, 7.55.Measured value: C, 56.11; H, 4.74; Br, 14.56; N, 7.49.Embodiment 244 (S)-4-[[2-[3-methoxyl group-4-[N '-(2-aminomethyl phenyl) urea groups] phenyl acetyl]-(2-aminobenzyl) amino]-the 1-propoxyl group] benzoic acid
Figure A0080951005261
(1.50g, 5.26mmol) (0.87g, (16ml 15: 1, adds NaBH in cold (0 ℃) solution of stirring v/v) to methanol-acetic acid 5.76mmol) with the 2-nitrobenzaldehyde to (S)-4-(2-amino-1-propoxyl group) benzyl benzoate 3(1.65g's CN 26.3mmol), spends the night the mixture stirring that obtains under room temperature.By adding this mixture of saturated sodium bicarbonate quencher.Use ethyl acetate extraction.With salt water washing extract, through dried over sodium sulfate and evaporation.Residue is through purification by silica gel column chromatography, with the chloroformic solution of chloroform to 5% methanol as eluant, obtain 931mg (42%) (S)-4-[2-(2-nitrobenzyl amino)-1-propoxyl group] benzyl benzoate, be yellow oil. 1H-NMR(CDCl 3)δ1.21(d，J＝6.4Hz，3H)，3.13-3.18(m，1H)，3.88-3.97(m，2H)，4.06-4.20(m，2H)，5.34(s，2H)，6.89-6.94(m，2H)，7.29-7.65(m，8H)，7.94-8.03(m，3H)；MS(FAB)m/z?421(M ++1)。
With 3-methoxyl group-4-[N '-(2-aminomethyl phenyl) urea groups] phenylacetic acid pentafluorophenyl esters (460mg, 0.96mmol), (S)-4-[2-(2-nitrobenzyl amino)-1-propoxyl group] benzyl benzoate (403mg, 0.96mmol) and triethylamine (200ml, 1.43mmol) mixture in DMF (8ml) stirs under room temperature and spends the night.Dilute this mixture with ethyl acetate, with 0.5N HCl, salt water washing, through dried over sodium sulfate and evaporation.Residue is through purification by silica gel column chromatography, with the chloroformic solution of 1% methanol as eluant, obtain 504mg (73%) (S)-4-[[2-[3-methoxyl group-4-[N '-(2-aminomethyl phenyl) urea groups] phenyl acetyl]-(2-nitrobenzyl) amino]-the 1-propoxyl group] benzyl benzoate, be brown amorphous solid; MS (FAB) m/z 717 (M ++ 1).
To (S)-4-[[2-[3-methoxyl group-4-[N '-(2-aminomethyl phenyl) urea groups] phenyl acetyl]-(2-nitrobenzyl) amino]-the 1-propoxyl group] benzyl benzoate (504mg; 0.70mmol) methanol-THF (11ml; 10: 1; v/v) (100mg, 20wt%) spend the night under 3 atmospheric pressure by hydrogenation through 5%Pd-C for agitating solution.Filter this mixture, to remove catalyst, evaporated filtrate.Residue is through preparation property TLC purification, as eluant, obtains 115mg (27%) 295 with the chloroformic solution of 5% methanol, is white powder.MS(FAB)m/z?597(M ++1)。Embodiment 245 (S)-4-[[2-[3-methoxyl group-4-[N '-(2-bromo phenyl) urea groups] phenyl acetyl]-(2-nitrobenzyl) amino]-the 1-propoxyl group] benzoic acid
Figure A0080951005271
(1.50g, 5.26mmol) (0.87g, (16ml 15: 1, adds NaBH in cold (0 ℃) solution of stirring v/v) to methanol-acetic acid 5.76mmol) with the 2-nitrobenzaldehyde to (S)-4-(2-amino-1-propoxyl group) benzyl benzoate 3(1.65g's CN 26.3mmol), spends the night the mixture stirring that obtains under room temperature.By adding this mixture of saturated sodium bicarbonate quencher.Use ethyl acetate extraction.With salt water washing extract, through dried over sodium sulfate and evaporation.Residue is through purification by silica gel column chromatography, with the chloroformic solution of chloroform to 5% methanol as eluant, obtain 931mg (42%) (S)-4-[2-(2-nitrobenzyl amino)-1-propoxyl group] benzyl benzoate, be yellow oil. 1H-NMR(CDCl 3)δ1.21(d，3H，J＝6.4Hz)，3.13-3.18(m，1H)，3.88-3.97(m，2H)，4.06-4.20(m，2H)，5.34(s，2H)，6.89-6.94(m，2H)，7.29-7.65(m，8H)，7.94-8.03(m，3H)；MS(FAB)m/z?421(M ++1)。
With 3-methoxyl group-4-[N '-(2-bromo phenyl) urea groups] phenylacetic acid (476mg, 1.26mmol), (S)-4-[2-(2-nitrobenzyl amino)-1-propoxyl group] benzyl benzoate (528mg, 1.26mmol), EDC (hydrochlorate) (361mg, 1.88mmol), HOBt (255mg, 1.89mmol) and DMAP (30mg, 0.25mmol) mixture in DMF (10ml) stirs under room temperature and spends the night and stirred 1 day in 60 ℃.Dilute this mixture with ethyl acetate, with 0.5N HCl, salt water washing, through dried over sodium sulfate and evaporation.Residue is through purification by silica gel column chromatography; with the chloroformic solution of chloroform to 2% methanol as eluant; obtain (S)-4-[[2-[3-methoxyl group-4-[N '-(2-bromo phenyl) urea groups] phenyl acetyl]-(2-nitrobenzyl) amino]-the 1-propoxyl group] benzyl benzoate, be grease.It can need not to be further purified and be used for subsequent reaction.
(10ml, 1: 1, adding 0.5N sodium hydroxide (10ml) in agitating solution v/v) heated the mixture that obtains 3 hours under refluxing the THF-methanol of crude product upward.In this mixture impouring ice-water, make alkaline water layer be acid (pH4.3) with 1N HCl.Collect solid, make the crude product solid, as eluant, obtain 162mg (two go on foot 19%) 296, be white amorphous solid with the chloroformic solution of 5% methanol through preparation property TLC purification.MS (FAB) m/z 692 (M ++ 1); C 33H 31BrN 4O 87/4H 2The analytical calculation value of O: C, 54.82; H, 4.81; N, 7.75.Measured value: C, 54.80; H, 4.61; N, 7.24.Embodiment 2464-[2-N-[3-methoxyl group-4-[N '-(2-aminomethyl phenyl) urea groups] the phenyl acetylamino] ethyoxyl] benzoic acid
Figure A0080951005281
With 5 minutes, to 2-(N-tert-butoxycarbonyl amino) ethanol (3.20g, 19.9mmol), the 4-methyl hydroxybenzoate (3.02g, 19.9mmol) and triphenyl phasphine (6.25g drips DIAD (4.69ml.23.8mmol) in cold (0 ℃) solution of the stirring of THF 23.8mmol) (50ml).With reaction mixture refluxed heating 3 hours.Evaporate this mixture, residue is dissolved among dichloromethane (30ml) and the TFA (30ml).Stirring this reactant mixture under room temperature spends the night.This mixture of vacuum evaporation is dissolved in chloroform and the water residue.Make this solution be alkalescence with saturated sodium bicarbonate, use chloroform extraction.With salt water washing extract, through dried over sodium sulfate and evaporation.Residue is used chloroform through purification by silica gel column chromatography: and methanol (30: 1, v/v) as eluant, obtain 4-(2-amino ethoxy) essence of Niobe (1.03g, two steps 34%), be colorless oil. 1H-NMR(CDCl 3)δ3.10-3.13(m，2H)，3.89(s，3H)，4.03-4.06(m，2H)，6.92-6.94(m，2H)，7.98-8.00(m，2H)。
With 3-methoxyl group-4-[N '-(2-aminomethyl phenyl) urea groups] phenylacetic acid (557mg, 1.77mmol), 4-(2-amino ethoxy) essence of Niobe (346mg, 1.77mmol), EDCHCl (408mg, 2.13mmol), HOBt (287mg, 2.12mmol) and DMAP (52mg, 0.43mmol) mixture in DMF (10ml) stirred 2 days under room temperature.Dilute this mixture with ethyl acetate and water.Collect the precipitation that produces, obtain 4-[2-N-[3-methoxyl group-4-[N '-(2-aminomethyl phenyl) urea groups] the phenyl acetylamino] ethyoxyl] essence of Niobe (598mg, 69%), be white crystalline powder. 1H-NMR (DMSO-d 6) δ 2.23 (s, 3H), 3.36 (s, 2H), 3.42-3.45 (m, 2H), 3.79 (s, 3H), 3.81 (s, 3H), 4.02-4.09 (m, 2H), 6.73-6.75 (m, 1H), 6.90-6.94 (m, 2H), 7.02-7.04 (m, 2H), and 7.09-7.15 (m, 2H), 7.77-7.79 (m, 1H), 7.88-7.90 (m, 2H), 7.95-7.98 (m, 1H), 8.23-8.24 (m, 1H), 8.44 (s, 1H), 8.53 (s, 1H); MS (FAB) m/z 492 (M ++ 1); C 27H 29N 3O 61/4H 2The analytical calculation value of O: C, 65.38; H, 5.99; N, 8.47.Measured value: C, 65.26; H, 5.99; N, 8.49.
To 4-[2-N-[3-methoxyl group-4-[N '-(2-aminomethyl phenyl) urea groups] the phenyl acetylamino] ethyoxyl] essence of Niobe (280mg, 0.57mmol) THF-methanol (10ml, 1: 1, v/v) add 0.5N sodium hydroxide (10ml) in the agitating solution, reactant mixture was heated 5 hours under refluxing.This mixture is cooled among room temperature and the impouring ice-1N HCl.The precipitation that collect to produce and from E-C-methanol recrystallization, obtain 297 (135mg, 50%), be white crystalline powder.MW 477.51 1H-NMR (DMSO-d 6) δ 2.24 (s, 3H), 3.38 (s, 2H), 3.43-3.47 (m, 2H), 3.82 (s, 3H), 4.06-4.09 (m, 2H), 6.76-6.78 (m, 1H), 6.92-7.17 (m, 6H), 7.79 (d, J=8.1Hz, 1H), 7.88-7.89 (m, 2H), 7.98 (d, J=8.1Hz, 1H), 8.24-8.27 (m, 1H), 8.45 (s, 1H), 8.55 (s, 1H); MS (FAB) m/z 478 (M ++ 1); C 26H 27N 3O 61/4H 2The analytical calculation value of O: C, 64.79; H, 5.75; N, 8.72.Measured value: C, 64.67; H, 5.63; N, 8.60.Embodiment 247 (S)-4-[2-N-[4-[N '-(2-bromo phenyl) urea groups]-3-methoxyphenyl acetylamino]-the 1-propoxyl group] benzoic acid
To (S)-2-(N-tert-butoxycarbonyl amino)-1-propanol (6.74g, 0.04mol), 4-hydroxy benzoic acid benzyl ester (8.78g, 0.04mol) and triphenyl phasphine (15.13g, add in cold (0 ℃) solution of the stirring of THF 0.06mol) (100ml) DIAD (11.4ml, 0.06mol).With the reaction mixture refluxed heated overnight.Evaporate this solution, residue is dissolved among dichloromethane (30ml) and the TFA (30ml).Under room temperature, stirred this solution 30 minutes and this solution of vacuum concentration.Residue is dissolved in the chloroform, with the saturated sodium bicarbonate washing, through dried over sodium sulfate and evaporation.Residue is through purification by silica gel column chromatography, with chloroform to chloroform: methanol (9: 1,, obtain (S)-4-(2-amino-1-propoxyl group) benzyl benzoate (6.63g, two steps, 60%) v/v) as eluant, be yellow oil. 1H-NMR(CDCl 3)δ1.18(d，J＝6.3Hz，3H)，3.35-3.39(m，1H)，3.71-3.75(m，1H)，3.90-3.93(m，1?H)，5.34(s，2H)，6.90-6.93(m，2H)，7.32-7.46(m，5H)，8.01-8.04(m，2H)。
With 4-[N '-(2-bromo phenyl) urea groups]-3-methoxybenzene guanidine-acetic acid (480mg, 1.27mmol), (S)-4-(2-amino-1-propoxyl group) benzyl benzoate (361mg, 1.27mmol), EDCHCl (364mg, 1.90mmol), HOBt (256mg, 1.89mmol) and DMAP (31mg, 0.25mmol) mixture in DMF (8ml) stirs under room temperature and spends the night.Dilute this mixture with ethyl acetate, with 0.5N HCl, salt water washing, through dried over sodium sulfate and evaporation.Residue is through purification by silica gel column chromatography, with the chloroformic solution of chloroform to 5% methanol as eluant, obtain (S)-4-[2-[4-[N '-(2-bromo phenyl) urea groups]-3-methoxyphenyl acetylamino]-the 1-propoxyl group] benzyl benzoate (476mg, 58%), be brown solid. 1H-NMR(CDCl 3)δ1.25-1.28(m，3H)，3.51(s，2H)，3.74(s，3H)，3.95-3.97(m，2H)，4.36-4.39(m，1H)，5.33(s，2H)，6.75-6.94(m，5H)，7.26-7.70(m，8H)，7.99-8.26(m，5H)。
To (S)-4-[2-[4-[N '-(2-bromo phenyl) urea groups]-3-methoxyphenyl acetylamino]-the 1-propoxyl group] benzyl benzoate (476mg, 1.39mmol) the agitating solution of THF (10ml) in add 0.5N sodium hydroxide (10ml), with reactant mixture heating 2 hours under refluxing.In this mixture impouring ice-1N HCl, collect the precipitation that produces.The crude product solid obtains 298 (240mg, 59%) through recrystallization purifying from methanol-chloroform-normal hexane, is white crystalline powder.MW556.41 1H-NMR (DMSO-d 6) δ 1.18 (d, J=6.8Hz, 3H), 3.37 (s, 2H), 3.82 (s, 3H), 3.92-4.00 (m, 2H), 4.03-4.15 (m, 1H), and 6.77-6.79 (m, 1H), 6.93-7.03 (m, 4H), 7.30-7.34 (m, 1H), 7.59-7.61 (m, 1H), 7.87-7.97 (m, 4H), 8.13-8.15 (m, 1H), 8.73 (s, 1H), 8.91 (s, 1H), 12.63 (bs, 1H); MS (FAB) m/z 557 (M ++ 1); C 26H 26BrN 3O 6The analytical calculation value: C, 56.12; H, 4.71; Br, 14.36; N, 7.55.Measured value: C, 56.11; H, 4.74; Br, 14.56; N, 7.49.Embodiment 248 (S)-4-[2-N-[[3-methoxyl group-4-[N '-(2-aminomethyl phenyl) urea groups] phenyl]-N-methyl acetylamino]-the 1-propoxyl group] benzoic acid
To (S)-2-[(N-tert-butoxycarbonyl) amino]-1-propanol (3.08g, 17.6mmol), 4-methyl hydroxybenzoate (2.67g, 17.6mmol) and triphenyl phasphine (5.53g, 21.1mmol) cold (0 ℃) solution of stirring of THF (35ml) in be added dropwise to DIAD (4.15ml, 21.1mmol).With the reaction mixture refluxed heated overnight.Evaporate this mixture, residue is used normal hexane through purification by silica gel column chromatography: ethyl acetate (5: 1,, obtain (S)-4-[2-[(N-tert-butoxycarbonyl v/v) as eluant) amino]-the 1-propoxyl group] essence of Niobe (1.24g, 23%), be white solid. 1H-NMR(CDCl 3)δ1.30(d，J＝6.8Hz，3H)，1.45(s，9H)，3.89(s，3H)，3.98-3.99(m，2H)，4.07(m，1H)，4.76(m，1H)，6.91-6.93(m，2H)，7.98-8.00(m，2H)；MS(FAB)m/z?310(M ++1)。
To (S)-4-[2-[(N-tert-butoxycarbonyl) amino]-the 1-propoxyl group] (1.24g adds TFA (10ml) to essence of Niobe in the agitating solution of dichloromethane 4.01mmol) (10ml), reactant mixture is stirred under room temperature spend the night.This mixture of vacuum concentration, with saturated sodium bicarbonate make be alkalescence.With this mixture of chloroform extraction, use the salt water washing, dry and evaporation obtains (S)-4-(2-amino-1-propoxyl group) essence of Niobe (790mg, 94%) through potassium carbonate, is yellow oil. 1H-NMR (CDCl 3) δ 1.19 (d, J=6.7Hz, 3H), 3.34-3.42 (m, 1H), 3.70-3.77 (m, 1H), 3.86-3.94 (a plurality of s and m, 4H altogether), 6.92 (d, J=9.0Hz, 2H), 7.98 (d, J=9.0Hz, 2H).
To (S)-4-(2-amino-1-propoxyl group) essence of Niobe (790mg, 3.78mmol) and triethylamine (630ml, 4.52mmol) cold (0 ℃) solution of THF (10ml) in add TFAA (640ml 4.53mmol), stir reactant mixture 4 days under room temperature.Dilute this mixture with 0.5N HCl, use chloroform extraction.With salt water washing extract, the dry and evaporation through potassium carbonate.Residue is recrystallization purifying from normal hexane-chloroform, obtains (S)-4-[2-(trifluoroacetamido)-1-propoxyl group] essence of Niobe (790mg, 69%), be white crystalline material. 1H-NMR (CDCl 3) δ 1.42 (d, J=6.8Hz, 3H), 3.89 (s, 3H), 4.01-4.13 (m, 2H), 4.44-4.50 (m, 1H), 6.57-6.61 (m, 1H), 6.92 (d, J=9.0Hz, 2H), 8.00 (d, J=9.0Hz, 2H); MS (FAB) m/z 306 (M ++ 1); C 13H 14F 3NO 4The analytical calculation value: C, 51.15; H, 4.62; F, 18.67; N, 4.59.Measured value: C, 51.14; H, 4.60; F, 18.50; N, 4.54.
To (S)-4-[2-(trifluoroacetamido)-1-propoxyl group] essence of Niobe (695mg, 2.28mmol) and potassium carbonate (630mg, 4.56mmol) the agitating solution of DMF (10ml) in add MeI (210ml 3.37mmol), stir reactant mixture 2 days under room temperature.This mixture of dilute with water is used ethyl acetate extraction.With salt water washing extract, through dried over sodium sulfate and evaporation.Residue is through purification by silica gel column chromatography, with ethyl acetate-chloroform (1: 19,, obtain (S)-4-[2-[(N-methyl v/v) as eluant) trifluoroacetamido]-the 1-propoxyl group] essence of Niobe (720mg, 99%), be white solid.Mp 73-75 ℃; 1H-NMR (CDCl 3) δ 1.36-1.39 (m, 3H), 2.96 and 3.10 (they respectively are s, are total to 3H), 3.89 (s, 3H), 3.99-4.14 (m, 2H), 4.84-4.92 (m, 1H), 6.88-6.92 (m, 2H), 7.98-8.00 (m, 2H); MS (FAB) m/z 320 (M ++ 1); C 14H 16F 3NO 4The analytical calculation value: C, 52.67; H, 5.05; F, 17.85; N, 4.39.Measured value: C, 52.76; H, 5.09; F, 17.53; N, 4.32.
To (S)-4-[2-[(N-methyl) trifluoroacetamido]-the 1-propoxyl group] essence of Niobe (710mg, methanol-water 2.22mmol) (1 0ml, 1: 1, v/v) add potassium carbonate (460mg in the agitating solution, 3.33mmol), under room temperature, the reactant mixture stirring is spent the night.The dilute with water mixture is used ethyl acetate extraction.With salt water washing extract,, obtain (S)-4-[2-(N-methylamino)-1-propoxyl group through dried over sodium sulfate and evaporation] essence of Niobe (430mg, 87%), be colorless oil. 1H-NMR (CDCl 3) δ 1.17 (d, J=6.6Hz, 3H), 2.48 (s, 3H), 2.98-3.06 (m, 1H), 3.86-3.96 (a plurality of s and m, 5H altogether), 6.92 (d, J=9.0Hz, 2H), 7.98 (d, J=9.0Hz, 2H).
To 3-methoxyl group-4-[N '-(2-aminomethyl phenyl) urea groups] phenylacetic acid (605mg, 1.93mmol), (S)-4-[2-(N-methylamino)-1-propoxyl group] essence of Niobe (430mg, 1.93mmol), EDCHCl (444mg, 2.32mmol), HOBt (313mg, 2.32mmol), (320ml, 2.30mmol) mixture in THF (13ml) stirs under room temperature and spends the night triethylamine.This mixture of dilute with water is used ethyl acetate extraction.With salt water washing extract, through dried over sodium sulfate and evaporation.Residue is through purification by silica gel column chromatography, with chloroform-methanol (100: 1 to 75: 1, v/v) as eluant, obtain (S)-4-[2-N-[[3-methoxyl group-4-[N '-(2-aminomethyl phenyl) urea groups] phenyl]-N-methyl acetylamino]-the 1-propoxyl group] essence of Niobe (953mg, 95%), is the white foam thing. 1H-NMR (CDCl 3) δ 1.12-1.13 and 1.25-1.27 (respectively be m, be total to 3H), 2.27 (s, 3H), 2.84 with 2.92 (they respectively are s, are total to 3H), 3.63 (s, 3H), 3.67 (s, 2H), 3.71-4.05 (a plurality of s and m, 5H altogether), 4.39-4.44 and 4.96-5.01 (respectively be m, altogether 1H), 6.66-6.85 (m, 5H), 7.09-7.27 (m, 4H), 7.53-7.55 (m, 1H), 7.92-7.98 (m, 2H), 8.04-8.08 (m, 1H); MS (FAB) m/z 520 (M ++ 1); C 29H 33N 3O 611/4H 2The analytical calculation value of O: C, 61.20; H, 6.82; N, 7.38.Measured value: C, 61.14; H, 5.86; N, 7.16.
To (S)-4-[2-N-[[3-methoxyl group-4-[N '-(2-aminomethyl phenyl) urea groups] phenyl]-N-methyl acetylamino]-the 1-propoxyl group] essence of Niobe (663mg, 1.28mmol) the agitating solution of THF (5ml) in add 0.5N sodium hydroxide (5ml), with reaction mixture refluxed heating 5 hours.After being cooled to room temperature, in this mixture impouring ice-1N HCl, the precipitation that produces is collected in decompression.Make the crude product solid be dissolved in the chloroform and evaporation.With ether wash residual thing, obtain 299 (465mg, 72%), be white amorphous solid.MW 505.56 1H-NMR (DMSO-d 6) δ 1.11-1.15 (m, 3H), 2.25 (s, 3H), 2.73 and 2.88 (they respectively are s, are total to 3H), 3.60-3.76 (m, 2H), 3.83 (s, 3H), 4.03-4.12 (m, 2H), 441-4.50 and 4.48-4.94 (respectively be m, altogether 1H), 6.71-6.76 (m, 1H), 6.84-6.86 (m, 1H), and 6.91-7.01 (m, 3H), 7.11-7.12 (m, 2H), 7.78-7.80 (m, 1H), 7.86-7.90 (m, 2H), 8.01-8.03 (m, 1H), 8.45 (s, 1H), 8.54 (s, 1H); MS (FAB) m/z 520 (M ++ 1); C 28H 31N 3O 63/2H 2The analytical calculation value of O: C, 63.15; H, 6.43; N, 7.89.Measured value: C, 63.09; H, 5.99; N, 7.64.Embodiment 2494-[2-N-[3-methoxyl group-4-[N '-(2-aminomethyl phenyl) urea groups] the phenyl acetylamino]-2-methyl isophthalic acid-propoxyl group] benzoic acid
Figure A0080951005341
Under nitrogen current, with 4-methyl hydroxybenzoate (3g, 19.72mmol), potassium carbonate (6.8g, 49.3mmol), 3-chloro neopentanoic acid (2.9g, 21.69mmol) and the KI (200mg) of catalytic amount stirring the mixture in DMF (70ml) in 100 ℃ of heating 14 days.In this mixture impouring frozen water, use ethyl acetate extraction.With salt water washing extract, through dried over sodium sulfate and evaporation.Residue is used chloroform through silica gel (50ml) chromatography: and ethanol (10: 1, v/v) as eluant, obtain title compound (1g, 23%), be amorphous solid. 1H-NMR(CDCl 3)δ1.37(s，6H)，3.89(s，3H)，4.03(s，2H)，6.92(d，J＝9Hz，2H)，7.98(d，J＝9Hz，2H)。
Under room temperature; to 2; 2-dimethyl-3-(4-methoxycarbonyl) phenoxy propionic acid (720mg; 2.85mmol) and triethylamine (0.46ml; 3.28mmol) the tert-butyl alcohol (10ml) and the stirring the mixture of benzene (10ml) in add diphenyl phosphoryl azide (870mg, benzene 3.14mmol) (3ml) solution.With the mixture reflux that obtains 20 hours.After the cooling, will ice and 1N HCl (5ml) adds in this mixture and uses methylbenzene extraction.With salt water washing extract, through dried over sodium sulfate and evaporation.Residue is used toluene through silica gel (50ml) chromatography: and ethyl acetate (10: 1,, obtain 4-[1-(2-amino-2-methyl) propoxyl group v/v) as eluant] essence of Niobe, be jelly (520mg) that it can need not to be further purified and be used for subsequent reaction. 1H-NMR (CDCl 3) δ 1.41 (s, 9H), 3.89 (s, 3H), 4.04 (s, 2H), 4.69 (br s, 1H), 6.94 (dd, J=2 and 7Hz, 2H), 7.98 (dd, J=2 and 7Hz).
