CN106146493A - Pyrazolo [3,4-b] pyridines and the preparation method of indazole compounds and purposes - Google Patents

Pyrazolo [3,4-b] pyridines and the preparation method of indazole compounds and purposes Download PDF

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CN106146493A
CN106146493A CN201510192164.3A CN201510192164A CN106146493A CN 106146493 A CN106146493 A CN 106146493A CN 201510192164 A CN201510192164 A CN 201510192164A CN 106146493 A CN106146493 A CN 106