Under room temperature, with 4-[1-(2-methyl-2-tert-butoxycarbonyl amino-) propoxyl group] essence of Niobe (520mg) and methyl phenyl ethers anisole (0.175ml, the solution stirring of dichloromethane 1.61mmol) (5ml) and TFA (3ml) 18 hours.Evaporate this mixture.Residue is dissolved in the dichloromethane, makes this mixture be alkalescence by adding saturated sodium bicarbonate.Through sodium sulfate and dry isolating dichloromethane layer of sodium carbonate and evaporation.Residue is through silica gel column chromatography, with chloroform-ethanol (10: 1,, obtain 4-[1-(2-amino-2-methyl) propoxyl group v/v) as eluant] essence of Niobe (250mg, two steps 39%), be jelly. 1H-NMR(CDCl 3)δ3.75(s，2H)，3.89(s，3H)，6.93(d，J＝8.8Hz，2H)，7.98(d，J＝8.8Hz，2H)。
Under room temperature, to 4-[1-(2-amino-2-methyl) propoxyl group] essence of Niobe (250mg, 1.12mmol), 3-methoxyl group-4-[N '-(2-aminomethyl phenyl) urea groups] phenylacetic acid (352mg, 1.12mmol), 4-DMAP (165mg, 1.34mmol) the stirring the mixture of DMF (10ml) in add EDCHCl (290mg, 1.51mmol).The mixture that obtains was stirred under room temperature 18 hours.In this mixture impouring ice-water.Collect solid, wash with water also air-dry.The crude product solid is through purification by silica gel column chromatography, use chloroform: ethanol (4: 1,, obtain 4-[2-N-[3-methoxyl group-4-[N '-(2-aminomethyl phenyl) urea groups v/v) as eluant] the phenyl acetylamino]-2-methyl isophthalic acid-propoxyl group] essence of Niobe (580mg, q.y.), be crystalline material.IR (KBr) 3350,3286,1712,1687,1637,1606cm -1 1H-NMR (CDCl 3) δ 1.39 (s, 6H), 2.33 (s, 3H), 3.41 (s, 2H), 3.63 (s, 3H), 3.88 (s, 3H), 4.05 (s, 2H), 5.44 (br s, 1H), 6.33 (br s, 1H), 6.79 (d, J=8.3Hz, 2H), 7.12 (s, 1H), 7.1 8 (t, J=7.5Hz, 1H), 7.53 (d, J=7.8Hz, 1H), 7.94 (d, J=7.8Hz, 2H), 8.14 (d, J=8.3Hz, 1H); MS (FAB) m/z 520 (M ++ 1); C 29H 33N 3O 6The analytical calculation value: C, 67.04; H, 6.40; N, 8.09.Measured value: C, 66.86; H, 6.36; N, 8.22.
Under room temperature, to 4-[2-[3-methoxyl group-4-[N '-(2-aminomethyl phenyl) urea groups] the phenyl acetylamino]-2-methyl-2-propoxyl group] essence of Niobe (510mg, and adding 0.25N sodium hydroxide in agitating solution 0.98mmol) (8ml, 2mmol).The mixture that obtains was stirred under ambient temperature 18 hours.In this mixture impouring ice-1N HCl (5ml).Collect solid, wash with water also air-dry.Crude product solid recrystallization from chloroform-alcohol-ether obtains 300 (480mg, 97%), is thin spicule.MW 505.56 IR (KBr) n 3346,3294,1687,1637,1604cm -1 1H-NMR (DMSO-d 6) δ 1.35 (s, 6H), 2.24 (s, 3H), 3.33 (s, 2H), 3.80 (s, 3H), 4.15 (s, 2H), 6.75 (d, J=8.3Hz, 1H), 6.88 (s, 1H), 6.95-6.99 (m, 3H), 7.11-7.17 (m, 3H), 7.80 (d, J=8.3Hz, 1H), 7.82 (s, 1H), 7.87 (d, J=8.8Hz, 2H), 7.98 (d, J=7.8Hz, 1H), 8.45 (s, 1H), 8.54 (s, 1H), 12.62 (br s, 1H); MS (FAB) m/z 506 (M ++ 1); C 28H 31N 3O 6The analytical calculation value: C, 66.52; H, 6.18; N, 8.31.Measured value: C, 66.22; H, 6.28; N, 8.11.Embodiment 2503-amino-4-[2-N-[4-[N '-(2-chlorophenyl) urea groups]-3-methoxyphenyl acetylamino]-2-methyl isophthalic acid-propoxyl group] benzoic acid
In 0-5 ℃, (10g adds concentrated sulphuric acid in the agitating solution of acetic acid 64.88mmol) (70ml) and DMSO (20ml) to 4-amino-2-nitrophenol.Being lower than under 20 ℃ the temperature, drip sodium nitrite (5.4g, water 77.9mmol) (5ml) solution in the solution with this stirring of clockwise in 10 fens.The mixture that obtains in 5 ℃ of restir 0.5 hour.(30g was 0.182mol) and in the agitating solution of the ice-water (200ml) of the Cu powder (200mg) of catalytic amount with this mixture impouring KI with 10 minutes.The mixture that restir obtains under room temperature 1 hour.With this mixture of dichloromethane extraction.Wash extract in proper order with saturated sodium thiosulfate and saline.Through dried over sodium sulfate organic layer and evaporation.Residue is used chloroform through silica gel (50ml) chromatography: and ethyl acetate (3: 1, v/v) as eluant, obtain 4-iodo-2-nitrophenol (2.5g, 15%), be the yellow crystal material. 1H-NMR (CDCl 3) δ 6.94 (d, J=8.8Hz, 1H), 7.82 (dd, J=2 and 8.8Hz, 1H), 8.42 (d, J=2Hz, 1H), 10.49 (s, 1H).
Under the ice-water bath cooling, to 4-iodo-2-nitrophenol (2g, 7.75mmol), the hydroxy new pentane acid methyl ester (1.05g, 7.92mmol) and triphenyl phasphine (2.3g, 8.68mmol) the agitating solution of THF (10ml) in drip DIAD (1.77g, THF 8.30mmol) (2ml) solution.Then with the mixture reflux that obtains 18 hours.After the cooling, evaporate this mixture.Residue is used toluene through silica gel (100ml) chromatography: and ethyl acetate (4: 1, v/v) as eluant, obtain 3-(4-iodo-2-nitro) phenoxy group-2, (2.9g q.y.), is crystalline material to the 2-dimethylated methyl propionate. 1H-NMR (CDCl 3) δ 1.34 (s, 6H), 3.71 (s, 3H), 4.08 (s, 2H), 6.86 (d, J=8.8Hz, 1H), 7.78 (dd, J=2 and 8.8Hz, 1H), 8.12 (d, J=2Hz, 1H).
Under room temperature, with 3-(4-iodo-2-nitro) phenoxy group-2, (2.8g, 7.38mmol) (60ml, 15mol) mixture in stirred 18 hours the 2-dimethylated methyl propionate at THF (15ml) and 0.25N sodium hydroxide.In this mixture impouring ice-1N HCl (20ml).Collect solid, wash with water also air-dry.The crude product solid obtains 3-(4-iodo-2-nitro) phenoxy group-2 through recrystallization from chloroform-ethanol-IPE, and 2-neopentanoic acid (2.0g, 74%) is crystalline material.Mp 165-182 ℃; IR (KBr) n 1716,1525,1344cm -1 1H-NMR (CDCl 3) δ 1.38 (s, 6H), 4.10 (s, 2H), 6.86 (d, J=8.8Hz, 1H), 7.79 (dd, J=2.2 and 8.8Hz, 1H), 8.12 (d, J=2Hz, 1H); MS (FAB) m/z 366 (M ++ 1); C 29H 33N 3O 6The analytical calculation value: C, 36.18; H, 3.18; N, 3.84.Measured value: C, 36.85; H, 3.35; N, 3.79.
Under room temperature; to 3-(4-iodo-2-nitro) phenoxy group-2; 2-neopentanoic acid (1.93g; 5.29mmol) and triethylamine (590mg; 5.81mmol) add diphenyl phosphoryl azide (1.53g, toluene 5.55mmol) (3ml) solution in the stirring the mixture in the tert-butyl alcohol (15ml) and toluene (15ml).Then with the mixture reflux that obtains 20 hours.After the cooling, will ice and 1N HCl (5ml) adds in this mixture and uses methylbenzene extraction.With salt water washing extract, through dried over sodium sulfate and evaporation.Residue is used toluene through silica gel (50ml) chromatography: and ethyl acetate (10: 1, v/v) as eluant, obtain 3-nitro-4-(2-tert-butoxycarbonyl amino-2-methyl-1-propoxyl group) iodobenzene (1.91g, 83%), be jelly. 1H-NMR (CDCl 3) δ 1.38 (s, 9H), 1.39 (s, 6H), 4.19 (s, 2H), 4.67 (br s, 1H), 6.88 (d, J=8.8Hz, 1H), 7.77 (dd, J=2.0 and 8.8Hz, 1H), 8.12 (d, J=2.0Hz, 1H).
In 70 ℃, CO (gas) flows down, with 3-nitro-4-(2-tert-butoxycarbonyl amino-2-methyl-1-propoxyl group) iodobenzene (1.9g, 4.36mmol), Pd (OAc) 2With 1, two (diphenylphosphino) propane (dppp) of 3-(90mg, 0.22mmol) triethylamine-methanol-DMSO (1: 2: 5, v/v, 48ml) mixture in stirred 6 hours.After the cooling, in this mixture impouring water, use ethyl acetate extraction.With salt water washing extract, through dried over sodium sulfate and evaporation.Residue is used toluene through silica gel (50ml) chromatography: and ethyl acetate (6: 1, v/v) as eluant, obtain 4-(2-tert-butoxycarbonyl amino-2-methyl-1-propoxyl group)-3-nitrobenzoic acid methyl ester (820mg, 51%), be jelly. 1H-NMR (CDCl 3) δ 1.38 (s, 9H), 1.42 (s, 6H), 3.93 (s, 3H), 4.29 (s, 2H), 4.67 (br s, 1H), 7.15 (d, J=8.8Hz, 1H), 8.18 (dd, J=1.7 and 8.8Hz, 1H), 8.52 (d, J=1.7Hz, 1H).
Under room temperature, with 4-(2-tert-butoxycarbonyl amino-2-methyl-1-propoxyl group)-3-nitrobenzoic acid methyl ester (350mg, 0.95mmol) and stir the mixture in normal pressure hydrogen under the hydrogenation 20 hour of 5%Pd-C (70mg) in ethanol (30ml).Insoluble Pd-catalyst is removed in suction, uses washing with alcohol.Evaporated filtrate obtains 4-(2-tert-butoxycarbonyl amino-2-methyl)-1-propoxyl group-3-Methyl anthranilate, is jelly, need not to be further purified and is used for subsequent reaction. 1H-NMR (CDCl 3) δ 1.41 (s, 9H), 1.43 (s, 6H), 3.86 (s, 3H), 4.07 (s, 2H), 4.67 (br s, 1H), 6.80 (d, J=8.5Hz, 1H), 7.39 (dd, J=2.2Hz, 1H), 7.44 (dd, J=2.2 and 8.5Hz, 1H).
In 0-5 ℃, to above 4-(2-tert-butoxycarbonyl amino-2-methyl)-1-propoxyl group-3-Methyl anthranilate and triethylamine (0.20ml, 1.43mmol) add trifluoroacetic anhydride (0.182ml, dichloromethane 1.28mmol) (3ml) solution in the stirring the mixture in dichloromethane (10ml).Under room temperature, the mixture that obtains was stirred 1 hour.To ice-saturated sodium bicarbonate joins in this mixture, uses dichloromethane extraction.With salt water washing extract, through dried over sodium sulfate and evaporation.Residue is dissolved in the dichloromethane (5ml), add methyl phenyl ethers anisole (0.105ml, 0.95mmol) and TFA (2ml).The mixture that obtains was stirred under room temperature 18 hours.This mixture of vacuum evaporation.Dilute residue with dichloromethane, make by the adding saturated sodium bicarbonate to be alkalescence.Through isolating dichloromethane layer of dried over sodium sulfate and evaporation.Residue is through silica gel (50ml) chromatography, with chloroform-ethanol (99: 1, v/v) as eluant, obtain 4-(2-amino-2-methyl-1-propoxyl group)-3-trifluoroacetamido essence of Niobe (631mg, three steps 63%), be jelly. 1H-NMR (CDCl 3) δ 1.29 (s, 9H, t-Bu), 3.86 (s, 2H, CH2), 3.91 (s, 3H), 6.99 (d, J=8.5Hz, 1H), 7.91 (dd, J=2.0 and 8.5Hz, 1H), 8.83 (d, J=8.5Hz, 1H).
Under room temperature, to 4-(2-amino-2-methyl)-1-propoxyl group-3-trifluoroacetamido essence of Niobe (200mg, 0.598mmol), 4-[N '-(2-chlorophenyl) urea groups]-3-methoxybenzene guanidine-acetic acid (210mg, 0.598mmol), 4-DMAP (90mg, 0.72mmol) add in the stirring the mixture in DMF (7ml) EDCHCl (160mg, 0.81mmol).The mixture that obtains was stirred under room temperature 20 hours.In this mixture impouring ice-water.Collect solid, wash with water also air-dry.The crude product solid is through purification by silica gel column chromatography, use chloroform: ethanol (98: 2, v/v) as eluant, obtain 4-[2-N-[4-[N '-(2-chlorophenyl) urea groups]-3-methoxyphenyl acetylamino]-2-methyl isophthalic acid-propoxyl group]-3-trifluoroacetamido essence of Niobe (580mg, q.y.), be crystalline material. 1H-NMR (CDCl 3) δ 1.42 (s, 6H), 3.42 (s, 2H), 3.69 (s, 3H), 3.89 (s, 3H), 4.23 (s, 2H), 5.33 (br s, 1H), 6.66 (s, 1H), 6.71 (m, 1H), 6.92 (d, J=8.5Hz, 1H), 7.02 (m, 1H), 7.09 (m, 2H), 7.29 (m, 1H), 7.37 (d, J=8.0Hz, 1H), 7.87 (dd, J=2 and 8.5Hz, 1H), 7.97 (d, J=8.0Hz, 1H), 8.19 (d, J=8.2Hz, 1H), 8.59 (br s, 1H), 8.74 (d, J=2.0Hz, 1H).
Under room temperature, to 4-[2-N-[4-[N '-(2-chlorophenyl) urea groups]-3-methoxyphenyl acetylamino]-2-methyl isophthalic acid-propoxyl group]-3-trifluoroacetamido essence of Niobe (320mg, 0.492mmol) the agitating solution of THF (2ml) in add the 0.25N sodium hydroxide (6ml, 1.5mmol).The mixture that obtains was stirred 20 hours.In this mixture impouring ice-1N HCl (2ml).Collect solid, wash with water also air-dry.Crude product solid recrystallization from chloroform-alcohol-ether obtains 301 (240mg, 89%), is thin spicule.MW 541.00 IR (KBr) n 3338,3296,1691,1641cm -1 1H-NMR (CDCl 3) δ 1H-NMR (DMSO-d 6) δ 1.37 (s, 6H), 3.35 (s, 2H), 3.77 (s, 3H), 4.05 (s, 2H), 4.96 (br s, 1H), 6.75 (br d, J=8.3Hz, 1H), 6.78 (d, J=8.3Hz, 1H), 6.87 (d, J=1.7Hz, 1H), 7.01 (m, 1H), 7.15 (dd, J=2 and 8.5Hz, 1H), 7.24 (d, J=2Hz, 1H), 7.27 (dt, J=2.0 and 8.5Hz, 1H), 7.43 (dd, J=2 and 8.0Hz, 1H), 7.78 (br s, 1H), 7.90 (d, J=8.0Hz, 1H), 8.08 (dd, J=2 and 8.3Hz, 1H), 8.85 (s, 1H), 8.89 (s, 1H), 12.23 (br s, 1H); MS (FAB) m/z 541 (M ++ 1); C 27H 29ClN 4O 6The analytical calculation value: C, 58.00; H, 5.59; N, 10.02.Measured value: C, 57.97; H, 5.39; N, 10.01.Embodiment 2512-acetylaminohydroxyphenylarsonic acid 4-[2-N-[3-methoxyl group-4-[N '-(2-aminomethyl phenyl) urea groups] phenyl]-N-methyl acetylamino] the ethylamino benzoic acid
Figure A0080951005391
In 0 ℃, to 2-acetylaminohydroxyphenylarsonic acid 4-nitrobenzoic acid (1.28g, benzene-methanol 5.71mmol) (4: 1, v/v, add in agitating solution 25ml) the trimethyl silyl Azimethylene. (hexane solution of 2.0M, 4.28ml, 8.56mmol).Under room temperature, continue to stir 18 hours.In reactant impouring hexane, filter and collect the precipitation that produces, obtain 2-acetylaminohydroxyphenylarsonic acid 4-nitrobenzoic acid methyl ester (1.32g, 97%), be white solid; The mp free of data; 1H-NMR (400MHz, CDCl 3) δ 2.30 (s, 3H), 4.00 (s, 3H), 7.88 (m, 1H), 8.18 (dd, J=2.0Hz, 8.8Hz, 1H), 9.60 (t, J=2.2Hz, 1H), 11.10 (s, 1H); MS (ESI) m/z 238 (M+).
Under room temperature, (1.31g adds the 5%Pd (195mg) that is carried on the carbon in methanol 5.50mmol) (30ml) solution, continue to stir 18 hours down in room temperature, hydrogen (3atm) to 2-acetylaminohydroxyphenylarsonic acid 4-nitrobenzoic acid methyl ester.Filtration catalizer evaporates this mixture.Make gained crude product solid recrystallization from chloroform-methanol-hexane, obtain 2-acetylaminohydroxyphenylarsonic acid 4-Methyl anthranilate (1.03g, 90%), be white solid. 1H-NMR(400MHz，CDCl 3)δ2.23(s，3H)，3.82(s，3H)，4.20(s，2H)，6.30(dd，J＝2.5Hz，8.8Hz，1H)，7.80(d，J＝8.8Hz，1H)，8.06(s，1H)，11.26(s，1H)；MS(FAB)m/z?208(M +)。
In 0 ℃, to 2-acetylaminohydroxyphenylarsonic acid 4-Methyl anthranilate (300mg, 1.44mmol) and N-tert-butoxycarbonyl-sweet ammonium aldehyde of N-methyl (499mg, 2.88mmol) 1, add NaBH (OAc) in the cooling solution of 2-dichloroethanes (30ml) 3(964mg 4.32mmol), continues to stir 64 hours in 0 ℃.In this mixture impouring saturated sodium bicarbonate, extract with chloroform (50ml * 3), use the salt water washing, through dried over mgso.After the solvent removed in vacuo, residue is through silica gel column chromatography (medium pressure chromatography system: YAMAZEN YFLC-5404-FC, hexane-ethyl acetate was from 9: 1 to 2: 1 linear gradient liquid), obtain 2-acetylaminohydroxyphenylarsonic acid 4-[2-(N-tert-butoxycarbonyl-N-methylamino) ethylamino] essence of Niobe (451mg, 86%), is colorless oil. 1H-NMR(CDCl 3，400MHz)δ1.48(s，9H)，2.22(s，3H)，2.90(s，3H)，3.32(m，2H)，3.50(m，2H)，3.80(s，3H)，6.20(dd，J＝2.2Hz，8.8Hz，1H)，7.80(m，1H)，7.95(m，1H)，11.30(brs，1H)；MS(FAB)m/z?366(M ++1)。
To 2-acetylaminohydroxyphenylarsonic acid 4-[2-(N-tert-butoxycarbonyl-N-methylamino) ethylamino] (450mg adds TFA (5ml) to essence of Niobe in the agitating solution of dichloromethane 1.23mmol) (5ml), continue to stir 18 hours under room temperature.After the solvent removed in vacuo, residue is dissolved in the chloroform (200ml), with saline, saturated sodium bicarbonate washing, through dried over mgso.Remove and desolvate, obtain 2-acetylaminohydroxyphenylarsonic acid 4-[2-(N-methylamino) ethylamino] essence of Niobe (298mg, 88%), be colorless oil. 1H-NMR(CDCl 3，400MHz)δ2.21(s，3H)，2.46(s，3H)，2.88(m，2H)，3.31(m，2H)，3.83(s，3H)，4.85(br，1H)，6.24(dd，J＝2.5Hz，8.8Hz，1H)，7.80(d，J＝8.8Hz，1H)，7.99(d，J＝2.5Hz，1H)；MS(FAB)，m/z?266(M ++1)。
With 2-acetyl-amino-4-[2-(N-methylamino) ethylamino] essence of Niobe (145mg; 0.55mmol), 3-methoxyl group-4-[N '-(2-aminomethyl phenyl) urea groups] phenylacetic acid (172mg; 0.55mmol), EDCHCl (158mg; 0.83mmol), HOBt (141mg; 1.05mmol) and DMAP (13mg, 0.11mmol) mixture in DMF (10ml) stirred 18 hours.(300ml) dilutes this mixture with ethyl acetate, uses the salt water washing, through dried over mgso.Except that after desolvating; residue is through silica gel column chromatography (medium pressure chromatography system: YAMAZEN YFLC-5404-FC; chloroform-methanol was from 100: 0 to 70: 30 linear gradient liquid); obtain 2-acetyl-amino-4-[2-N-[3-methoxyl group-4-[N '-(2-aminomethyl phenyl) urea groups] phenyl]-N-methyl acetylamino] ethylamino essence of Niobe (309mg; 100%), is the amorphous foam thing. 1H-NMR(CDCl 3)δ2.22(s，3H)，2.30(s，3H)，3.02(s，3H)，3.35(m，2H)，3.58(s，3H)，3.50-3.74(m，4H)，3.85(s，3H)，6.20(m，1H)，6.58(s，1H)，6.65-6.75(m，3H)，7.13(m，2H)，7.40-7.50(m，2H)，7.75(m，2H)，7.90(m，1H)，8.00(m，1H)，11.32(s，1H)；MS(FAB)m/z?562(M ++1)。
Under room temperature; to 2-acetyl-amino-4-[2-N-[3-methoxyl group-4-[N '-(2-aminomethyl phenyl) urea groups] phenyl]-N-methyl acetylamino] ethylamino essence of Niobe (309mg; 0.55mmol) THF-methanol (1: 1; v/v; add in solution 9ml) the 0.25N sodium hydroxide (4.4ml, 1.1mmol) and be heated to backflow.Under refluxing, continue to stir 18 hours.In reactant mixture impouring water, be acidified to pH 5.0 with 1.0N HCl.The precipitation that obtains obtains 302 (175mg, 58%) with hexane-ether recrystallization, is the light red powder.MW 547.60 1H-NMR (CD 3OD) δ 2.16 (d, J=4.8Hz, 3H), 2.28 (d, J=4.2Hz, 3H), 2.98 and 3.10 (2s, 3H altogether), 3.35 (m, 2H), 3.68 (m, 4H), 3.81 and 3.84 (2s, 3H altogether), 6.30 (m, 1H), 6.60-6.82 (m, 2H), 7.00 (m, 1H), 7.15 (m, 2H), 7.53 (m, 1H), 7.77-7.96 (m, 3H); MS (FAB) m/z 548 (M ++ 1); C 29H 33N 5O 60.5H 2The analytical calculation value of O: C, 62.58; H, 6.61; N, 12.58.Measured value: C, 62.55; H, 6.31; N, 12.15.Embodiment 2522-acetyl-amino-4-[2-N-[4-[N '-(2-bromo phenyl) urea groups]-the 3-methoxyphenyl]-N-methyl acetylamino] the ethylamino benzoic acid
With 2-acetyl-amino-4-(2-N-methylamino-1-ethylamino) essence of Niobe (145mg; 0.55mmol), 3-methoxyl group-4-[N '-(2-bromo phenyl) urea groups] phenylacetic acid (209mg; 0.55mmol), EDCHCl (158mg; 0.83mmol), HOBt (141mg; 1.05mmol) and DMAP (13mg, 0.11mmol) mixture in DMF (10ml) stirred 18 hours.(300ml) dilutes this mixture with ethyl acetate, uses the salt water washing, through dried over mgso.Except that after desolvating; residue is through silica gel column chromatography (medium pressure chromatography system: YAMAZEN YFLC-5404-FC; chloroform-methanol was from 100: 0 to 70: 30 linear gradient liquid); obtain 2-acetyl-amino-4-[2-N-[4-[N '-(2-bromo phenyl) urea groups]-the 3-methoxyphenyl]-N-methyl acetylamino] ethylamino essence of Niobe (294mg; 85%), is the amorphous foam thing. 1H-NMR(CDCl 3)δ2.21(s，3H)，3.05(s，3H)，3.40(m，2H)，3.65-3.70(m，4H)，3.78(s，3H)，3.86(s，3H)，6.21(m，1H)，6.79(m，2H)，6.93(m，1H)，7.10(d，J＝10.3Hz，1H)，7.30(m，1H)，7.42(m，1H)，7.61(m，1H)，7.78-7.85(m，2H)，7.93(m，2H)，8.13(m，1H)；MS(FAB)m/z?627(M +)。
Under room temperature; to 2-acetyl-amino-4-[3-methoxyl group-4-[N '-(2-bromo phenyl) urea groups] the phenyl acetylamino]-2-N-methylamino-1-ethylamino essence of Niobe (294mg; 0.47mmol) THF-methanol (1: 1; v/v; add 0.25N sodium hydroxide (3.8ml in solution 8ml); 0.94mmol), and be heated to backflow.Under refluxing, continue to stir 18 hours.In reactant mixture impouring water, be acidified to pH 5.0 with 1.0N HCl.The precipitation that obtains obtains 303 with hexane-ether recrystallization, is white powder (210mg, 73%).MW 612.47 mp155-160 ℃; 1H-NMR (CD 3OD) δ 22.18 (d, J=5.5Hz, 3H), 3.00 and 3.12 (2s, altogether 3H), 3.39 (m, 1H), 3.60 (m., 4H), 3.70 (s, 1H), 3.85 and 3.86 (2s, 3H altogether), 6.31 (m, 1H), 6.68 (m, 1H), 6.78 (m, 1H), 6.78 and 6.85 (2m, be total to 1H), 6.97 (m, 1H), 7.30 (m, 1H), 7.56 (m, 1H), 7.80-7.95 (m, 4H); MS (ESI) m/z613 (M +); C 28H 31Br 1N 5O 60.75H 2The analytical calculation value of O: C, 53.64; H, 5.22; N, 11.17.Measured value: C, 53.89; H, 5.23; N, 10.69.Embodiment 2534-[2-N-[[4-[N '-(2-chlorophenyl) urea groups]-the 3-methoxyphenyl]-N-phenyl acetylamino] ethyoxyl] benzoic acid
Under room temperature, to 4-[2-(mesyloxy) ethyoxyl] essence of Niobe (2.74g, add in the solution of MeCN 10mmol) (50ml) aniline (9.1ml, 100mmol).Reflux down, reactant was stirred 64 hours.In this mixture impouring water (200ml), extract with ethyl acetate (100ml * 2), through dried over mgso.After the solvent removed in vacuo, by in 80 ℃ with toluene (10ml * 3) coevaporation, vacuum is removed unreacted aniline.Residue obtains 4-[2-(N-phenyl amino) ethyoxyl through silica gel column chromatography (medium pressure chromatography system: YAMAZEN YFLC-5404-FC, f50mm * 150mm, chloroform)] essence of Niobe (2.23g, 82%), be colorless oil. 1H-NMR (CDCl 3) δ 3.56 (t, J=5.1Hz, 2H), 3.90 (s, 3H), 4.21 (t, J=5.1Hz, 2H), 6.68 (dd, J=1.0Hz, 8.6Hz, 2H), 6.75 (t, J=7.3Hz, 1H), 7.20 (AB type d, J=7.3Hz, 2H), 8.00 (d, J=9.1Hz, 2H); MS (ESI) m/z 272 (M ++ 1).
With 4-[2-(N-phenyl amino) ethyoxyl] essence of Niobe (136mg, 0.5mmol), 3-methoxyl group-4-[N '-(2-chlorophenyl) urea groups] phenylacetic acid (167mg, 0.5mmol) and PyBOP (781mg, 0.75mmol), i-PrNEt 2(261ml, 0.96mmol) mixture in DMF (10ml) stirred 18 hours.(100ml) dilutes this mixture with ethyl acetate, with 1N HCl, salt water washing, through dried over mgso.Make residue and toluene (10ml * 3) coevaporation to remove DMF.Residue is through TLC chromatography (MERCK, silica gel 60,2mm, 2 blocks of plates, chloroform-methanol, 20: 1), obtain 4-[2-N-[[4-[N '-(2-chlorophenyl) urea groups]-the 3-methoxyphenyl]-N-phenyl acetylamino] ethyoxyl] essence of Niobe (129mg, 44%), be white amorphous foam thing. 1H-NMR(CDCl 3)δ3.42(s，1H)，3.69(d，J＝8.3Hz，1H)，3.74(s，3H)，3.87(s，3H)，4.10(m，2H)，4.23(m，2H)，6.48-7.44(m，13H)，7.93(d，J＝9.3Hz，2H)，8.18(dd，J＝1.5Hz，8.3Hz，1H)；MS(ESI)m/z?588(M +)。
Under room temperature, to 4-[2-N-[[4-[N '-(2-chlorophenyl) urea groups]-the 3-methoxyphenyl]-N-phenyl acetylamino] ethyoxyl] essence of Niobe (124mg, 0.19mmol) THF-methanol (6: 1, v/v, add 0.5N sodium hydroxide (2ml in solution 3ml), 1mmol), and in an air-tight bottle be heated to backflow.Under refluxing, continue to stir 15 hours.In reactant mixture impouring water,, extract with chloroform-methanol (2: 1,20ml * 3), through dried over mgso with 1.0N HCl acidify.Except that after desolvating, make residue crystallization from chloroform-hexane-ether, obtain 304 (77mg, 64%), be white powder.MW 574.02 1H-NMR (CD 3OD) δ 3.45 (s, 2H), 3.79 (s, 3H), 4.12 (m, 2H), 4.22 (m, 2H), 6.48 (dd, J=2.0Hz, 8.3Hz, 1H), 6.61 (d, J=2.0Hz, 1H), 6.87 (d, J=8.8Hz, 2H), 7.00 (m, 1H), 7.22 (m, 3H), 7.36 (m, 1H), 7.43 (m, 3H), 7.90 (d, J=8.3Hz, 1H), 7.95 (d, J=8.8Hz, 2H), 8.02 (dd, J=1.5Hz, 8.3Hz, 1H); MS (ESI) m/z 574 (M +); C 31H 28ClN 3O 60.5H 2The analytical calculation value of O: C, 63.86; H, 5.01; N, 7.21.Measured value: C, 63.67; H, 4.91; N, 6.99.Embodiment 254 (S)-4-[2-N-[[3-methoxyl group-4-[N '-(2-aminomethyl phenyl) urea groups] phenyl]-N-(2-aminobenzyl) acetylamino]-the 1-propoxyl group] benzoic acid
Figure A0080951005441
(1.50g, 5.26mmol) (0.87g, (16ml 15: 1, adds NaBH in cooling v/v) (0 ℃) solution to MeOH-AcOH 5.76mmol) with the 2-nitrobenzaldehyde to (S)-4-(2-amino-1-propoxyl group) benzyl benzoate 3(1.65g's CN 26.3mmol) and under room temperature spends the night the reactant mixture stirring.Through this mixture of saturated sodium bicarbonate quencher.Use ethyl acetate extraction.With salt water washing extract, through dried over sodium sulfate and evaporation.Residue is through purification by silica gel column chromatography, as eluant, obtains (S)-4-[2-(2-nitrobenzyl amino)-1-propoxyl group with the chloroformic solution of chloroform to 5% methanol] benzyl benzoate (931mg, 42%), be yellow oil. 1H-NMR(CDCl 3)δ1.21(d，J＝6.4Hz，3H)，3.13-3.18(m，1H)，3.88-3.97(m，2H)，4.06-4.20(m，2H)，5.34(s，2H)，6.89-6.94(m，2H)，7.29-7.65(m，8H)，7.94-8.03(m，3H)；MS(FAB)m/z?421(M ++1)。
Under room temperature, with 3-methoxyl group-4-[N '-(2-aminomethyl phenyl) urea groups] phenylacetic acid pentafluorophenyl esters (460mg, 0.96mmol), (S)-4-[2-(2-nitrobenzyl amino)-1-propoxyl group] benzyl benzoate (403mg, 0.96mmol) and triethylamine (200ml, 1.43mmol) mixture in DMF (8ml) stirs and to spend the night.Dilute this mixture with ethyl acetate, with 0.5N HCl, salt water washing, through dried over sodium sulfate and evaporation.Residue is through purification by silica gel column chromatography, with the chloroformic solution of 1% methanol as eluant, obtain (S)-4-[2-N-[[3-methoxyl group-4-[N '-(2-aminomethyl phenyl) urea groups] phenyl]-N-(2-nitrobenzyl) acetylamino]-the 1-propoxyl group] benzyl benzoate (504mg, 73%), is brown amorphous solid; MS (FAB) m/z 717 (M ++ 1).
With (S)-4-[2-N-[[3-methoxyl group-4-[N '-(2-aminomethyl phenyl) urea groups] phenyl]-N-(2-nitrobenzyl) acetylamino]-the 1-propoxyl group] benzyl benzoate (504mg, 0.70mmol) MeOH-THF (11ml, 10: 1, v/v) (100mg, 20wt%) spend the night under 3 atmospheric pressure by hydrogenation with 5%Pd-C for agitating solution.Filter this mixture to remove catalyst, evaporated filtrate.Residue is through the preparation of lamina chromatography purification, as eluant, obtains 305 (115mg, 27%) with the chloroformic solution of 5% methanol, is white powder.MW?596.67?MS(FAB)，m/z?597(M ++1)。Embodiment 255 (S)-4-[2-N-[[4-[N '-(2-bromo phenyl) urea groups]-the 3-methoxyphenyl]-N-(2-nitrobenzyl) acetylamino]-the 1-propoxyl group] benzoic acid
Figure A0080951005451
(1.50g, 5.26mmol) (0.87g, (16ml 15: 1, adds NaBH in cooling v/v) (0 ℃) solution to MeOH-AcOH 5.76mmol) with the 2-nitrobenzaldehyde to (S)-4-(2-amino-1-propoxyl group) benzyl benzoate 3(1.65g's CN 26.3mmol), spends the night the reactant mixture stirring under room temperature.Through this mixture of saturated sodium bicarbonate quencher.Use ethyl acetate extraction.With salt water washing extract, through dried over sodium sulfate and evaporation.Residue is through purification by silica gel column chromatography, as eluant, obtains (S)-4-[2-(2-nitrobenzyl amino)-1-propoxyl group with the chloroformic solution of chloroform to 5% methanol] benzyl benzoate (931mg, 42%), be yellow oil. 1H-NMR(CDCl 3)δ1.21(d，J＝6.4Hz，3H)，3.13-3.18(m，1H)，3.88-3.97(m，2H)，4.06-4.20(m，2H)，5.34(s，2H)，6.89-6.94(m，2H)，7.29-7.65(m，8H)，7.94-8.03(m，3H)；FAB-MS?m/z?421(M ++1)。
Under room temperature, with 4-[N '-(2-bromo phenyl) urea groups]-3-methoxybenzene guanidine-acetic acid (476mg, 1.26mmol), (S)-4-[2-(2-nitrobenzyl amino)-1-propoxyl group] benzyl benzoate (528mg, 1.26mmol), EDCHCl (361mg, 1.88mmol), HOBt (255mg, 1.89mmol) and DMAP (30mg, 0.25mmol) mixture in DMF (10ml) stirs and to spend the night.This reaction is incomplete, so stirred these reactant mixtures 1 day in 60 ℃.Dilute this mixture with ethyl acetate, with 0.5N HCl, salt water washing, through dried over sodium sulfate and evaporation.Residue is through purification by silica gel column chromatography, as eluant, obtains title compound with the chloroformic solution of chloroform to 2% methanol, is crude product oil.(10ml, 1: 1, adding 0.5N sodium hydroxide (10ml) heated this reactant mixture 3 hours under refluxing in agitating solution v/v) to the THF-MeOH of this crude product.In mixture impouring ice-water, use 1N HCl with alkaline water layer acidify (pH4.3).The precipitation that collect to produce, crude product solid be through the preparation of lamina chromatography purification, as eluant, obtains 306 (162mg, two go on foot 19%) with the chloroformic solution of 5% methanol, is white amorphous solid.MW 691.53MS (FAB) m/z 692 (M ++ 1); C 33H 31BrN 4O 87/4H 2The analytical calculation value of O: C, 54.82; H, 4.81; N, 7.75.Measured value: C, 54.80; H, 4.61; N, 7.24.Embodiment 2564-[2-N-cyclopropyl-N-[4-[N '-(2-chlorophenyl) urea groups]-the 3-methoxyphenyl] acetylamino] ethoxybenzoic acid
With 4-(2-cyclopropyl amino ethoxy) essence of Niobe (290mg, 1.23mmol), 4-[N '-(2-chlorophenyl) urea groups]-3-methoxybenzene guanidine-acetic acid (412mg, 1.23mmol), EDCHCl (354mg, 1.85mmol), HOBt (catalytic amount) and DMAP (catalytic amount) mixture in DMF (10ml) stirs and spend the night.This mixture is allocated between ethyl acetate (300ml) and the water (100ml).Separate organic facies, (2 * 100ml) washings are through dried over mgso and evaporation with saline.Residue is through silica gel column chromatography, with chloroform-methanol (20: 1) as eluant, obtain 4-[2-N-cyclopropyl-N-[4-[N '-(2-chlorophenyl) urea groups]-the 3-methoxyphenyl] acetylamino] ethoxy-benzoic acid methyl ester (506mg, 75%), be the oil of yellow viscosity. 1H-NMR(CDCl 3)δ0.90-0.97(m，4H)，2.75(m，1H)，3.61(s，3H)，3.79(t，J＝5.4Hz，2H)，3.87(s，3H)，3.88(s，2H)，4.16(t，J＝5.4Hz，2H)，6.76-6.80(m，4H)，6.95(dt，J＝7.8，1.5Hz，1H)，7.21-7.31(m，2H)，7.53(s，1H)，7.56(s，1H)，7.93(d，J＝8.3Hz，3H)，8.19(dd，J＝8.3，1.5Hz，1H)。
To 4-[2-N-cyclopropyl-N-[4-[N '-(2-chlorophenyl) urea groups]-the 3-methoxyphenyl] acetylamino] ethoxy-benzoic acid methyl ester (506mg, 0.917mmol) the agitating solution of THF (7ml) in add the 0.25N sodium hydroxide (7.3ml, 1.83mmol).After stirring was spent the night, in mixture impouring 1NHCl (50ml), (4: 1,2 * 200ml) extracted with chloroform-methanol.The extract that merges through dried over mgso also evaporates.Residue as eluant, obtains 307 (403mg, 82%) with chloroform-methanol (20: 1 to 10: 1) through silica gel column chromatography, is colourless amorphous solid.MW 537.99 1H-NMR (DMSO) δ 0.86-0.91 (m, 4H), 2.75 (m, 1H), 3.69 (t, J=5.5Hz, 2H), 3.81 (s, 3H), 3.84 (s, 2H), 4.16 (t, J=5.5Hz, 2H), 6.76 (d, J=8.3Hz, 1H), 6.88 (s, 1H), and 6.97-7.04 (m, 3H), 7.28 (t, J=7.8Hz, 1H), 7.44 (d, J=7.8Hz, 1H), 7.88 (d, J=8.8Hz, 2H), 7.96 (d, J=8.1Hz, 1H), 8.10 (d, J=8.3Hz, 1H), 8.89 (s, 1H), 8.93 (s, 1H), 12.65 (s, br s); MS (FAB) m/z 538 (M ++ 1); C 28H 28ClN 3O 6The analytical calculation value: C, 62.51; H, 5.25; N, 7.81.Measured value: C, 61.85; H, 5.42; N, 7.41.Embodiment 2574-[2-N-cyclohexyl-N-[3-methoxyl group-4-[N '-(2-aminomethyl phenyl) urea groups] phenyl] acetylamino] ethoxybenzoic acid
Under room temperature, to the 4-[(2-mesyloxy)-the 1-ethyoxyl] essence of Niobe (2.74g, add in the solution of MeCN 10mmol) (50ml) cyclohexylamine (5.72ml, 50mmol).Reflux down, reactant was stirred 18 hours.In this mixture impouring water (200ml), extract with ethyl acetate (100ml * 2), through dried over mgso.Except that after desolvating, residue is through silica gel column chromatography (medium pressure chromatography system: YAMAZEN YFLC-5404-FC, f50mm * 150mm, chloroform-ethyl acetate was from 10: 0 to 1: 1 linear gradient liquid), obtain 4-(2-N-cyclohexyl amino) ethoxy-benzoic acid methyl ester (2.43g, 88%), is colorless oil. 1H-NMR(CDCl 3)δ1.10(m，2H)，1.25(m，2H)，1.60(br，2H)，1.73(m，2H)，1.90(br，2H)，2.49(m，1H)，3.02(t，J＝5.2Hz，2H)，3.88(s，3H)，4.12(t，J＝5.2Hz，2H)，6.90(d，J＝6.90Hz，2H)，7.99(d，J＝7.99Hz，2H)；MS(ESI)m/z?278(M ++1)。
With 4-(2-N-cyclohexyl amino) ethoxy-benzoic acid methyl ester (139mg, 0.5mmol), 3-methoxyl group-4-[N '-(2-aminomethyl phenyl) urea groups] phenylacetic acid (157mg, 0.5mmol), EDCHCl (144mg, 0.75mmol), HOBt (128mg, 0.95mmol) and DMAP (12mg, 0.1mmol) mixture in DMF (2.5ml) stirred 18 hours.(200ml) dilutes this mixture with ethyl acetate, with 1N HCl and salt water washing, through dried over mgso.Except that after desolvating, residue is through silica gel column chromatography (medium pressure chromatography system: YAMAZEN YFLC-5404-FC, chloroform-ethyl acetate was from 10: 0 to 1: 4 linear gradient liquid), obtain 4-[2-N-cyclohexyl-N-[3-methoxyl group-4-[N '-(2-aminomethyl phenyl) urea groups] phenyl] acetylamino] ethoxy-benzoic acid methyl ester (247mg, 86%), is the amorphous foam thing. 1H-NMR(CDCl 3)δ1.08-1.80(m，10H)，2.30(s，3H)，3.60-3.79(m，8H)，3.88(s，3H)，4.16(m，2H)，6.30(s，1H)，6.70-6.83(m，2H)，6.88(d，2H，J＝9.0Hz)，7.12(m，2H)，7.23(m，1H)，7.60(d，1H，J＝8.3Hz)，7.92(d，2H，J＝9.0Hz)，8.10(d，1H，J＝8.0Hz)；MS(ESI)m/z?574(M ++1)。
Under room temperature, to 4-[2-N-cyclohexyl-N-[3-methoxyl group-4-[N '-(2-aminomethyl phenyl) urea groups] phenyl] acetylamino] ethoxy-benzoic acid methyl ester (247mg, 0.43mmol) THF-methanol (6: 1, v/v, add 0.5N sodium hydroxide (3.4ml in solution 7ml), 0.84mmol), and in an air-tight bottle, be heated to backflow.Under refluxing, continue to stir 18 hours.In reactant mixture impouring water,, extract with chloroform-methanol (2: 1,20ml * 3), through dried over mgso with 1.0N HCl acidify.Except that after desolvating, make residue crystallization from chloroform-hexane-ether, obtain 308 (196mg, 81%), be white powder.MW 559.62 1H-NMR (CD 3OD) δ 0.90-1.82 (m, 10H), 2.29 (s, 3H), 3.62 (m, 2H), 3.78 (s, 3H), 3.80 (m, 3H), 4.12 (m, 2H), 6.82 (m, 2H), 6.96 (m, 3H), 7.16 (m, 2H), 7.58 (d, J=7.7Hz, 1H), 7.92 (m, 3H); MS (ESI) m/z 560 (M ++ 1); C 32H 37N 3O 60.5H 2The analytical calculation value of O: C, 67.59; H, 6.74; N, 7.39.Measured value: C, 67.83; H, 6.80; N, 7.13.Embodiment 2584-[2-N-[4-[N '-(2-chlorophenyl) urea groups]-the 3-methoxyphenyl]-N-propargyl acetylamino] ethoxybenzoic acid
Under room temperature, to 44 (2-mesyloxy)-1-ethyoxyls] essence of Niobe (2.74g, and adding propargyl amine in the solution of MeCN 10mmol) (50ml) (3.43ml, 50mmol).Reflux down, reactant was stirred 18 hours.In this mixture impouring water (200ml), extract with ethyl acetate (100ml * 2), through dried over mgso.After the solvent removed in vacuo, residue is through silica gel column chromatography (medium pressure chromatography system: YAMAZEN YFLC-5404-FC, f50mm * 150mm, chloroform-ethyl acetate was from 10: 0 to 9: 1 linear gradient liquid), obtain 4-(2-N-propargyl amino) ethoxy-benzoic acid methyl ester (2.33g, 100%), is colorless oil. 1H-NMR(CDCl 3)δ2.28(d，J＝2.4Hz，1H)，3.11(t，J＝5.1Hz，2H)，3.52(d，J＝2.4Hz，2H)，3.88(s，3H)，4.15(t，J＝5.1Hz，2H)，6.90(d，J＝8.8Hz，2H)，7.98(d，J＝8.8Hz，2H)；MS(ESI)m/z?234(M ++1)。
With 4-(2-N-propargyl amino) ethoxy-benzoic acid methyl ester (117mg, 0.5mmol), 3-methoxyl group-4-[N '-(2-chlorophenyl) urea groups] phenylacetic acid (1 67mg, 0.5mmol), EDCHCl (144mg, 0.75mmol), HOBt (128mg, 0.96mmol) and DMAP (12mg, 0.1mmol) mixture in DMF (10ml) stirred 18 hours.(100ml) dilutes this mixture with ethyl acetate, with 1N HCl, salt water washing, through dried over mgso.Make residue and toluene (10ml * 3) coevaporation to remove DMF.Residue is through TLC chromatography (MERCK, silica gel 60,2mm, 2 blocks of plates, chloroform-methanol, 20: 1), obtain 4-[2-N-[4-[N '-(2-chlorophenyl) urea groups]-the 3-methoxyphenyl]-N-propargyl acetylamino] ethoxy-benzoic acid methyl ester (244mg, 89%), be white amorphous foam thing. 1H-NMR (CDCl 3) δ 2.20 and 2.32 (2m, 1H altogether), 3.72 (s, 2H), 3.83 (m, 5H), 3.88 (s, 3H), 4.09-4.35 (m, 4H), 6.77-6.86 (m, 4H), 6.99 (m, 1H), 7.11 (m, 2H), 7.24 (m, 1H), 7.34 (d, J=7.9Hz, 1H), 7.96 (m, 3H), 8.18 (dd, J=1.5Hz, 8.3Hz, 1H); MS (ESI) m/z 550 (M +).
Under room temperature, to 4-[2-N-[4-[N '-(2-chlorophenyl) urea groups]-the 3-methoxyphenyl]-N-propargyl acetylamino] ethoxy-benzoic acid methyl ester (240mg, 0.44mmol) THF-methanol-water (2: 2: 1, v/v, add in solution 10ml) sodium hydroxide (500mg, 12.5mmol).Under room temperature, continue to stir 2 hours.In reactant mixture impouring water,, extract with chloroform-methanol (2: 1,20ml * 3), through dried over mgso with 1.0N HCl acidify.Residue obtains 309 (202mg, 86%) through TLC chromatography (Whatman, 1mm, 3 blocks of plates, chloroform-methanol, 92: 8), is white solid.MW 535.98 1H-MR (CD 3OD) δ 2.60 and 2.81 (2d, J=2.5Hz, 1H altogether), and 3.79-3.94 (m, 4H), 3.85 (s, 3H), 4.15 (m, 1H), 4.24 (m, 1H), 4.32 (m, 2H), 6.80 (d, J=8.3Hz, 1H), 6.85 (d, J=4.3Hz, 1H), 6.94 (m, 2H), 7.02 (m, 1H), 7.25 (m, 1H), 7.38 (m, 1H), 7.87-8.02 (m, 4H); MS (ESI) m/z 536 (M ++ 1); C 28H 26ClN 3O 62.25H 2The analytical calculation value of O: C, 58.33; H, 5.33; N, 7.29.Measured value: C, 58.23; H, 4.77; N, 6.91.Embodiment 259 and 2604-[2-N-pi-allyl-N-[4-[N '-(2-chlorophenyl) urea groups]-the 3-methoxyphenyl] acetylamino] ethoxybenzoic acid
With 4-(2-N-allyl amino) ethoxy-benzoic acid methyl ester (118mg, 0.5mmol), 3-methoxyl group-4-[N '-(2-chlorophenyl) urea groups] phenylacetic acid (167mg, 0.5mmol), EDCHCl (144g, 0.75mmol), HOBt (128mg, 0.95mmol) and DMAP (12mg, 0.1mmol) mixture in DMF (2.5ml) stirred 18 hours.(300ml) dilutes this mixture with ethyl acetate, with 1N HCl, salt water washing, through dried over mgso.Except that after desolvating, residue is through silica gel column chromatography (medium pressure chromatography system: YAMAZEN YFLC-5404-FC, chloroform-ethyl acetate was from 100: 0 to 85: 15 linear gradient liquid), obtain 4-[2-N-pi-allyl-N-[4-[N '-(2-chlorophenyl) urea groups]-the 3-methoxyphenyl] acetylamino] ethoxy-benzoic acid methyl ester (253mg, 92%), is the amorphous foam thing. 1H-NMR(CDCl 3)δ3.65-3.85(m，4H)，3.73(s，3H)，3.88(s，3H)，5.08(m，2H)，4.22(m，2H)，5.10-5.24(m，2H)，5.76(m，1H)，6.77(m，2H)，6.85(m，2H)，6.99(m，1H)，7.06(m，2H)，7.26(m，1H)，7.34(d，1H，J＝8.1Hz)，7.94(d，2H，J＝8.8Hz)，7.98(m，1H)，8.18(d，1H，J＝6.9Hz)；MS(FAB)m/z?552(M +)。
Under room temperature, to 4-[2-N-pi-allyl-N-[4-[N '-(2-chlorophenyl) urea groups]-the 3-methoxyphenyl] acetylamino] ethoxy-benzoic acid methyl ester (250mg, 0.45mmol) THF-methanol (1: 1, v/v, add in solution 8ml) the 0.25N sodium hydroxide (3.6ml, 0.91mmol) and be heated to backflow.Under refluxing, continue to stir 18 hours.In reactant mixture impouring water, with 1.0N HCl acidify.Filter and collect the precipitation that obtains.Make the precipitate recrystallization with hexane-ether, obtain 310, be white powder (195mg, 80%).MW 537.99 1H-NMR (CD3OD) δ 3.61 (s, 1H), 3.76 (s, 3H), 3.82 (m, 1H), 3.85 (s, 1H), 3.88 (m, 1H), 4.11-4.25 (m, 4H), 5.12-5.25 (m, 2H), 5.81 (m, 1H), 6.78 (d, 1H, J=8.3Hz), 6.82 (m, 1H), 6.92 (m, 2H), 7.01 (m, 1H), 7.26 (m, 1H), 7.37 (m, 1H), 7.96 (m, 3H), 8.02 (m, 1H); MS (FAB) m/z 537 (M +); C 28H 28ClN 3O 61/4H 2The analytical calculation value of O: C, 61.99; H, 5.30; N, 7.75.Measured value: C, 62.00; H, 5.56; N, 7.76.Embodiment 2614-[2-N-pi-allyl-N-[4-[N '-(2-bromo phenyl) urea groups]-the 3-methoxyphenyl] acetylamino] ethoxybenzoic acid
Figure A0080951005521
With 4-(2-N-allyl amino) ethoxy-benzoic acid methyl ester (118mg, 0.5mmol), 4-[N '-(2-bromo phenyl) urea groups]-3-methoxybenzene guanidine-acetic acid (1 90mg, 0.5mmol), EDCHCl (144mg, 0.75mmol), HOBt (128mg, 0.95mmol) and DMAP (12mg, 0.1mmol) mixture in DMF (2.5ml) stirred 18 hours.(300ml) dilutes this mixture with ethyl acetate, with 1N HCl, salt water washing, through dried over mgso.Except that after desolvating, residue is through silica gel column chromatography (medium pressure chromatography system: YAMAZEN YFLC-5404-FC, chloroform-ethyl acetate was from 100: 0 to 70: 30 linear gradient liquid), obtain 4-[2-N-pi-allyl-N-[4-[N '-(2-bromo phenyl) urea groups]-the 3-methoxyphenyl] acetylamino] ethoxy-benzoic acid methyl ester (251mg, 84%), is the amorphous foam thing. 1H-NMR(CDCl 3)δ3.65-3.85(m，4H)，3.73(s，3H)，3.88(s，3H)，4.08(m，2H)，4.22(m，2H)，5.10-5.25(m，2H)，5.78(m，1H)，6.79(m，1H)，6.85(m，3H)，6.93(m，1H)，7.02(m，2H)，7.30(m，1H)，7.51(m，1H)，7.94(d，2H，J＝8.8Hz)，7.97(m，1H)，8.14(m，1H)；MS(FAB)m/z?596?M +)。
Under room temperature, to 4-[2-N-pi-allyl-N-[4-[N '-(2-bromo phenyl) urea groups]-the 3-methoxyphenyl] acetylamino] ethoxy-benzoic acid methyl ester (251mg, 0.42mmol) THF-methanol (1: 1, v/v, add in solution 8ml) the 0.25N sodium hydroxide (3.4ml, 0.84mmol) and be heated to backflow.Under refluxing, continue to stir 18 hours.In reactant mixture impouring water, with 1.0N HCl acidify.Filter and collect the precipitation that obtains.Make the precipitate recrystallization with hexane-ether, obtain 311 (192mg, 78%), be white powder.MW 582.44 1H-NMR (CD 3OD) δ 3.61 (s, 3H), 3.77 (s, 3H), 3.80 (m, 1H), 3.85 (s, 1H), 3.88 (s, 1H), 4.12-4.25 (m, 4H), 5.12-5.23 (m, 2H), 5.81 (m, 1H), 6.76 (m, 1H), 6.82 (m, 1H), 6.93 (m, 3H), 7.29 (m, 1H), 7.56 (m, 1H), 7.94 (m, 4H); MS (FAB) m/z 582 (M +); C 28H 28BrN 3O 6The analytical calculation value: C, 57.74; H, 4.85; N, 7.21.Measured value: C, 57.40; H, 5.07; N, 7.04.Hydrochlorate for 311: C 28H 27BrN 3O 60.25H 2The analytical calculation value of O: C, 55.23; H, 4.55; N, 6.90.Measured value: C, 54.98; H, 4.71; N, 6.53.Embodiment 2624-[2-N-pi-allyl-N-[3-methyl-4-[N '-(2-aminomethyl phenyl) urea groups] phenyl] acetylamino] ethoxybenzoic acid
Figure A0080951005531
With 4-(2-N-allyl amino-1-ethyl) ethoxy-benzoic acid methyl ester (87mg, 0.37mmol), 3-methyl-4-[N '-(2-aminomethyl phenyl) urea groups] phenylacetic acid (100mg, 0.37mmol), EDCHCl (105mg, 0.56mmol), HOBt (95mg, 0.70mmol) and DMAP (9mg, 0.07mmol) mixture in DMF (7.4ml) stirred 18 hours.(100ml) dilutes this mixture with ethyl acetate, with 1N HCl, salt water washing, through dried over mgso.Make residue and toluene (10ml * 3) coevaporation to remove DMF.Residue is through TLC chromatography (Whatman, PLK-5F, 2 blocks of plates, chloroform-methanol, 97: 3), obtain 4-[2-N-pi-allyl-N-[3-methyl-4-[N '-(2-aminomethyl phenyl) urea groups] and phenyl] acetylamino] ethoxy-benzoic acid methyl ester (190mg, 100%), is white amorphous foam thing. 1H-NMR (CDCl 3) δ 2.05 and 2.09 (2s, 3H altogether), 2.20 and 2.21 (s, 3H altogether), 3.62 (s, 2H), 3.76 (m, 2H), 3.90 (s, 3H), 3.88 (m, 1H), 4.08 (m, 2H), 4.11 (m, 1H), 6.28 (m, 2H), 5.78 (m, 1H), 6.88 (d, J=8.8Hz, 2H), 7.05 (m, 1H), 7.12 (m, 1H), 7.21 (m, 1H), 7.58 (m, 2H), 7.95 (d, J=8.8Hz, 2H), 8.02 (m, 1H); MS (FAB) m/z 516 (M ++ 1).
Under room temperature, to 4-[2-N-pi-allyl-N-[3-methyl-4-[N '-(2-aminomethyl phenyl) urea groups] phenyl] acetylamino] ethoxy-benzoic acid methyl ester (217mg, 0.43mmol) THF-methanol (6: 1, v/v, add in solution 7ml) the 0.5N sodium hydroxide (1.9ml, 0.86mmol) and be heated to backflow.In an air-tight bottle, under refluxing, continue to stir 2 hours.In reactant mixture impouring water,, extract with chloroform-methanol (2: 1,20ml * 3), through dried over mgso with 1.0N HCl acidify.Except that after desolvating, make residue crystallization from chloroform-hexane-ether, obtain 312 (84mg, 38%), be white powder.MW 501.57 1H-NMR (CD 3OD) δ 2.18 and 2.24 (s, 3H altogether), 2.30 (d, J=4.9Hz, 3H), 3.70 (s, 1H), 3.78 (m, 2H), 3.88 (s, 1H), 4.12 (m, 4H), 5.20 (m, 2H), 5.81 (m, 1H), 6.92-7.20 (m, 7H), 7.58 (m, 2H), 7.96 (m, 2H); MS (ESI) m/z 502 (M +); C 29H 31N 3O 5The analytical calculation value: C, 69.44; H, 6.23; N, 8.38.Measured value: C, 68.99; H, 6.39; N, 8.03.Embodiment 2634-[2-N-pi-allyl-N-[3-chloro-4-[N '-(2-aminomethyl phenyl) urea groups] phenyl] acetylamino] ethoxybenzoic acid
Figure A0080951005541
To 4-(2-N-allyl amino ethyoxyl) essence of Niobe (141mg, 0.60mmol) and 3-chloro-4-[N '-(2-aminomethyl phenyl) urea groups] phenylacetic acid (191mg, 0.60mmol) the agitating solution of DMF (5ml) in add EDCHCl (172.5mg, 0.90mmol), HOBt (154mg, 1.14mmol) and DMAP (15mg, 0.12mmol), and under room temperature, continue to stir to spend the night.(50ml) dilutes this mixture with ethyl acetate, with 1M HCl (* 3), 1M sodium hydroxide (* 1) and salt water washing.Through this mixture of anhydrous magnesium sulfate drying, concentrating under reduced pressure obtains 4-[2-N-pi-allyl-N-[3-chloro-4-[N '-(2-aminomethyl phenyl) urea groups] phenyl] acetylamino] ethoxy-benzoic acid methyl ester (350mg, 109%), be white powder. 1H-NMR(CDCl 3)δ2.35(s，3H)，3.60(s，1H)，3.75(m，2H)，3.90(s，3H)，4.10(m，4H)，4.21(m，1H)，5.20(m，2H)，5.80(m，1H)，6.50(s，1H)，6.85(m，2H)，7.08(m，2H)，7.20(m，4H)，7.50(d，J＝8.1Hz，1H)，7.95(d，J＝8.1Hz，2H)，8.12(d，J＝8.1Hz，1H)；MS(ESI)m/z?536(M ++H)。
Under room temperature, to 4-[2-N-pi-allyl-N-[3-chloro-4-[N '-(2-aminomethyl phenyl) urea groups] phenyl] acetylamino] ethoxy-benzoic acid methyl ester (321mg, 0.6mmol) THF-methanol-water (2: 2: 1, v/v, add in agitating solution 30ml) sodium hydroxide (500mg, 12.5mmol).Under room temperature, continue to stir 18 hours.In reactant mixture impouring water,,, extract with chloroform-methanol (2: 1,20ml * 3), through anhydrous magnesium sulfate drying and concentrating under reduced pressure with 1M HCl acidify with the ether washing.With chloroform/normal hexane residue is solidified, obtain 313 (283mg, 83%), be white solid.IR (KBr) 3345,1581,1529,1243,1167cm -1 1H-NMR (CD 3OD) δ 2.30 (s, 3H), 3.71 (s, 1H), 3.78 (m, 1H), 3.82 (m, 1H), 3.89 (s, 1H), 4.10 (m, 1H), 4.19 (m, 2H), 4.21 (t, J=5.4Hz, 1H), 5.20 (m, 2H), 5.82 (m, 1H), 6.95 (m, 2H), 7.03 (m, 1H), 7.18 (m, 3H), 7.28 (s, 1H), 7.60 (d, J=8.1Hz, 1H), 7.99 (m, 3H); MS (ESI) m/z 522 (M ++ 1); C 28H 28ClN 3O 51.75H 2The analytical calculation value of O: C, 60.76; H, 5.74; N, 7.59.Measured value: C, 60.43; H, 5.34; N, 7.17.Embodiment 2644-[2-N-[2-(4-morpholinyl) ethyl]-N-[3-methoxyl group-4-[N '-(2-aminomethyl phenyl) urea groups] phenyl] acetylamino] ethoxy-benzoic acid methyl ester
To 4-[2-N-(2,3-dihydroxy-1-propyl group)-[3-methoxyl group-4-[N '-(2-aminomethyl phenyl) urea groups] phenyl acetylamino] ethyoxyl] essence of Niobe (1.83g, 3.24mmol) THF-methanol-water (1: 1: 1, v/v/v, add sodium metaperiodate (2.08g in agitating solution 15ml), 9.71mmol), under room temperature, stirred 18 hours.(50ml) adds in this reactant mixture with saturated sodium thiosulfate, stirs this mixture 1 hour.Extract this mixture with ethyl acetate (100ml * 3), use the salt water washing.Through dried over mgso.Removing desolvates obtains title compound (1.73g, 100%), is the amorphous foam thing. 1H-NMR (CDCl 3) δ 2.32 (t, 3H, J=2.8Hz), 3.33-4.30 (m, 8H), 3.72 (s, 3H), 3.86 (s, 3H), 6.20 (m, 1H), 6.70 (m, 1H), 6.80 (m, 4H), 7.06 (m, 1H), 7.18 (m, 1H), 7.26 (m, 1H), 7.49 (d, 1H, J=7.4Hz), 7.96 (m, 2H), 8.10 (m, 1H), 9.57 and 9.63 (2s, 1H altogether); MS (FAB) m/z 534 (M ++ 1).
Under room temperature; to 4-[2-N-formoxyl methyl-N-[3-methoxyl group-4-[N '-(2-aminomethyl phenyl) urea groups] the phenyl acetylamino] ethyoxyl] essence of Niobe (400mg; 0.75mmol) the agitating solution of ethanol (7.5ml) in add morpholine (654ml; 7.5mmol) and acetic acid (429ml, 7.5mmol).Reaction stirred is 5 minutes under room temperature, is cooled to 0 ℃ then.In this refrigerative solution, add NaBH 3(471mg 7.5mmol), continues to stir 1 hour in room temperature CN.In this mixture impouring saturated sodium bicarbonate, extract with ethyl acetate (50ml * 3), use the salt water washing, through dried over mgso.After the solvent removed in vacuo, residue obtains 314 (346mg, 76%) through silica gel column chromatography (medium pressure chromatography system: YAMAZEN YFLC-5404-FC, toluene-acetone was from 100: 0 to 1: 1 linear gradient liquid), is white amorphous foam thing. 1H-NMR (CDCl 3, 400MHz) δ 2.31 (s, 3H), 2.46 (m, 4H), 3.52-3.79 (m, 12H), 3.70 (d, 3H, J=2.7Hz), 4.05 and 4.22 (m, be total to 2H), 6.24 and 6.29 (s, 1H altogether), 6.73 (m, 2H), 6.85 (m, 2H), 7.07 (s, 1H), 7.17 (m, 1H), 7.25 (m, 2H), 7.50 (t, 1H, J=7.3Hz), 7.96 (m, 2H), 8.08 (m, 1H); MS (FAB) m/z 605 (M ++ 1); C 33H 40N 4O 71/2H 2The analytical calculation value of O: C, 64.58; H, 6.73; N, 9.13.Measured value: C, 64.95; H, 6.88; N, 8.82.314 hydrochlorate: C 33H 41ClN 4O 72.5H 2The analytical calculation value of O: C, 57.76; H, 6.76; N, 8.16; Cl, 5.17.Measured value: C, 58.29; H, 6.81; N, 7.42; Cl, 5.05.Embodiment 2654-[2-N-[2-(4-morpholinyl) ethyl]-N-[3-methoxyl group-4-[N '-(2-aminomethyl phenyl) urea groups] phenyl] acetylamino] ethoxybenzoic acid
Figure A0080951005561
Under room temperature, to 4-[2-N-[2-(4-morpholinyl) ethyl]-N-[3-methoxyl group-4-[N '-(2-aminomethyl phenyl) urea groups] phenyl] acetylamino] ethoxy-benzoic acid methyl ester (146mg, 0.24mmol) THF-methanol (1: 1, v/v, add in solution 6ml) the 0.25N sodium hydroxide (1.9ml, 0.48mmol) and be heated to backflow.Under refluxing, continue to stir 18 hours.In reactant mixture impouring water, with 1.0N HCl acidify.Extract this mixture with chloroform-methanol (3: 1, v/v, 30ml * 5).Extract through the dried over mgso merging.Except that after desolvating, make the residue crystallization with ether, obtain 315 (102mg, 71%), be white powder. 1H-NMR (CD 3OD) δ 2.28 (d, J=3.0Hz, 3H), 2.46 (m, 1H), 2.40 (m, 1H), 2.56 (m, 1H), 2.63 (m, 1H), 3.62-3.80 (m, 12H), 3.85 (s, 3H), 4.12 (m, 1H), 4.26 (m, 1H), 6.82 (m, 2H), 6.96 (m, 2H), 7.01 (m, 1H), 7.17 (m, 2H), 7.58 (d, J=7.8Hz, 1H), 7.93 (m, 3H); MS (FAB) m/z 591 (M +); C 32H 38N 4O 71.0H 2The analytical calculation value of O: C, 63.14; H, 6.62; N, 9.20.Measured value: C, 63.48; H, 6.66; N, 8.79.Embodiment 2664-[2-N-cyclopropyl-N-[3-methoxyl group-4-[N '-(2-aminomethyl phenyl) urea groups] phenyl] acetylamino] ethoxybenzoic acid
To 4-(2-hydroxyl-oxethyl) essence of Niobe (5.00g, 25.5mmol), DMSO (18.1ml, 255mmol), (17.7ml adds SO in the agitating solution of dichloromethane 127.5mmol) (200ml) to triethylamine 3Py (12.2g, 76.5mmol).Stir after 5 hours this mixture of vacuum concentration, water (100ml) dilution residue.(2 * 200ml) extract this mixture with ethyl acetate.Extract with saline (100ml) washing merges also evaporates through dried over mgso.Residue with hexane-ethyl acetate (4: 1) eluting, obtains 4: 1 the mixture (2.0g) of 4-formoxyl methoxyl methyl benzoate and 4-methyl hydroxybenzoate (2.00g) through silica gel column chromatography, is white solid. 1H-NMR(CDCl 3)δ3.90(s，3H)，4.64(d，J＝1.0Hz，2H)，6.92(d，J＝9.0Hz，2H)，8.02(d，J＝9.0Hz，2H)，9.86(d，J＝1.0Hz，1H)。
To 4: 1 the mixture (1.00g) of 4-formoxyl methoxyl methyl benzoate and 4-methyl hydroxybenzoate and cyclopropylamine (425ml, 6.18mmol) MeOH-AcOH (10: 1,11ml) add NaBH in the agitating solution in 3CN (681mg, 10.3mmol).After stirring is spent the night, by adding this mixture of saturated sodium bicarbonate (50ml) quencher.(2 * 200ml) extract with chloroform.The extract that merges through dried over mgso also evaporates.Residue with chloroform-methanol (20: 1) eluting, obtains 4-(2-cyclopropyl amino ethoxy) essence of Niobe (595mg, 49%) through silica gel column chromatography, is colorless oil. 1H-NMR(CDCl 3)δ0.37-0.49(m，4H)，1.91(m，1H)，2.18-2.23(m，1H)，3.11(t，J＝5.2Hz，2H)，3.88(s，3H)，4.12(t，J＝5.2Hz，2H)，6.92(d，J＝8.8Hz，2H)，7.98(d，J＝8.8Hz，2H)。
With 4-(2-cyclopropyl amino ethoxy) essence of Niobe (290mg, 1.23mmol), 3-methoxyl group-4-[N '-(2-aminomethyl phenyl) urea groups] phenylacetic acid (387mg, 1.23mmol), EDCHCl (354mg, 1.85mmol), HOBt (catalytic amount) and DMAP (catalytic amount) mixture in DMF (10ml) stirs and spend the night.This mixture is allocated between ethyl acetate (300ml) and the water (100ml).Separate organic facies, (2 * 100ml) washings are through dried over mgso and evaporation with saline.Residue as eluant, obtains title compound (426mg, 65%) with chloroform-methanol (50: 1) through silica gel column chromatography, is the oil of yellow viscosity. 1H-NMR(CDCl 3)δ0.90-0.97(m，4H)，2.28(s，3H)，3.60(s，3H)，3.77(t，J＝5.4Hz，2H)，3.85(s，2H)，3.87(s，3H)，4.15(t，J＝5.4Hz，2H)，6.60-6.81(m，5H)，7.09-7.23(m，4H)，7.57(d，J＝8.3Hz，1H)，7.92-7.95(m，2H)，8.04(d，J＝8.3Hz，1H)。
To 4-[2-[N-cyclopropyl-[3-methoxyl group-4-[N '-(2-aminomethyl phenyl) urea groups] phenyl] acetylamino] ethyoxyl] essence of Niobe (426mg, 0.801mmol) the agitating solution of THF (7ml) in add the 0.25N sodium hydroxide (6.4ml, 1.60mmol).After stirring was spent the night, in mixture impouring 1NHCl (50ml), (4: 1,2 * 200ml) extracted with chloroform-methanol.The extract that merges through dried over mgso also evaporates.Residue as eluant, obtains 316 (333mg, 80%) with chloroform-methanol (20: 1 to 10: 1) through silica gel column chromatography, is colourless amorphous solid. 1H-NMR (DMSO) δ 0.86-0.91 (m, 4H), 2.25 (s, 3H), 2.74 (m, 1H), 3.69 (t, J=5.5Hz, 2H), 3.81 (s, 3H), 3.83 (s, 2H), 4.15 (t, J=5.5Hz, 2H), 6.75 (d, J=8.3Hz, 1H), 6.87 (s, 1H), 6.92-6.99 (m, 3H), 7.11-7.17 (m, 2H), 7.81 (d, J=8.1Hz, 1H), 7.88 (d, J=8.5Hz, 2H), 8.01 (d, J=8.1Hz, 1H), 8.47 (s, 1H), 8.55 (s, 1H), 12.96 (s, br s); MS (FAB) m/z 518 (M ++ 1); C 29H 31N 3O 6The analytical calculation value: C, 67.30; H, 6.04; N, 8.12.Measured value: C, 66.71; H, 6.26; N, 7.82.Embodiment 2674-[2-N-[2-(N ', N '-dimethylamino)-the 1-ethyl)-N-[-4-[N "-(2-chlorophenyl) urea groups]-the 3-methoxyphenyl] acetylamino] ethoxybenzoic acid
Figure A0080951005591
Under room temperature, to 4-[2-N-[2-(N ', N '-dimethylamino)-the 1-ethyl)-N-[-4-[N "-(2-chlorophenyl) urea groups]-the 3-methoxyphenyl] acetylamino] ethoxy-benzoic acid methyl ester (100mg; THF-methanol 0.17mmol) (1: 1; v/v; add in solution 3ml) the 0.25N sodium hydroxide (2.0ml, 0.5mmol) and be heated to backflow.Under refluxing, continue to stir 3 hours.In reactant mixture impouring water, be acidified to pH5.0 with 1.0N HCl.After vacuum was removed organic solvent, the mixture that obtains obtained 317 (63mg, 64%) through HP-20 chromatography (water-methanol 100: 0 to 0: 100), is white powder. 1H-NMR (CD 3OD) δ 2.41 (s, 2H), 2.65 (s, 3H), 2.69 (s, 3H), 3.02 (m, 2H), 3.62-3.85 (m, 4H), 3.84 (s, 3H), 4.05 and 4.22 (m, 2H altogether), 6.82-6.88 (m, 4H), 7.02 (m, 1H), 7.25 (m, 1H), 7.38 (m, 1H), 7.92 (m, 2H), 8.01 (m, 2H); MS (FAB), m/z 569 (M +); C 29H 33ClN 4O 63.0H 2The analytical calculation value of O: C, 55.90; H, 6.31; N, 8.99.Measured value: C, 56.40; H, 6.50; N, 8.08.Embodiment 2684-[2-N-[2-(4-morpholinyl) ethyl]-N-[3-methoxyl group-4-[N '-(2-aminomethyl phenyl) urea groups] phenyl] acetylamino] the ethoxy benzonitrile isopropyl propionate
Under room temperature, to 4-[2-N-[2-(4-morpholinyl) ethyl]-N-[3-methoxyl group-4-[N '-(2-aminomethyl phenyl) urea groups] phenyl] acetylamino] ethoxybenzoic acid (250mg, 0.42mmol) the agitating solution of DMF (2ml) in add isopropyl iodide (264ml, 2.53mmol) and potassium carbonate (88mg, 0.64mmol).In 50 ℃ of reaction stirred 2 hours.In this mixture impouring saline, extract with chloroform (50ml * 3), use the salt water washing, through dried over mgso.After the solvent removed in vacuo, residue obtains 318 (261mg, 97%) through silica gel column chromatography (medium pressure chromatography system: YAMAZEN YFLC-5404-FC, toluene-acetone was from 100: 0 to 40: 60 linear gradient liquid), is white amorphous foam thing. 1H-NMR (CDCl 3, 400MHz) δ 1.35 (s, 3H), 1.36 (s, 3H), 2.31 (s, 3H), 2.45 (m, 4H), 2.50 (m, 2H), and 3.55-3.78 (m, 10H), 3.70 (s, 3H), 4.05 and 4.20 (t, J=5.2Hz, 2H altogether), 5.22 (m, 1H), 6.24 and 6.33 (2s, 1H altogether), 6.70-6.83 (m, 4H), 7.08 (s, 1H), 7.15 (m, 1H), 7.22 (m, 1H), 7.40 (t, J=9.0Hz, 1H), 7.93 (d, J=8.8Hz, 1H), 7.97 (d, J=8.8Hz, 1H), 8.07 (t, J=7.8Hz, 1H); MS (FAB) m/z 633 (M ++ 1); C 35H 44N 4O 70.75H 2The analytical calculation value of O: C, 65.05; H, 7.10; N, 8.67.Measured value: C, 65.19; H, 7.09; N, 8.50.Embodiment 2694-[2-N-[2-(3,3-two fluoro-1-pyrrolidinyls) ethyl]-N-[4-[N '-(2-bromo phenyl) urea groups]-the 3-methoxyphenyl] acetylamino] the ethoxy benzonitrile acid sodium-salt
Under room temperature; to 4-[2-N-formoxyl methyl-N-[3-methoxyl group-4-[N '-(2-bromo phenyl) urea groups] phenyl] acetylamino] ethoxy-benzoic acid methyl ester (300mg; 0.5mmol) 1; add 3 in the agitating solution of 2-dichloroethanes (3.6ml); 3-difluoro pyrrolidine acetic acid salt (420mg, 2.5mmol).Under room temperature, stir this reactant 5 minutes, be cooled to 0 ℃ then.In this refrigerative solution, add NaBH (OAc) 3(530mg 2.5mmol), continues to stir 4 hours in room temperature.In this mixture impouring saturated sodium bicarbonate, extract with chloroform (50ml * 3), use the salt water washing, through dried over mgso.After the solvent removed in vacuo, residue is through silica gel column chromatography (medium pressure chromatography system: YAMAZEN YFLC-5404-FC, chloroform-methanol was from 10: 0 to 97: 3 linear gradient liquid), obtain 4-[2-N-[2-(3,3-two fluoro-1-pyrrolidinyls) ethyl]-N-[4-[N '-(2-bromo phenyl) urea groups]-the 3-methoxyphenyl] acetylamino] ethoxy-benzoic acid methyl ester (345mg, 100%), is white amorphous foam thing. 1H-NMR(CDCl 3，400MHz)δ2.20-2.80(m，6H)，3.48(m，2H)，3.70-3.80(m，9H)，3.89(s，3H)，4.09(m，1H)，4.21(m，1H)，6.79-6.85(m，4H)，6.93(m，1H)，7.08(m，2H)，7.30(m，1H)，7.50(m，1H)，7.96(m，3H)，8.13(dd，J＝1.5Hz，8.3Hz，1H)；MS(ESI)m/z?689(M +)。
Under room temperature, to 4-[2-N-[2-(3,3-two fluoro-1-pyrrolidinyls) ethyl]-N-[4-[N '-(2-bromo phenyl) urea groups]-the 3-methoxyphenyl] acetylamino] ethoxy-benzoic acid methyl ester (345mg, 0.5mmol) THF-MeOH (1: 1, v/v, add in solution 8ml) the 0.25N sodium hydroxide (4ml, 1.0mmol) and be heated to backflow.Under refluxing, continue to stir 6 hours.Remove and desolvate, residue obtains 319 (306mg, 91%) through HP-20 chromatography (water-methanol, 0: 100 to 100: 0), is the pale red powder. 1H-NMR (CD 3OD, 400MHz) δ 2.20-2.90 (m, 6H), 3.60 (m, 2H), 3.70-3.92 (m, 9H), 4.10 (m, 1H), 4.22 (m, 1H), 6.84 (m, 4H), 6.96 (m, 1H), 7.30 (m, 1H), 7.55 (m, 1H), 7.93 (m, 3H), 7.97 (m, 1H); MS (ESI) m/z 676 (M ++ 1); C 31H 32BrF 2N 4O 62.5H 2The analytical calculation value of O: C, 52.85; H, 5.29; N, 7.95.Measured value: C, 52.67; H, 5.20; N, 8.11.Embodiment 2704-[2-N-(N '-methoxyl group-N '-methylamino) ethyl-N-[3-methoxyl group-4-[N "-(2-aminomethyl phenyl) urea groups] phenyl] acetylamino] the ethoxy benzonitrile acid sodium-salt
Figure A0080951005611
Under room temperature; to 4-[2-N-formoxyl methyl-[3-methoxyl group-4-[N '-(2-aminomethyl phenyl) urea groups] phenyl acetylamino] ethyoxyl] essence of Niobe (350mg; 0.66mmol) the agitating solution of ethanol (13ml) in add N-methoxyl group-N-methylamine hydrochloride (637mg, 6.6mmol).This reactant of sonication is 5 minutes under room temperature, is cooled to 0 ℃ then.In this refrigerative solution, add NaBH 3(105mg 1.65mmol), continues to stir 18 hours in room temperature CN.In this mixture impouring saturated sodium bicarbonate, extract with chloroform (50ml * 3), use the salt water washing, through dried over mgso.After the solvent removed in vacuo, residue obtains title compound (344mg, 91%) through silica gel column chromatography (medium pressure chromatography system: YAMAZENYFLC-5404-FC, toluene-acetone was from 9: 1 to 2: 3 linear gradient liquid), is white amorphous foam thing. 1H-NMR (CDCl 3, 400MHz) δ 2.29 (s, 3H), 2.52 and 2.58 (s, altogether 3H), 2.79 (m, 2H), 3.49-3.76 (m, 9H), 3.88 (s, 3H), 4.05 and 4.20 (m, 2H altogether), 6.69-6.86 (m, 5H), 7.10 (m, 1H), 7.20 (m, 2H), 7.29 (m, 1H), 7.44 (m, 1H), 7.94 and 7.99 (d, J=8.6Hz, 2H altogether), 8.06 (m, 1H); MS (FAB) m/z 579 (M ++ 1).C 31H 38N 4O 72.5H 2The analytical calculation value of O: C, 59.70; H, 6.95; N, 8.98.Measured value: C, 59.58; H, 6.65; N, 8.90.
Under room temperature, to 4-[2-N-(N '-methoxyl group-N '-methylamino) ethyl-N-[3-methoxyl group-4-[N "-(2-aminomethyl phenyl) urea groups] phenyl] acetylamino] ethoxy-benzoic acid methyl ester (138mg; THF-MeOH 0.24mmol) (1: 1; v/v; add in solution 4ml) the 0.25N sodium hydroxide (1.9ml, 0.48mmol) and be heated to backflow.Under refluxing, continue to stir 18 hours.Remove and desolvate, residue obtains 320 (140mg, 100%) through HP-20 chromatography (water-methanol, 100: 0 to 0: 100), is white powder. 1H-NMR (CD 3OD) δ 2.29 (s, 3H), 2.54 and 2.56 (2s, altogether 3H), 2.82 (m, 2H), 3.48 (m, 2H), 3.65-5.80 (m, 9H), 4.09 with 4.21 (2m, 2H altogether), 6.80 (m, 4H), 7.00 (t, J=7.5Hz, 1H), 7.18 (m, 2H), 7.57 (d, J=7.8Hz, 1H), 7.88 (m, 2H), 7.99 (m, 1H); MS (FAB) m/z 565 (M ++ 1).C 30H 35N 4O 7Na1.0H 2The analytical calculation value of O: C, 59.59; H, 6.17; N, 9.27.Measured value: C, 59.10; H, 6.28; N, 8.86.Embodiment 2714-[2-N-(N '-methoxyl group-N '-methylamino) ethyl-N-[4-[N '-(2-bromo phenyl) urea groups]-3-methoxyphenyl acetylamino] ethyoxyl] benzoic acid
Under room temperature; to 4-[2-N-formoxyl methyl-N-[3-methoxyl group-4-[N '-(2-bromo phenyl) urea groups] phenyl] acetylamino] ethoxy-benzoic acid methyl ester (209mg; 0.35mmol) the agitating solution of ethanol (7ml) in add N-methoxyl group-N-methylamine hydrochloride (341mg, 3.5mmol).This reactant of sonication is 5 minutes under room temperature, is cooled to 0 ℃ then.In this refrigerative solution, add NaBH (OAc) 3(370mg 1.75mmol), continues to stir 18 hours in room temperature.In this mixture impouring saturated sodium bicarbonate, extract with chloroform (50ml * 3), through dried over mgso.Except that after desolvating, residue is through TLC chromatography (Whatman, PLK-5F, 2 blocks of plates, chloroform-methanol, 98: 2), obtain 4-[2-N-(N '-methoxyl group-N '-methylamino) ethyl-N-[4-[N '-(2-bromo phenyl) urea groups]-3-methoxyphenyl acetylamino] ethyoxyl] essence of Niobe (89mg, 40%), is white amorphous foam thing. 1H-NMR (CDCl 3, 400MHz) δ 2.52 and 2.60 (2s, 3H), 2.80 (m, 2H), 3.48 and 3.50 (2s, be total to 3H), 3.65 (m, 2H), 3.72 (s, 3H), 3.77 (m, 4H), 3.90 (s, 3H), 4.08 (m, 1H), 4.22 (m, 1H), 6.82 (m, 5H), 7.12 (s, 2H), 7.30 (m, 1H), 7.52 (d, J=8.1Hz, 1H), 7.94 (m, 3H), 8.15 (d, J=8.3Hz, 1H); MS (ESI) m/z 643 (M +).
Under room temperature, to 4-[2-N-(N '-methoxyl group-N '-methylamino) ethyl-N-[4-[N '-(2-bromo phenyl) urea groups]-3-methoxyphenyl acetylamino] ethyoxyl] essence of Niobe (89mg, 0.14mmol) THF-MeOH (5: 1, v/v, (1.4ml 0.7mmol) and in an air-tight bottle is heated to backflow to add the 0.5N sodium hydroxide in solution 6ml).Under refluxing, continue to stir 3 hours.In reactant impouring water, be acidified to pH 5 with 1N HCl, extract with chloroform-methanol (2: 1, v/v, 30ml * 3), through dried over mgso.Obtain 321 (53mg, 60%) after the solvent removed in vacuo, be white powder. 1H-NMR (CD 3OD, 400MHz) δ 2.62 and 2.64 (2s, 3H altogether), 2.80 (m, 2H), 3.50 (d, J=7.3Hz, 3H), 3.63-3.88 (m, 6H), 4.12 (m, 1H), 4.25 (m, 1H), 6.82-7.00 (m, 5H), 7.30 (m, 1H), 7.58 (m, 1H), 7.95 (m, 4H); MS (FAB), m/z 629 (M +).C 29H 33BrN 4O 70.25H 2The analytical calculation value of O: C, 54.94; H, 5.33; N, 8.84.Measured value: C, 55.39; H, 5.53; N, 8.23.Embodiment 2724-[2-N-(N ', N '-diallyl) ethyl-N-[3-methoxyl group-4-[N "-(2-aminomethyl phenyl) urea groups] phenyl] acetylamino] the ethoxy benzonitrile acid sodium-salt
Figure A0080951005631
Under room temperature; to 4-[2-N-formoxyl methyl-N-[3-methoxyl group-4-[N '-(2-aminomethyl phenyl) urea groups] phenyl] acetylamino] ethoxy-benzoic acid methyl ester (400mg; 0.75mmol) the agitating solution of ethanol (15ml) in add diallylamine (926ml; 7.5mmol) and acetic acid (429ml, 7.5mmol).Under room temperature, stir this reactant 5 minutes, be cooled to 0 ℃ then.In this refrigerative solution, add NaBH 3(118mg 1.9mmol), continues to stir 1 hour in room temperature CN.In this mixture impouring saturated sodium bicarbonate, extract with ethyl acetate (50ml * 3), use the salt water washing, through dried over mgso.After the solvent removed in vacuo, residue is through silica gel column chromatography (medium pressure chromatography system: YAMAZEN YFLC-5404-FC, toluene-acetone was from 9: 1 to 1: 1 linear gradient liquid), obtain 4-[2-N-(N ', N '-diallyl) ethyl-N-[3-methoxyl group-4-[N '-(2-aminomethyl phenyl) urea groups] phenyl] acetylamino] ethoxy-benzoic acid methyl ester (385mg, 84%), is white amorphous foam thing. 1H-NMR (CDCl 3, 400MHz) δ 2.30 (s, 3H), 2.60 (m, 2H), 3.09 (m, 4H), 3.49 (m, 2H), 3.60 (s, 2H), 3.70 (m, 5H), 3.89 (s, 3H), 4.02 and 4.19 (2m, 2H altogether), 5.01-5.20 (m, 4H), 4.79 (m, 2H), 6.30 with 6.32 (2s, 1H altogether), 6.70-6.84 (m, 5H), 7.08 (m, 1H), 7.14 (m, 1H), 7.25 (m, 1H), 7.60 (m, 1H), 7.93 and 7.98 (2d, J=8.8Hz, 2H), 8.03 and 8.06 (2d, J=8.3Hz, 1H); MS (FAB) m/z 615 (M ++ 1).
Under room temperature, to 4-[2-N-(N ', N '-diallyl) ethyl-N-[3-methoxyl group-4-[N "-(2-aminomethyl phenyl) urea groups] phenyl] acetylamino] ethoxy-benzoic acid methyl ester (385mg, and adding 1N HCl in the agitating solution of MeOH 0.63mmol) (3ml) (756ml, 0.76mmol).Under room temperature, stirred this reactant 5 minutes, then the evaporation obtain 4-[2-N-(N ', N '-diallyl) ethyl-N-[3-methoxyl group-4-[N "-(2-aminomethyl phenyl) urea groups] phenyl] acetylamino] ethoxy-benzoic acid methyl ester hydrochlorate (385mg, 99%), be the amorphous foam thing.C 35H 43ClN 4O 60.5H 2The analytical calculation value of O: C, 63.67; H, 6.72; N, 8.49.Measured value: C, 63.67; H, 6.69; N, 8.43.
Under room temperature, to 4-[2-N-(N ', N '-diallyl) ethyl-N-[3-methoxyl group-4-[N "-(2-aminomethyl phenyl) urea groups] phenyl] acetylamino] the ethoxy benzonitrile acid esters (175mg; THF-MeOH 0.29mmol) (1: 1; v/v; add in solution 20ml) the 0.25N sodium hydroxide (2.5ml, 0.63mmol) and be heated to backflow.Under refluxing, continue to stir 1 hour.Remove and desolvate, residue obtains 322 (160mg, 94%) through through HP-20 chromatography (water-methanol, 100: 0 to 0: 100), is white powder. 1H-NMR(CD 3OD)δ2.29(s，3H)，2.60(t，J＝6.9Hz，1H)，2.67(t，J＝7.0Hz，1H)，3.10(d，J＝6.6Hz，2H)，3.14(d，J＝6.6Hz，2H)，3.59(m，2H)，3.69-3.80(m，4H)，3.80(s，3H)，4.06(t，J＝5.2Hz，4.21(t，J＝5.1Hz，1H)，5.15(m，4H)，5.80(m，2H)，6.79(m，2H)，6.84(d，J＝8.8Hz，2H)，7.00(t，J＝7.5Hz，1H)，7.14(m，2H)，7.48(m，1H)，7.91(dd，J＝6.1Hz，8.8Hz，2H)，8.00(m，1H)；MS(FAB)，m/z?601(M +)。C 34H 39N 4O 6Na0.5H 2The analytical calculation value of O: C, 64.65; H, 6.38; N, 8.87.Measured value: C, 64.53; H, 6.58; N, 8.78.Embodiment 2734-[2-N-(N ', N '-diallyl) ethyl-N-[4-[N '-(2-chlorophenyl) urea groups]-3-methoxyphenyl acetylamino] ethyoxyl] the benzoic acid sodium salt
Figure A0080951005651
Under room temperature; to 4-[2-N-formoxyl methyl-N-[4-[N '-(2-chlorophenyl) urea groups]-3-methoxyphenyl acetylamino] ethyoxyl] essence of Niobe (100mg; 0.18mmol) the agitating solution of ethanol (3.6ml) in add diallylamine (223ml, 1.81mmol).Under room temperature, stir this reactant 5 minutes, be cooled to 0 ℃ then.In this refrigerative solution, add AcOH (104ml, 1.81mmol) and NaBH 3(28mg 0.45mmol), continues to stir 18 hours in room temperature CN.In this mixture impouring saturated sodium bicarbonate, extract with chloroform (30ml * 3), use the salt water washing, through dried over mgso.After the solvent removed in vacuo, residue is through silica gel column chromatography (medium pressure chromatography system: YAMAZEN YFLC-5404-FC, chloroform-methanol was from 10: 0 to 20: 1 linear gradient liquid), obtain 4-[2-N-(N ', N '-diallyl) ethyl-N-[4-[N '-(2-chlorophenyl) urea groups]-3-methoxyphenyl acetylamino] ethyoxyl] essence of Niobe (96mg, 83%), is white amorphous foam thing. 1H-NMR (CDCl 3, 400MHz) δ 2.60 (m, 2H), 3.09 (d, J=6.4Hz, 1H), 3.11 (d, J=6.4Hz, 3H), 3.52 (m, 2H), 3.61 (s, 2H), 3.70-3.80 (m, 5H), 3.86 (s, 3H), 4.05 and 4.20 (2m, 2H altogether), 5.06-5.21 (m, 4H), 5.80 (m, 2H), 6.71-6.85 (m, 4H), 6.98 (m, 1H), 7.22 (m, 1H), 7.32 (m, 3H), 7.93 (d, J=7.8Hz, 2H), 7.98 (m, 1H), 8.18 (dd, J=1.5Hz, 8.2Hz, 1H); MS (ESI) m/z635 (M +).
Under room temperature, to 4-[2-N-(N ', N '-diallyl) ethyl-N-[4-[N '-(2-chlorophenyl) urea groups]-3-methoxyphenyl acetylamino] ethyoxyl] essence of Niobe (96mg, 0.15mmol) THF-MeOH (1: 1, v/v, add in solution 8ml) the 0.25N sodium hydroxide (3.91ml, 0.98mmol) and be heated to 50 ℃.Under refluxing, continue to stir 6 hours.Remove and desolvate, residue obtains 323 (88mg, 94%) through HP-20 chromatography (water-methanol, 0: 100 to 100: 0), is white powder. 1H-NMR(CD 3OD，400MHz)δ2.61(m，2H)，3.10(s，2H)，3.12(s，2H)，3.59(m，2H)，3.70(s，2H)，3.78(m，2H)，3.82(s，3H)，4.10(m，1H)，4.21(m，1H)，5.18(m，4H)，5.81(m，2H)，6.82(m，4H)，7.01(t，J＝7.8Hz，1H)，7.25(m，1H)，7.39(d，J＝7.8Hz，1H)，7.90(m，2H)，8.02(m，2H)；MS(ESI)m/z?621(M +)。C 33H 36ClN 4O 6Na1.25H 2The analytical calculation value of O: C, 59.55; H, 5.83; N, 8.42.Measured value: C, 59.90; H, 5.74; N, 7.96.Embodiment 2744-[2-N-[3-methoxyl group-4-[N '-(2-aminomethyl phenyl) urea groups] phenyl]-N-methyl acetylamino] ethyl-1-piperazinyl acetic acid
Figure A0080951005661
In 0 ℃, to 1-(2-ethoxy) piperazine (5.21g, 40.0mmol) and potassium carbonate (8.76g, add in the stirred suspension of acetonitrile 63.4mmol) (100ml) bromoethyl acetate (5.60ml, 50.5mmol).This reactant mixture was heated 5 hours under refluxing, with ethyl acetate dilution, water and salt water washing.Through the dried over sodium sulfate extract, be concentrated into driedly, obtain 4-(2-ethoxy)-1-piperazine ethyl acetate (9.65g, 100%), be yellow oil. 1H-NMR(CDCl 3)δ1.23(t，3H，J＝7.3Hz)，2.51-2.61(m，11H)，3.22(s，2H)，3.61(t，2H，J＝5.4Hz)，4.19(q，2H，J＝7.3Hz)。
In 0 ℃, to 2, the 4-dinitrophenyl chloride (1.0g, 3.75mmol) and pyridine (0.34ml, drip in THF 4.20mmol) (19ml) solution methylamine (2.0M THF solution, 2.3ml, 4.60mmol).Stirred this reactant mixture 1 hour, and, used ethyl acetate extraction by adding the quencher of 1N HCl solution.With saturated sodium bicarbonate and salt water washing extract, through dried over sodium sulfate and be concentrated into dried.Residue obtains methyl 2 through recrystallization from ethyl acetate-ether, and 4-dinitro benzene sulfonamide (546mg, 56%) is colorless solid. 1H-NMR(DMSO)δ2.60(d，3H，J＝4.9Hz)，8.22(d，1H，J＝8.8Hz)，8.31(q，1H，J＝4.9Hz)，8.66(dd，1H，J＝8.8，2.0Hz)，8.90(d，1H，J＝2.0Hz)。
In 0 ℃, to 4-(2-ethoxy)-1-piperazine ethyl acetate (452mg, 2.09mmol), methyl 2,4-dinitro benzene sulfonamide (546mg, 2.09mmol) and triphenyl phasphine (658mg, add in THF solution 2.51mmol) DIAD (0.50ml, 251mmol).After stirring 17 hours under the room temperature, be concentrated into this reactant mixture dried.The residue ethyl acetate: methanol (10: 1) chromatography obtains 4-[2-[N-(2,4-dinitro benzene sulfonyl)-N-methylamino] ethyl]-1-piperazinyl ethyl acetate (864mg, 90%), be red oil. 1H-NMR(CDCl 3)δ1.27(t，3H，J＝6.8Hz)，2.35-2.63(m，10H)，2.98(s，3H)，3.20(s，2H)，3.41(t，2H，J＝6.8Hz)，4.17(q，2H，J＝6.8Hz)，8.33(d，1H，J＝8.3Hz)，8.46(d，1H，J＝2.0Hz)，8.50(dd，1H，J＝8.3，2.0Hz)。
Under room temperature; with 4-[2-[N-(2; 4-dinitro benzene sulfonyl)-and the N-methylamino] ethyl]-1-piperazinyl ethyl acetate (864mg; 1.88mmol), TGA (0.17ml; 2.44mmol) and triethylamine (0.53ml, 3.76mmol) solution stirring in dichloromethane (25ml) is 3 hours.Obtain reactant mixture 4-(2-methylamino ethyl)-1-piperazinyl ethyl acetate (388mg, 90%), be red oil. 1H-NMR(CDCl 3)δ1.27(t，3H，J＝6.8Hz)，2.50(s，3H)，2.53-2.60(m，8H)，2.75(t，2H，J＝5.9Hz)，3.20(s，2H)，4.18(q，2H，J＝6.8Hz)。
Under room temperature, with 4-(2-methylamino ethyl)-1-piperazinyl ethyl acetate (388mg, 1.69mmol), triethylamine (0.32ml, 2.25mmol) and DMAP (46mg, DMF 0.38mmol) (15ml) solution stirring 15 minutes.Then will (532mg, 1.69mmol), HOBt (103mg, 0.76mmol) and EDCHCl (486mg, 2.53mmol) adding reactant mixture in, it was stirred under room temperature 15 hours.Use the ethyl acetate diluted reaction mixture, use the salt water washing, through dried over sodium sulfate and be evaporated to dried.The residue chloroform: methanol (10: 1, v/v) chromatography obtains 4-[2-N-[3-methoxyl group-4-[N '-(2-aminomethyl phenyl) urea groups] phenyl]-N-methyl acetylamino] ethyl-1-piperazinyl ethyl acetate (889mg, the mixture of DMF), be red oil. 1H-NMR (CDCl 3) δ 1.25-1.29 (m, 3H), 2.29 (s, 3H), 2.42-2.63 (m, 10H), 3.20,3.18 (they respectively are s, are total to 3H), and 3.55,3.40 (respectively are t, are total to 2H, J=6.8Hz), 3.65,3.69 (they respectively are s, are total to 2H), 3.72 (s, 3H), 4.15-4.21 (m, 2H), 6.50 (m, 1H), 6.77-6.81 (m, 8H), 7.11-7.24 (m, 3H), 7.53 (d, 1H, J=8.3Hz), 8.02 (s, 1H), 8.06 (d, 1H, J=7.8Hz).
To 4-[2-N-[3-methoxyl group-4-[N '-(2-aminomethyl phenyl) urea groups] phenyl]-N-methyl acetylamino] ethyl-1-piperazinyl ethyl acetate (889mg, 1.69mmol) THF-EtOH (5: 1, v/v, and adding 4N sodium hydroxide in agitating solution 18ml) (0.84ml, 3.38mmol).Under room temperature, reactant mixture was stirred 4 hours, transfer to pH7.5 with 1N HCl, with chloroform-methanol (4: 1.v/v) extract.Through the extract that dried over mgso merges, concentrate and obtained for 324 (218mg, two steps 26%), be brown amorphous foam thing.IR (KBr) n 3299,3004,1700,1627,1598,1536cm -1 1H-NMR (DMSO) δ 2.25 (s, 3H), 2.36-2.62 (m, 10H), 2.84,2.99 (they respectively are s, are total to 3H), 3.13,3.14 (they respectively are s, are total to 2H), and 3.38-3.45 (m, 2H), 3.61, (3.65 respectively be s, be total to 2H), 3.86 (s, 3H), 6.74 (t, 1H, J=7.8Hz), 6.87 (s, 1H), 6.93 (t, 1H, J=7.8Hz), 7.11-7.17 (m, 2H), 7.79 (d, 1H, J=7.8Hz), 8.01 (d, 1H, J=7.8Hz), 8.47 (s, 1H), 8.57 (s, 1H); MS (FAB) m/z 498 (M ++ 1); C 26H 35N 5O 52HClH 2The analytical calculation value of O: C, 53.06; H, 6.67; N, 11.89.Measured value: C, 53.04; H, 6.15; N, 11.09.Embodiment 2751-[2-[N-methyl-N-[3-methoxyl group-4-[N '-(2-aminomethyl phenyl) urea groups] phenyl] acetylamino] ethyl]-4-piperidyl acetic acid
(2.07g, 10.0mmol) (1.69g, (10: 1, v/v added NaBH in agitating solution 22ml) to MeOH-AcOH 10.0mmol) with 4-piperidylidene ethyl acetate to 2-(N-benzyloxycarbonyl-N-methylamino) acetaldehyde 3(1.32g 20mmol) also continues to stir to spend the night CN.By adding this mixture of saturated sodium bicarbonate (200ml) quencher, (3 * 150ml) extract with chloroform.The extract that merges through dried over mgso also evaporates.Residue is through silica gel column chromatography, with chloroform-ethanol (40: 1, v/v) eluting obtains 1-[2-(N-benzyloxycarbonyl-N-methylamino) ethyl]-4-piperidylidene ethyl acetate (1.71g, 47%), be colorless oil. 1H-NMR(CDCl 3)δ1.25(t，J＝7.3Hz，3H)，2.16(m，2H)，2.57(m，4H)，2.95(m，7H)，3.44(m，2H)，4.13(q，J＝7.3Hz，2H)，5.12(s，2H)，5.49-5.53(m，1H)，7.35(m，5H)。
Stir down, with 1-[2-(N-benzyloxycarbonyl-N-methylamino) ethyl]-4-piperidylidene ethyl acetate (1.70g, and EtOH-AcOH 4.72mmol) (20: 1, v/v, solution 21ml) was through 5%Pd/C (2g) hydrogenation 3 days.Filter this mixture, vacuum concentrated filtrate.Make residue be alkalescence with saturated sodium bicarbonate, extract with chloroform (300ml).Through sodium carbonate dry extract and evaporation, obtain 1-(2-methylamino ethyl)-4-piperidyl ethyl acetate (813mg, 75%), be yellow oil. 1H-NMR(CDCl 3)δ1.25(t，J＝7.3Hz，3H)，1.68-1.81(m，5H)，1.97(t，J＝11.2Hz，2H)，2.22(t，J＝7.3Hz，2H)，2.43-2.47(m，5H)，2.66(t，J＝6.4Hz，2H)，2.85-2.90(m，2H)，4.13(q，J＝7.3Hz，2H)。
To 3-methoxyl group-4-[N '-(2-aminomethyl phenyl) urea groups] phenylacetic acid (550mg, 1.75mmol) and 1-(2-methylamino ethyl)-4-piperidyl ethyl acetate (400mg, 1.75mmol) the agitating solution of DMF (10ml) in add EDCHCl (503mg, 2.63mmol), HOBt (catalytic amount) and DMAP (catalytic amount) and continue to stir and spend the night.(300ml) dilutes this mixture with ethyl acetate, with saline (200ml) washing, through dried over mgso and evaporation.Residue is through silica gel column chromatography, with chloroform-ethanol (10: 1, v/v) eluting obtains 1-[2-[N-methyl-N-[3-methoxyl group-4-[N '-(2-aminomethyl phenyl) urea groups] phenyl] acetylamino] ethyl]-4-piperidyl ethyl acetate (697mg, 76%), is yellow jelly. 1H-NMR (CDCl 3) δ 1.19-2.06 (a plurality of m, 12H), 2.21 (t, J=7.8Hz, 2H), 2.28 (s, 3H), 2.41 (t, J=7.3Hz, 1H), 2.46 (t, J=7.3Hz, 1H), and 2.80-2.89 (m, 2H), 2.95 and 3.01 (respectively is s, be total to 3H), 3.40 and 3.50 (they respectively being t, J=6.8Hz, 2H altogether), 3.64-3.75 (m, 5H), 4.09-4.16 (m, 2H), 6.59 (s, 1H), 6.77-6.79 (m, 2H), 7.12 (t, J=7.3Hz, 1H), 7.21-7.27 (m, 3H), 7.54 (d, J=8.3Hz, 1H), 8.06 (dd, J=8.3,2.4Hz, 1H).
To 1-[2-[N-methyl-N-[3-methoxyl group-4-[N '-(2-aminomethyl phenyl) urea groups] phenyl] acetylamino] ethyl]-4-piperidyl ethyl acetate (690mg, 1.32mmol) the agitating solution of THF (11ml) in add the 0.25N sodium hydrate aqueous solution (11ml, 2.75mmol) and continue to stir and spend the night.Water (50ml) diluted mixture thing, usefulness 1N HCl neutralization, the usefulness chloroform-methanol (2: 1, v/v, 3 * 100ml) extract.The extract that merges through dried over mgso also evaporates.Residue is dissolved in the methanol (50ml), and (2g) adds in this solution with active carbon.Stir down, make suspension returning 30 minutes and pass through diatomite filtration.Evaporated filtrate grinds residue by absorbing in the chloroform, add hexane until forming precipitation.Collecting precipitation and vacuum drying obtain 325 (75mg, 11%), are white amorphous solid. 1H-NMR (DMSO) δ 1.19-2.99 (a plurality of m, 17H altogether), and 3.32-3.43 (m, 4H), 3.62-3.65 (m, 2H), 3.86 (s, 3H), 6.73 (t, J=8.3Hz, 1H), 6.87 (s, 1H), 6.93 (t, J=7.8Hz, 1H), and 7.11-7.17 (m, 2H), 7.79 (d, J=8.3Hz, 1H), 8.01 (d, J=8.3Hz, 1H), 8.47 (s, 1H), 8.57 (s, 1H); MS-FAB m/z 497 (M ++ 1); C 27H 36N 4O 5The analytical calculation value of HCl: C, 60.84; H, 7.00; N, 10.51.Measured value: C, 60.97; H, 7.14; N, 10.17.Embodiment 2761-[2-[N-methyl-N-[4-[N '-(2-aminomethyl phenyl) urea groups] phenyl] acetylamino] ethyl]-4-piperidyl acetic acid
To 1-(2-methylamino ethyl)-4-piperidyl ethyl acetate (400mg, 1.75mmol) and triethylamine (366 μ l, 2.63mmol) the agitating solution of DMF (10ml) in add 4-(N '-(2-aminomethyl phenyl) urea groups) phenylacetic acid pentafluorophenyl esters (788mg, 1.75mmol) and continue to stir and spend the night.(300ml) dilutes this mixture with ethyl acetate, with saline (200ml) washing, through dried over mgso and evaporation.Residue is through silica gel column chromatography, with chloroform-ethanol (10: 1, v/v) eluting obtains 1-[2-[N-methyl-N-[4-[N '-(2-aminomethyl phenyl) urea groups] phenyl] acetylamino] ethyl]-4-piperidyl ethyl acetate (630mg, 73%), be colorless oil.
To 1-[2-[N-methyl-N-[4-[N '-(2-aminomethyl phenyl) urea groups] phenyl] acetylamino] ethyl]-(630mg adds 0.25N sodium hydrate aqueous solution (10ml) to 4-piperidyl ethyl acetate in the agitating solution of THF 1.27mmol) (10ml) and the continuation stirring is spent the night.Water (100ml) diluted reaction mixture, usefulness 1N HCl neutralization, the usefulness chloroform-methanol (2: 1, v/v, 3 * 100ml) extract.The extract that merges through dried over mgso also evaporates.Be dissolved in and grind residue in the chloroform, add hexane until forming precipitation.Collecting precipitation and vacuum drying obtain 326 (20mg, 3%), are white amorphous solid. 1H-NMR (DMSO) δ 1.69 (m, 5H), 2.15 (m, 4H), 2.24 (s, 2H), 2.50 (m, 2H), 2.83 and 2.99 (they respectively are s, are total to 3H), 3.32-3.49 (m, 4H), 3.63 (d, J=6.8Hz, 2H), 6.91-6.95 (m, 1H), 7.13 (m, 4H), 7.39 (d, J=8.3Hz, 2H), 7.83 (d, J=7.3Hz, 1H), 7.97 (m, 1H), 9.12 (m, 1H); MS (FAB) m/z 467 (M ++ 1).Embodiment 2774-[2-N-[4-[N '-(2-aminomethyl phenyl) urea groups] phenyl]-N-methyl acetylamino] ethyl-1-piperazinyl acetic acid
Figure A0080951005711
Under room temperature, with 4-(2-methylamino ethyl)-1-piperazinyl ethyl acetate (700mg, 3.05mmol), triethylamine (0.64ml, 4.58mmol) and DMAP (75mg, THF 0.61mmol) (15ml) solution stirring 30 minutes.Then, with 4-[N '-(2-aminomethyl phenyl) urea groups] phenylacetic acid (917mg, 3.05mmol), HOBt (82mg, 0.61mmol) and EDCHCl (879mg 4.58mmol) joins in this reactant mixture, stirs 12 hours under room temperature.Dilute this reactant mixture with ethyl acetate, use the salt water washing, through dried over sodium sulfate and be concentrated into dried.The residue chloroform-methanol (10: 1, v/v) chromatography obtains 4-[2-N-[4-[N '-(2-aminomethyl phenyl) urea groups] phenyl]-N-methyl acetylamino] ethyl-1-piperazinyl ethyl acetate (996mg, 66%), be yellow amorphous foam thing. 1H-NMR (CDCl 3) δ 1.25-1.29 (m, 3H), 2.20 (s, 3H), 2.47-2.58 (m, 10H), 2.97,3.05 (respectively are s, are total to 3H), 3.17, (3.20 respectively be s, be total to 2H), 3.45 and 3.52 (respectively are d, are total to 2H, J=6.8Hz), 3.64,3.68 (respectively is s, 2H), 4.15-4.21 (m, 2H), 7.01-7.19 (m, 8H), 7.48 (m, 1H), 7.64 (m, 1H); MS (FAB) m/z 496 (M ++ 1).
To 4-[2-N-[4-[N '-(2-aminomethyl phenyl) urea groups] phenyl]-N-methyl acetylamino] ethyl-1-piperazinyl ethyl acetate (996mg, 2.01mmol) THF-EtOH (5: 1, adding 4N sodium hydroxide in agitating solution 12ml) (1.0ml, 4.00mmol).Under room temperature, reactant mixture was stirred 4 hours, transfer to pH 7.5 with 1N HCl, with chloroform-methanol (4: 1.v/v) extract.Extract through dried over mgso merges concentrates and obtains 327 (73mg, 8%), is yellow amorphous foam thing.IR (KBr) n 3338,2925,2850,2821,1704,1627,1540cm -1 1H-NMR (DMSO) δ 2.24 (s, 3H), 2.33-2.61 (m, 10H), 2.82,2.95 (respectively is s, be total to 3H), 3.00,3.02 (they respectively are s, are total to 2H), 3.39 (t, 2H, J=6.8Hz), 3.60,3.62 (respectively are s, are total to 2H), 6.92 (t, 1H, J=7.8Hz), and 7.09-7.16 (m, 4H), 7.41-7.44 (m, 2H), 7.76,7.77 (respectively be d, 2H, J=7.8Hz), 8.46,8.53 (respectively be s, 1H), 9.54,9.59 (respectively is s, 1H); MS (FAB) m/z 468 (M ++ 1); C 25H 33N 5O 5The analytical calculation value of 2HCl: C, 55.56; H, 6.53; N, 12.96.Measured value: C, 54.99; H, 6.45; N, 11.58.Embodiment 2784-[2-N-[4-[N '-(2-fluoro phenyl) urea groups]-the 3-methoxyphenyl]-N-methyl acetylamino] ethyl-1-piperazinyl acetic acid
Under room temperature, with 4-(2-methylamino ethyl)-1-piperazinyl ethyl acetate (695mg, 3.03mmol), triethylamine (0.64ml, 4.58mmol) and DMAP (75mg, DMF 0.61mmol) (15ml) solution stirring 15 minutes.Then, with 4-[N '-(2-fluoro phenyl) urea groups]-3-methoxybenzene guanidine-acetic acid (965mg, 3.03mmol), HOBt (82mg, 0.61mmol) and EDCHCl (872mg 4.54mmol) joins in this reactant mixture, stirs 12 hours under room temperature.Dilute this reactant mixture with ethyl acetate, use the salt water washing, through dried over sodium sulfate and be concentrated into dried.The residue chloroform-methanol (10: 1, v/v) chromatography obtains 4-[2-N-[4-[N '-(2-fluoro phenyl) urea groups]-the 3-methoxyphenyl]-N-methyl acetylamino] ethyl-1-piperazinyl ethyl acetate (1.21g, the mixture of DMF), be the dark oil thing. 1H-NMR (CDCl 3) δ 1.24-1.29 (m, 3H), 2.45-2.59 (m, 10H), 2.98, (3.05 respectively be s, be total to 3H), 3.17,3.20 (they respectively are s, are total to 2H), 3.44,3.52 (respectively be t, be total to 2H, J=6.8Hz), 3.66 (s, 2H), and 4.15-4.21 (m, 2H), 6.77-6.78 (m, 2H), 6.79-7.11 (m, 3H), 7.68-7.95 (m, 2H), 7.64 (wide s, 1H), 8.20 (t, 1H, J=7.8Hz); MS (FAB) m/z 530 (M ++ 1).
To 4-[2-N-[4-[N '-(2-fluoro phenyl) urea groups]-the 3-methoxyphenyl]-N-methyl acetylamino] ethyl-1-piperazinyl ethyl acetate (1.21g, the mixture of DMF) THF-EtOH (5: 1, v/v, and adding 4N sodium hydroxide in agitating solution 12ml) (1.0ml, 4.00mmol).Under room temperature, reactant mixture was stirred 4 hours, transfer to pH7.5 with 1N HCl, with chloroform-methanol (4: 1.v/v) extract.Through the extract that dried over mgso merges, concentrate and obtained for 328 (78mg, two steps 5%), be brown amorphous foam thing.IR (KBr) n 3299,2940,2830,1704,1627,1598,1536cm -1 1H-NMR (DMSO) δ 2.36-2.61 (m, 10H), 2.83,2.98 (they respectively are s, are total to 3H), 3.11 (s, 2H),, 3.37-3.43 (m, 2H), 3.62,3.65 (they respectively are s, are total to 2H), 3.85 (s, 3H), 6.75 (m, 1H), 6.87 (s, 1H), 6.98 (s, 1H), 7.12 (J=7.8Hz), 7.20,7.23 (respectively is d for t, 1H, 2H, J=7.8Hz), 8.01 (d, 1H, J=7.8Hz), 8.17 (t, 1H, J=7.8Hz), 8.72 (s, 1H), 9.19 (s, 1H); MS (FAB) m/z 502 (M ++ 1); C 25H 32FN 5O 52HCl0.5H 2The analytical calculation value of O: C, 51.46; H, 6.05; N, 12.00.Measured value: C, 51.08; H, 5.69; N, 11.27.Embodiment 2793-fluoro-1-[2-N-methyl-N-[3-methoxyl group-4-[N '-(2-aminomethyl phenyl) urea groups] phenyl] acetylamino] ethyl-4-piperidyl acetic acid
Under room temperature, (14.9g adds TMSCI (11.4ml in the agitating solution of DMF 74.8mmol) (35ml) to 1-tert-butoxycarbonyl-4-piperidones, 89.7mmol), (25.0ml 179mmol), heats this reactant mixture 18 hours in 80 ℃ to drip triethylamine then under room temperature.Hexane is joined in this reactant mixture, and the mixture that produces with saturated sodium bicarbonate and salt water washing then is through dried over sodium sulfate and be concentrated into dried.Residue is through chromatography, with hexane-ethyl acetate (5: 1, v/v) as eluant, obtain 1-tert-butoxycarbonyl-1,2,3,6-tetrahydrochysene-4-(trimethylsiloxy) pyridine (20.4g, 99%) is yellow oil. 1H-NMR (CDCl 3) δ 0.19 (s, 9H), 1.46 (s, 9H), 2.05-2.15 (m, 2H), 3.48-3.56 (m, 2H), 3.83-3.91 (m, 2H), 4.79 (wide s, 1H).
Under room temperature, to 1-tert-butoxycarbonyl-1,2,3, (20.4g, (29.2g 82.5mmol), stirs reactant mixture 2 hours 6-tetrahydrochysene-4-(trimethylsiloxy) pyridine to add SelectfluorTM in acetonitrile 75.0mmol) (500ml) solution.Ethyl acetate is joined in the reactant mixture, use the saline purging compound, through dried over sodium sulfate and be concentrated into dried.Residue is through chromatography, with chloroform-methanol (6: 1, v/v) as eluant, obtain 1-tert-butoxycarbonyl-3-fluoro-4-piperidones (14.5g, 89%), be colorless oil. 1H-NMR(CDCl 3)δ1.50(s，9H)，2.44(t，J＝6.9Hz，1H)，2.48-2.63(m，2H)，3.26(ddd，J＝13.5，10.5，3.9Hz，1H)，3.72(t，J＝6.9Hz，1H)，4.16(m，1H)，4.42(m，1H)，4.83(dt，49.2，6.9Hz，1H)。
In-78 ℃, to phosphine acyl acetic acid three ethyl (3.72g, add in THF 16.6mmol) (70ml) solution two (trimethyl silyl) lithamides (1.0M THF solution, 15.5ml, 15.5mmol).After uniform temp stirred 1 hour down, (3.02g 13.9mmol) joined in the reactant mixture with 1-tert-butoxycarbonyl-3-fluoro-4-piperidones.Stirred this mixture 30 minutes down in uniform temp,, use ethyl acetate extraction by adding the saturated ammonium chloride solution quencher.With salt water washing extract, through dried over sodium sulfate and be concentrated into dried.Residue is through chromatography, with hexane-ethyl acetate (8: 1,, obtain (1-butoxy carbonyl-3-fluorine piperidines-4-subunit) ethyl acetate (3.23g, 81%) v/v) as eluant, be colorless solid. 1H-NMR δ 1.30 (t, J=7.1Hz, 3H), 1.48 (s, 9H), 2.10 (m, 1H), 2.56 (m, 1H), 2.77 (m, 1H), 3.13-3.54 (m, 2H), 3.70 (m, 1H), 4.17,4.18 (respectively is q, J=7.1Hz, 2H altogether), 5.82,5.98 (respectively is s, be total to 1H), 6.41 (they respectively being d, J=46.9Hz, 1H altogether); MS (FAB) m/z 288 (M ++ 1).
Under room temperature, (1.32g, THF 4.59mmol) (30ml) solution is through Pd-C (TMEDA complex, 66.0mg) hydrogenation 2 hours under nitrogen atmosphere with (1-butoxy carbonyl-3-fluorine piperidines-4-subunit) ethyl acetate.Remove by filter catalyst, filtrate is concentrated into dried.The chromatography residue, and usefulness hexane-ethyl acetate (9: 1, v/v) as eluant, obtain 1-tert-butoxycarbonyl-3-fluoro-4-piperidyl ethyl acetate (653mg, 73%), be colorless oil. 1H-NMR(CDCl 3)δ1.26(t，J＝7.1Hz，3H)，1.45(s，9H)，1.53-1.79(m，2H)，1.92-2.09(m，2H)，2.31(dd，J＝16.4，6.9Hz，1H)，2.52(dd，J＝16.4，7.3Hz，1H)，2.61-3.06(m，2H)，4.14(q，J＝7.1Hz，2H)，4.28-4.77(m，2H)； 13C?NMR(CDCL 3)δ14.29，25.80，28.42，35.99，36.20，60.55，79.78，86.72，88.48，154.94，171.93；FAB-MS?m/z?290(M ++1)。
In 0 ℃, to the 1-tert-butoxycarbonyl-(653mg adds TFA (5ml) to 3-fluoro-4-piperidyl ethyl acetate in the solution of dichloromethane 2.26mmol) (10ml).After stirring 4 hours under the room temperature, concentrate this reactant mixture.Handle residue with saturated sodium bicarbonate solution, and usefulness THF-EtOAc (1: 1, v/v) extract.Through the dried over mgso extract, concentrate and obtain 3-fluoro-4-piperidyl ethyl acetate (420mg, 98%), be yellow oil. 1H-NMR (CDCl 3) δ 1.26 (t, J=7.4Hz, 3H), 1.68-1.80 (m, 2H), 2.18 (m, 1H), 2.32 (dd, J=16.6,6.8Hz, 1H), 2.54 (dd, J=16.6,7.5Hz, 1H), 2.82 (m, 1H), 2.90,3.00 (respectively being d, J=14.4Hz, 1H altogether), 3.31 (m, 1H), 3.51 (m, 1H), 4.15 (q, J=7.4Hz, 2H), 4.82,4.71 (they respectively are wide s, are total to 1H).
Under room temperature, to 3-fluoro-4-piperidyl ethyl acetate (230mg, 1.22mmol) and 2-(N-tert-butoxycarbonyl-N-methylamino) acetaldehyde (211mg adds NaBH (OAc) in the solution of THF 1.22mmol) (5ml) 3(386mg, 1.82mmol) and acetic acid (70.0ml, 1.22mmol).Stir after 24 hours,, use ethyl acetate extraction by adding this reactant mixture of saturated sodium bicarbonate quencher.With salt water washing extract, dry and be concentrated into dried through sodium carbonate.Residue is through chromatography, with chloroform-methanol (6: 1,, obtain 3-fluoro-1-[2-(N-tert-butoxycarbonyl-N-methylamino) ethyl v/v) as eluant]-4-piperidyl ethyl acetate (236mg, 56%), be red oil. 1H-NMR (CDCl 3) δ 1.25 (t, J=7.1Hz, 3H), 1.45 (s, 9H), 1.53,1.79 (m, 2H), 1.90-2.09 (m, 2H), 2.15 (dd, J=16.4,7.1Hz, 1H), 2.45-2.58 (m, 2H), 2.87 (s, 3H), 2.90-2.99 (m, 2H), 3.20 (m, 1H), 3.24-3.45 (m, 2H), 4.14 (q, J=7.1Hz, 2H), 4.61,4.73 (respectively is wide s, 1H); ESI-MS m/z 347.
In 0 ℃, to 3-fluoro-1-[2-(N-tert-butoxycarbonyl-N-methylamino) ethyl]-(236mg adds TFA (5ml) in dichloromethane 0.68mmol) (10ml) solution to 4-piperidyl ethyl acetate.After stirring 4 hours under the room temperature, concentrate this reactant mixture.Handle residue with saturated sodium bicarbonate solution, use chloroform extraction.Through the dried over mgso extract, concentrate and obtain 3-fluoro-1-[2-(N-methylamino) ethyl]-4-piperidyl ethyl acetate (117mg, 70%), be red oil. 1H-NMR(CDCl 3)δ1.26(t，J＝7.1Hz，3H)，1.58(m，1H)，1.70(m，1H)，1.99(m，1H)，2.14(m，1H)，2.27-2.33(m，1H)，2.47(s，3H)，2.48-2.56(m，4H)，2.72(t，J＝6.3Hz，2H)，2.90(m，1H)，3.16(m，1H)，4.14(q，J＝7.1Hz，2H)，4.67(d，J＝48.3Hz，1H)；ESI-MSm/z?247(M ++1)。
To 3-methoxyl group-4-[N '-(2-aminomethyl phenyl) urea groups] phenylacetic acid (164mg, 0.52mmol), 3-fluoro-1-[2-(N-methylamino) ethyl]-4-piperidyl ethyl acetate (117mg, 0.47mmol), triethylamine (0.10ml, 0.71mmol) and HOBt (13.0mg, 0.09mmol) the solution of THF (5ml) in add EDCHCl (137mg, 0.71mmol).After stirring 8 hours under the room temperature, this reactant mixture of dilute with water is used ethyl acetate extraction.With salt water washing extract, through dried over sodium sulfate and be evaporated to dried.Residue is through chromatography, with toluene-acetone (1: 2,, obtain 3-fluoro-1-[2-N-methyl-N-[3-methoxyl group-4-[N '-(2-aminomethyl phenyl) urea groups v/v) as eluant] phenyl] acetylamino] ethyl-4-piperidyl ethyl acetate (160mg, 62%), is yellow amorphous solid. 1H-NMR (CDCl 3) δ 1.24-1.28 (m, 3H), 1.51-2.23 (m, 7H), 2.26 (s, 3H), 2.35 (s, 2H), 2.39-2.56 (m, 3H), 2.88 (m, 1H), 2.95,3.03 (respectively is s, be total to 3H), 3.11 (m, 1H), 3.44 (m, 1H), 3.56 (m, 1H), 3.64 (s, 2H), 3.68 (s, 3H), 4.11-4.17 (m, 2H), 4.57, (4.69 respectively be s, be total to 1H), and 6.72-6.82 (m, 2H), 7.10-7.33 (m, 5H), 7.54 (d, J=8.1Hz, 1H), 8.06 (d, J=8.1Hz, 1H); ESI-MS m/z543 (M ++ 1).
To 3-fluoro-1-[2-N-methyl-N-[3-methoxyl group-4-[N '-(2-aminomethyl phenyl) urea groups] phenyl] acetylamino] ethyl-4-piperidyl ethyl acetate (160mg, 0.29mmol) the solution of THF (3ml) in add the 0.25N sodium hydroxide (1.30ml, 0.32mmol).After stirring 1 hour under the room temperature, concentrate this reactant mixture.The dilute with water residue neutralizes with 1N HCl in 0 ℃.Concentrate this mixture,, obtain 329 (110mg, 74%), be yellow amorphous solid by ion exchange resin (HP-20, Mitsubishi Chemical) purification.IR (KBr) 3343,2937,1700,1617,1589,1535,1486,1455,1417cm -1 1H-NMR (CD 3OD) δ 1.57-1.72 (m, 2H), 1.98 (m, 1H), 2.26 (m, 1H), 2.28 (s, 3H), 2.37-2.59 (m, 3H), 2.66-2.89 (m, 2H), 2.95,3.09 (they respectively are s, are total to 3H), 3.14 (m, 1H), 3.40 (m, 1H), 3.51 (m, 1H), 3.59 (m, 1H), 3.71 (s, 2H), 3.78 (m, 1H), 3.89 (s, 3H), 4.69, (4.81 respectively be s, be total to 1H), 6.79 (dd, J=8.1,1.5Hz, 1H), 6.90 (wide s, 1H), 7.01 (t, J=7.8Hz, 1H), and 7.13-7.19 (m, 2H), 7.58 (d, J=7.8Hz, 1H), 8.00 (d, J=8.1Hz, 1H); ESI-MS m/z 515 (M ++ 1); C 27H 35FN 4O 5H 2The analytical calculation value of O: C, 60.89; H, 7.00; N, 10.52.Measured value: C, 61.09; H, 6.80; N, 9.87.Embodiment 2804-[2-N-[2-(4-fluorinated phenoxy) ethyl]-N-[3-methoxyl group-4-[N '-(2-aminomethyl phenyl) urea groups] phenyl] acetylamino] ethyl piperazidine base-1-acetic acid
Figure A0080951005771
Under room temperature, to 2-(N-benzyl-N-tert-butoxycarbonyl amino) ethanol (6.85g, 27.3mmol), the 4-fluorophenol (3.07g, 27.3mmol) and triphenyl phasphine (7.83g, 30.0mmol) THF (100ml) solution in add DIAD (6.00ml, 30.0mmol).Stir after 3 hours, concentrate this reactant mixture.Residue is used hexane through chromatography: ethyl acetate (8: 1, v/v) eluting obtains 1-[2-(N-benzyl-N-tert-butoxycarbonyl amino) ethyoxyl]-4-fluorobenzene (8.19g, 64%), be yellow oil. 1H-NMR(CDCl 3)δ1.42-1.50(m，9H)，3.41-3.67(m，2H)，3.92-4.11(m，2H)，4.51-4.63(m，2H)，6.73-6.85(m，2H)，6.89-6.98(m，2H)，7.24(m，5H)；FAB-MSm/z?346(M ++1)。
In 0 ℃, to 1-[2-(N-benzyl-N-tert-butoxycarbonyl amino) ethyoxyl]-(8.19g adds TFA (40ml) in dichloromethane 23.7mmol) (50ml) solution to the 4-fluorobenzene.After stirring 1 hour under the room temperature, concentrate this reactant mixture.Handle residue with saturated sodium bicarbonate solution, use chloroform extraction.With salt water washing extract, through dried over sodium sulfate and concentrated, obtain 1-(2-N-benzylamino ethyoxyl)-4-fluorobenzene (4.38g, 75%), be red oil. 1H-NMR(CDCl 3)δ3.00(t，J＝5.2Hz，2H)，3.87(s，2H)，4.04(t，J＝5.2Hz，2H)，6.80-6.85(m?2H)，6.92-6.98(m，2H)，7.23-7.36(m，5H)；FAB-MSm/z?246(M ++1)。
With 1-(2-N-benzylamino ethyoxyl)-4-fluorobenzene (1.08g, 4.40mmol), 4-(2-bromoethyl) piperazinyl-1-ethyl acetate (1.23g, 4.40mmol) and potassium carbonate (0.61g, the 17.9mmol) heating 8 hours under refluxing of the mixture in acetonitrile (50ml).The mixture that filtration obtains also is concentrated into filtrate dried.Residue is through chromatography, with toluene-acetone (3: 1,, obtain 4-[2-N-benzyl-N-[2-(4-fluorinated phenoxy) ethyl v/v) as eluant] amino] ethyl piperazidine base-1-ethyl acetate (1.51g, 77%), be red oil. 1H-NMR(CDCl 3)δ1.27(t，J＝7.1Hz，3H)，2.31-2.66(m，10H)，2.75(t，J＝6.6Hz，2H)，2.90(t，J＝6.1Hz，2H)，3.18(s，2H)，3.72(s，2H)，3.97(t，J＝6.1Hz，2H)，4.17(q，J＝7.1Hz，2H)，6.75-6.79(m，2H)，6.91-6.96(m，2H)，7.21-7.35(m，5H)；FAB-MSm/z?444(M ++1)。
With 4-[2-N-benzyl-N-[2-(4-fluorinated phenoxy) ethyl] amino] (1.50g, ethanol 3.38mmol) (30ml) solution is through 5%Pd-C (53.1% weight in wet base, 0.73g) hydrogenation 4 hours under nitrogen atmosphere for ethyl piperazidine base-1-ethyl acetate.Remove by filter catalyst, concentrated filtrate.Handle residue with saturated sodium bicarbonate solution, use chloroform extraction.With salt water washing extract,, obtain 4-[2-N-[2-(4-fluorinated phenoxy) ethyl through dried over sodium sulfate and concentrated] amino] ethyl piperazidine base-1-ethyl acetate (1.12g, 94%), be red oil. 1H-NMR (CDCl 3) δ 1.27 (t, J=7.1Hz, 3H), 1.81 (wide s, 1H), 2.47-2.66 (m, 12H), 3.00 (t, J=5.4Hz, 2H), 3.20 (s, 2H), 4.03 (t, J=5.4Hz, 2H), 4.18 (q, J=7.1Hz, 2H), 6.81-6.85 (m, 2H), 6.90-6.99 (m, 2H); FAB-MS m/z 354 (M ++ 1).
Under room temperature, to 3-methoxyl group-4-[N '-(2-aminomethyl phenyl) urea groups] phenylacetic acid (485mg, 1.54mmol), 4-[2-N-[2-(4-fluorinated phenoxy) ethyl] amino] ethyl piperazidine base-1-ethyl acetate (545mg, 1.54mmol), triethylamine (0.32ml, 2.32mmol) and HOBt (41.5mg, 0.31mmol) the solution of THF (15ml) in add EDCHCl (883mg, 2.32mmol).Stir after 24 hours, the dilute with water reactant mixture, the usefulness chloroform-methanol (10: 1, v/v) extract.With salt water washing extract, through dried over sodium sulfate and be evaporated to dried.Residue is through chromatography, with chloroform-methanol (4: 1, v/v) as eluant, obtain 4-[2-N-[2-(4-fluorinated phenoxy) ethyl]-N-[3-methoxyl group-4-[N '-(2-aminomethyl phenyl) urea groups] phenyl] acetylamino] ethyl piperazidine base-1-ethyl acetate (532mg, 53%), is yellow amorphous solid. 1H-NMR (CDCl 3) δ 1.26,1.27 (respectively being t, J=7.1Hz, 3H altogether), 2.27 (s, 3H), and 2.51-2.63 (m, 10H), 3.16,3.19 (respectively is s, be total to 2H), and 3.51-3.56 (m, 2H), 3.63,3.67 (respectively is s, be total to 2H), and 3.69-3.81 (m, 5H), 3.95,4.10 (respectively is t, J=5.2Hz, 2H altogether), 4.16,4.18 (respectively is q, J=7.1Hz, 2H altogether), 6.56 (d, J=8.1Hz, 1H), and 6.72-6.78 (m, 4H), 6.89-6.98 (m, 2H), 7.12 (t, J=7.6Hz, 1H), 7.20-7.23 (m, 3H), 7.51 (d, J=7.6Hz, 1H), 8.04 (d, J=8.1Hz, 1H); FAB-MS m/z 650 (M ++ 1).
Under room temperature, to 4-[2-N-[2-(4-fluorinated phenoxy) ethyl]-N-[3-methoxyl group-4-[N '-(2-aminomethyl phenyl) urea groups] phenyl] acetylamino] ethyl piperazidine base-1-ethyl acetate (532mg, 0.82mmol) the solution of dioxane (8ml) in drip 0.25N sodium hydroxide (5.00ml, 1.25mmol), stirred reaction mixture 1 hour.Concentrate the mixture that obtains, dilute with water neutralizes with 1N HCl in 0 ℃.With chloroform-methanol (4: 1, v/v) extract this mixture.With salt water washing extract, through dried over sodium sulfate and be evaporated to dried.Residue is through chromatography, with chloroform-methanol (3: 1, v/v) as eluant, obtain 330 (168mg, 33%), be faint yellow amorphous solid.IR (KBr) 3338,2938,2829,1635,1533,1506,1454,1415cm -1 1H-NMR (DMSO-d 6) δ 2.25 (s, 3H), 2.37-2.48 (m, 6H), 2.53-2.67 (m, 6H), 3.09 (m, 2H), 3.44-3.49 (m, 2H), 3.64-3.69 (m, 2H), 3.72 (s, 2H), 3.80,3.84 (they respectively are s, are total to 3H), and 4.06-4.09 (m, 2H), 6.73 (m, 1H), 6.85 (s, 1H), 6.91-6.97 (m, 3H), 7.07-7.18 (m, 4H), 7.78 (d, J=8.1Hz, 1H), 8.00 (dd, J=8.1,2.7Hz, 1H), 8.53 (m, 1H), 8.59 (m, 1H); FAB-MSm/z 622 (M ++ 1); C 33H 40FN 5O 6The analytical calculation value of 2HCl: C, 57.06; H, 6.09; N, 10.08.Measured value: C, 56.83; H, 6.05; N, 9.90.Embodiment 2814-[2-N-[2-(4-acetyl group phenoxy group) ethyl]-N-[3-methoxyl group-4-[N '-(2-aminomethyl phenyl) urea groups] phenyl] acetylamino] ethyl piperazidine base-1-acetic acid
Under room temperature, to 2-(N-benzyl-N-tert-butoxycarbonyl amino) ethanol (5.90g, 23.5mmol), the 4-hydroxy acetophenone (3.18g, 23.5mmol) and triphenyl phasphine (6.74g, 25.8mmol) THF (100ml) solution in add DIAD (5.20ml, 25.8mmol).With the heating 4 hours and concentrated under refluxing of this reactant mixture.Residue is used hexane through chromatography: and ethyl acetate (5: 1,, obtain 4-[2-(N-benzyl-N-tert-butoxycarbonyl amino) ethyoxyl v/v) as eluant] 1-Phenylethanone. (3.64g, 42%), be yellow oil. 1H-NMR(CDCl 3)δ1.43-1.65(m，9H)，2.55(s，3H)，3.41-3.69(m，2H)，4.02-4.24(m，2H)，4.49-4.66(m，2H)，6.81-6.93(m，2H)，7.19-7.37(m，5H)，7.91(d，J＝8.8Hz，2H)；FAB-MS?m/z370(M ++1)。
In 0 ℃, to 4-[2-(N-benzyl-N-tert-butoxycarbonyl amino) ethyoxyl] (3.05g adds TFA (20ml) in dichloromethane 8.26mmol) (30ml) solution to 1-Phenylethanone..After stirring 2 hours under the room temperature, concentrate this reactant mixture.Handle residue with saturated sodium bicarbonate solution, use chloroform extraction.With salt water washing extract, through dried over sodium sulfate and concentrated, obtain 4-(2-N-benzylamino ethyoxyl) 1-Phenylethanone. (2.21g, 99%), be red oil. 1H-NMR (CDCl 3) δ 2.04 (wide s, 1H), 2.55 (s, 3H), 3.05 (t, J=5.4Hz, 2H), 3.88 (s, 2H), 4.15 (t, J=5.4Hz, 2H), 6.92 (d, J=8.9Hz, 2H), 7.24-7.36 (m, 5H), 7.91 (d, J=8.9Hz, 2H); FAB-MSm/z 270 (M ++ 1).
In 0 ℃, to 4-(2-ethoxy) piperazinyl-1-ethyl acetate (11.3g, 52.1mmol) and carbon tetrabromide (20.7g adds triphenyl phasphine (19.2g in dichloromethane 62.5mmol) (200ml) solution in batches, 73.0mmol), stir this reactant mixture 30 minutes, hexane is joined in this reactant mixture, filter out precipitation, concentrated filtrate, obtain 4-(2-bromoethyl) piperazinyl-1-ethyl acetate (13.7g, 94%), be yellow oil. 1H-NMR(CDCl 3)δ1.27(t，J＝7.1Hz，3H)，2.61-2.78(m，8H)，2.81(t，J＝7.6Hz，2H)，3.20(s，2H)，3.42(t，J＝7.6Hz，3H)，4.18(q，J＝7.1Hz，2H)。
With 4-(2-N-benzylamino ethyoxyl) 1-Phenylethanone. (681mg, 2.52mmol), 4-(2-bromoethyl) piperazinyl-1-ethyl acetate (705mg, 2.52mmol) and potassium carbonate (349g, the 2.52mmol) heating 22 hours under refluxing of the mixture in acetonitrile (10ml).The mixture that filtration obtains also is concentrated into filtrate dried.Residue is through chromatography, with toluene-acetone (1: 1,, obtain 4-[2-[N-benzyl-N-[2-(4-acetyl group phenoxy group) ethyl v/v) as eluant] amino] ethyl piperazidine base-1-ethyl acetate (920mg, 78%), be red oil. 1H-NMR(CDCl 3)δ1.27(t，J＝7.1Hz，3H)，2.45-2.53(m，10H)，2.55(s，3H)，2.76(t，J＝6.8Hz，2H)，2.94(t，J＝6.1Hz，2H)，3.18(s，2H)，3.73(s，2H)，4.06(t，J＝6.1Hz，2H)，4.17(q，J＝7.1Hz，2H)，6.85(d，J＝7.1Hz，2H)，7.22-7.35(m，5H)，7.90(d，J＝7.1Hz，2H)；FAB-MS?m/z?468(M ++1)。
With 4-[2-[N-benzyl-N-[2-(4-acetyl group phenoxy group) ethyl] amino] ethyl piperazidine base-1-ethyl acetate (and 920mg, ethanol 1.97mmol) (30ml) solution through 5%Pd-C (53.1%, weight in wet base, 510mg) hydrogenation 4 hours under nitrogen atmosphere.Remove by filter catalyst, concentrated filtrate.Handle residue with saturated sodium bicarbonate solution, use chloroform extraction.With salt water washing extract,, obtain 4-[2-N-[2-(4-acetyl group phenoxy group) ethyl through dried over sodium sulfate and concentrated] amino] ethyl piperazidine base-1-ethyl acetate (690mg, 93%), be red oil. 1H-NMR(CDCl 3)δ1.27(t，J＝7.1Hz，3H)，2.55(s，3H)，2.46-2.54(m，10H)，2.78(t，J＝6.2Hz，2H)，3.04(t，J＝5.2Hz，2H)，3.20(s，2H)，4.13(t，J＝5.2Hz，2H)，4.18(q，J＝7.1Hz，2H)，6.92(d，J＝8.8Hz，2H)，7.92(d，J＝8.8Hz，2H)；FAB-MS?m/z378(M ++1)。
Under room temperature; to 3-methoxyl group-4-[N '-(2-aminomethyl phenyl) urea groups] phenylacetic acid (558mg; 1.77mmol), 4-[2-N-[2-(4-acetyl group phenoxy group) ethyl] amino] ethyl piperazidine base-1-ethyl acetate (670mg; 1.77mmol), triethylamine (0.38ml; 2.66mmol) and HOBt (50.0mg; 0.35mmol) the solution of THF (10ml) in add EDCHCl (883mg, 2.32mmol).Stir after 15 hours, the dilute with water reactant mixture, the usefulness chloroform-methanol (10: 1, v/v) extract.With salt water washing extract, through dried over sodium sulfate and be concentrated into dried.Residue is through chromatography; with chloroform-methanol (4: 1; v/v) as eluant; obtain 4-[2-N-[2-(4-acetyl group phenoxy group) ethyl]-N-[3-methoxyl group-4-[N '-(2-aminomethyl phenyl) urea groups] phenyl] acetylamino] ethyl piperazidine base-1-ethyl acetate (724mg; 61%), is yellow amorphous solid. 1H-NMR (CDCl 3) δ 1.25,1.27 (respectively being t, J=7.1Hz, 3H altogether), 2.29 (s, 3H), 2.45-2.51 (m, 8H), 2.54 (s, 3H), 2.55-2.63 (m, 2H), 3.17,3.19 (they respectively are s, are total to 2H), 3.51-3.55 (m, 2H), 3.60 (s, 2H), 3.69 (s, 3H), and 3.71-3.78 (m, 2H), 4.04-4.23 (m, 4H), 6.47 (m, 1H, J=8.1Hz), 6.72-6.76 (m, 2H), 6.85 (d, J=8.8Hz, 2H), 7.12-7.19 (m, 2H), and 7.20-7.24 (m, 2H), 7.51 (d, 8.1Hz, 1H), 7.88 (d, J=8.8Hz, 2H), 8.04 (d, J=8.1Hz, 1H); FAB-MS m/z 674 (M ++ 1).
Under room temperature; to 4-[2-N-[2-(4-acetyl group phenoxy group) ethyl]-N-[3-methoxyl group-4-[N '-(2-aminomethyl phenyl) urea groups] phenyl] acetylamino] ethyl piperazidine base-1-ethyl acetate (724mg; 1.07mmol) the solution of THF (10ml) in drip 0.25N sodium hydroxide (6.50ml; 1.61mmol), stirred reaction mixture 1 hour.Concentrate the mixture that obtains, dilute with water neutralizes with 1N HCl in 0 ℃.With chloroform-methanol (4: 1, v/v) extract this mixture.With salt water washing extract, through dried over sodium sulfate and be evaporated to dried.Residue is through chromatography, with chloroform-methanol (3: 1, v/v) as eluant, obtain 331 (450mg, 65%), be faint yellow amorphous solid.IR (KBr) 3345,2938,2821,1673,1631,1598,1533,1455,1417cm -1 1H-NMR (DMSO-d 6) δ 2.25 (s, 3H), 2.35-2.47 (m, 8H), 2.51 (s, 3H), 2.53-2.58 (m, 2H), 2.96 (s, 2H), 3.46-3.50 (m, 2H), 3.69 (s, 2H), 3.73-3.77 (m, 2H), 3.80,3.83 (respectively are s, are total to 3H), 4.19-4.22 (m, 2H), 6.73 (m, 1H), 6.85 (s, 1H), 6.92 (t, J=7.3Hz, 1H), 7.30 (d, J=7.3Hz, 2H), 7.10-7.16 (m, 2H), 7.79 (d, J=8.3Hz, 1H), 7.90-7.95 (m, 2H), 8.00 (t, J=8.3Hz, 1H), 8.55 (m, 1H), 8.61 (m, 1H); FAB-MS m/z 646 (M ++ 1); C 35H 43N 5O 72HCl1H 2The analytical calculation value of O: C, 57.06; H, 6.43; N, 9.51.Measured value: C, 59.17; H, 6.32; N, 9.61.Embodiment 2824-[2-N-[4-[N '-(2-bromo phenyl) urea groups]-the 3-methoxyphenyl]-N-benzyl acetylamino] ethyl-piperidino acetic acid
Figure A0080951005821
4-[2-N-[4-[N '-(2-bromo phenyl) urea groups]-the 3-methoxyphenyl]-N-benzyl acetylamino] ethyl-piperidino methyl acetate
Figure A0080951005822
Stir down, with piperidines ethanol (10.0g, 77.4mmol), (21.4g, 155mmol), the mixture in acetonitrile (200ml) heated 2 hours under refluxing for 2-monobromo-acetic acid benzyl ester (17.8g, 77.6 mmol) and potassium carbonate.Remove by filter insoluble solid, vacuum concentrated filtrate.Among the 1N HCl that the residue impouring is ice-cooled and use chloroform extraction.Wash organic layer with water, through anhydrous sodium sulfate drying and vacuum concentration.Residue as eluant, obtains 4-(2-ethoxy)-piperidino benzyl acetate (16.3g, 76%) with ethyl acetate through silica gel column chromatography, is colorless oil. 1H-NMR(CDCl 3)δ1.31-1.40(m，3H)，1.52(dd，J＝12.8，6.3Hz，2H)，1.68(brd，J＝10.0Hz，3H)，2.16(t，J＝11.6Hz，2H)，2.92(brd，J＝11.6Hz，2H)，3.24(s，2H)，3.69(t，J＝6.8Hz，2H)，5.16(s，2H)，7.35(m，5H)。
To 4-(2-ethoxy)-piperidino benzyl acetate (14.9g, add in the agitating solution of dichloromethane 53.8mmol) (150ml) triethylamine (37.5ml, 269mmol) and DMSO (41.6ml, 538mmol).This reactant mixture is cooled to 0 ℃, and gradation adds SO 3(25.7g's Py 161mmol), spends the night the mixture stirring that obtains under room temperature.This mixture of vacuum concentration, the dilute with water residue is then used ether extraction.Wash organic layer with water, through dried over anhydrous sodium carbonate and vacuum concentration.Residue as eluant, obtains (4-N-carbobenzoxy group methyl piperidine base) acetaldehyde (4.63g, 31%) with ethyl acetate-hexane (2: 1) through silica gel column chromatography, is colorless oil. 1H-NMR(CDCl 3)δ1.36-1.46(m，2H)，1.69(br?s，2H)，1.72(brs，1H)，1.90(m，1H)，2.21(dt，J＝11.6，2.4Hz，2H)，2.37(dd，J＝6.8，1.6Hz，2H)，2.92(d，J＝11.6Hz，2H)，3.25(s，2H)，5.16(s，2H)，7.35(m，5H)，9.77(t，J＝2.0Hz，1H)。
(1.06l, (560ml, 9.75mmol) (1.79g, (5ml) solution of methanol 6.50mmol) also is cooled to 0 ℃ with (4-N-carbobenzoxy group methyl piperidine base) acetaldehyde to add AcOH in the agitating solution of methanol 9.75mmol) (20ml) to the N-benzylamine.Disposable adding NaBH 3CN (645mg, 9.75 mmol) stirs the mixture overnight that obtains under room temperature.This mixture of vacuum concentration in residue impouring sodium bicarbonate aqueous solution, is used chloroform extraction then.Wash organic layer with water, through anhydrous sodium sulfate drying and vacuum concentration.Residue as eluant, obtains N-benzyl-2-[4-(N-carbobenzoxy group methyl piperidine base) ethamine (872mg, 37%) with chloroform-methanol-ethyl acetate (10: 1: 1) through silica gel column chromatography, is colorless oil. 1H-NMR(CDCl 3)δ1.36-1.46(m，2H)，1.69(br?s，2H)，1.72(br?s，1H)，1.90(m，1H)，2.21(dt，J＝11.6，2.4Hz，2H)，2.37(dd，J＝6.8，1.6Hz，2H)，2.92(d，J＝11.6Hz，2H)，3.25(s，2H)，5.16(s，2H)，7.35(m，5H)，9.77(t，J＝2.0Hz，1H)。
Under room temperature, to N-benzyl-2-[4-(N-carbobenzoxy group methyl piperidine base) ethamine (356mg, 0.972mmol), 4-[N '-(2-bromo phenyl) urea groups]-3-methoxybenzene guanidine-acetic acid (369mg, 0.972mmol) and N, N-dimethyl aminopyridine (119mg, 0.972mmol) the agitating solution of DMF (15ml) in add EDCHCl (372mg 1.94mmol), stir the mixture that obtains and spends the night.In this reactant mixture impouring water, use ethyl acetate extraction.With salt water washing organic layer, through anhydrous sodium sulfate drying, vacuum concentration then.Residue is through silica gel column chromatography, with chloroform-methanol (10: 1) as eluant, obtain 4-[2-N-[4-[N '-(2-bromo phenyl) urea groups]-the 3-methoxyphenyl]-N-benzyl acetylamino] ethyl-piperidino benzyl acetate (611mg, 86%), be colorless oil. 1H-NMR (CDCl 3) rotamer mixture δ 1.15-2.09 (a plurality of m, 7H), 2.06-2.14 (m, 2H), 2.84-2.96 (m, 2H), (3.17-3.23 s, 2H altogether), 3.21 and 3.23 (s, 2H altogether), 3.38-3.42 (m, 1H), 3.65 with 3.73 (s, 2H altogether), 3.83 and 3.84 (s, 3H altogether), 4.49 (s, 1H), 4.60 (s, 1H), 5.15 (d, J=2.8Hz, 2H), and 6.74-7.83 (a plurality of m, 16H), 7.51-7.53 (m, 1H), 7.94-8.02 (m, 1H), 8.18 (d, J=8.4Hz, 2H); MS (FAB) m/z 727 (M +), 729 (M ++ 2).
Under room temperature, to 4-[2-N-[4-[N '-(2-bromo phenyl) urea groups]-the 3-methoxyphenyl]-N-benzyl acetylamino] ethyl-piperidino benzyl acetate (347mg, 0.477mmol) methanol-water (5: 1, add Lithium hydrate (13.6mg in agitating solution 6ml), 0.57mmol), the mixture stirring that obtains is spent the night.In this reactant mixture impouring water,, use chloroform extraction then with the neutralization of 1N HCl aqueous solution.With salt water washing organic layer, through anhydrous sodium sulfate drying, vacuum concentration then.Residue as eluant, obtains being 332 (97.3mg, 32%) of colourless amorphous solid and 333 (168mg, 54%) of colourless amorphous solid with chloroform-methanol (10: 1) respectively through silica gel column chromatography.332 1H-NMR (CD 3OD), and the mixture δ 1.29-1.90 of rotamer (a plurality of m, 7H), 2.75-2.86 (m, 2H), 3.28-3.51 (a plurality of m, 6H), 3.74 and 3.78 (s, 2H altogether), 3.87 and 3.89 (s, be total to 3H), 4.66 (s, 1H), 4.85 (s, 1H), 6.79-7.00 (a plurality of m, 3H), 7.16 (d, J=7.2Hz, 1H), 7.23-7.37 (m, 5H), 7.57 (dd, J=8.4,1.2Hz, 1H), and 7.88-8.00 (a plurality of m, 2H); MS (FAB) m/z 637 (M +), 639 (M ++ 2).333 1H-NMR (CDCl 3), the mixture δ 1.15-2.11 of rotamer (a plurality of m, 7H), 2.89-2.97 (m, 2H), 3.17-3.49 (a plurality of m, 6H), 3.17 and 3.18 (s, 3H altogether), 3.70 with 3.71 (s, 3H altogether), 3.83 and 3.84 (s, be total to 2H), 4.49,4.60 and 4.71 (s, be total to 2H), and 6.75-8.16 (a plurality of m, 14H); MS (FAB) m/z 651 (M +), 653 (M ++ 2).Embodiment 2834-[2-N-[4-[N '-(2-chlorophenyl)-3-methoxyl group urea groups] phenyl]-N-benzyl acetylamino] ethyl-piperidino acetic acid
Figure A0080951005851
Under room temperature, to N-benzyl-2-[4-(N-carbobenzoxy group methyl piperidine base) ethamine (372mg, 1.11mmol), 4-[N '-(2-chlorophenyl) urea groups]-the 3-methoxyphenyl] acetic acid (758mg, 1.11mmol) and N, N-dimethyl aminopyridine (136mg, 1.11mmol) the agitating solution of DMF (15ml) in add EDCHCl (426mg 2.22mmol), stir the mixture that obtains 12 hours.In this reactant mixture impouring water, use ethyl acetate extraction.With salt water washing organic layer, through anhydrous sodium sulfate drying, vacuum concentration then.Residue is through silica gel column chromatography, with chloroform-methanol (10: 1) as eluant, obtain 4-[2-N-[4-N '-(2-chlorophenyl)-3-methoxyl group urea groups] phenyl]-N-benzyl acetylamino] ethyl-piperidino benzyl acetate (668mg, 88%), be faint yellow oily thing. 1H-NMR (CDCl 3) rotamer mixture δ 1.15-1.83 (a plurality of m, 7H), 2.04-2.13 (m, 2H), 2.83-2.95 (m, 2H), 2.88 and 2.99 (s, 2H altogether), 3.20 with 3.23 (s, 2H altogether), and 3.20-3.23 (overlapping, 1H), 3.38-3.42 (m, 1H), 3.65-3.74 (m, 3H), 4.50 (s, 1H), 4.61 (s, 1H), 5.14 (d, J=4.4Hz, 2H), and 6.72-6.82 (a plurality of m, 2H), 6.94-6.99 (m, 1H), 7.13-7.50 (a plurality of m, 14H), and 7.94-8.02 (m, 1H), 8.18 (d, J=8.0Hz, 2H); MS (FAB) m/z 683 (M ++ H).
Under room temperature, to 4-[2-N-[4-[N '-(2-chlorophenyl)-3-methoxyl group urea groups] phenyl]-N-benzyl acetylamino] ethyl-piperidino benzyl acetate (633mg, 0.93mmol) methanol-water (10: 1, add Lithium hydrate (24.4mg in agitating solution 10ml), 1.02mmol), the mixture stirring that obtains is spent the night.In this reactant mixture impouring water,, use chloroform extraction then with the neutralization of 1N HCl aqueous solution.With salt water washing organic layer, through anhydrous sodium sulfate drying, vacuum concentration then.Residue is through silica gel column chromatography, with chloroform-methanol (20: 1) as eluant, obtain 4-[2-N-[4-[N '-(2-chlorophenyl)-3-methoxyl group urea groups] phenyl]-N-benzyl acetylamino] ethyl-piperidino methyl acetate (504mg, 89%), be colourless amorphous solid. 1H-NMR (CDCl 3), the mixture δ 1.26-1.64 of rotamer (a plurality of m, 7H), 2.02-2.10 (m, 2H), 2.86 (t, J=10.4Hz, 2H), 3.17 (d, J=8.0Hz, 2H), 3.16-3.22 (m, overlapping, 1H), 3.40 (t, J=7.6Hz, 1H), 3.48 (s, 1H), 3.65 and 3.73 (s, be total to 2H), 3.70 and 3.71 (s, 3H altogether), 3.79 and 3.80 (s, be total to 3H), 4.50,4.60 and 4.70 (s, 2H altogether), 6.73-6.85 (a plurality of m, 2H), 6.98 (t, J=7.6Hz, 1H), and 7.13-7.37 (a plurality of m, 9H), 7.96 (m, 1H), 8.18 (d, J=8.0Hz, 2H); MS (FAB) m/z 607 (M ++ H).
To 4-[2-N-[4-[N '-(2-chlorophenyl)-3-methoxyl group urea groups] phenyl]-N-benzyl acetylamino] ethyl-piperidino methyl acetate (177mg, 0.292mmol) methanol-water (5: 1, add Lithium hydrate (21.6mg in agitating solution 6ml), 0.90mmol), under room temperature, the mixture that obtains was stirred 4 hours.In this mixture impouring water,, use chloroform extraction then with 1N HCl aqueous solution this solution that neutralizes.With salt water washing organic layer, through anhydrous sodium sulfate drying, concentrate then and obtain 334 (155mg, 92%), be colourless amorphous solid. 1H-NMR (CD 3OD), the mixture δ 1.28-1.90 of rotamer (a plurality of m, 7H), 2.84 (brs, 2H), 3.30-3.52 (a plurality of m, 6H), 3.74 and 3.82 (s, 2H altogether), 3.87 and 3.88 (s, be total to 3H), 4.63 (s, 1H), 4.66 (s, 1H), and 6.79-7.05 (a plurality of m, 3H), 7.16 (d, J=7.2Hz, 1H), 7.24-7.40 (m, 5H), 7.88 (s, 1H), 7.98-8.04 (a plurality of m, 2H); MS (ESI) m/z 593,615 (M ++ Na +).
Should be appreciated that when this paper describes when of the present invention according to the particular of elaboration, these embodiments propose by illustrating General Principle, the present invention also needn't make restriction to this.Under the situation of marrow that does not deviate from following claims and scope, for a person skilled in the art, any given material or the modification and the change of processing step all are conspicuous, and all such modifications include within the scope of the invention.
19. the compound or its salt of claim 2, wherein R 5Be lower alkoxy.
20. a method that suppresses cell adhesion in mammal, described method comprise among the claim 1-19 that gives described mammal effective dose each chemical compound.
21. the method for treatment and the cell adhesion diseases associated of VLA-4 mediation in a mammal, described method comprise among the claim 1-19 that gives described mammal effective dose each chemical compound.
22. the method for claim 21, wherein said cell adhesion diseases associated with the VLA-4 mediation is selected from inflammatory reaction, autoimmunity reaction and neoplasm metastasis.
23. the method for claim 21, wherein said cell adhesion diseases associated with the VLA-4 mediation is selected from asthma, diabetes, arthritis, psoriasis, multiple sclerosis, inflammatory bowel disease and graft-rejection.
24. the purposes of each chemical compound in treatment among the claim 1-19.
25. each chemical compound is used for the treatment of purposes in the medicine with the cell adhesion diseases associated of VLA-4 mediation in preparation among the claim 1-19.
26. a Pharmaceutical composition, it comprises as each chemical compound and pharmaceutically acceptable carrier or excipient among the claim 1-19 of therapeutic agent.
27. the Pharmaceutical composition of claim 26, it also comprises one or more other therapeutic agent.
28. the Pharmaceutical composition of claim 27, wherein said one or more other therapeutic agent is selected from anti-inflammatory agent, antirheumatic, corticosteroid, immunosuppressant, psoriasis medicine, bronchodilator, antasthmatic and antidiabetic drug.
29. the Pharmaceutical composition of claim 28, a kind of in wherein said one or more other the therapeutic agent is anti-inflammatory agent.
30. the Pharmaceutical composition of claim 29, wherein said anti-inflammatory agent is selected from steroid and NSAID.

Claims (19)

1. compound or its salt by formula I representative,
Figure A0080951000021
Wherein W is selected from the heteroaryl of aryl, heteroaryl and the replacement of aryl, replacement; W 1Be selected from the inferior heteroaryl of arlydene, inferior heteroaryl and the replacement of arlydene, replacement; A is selected from=O ,=S and=NH; R be selected from a direct key, alkylene group and-(CH 2) n-;
Wherein
N is selected from 1 and 2; X is selected from-C (O)-,-CH 2-and S (O) 2M is selected from
Wherein Be 4-, 5-, 6-or the 7-unit heterocyclic moiety of bivalence,
Wherein nitrogen-atoms is the junction point with X; R 1, R 2And R 3Independently be selected from-H ,-OH ,-NH 2, halogen atom, alkyl, replacement
The alkoxyl of the aryl of alkyl, aryl, replacement, alkoxyl, replacement, an alkyl
The dialkyl amido of amino, an alkyl amino that replaces, dialkyl amido, replacement,
The cycloalkyl amino of cycloalkyl amino, replacement, alkyl sulfonyl-amino, replacement
The aryl sulfonyl ammonia of alkyl sulfonyl-amino, arlysulfonylamino, replacement
The heteroaryloxy of the aryloxy group of base, aryloxy group, replacement, heteroaryloxy, replacement,
The benzyloxy of benzyloxy and replacement, or R 1, R 2And R 3In two combine and can form by independently being selected from following 1-3
Optional 3-, the 4-that replaces of individual substituent group, 5-, 6-or 7-unit's carbocyclic ring or heterocycle are residual
Base :-OH, halogen atom ,-NH 2, alkyl, alkoxyl, aryl, aryloxy group,
Alkyl amino, benzyloxy and heteroaryl; R 4Be selected from-H and low alkyl group; Y is a direct key or be selected from-C (O)-,-C (O) NH-, alkylene group, inferior alkynyl group
With-(CH 2) kY 2Divalent group,
Wherein
K is selected from 1,2 and 3; With
Y 2Be a direct key or be selected from-O-,-S-,-S (O)-,-S (O) 2-and-NY 3-
Divalent group,
Wherein
Y 3Be selected from-H and low alkyl group;
It is assorted that Z is selected from the Asia of the arlydene of arlydene, replacement, inferior heterocyclic radical, replacement
The cycloalkylidene of cyclic group, cycloalkylidene and replacement; A 1Be a direct key or be selected from alkylene group, inferior alkynyl group ,-(CH 2) t-and-O (CH 2) v
Divalent group,
Wherein
T is selected from 1,2 and 3; With
V is selected from 0,1,2 and 3; And R 5Be selected from-OH, lower alkoxy ,-N (H) OH,
Figure A0080951000031
Wherein Be 4-, 5-, 6-or the 7-unit heterocyclic moiety of bivalence,
Wherein nitrogen-atoms is the junction point with X; R 6And R 7Independently be selected from-H ,-OH, halogen atom, alkyl and alkoxyl; Y 1For be selected from-O-,-S-,-S (O)-,-S (O) 2-and-NY 4-divalent group,
Wherein
Y 4Be selected from-H and low alkyl group; Z 1For mixing the Asia of the arlydene that is selected from arlydene, replacement, inferior heterocyclic radical, replacement
The divalent group of the cycloalkylidene of cyclic group, cycloalkylidene and replacement; A 2Be a direct key or be selected from alkylene group, inferior alkynyl group and-(CH 2) e-two
The valency group,
Wherein
E is selected from 1,2 and 3; And R 8Be selected from-OH, lower alkoxy ,-N (H) OH,
Figure A0080951000042
Wherein
Figure A0080951000043
Wherein Be 4-, 5-, 6-or the 7-unit heterocyclic moiety of bivalence, optional by independently being selected from following 1-3 substituent group replacement: alkyl, alkoxyl, hydroxyalkyl ,-OH, benzyloxy ,-NH 2, halogen atom, aryl and heteroaryl, described part can condense with 1 or 2 other carbocyclic ring or heterocycle residue, and a following 1-3 substituent group is optional to be replaced by independently being selected from for described carbocyclic ring or heterocycle residue: alkyl, aryloxy group, alkoxyl, hydroxyalkyl ,-OH, benzyloxy ,-NH 2, halogen atom, aryl and heteroaryl; M and q independently are selected from 0,1,2 and 3; X 1Be selected from-CH=and-N=; R 9Be selected from-H and low alkyl group; R 10Be selected from-COOH, elementary alkoxy carbonyl, With And Z 2Be selected from-H, COOH and elementary alkoxy carbonyl; With
R wherein 11Be selected from-O-,
Figure A0080951000054
With-NR 12-
Wherein
R 12Be selected from-H, the cycloalkanes of the alkyl of alkyl, replacement, cycloalkyl, replacement
The benzyl of the aryl of base, aryl, replacement, benzyl, replacement, rudimentary
The low-grade alkenyl of alkenyl, replacement and alkynyl of low-grade chain;
The left hand key for the junction point right hand key with-X-is and-Z 3Junction point; Z 3Be selected from a direct key, have the divalent aliphatic hydrocarbon part of 1-12 carbon atom,
Wherein
One or more carbon atoms can by-O-or-NR 13-displacement,
Wherein
R 13Be selected from-H and low alkyl group and
The one or more hydrogen atoms that are connected in aliphatic carbon atom can be by lower alkyl
The base displacement; With
Figure A0080951000055
Wherein
X is selected from 0 and 1; Y is selected from 1,2 and 3; And R 14Be selected from-H ,-OH and halogen atom,
Figure A0080951000061
With, work as R 11For-NR 12,
Figure A0080951000062
Z wherein 4Be selected from
Figure A0080951000063
Wherein
R 14aBe selected from-H ,-OH, low alkyl group and halogen atom; With Wherein
The left hand key is and R 11Junction point and right hand key is and Q 2
Junction point; Q 2For mixing the Asia of the arlydene that is selected from arlydene, replacement, inferior heterocyclic radical, replacement
The divalent group of the cycloalkylidene of cyclic group, cycloalkylidene, replacement, R wherein 15And R 16Independently be selected from-H, halogen atom
And low alkyl group; With R wherein 17And R 18Independently be selected from-H, low alkyl group,
The low alkyl group and the low-grade alkenyl that replace; And L 1Be selected from-COOH and-COOR 19
Wherein
R 19Be low alkyl group.
2. the compound or its salt of claim 1, wherein M is
Figure A0080951000068
3. the compound or its salt of claim 1, wherein M is
Figure A0080951000071
4. the compound or its salt of claim 1, wherein M is
5. the compound or its salt of claim 1, wherein M is
6. the compound or its salt of claim 2, wherein R 1, R 2And R 3In at least one group be-OH or halogen atom.
7. the compound or its salt of claim 6, wherein A is=O, R is-(CH 2) n-with X be-C (O)-.
8. the compound or its salt of claim 7, wherein Y be selected from alkylene group, inferior alkynyl group and-(CH 2) kY 2Y 2Be selected from a direct key ,-O-,-S (O) and-NY 3-, and Y 3For-H.
9. the compound or its salt of claim 8, wherein Y be selected from-O-and-NY 3-.
10. the compound or its salt of claim 2, wherein W is unsubstituted phenyl or has 1 or 2 substituent phenyl that is selected from low alkyl group and halogen atom at its ortho position.
11. the compound or its salt of claim 10, wherein W 1Has a substituent phenylene that is selected from methoxyl group, low alkyl group and halogen atom for unsubstituted phenylene or at its ortho position with respect to-NH-.
12. the compound or its salt of claim 2, wherein W 1For having a substituent group that is selected from methoxyl group, low alkyl group and halogen atom and have 1-3 substituent phenylene that is selected from low alkyl group and halogen atom at its ortho position with respect to-NH-.
13. the compound or its salt of claim 2, wherein A 1Be a direct key or-(CH 2) t-.
14. the compound or its salt of claim 13, wherein A 1It is a direct key.
15. the compound or its salt of claim 14, wherein R 5For-OH.
16. the compound or its salt of claim 2, wherein W is unsubstituted phenyl or has 1 or 2 substituent phenyl that is selected from low alkyl group and halogen atom at its ortho position; W 1Has a substituent phenylene that is selected from methoxyl group, low alkyl group and halogen atom for unsubstituted phenylene or at its ortho position with respect to-NH-; R is-CH 2-; X is-C (O)-and R 5For-OH.
17. the compound or its salt of claim 16, it is selected from
Figure A0080951000081
Figure A0080951000141
Figure A0080951000151
Figure A0080951000171
Figure A0080951000181
18. the compound or its salt of claim 17, it is selected from
Figure A0080951000191
Figure A0080951000211
Figure A0080951000251
19. the compound or its salt of claim 2, wherein R 5Be lower alkoxy.
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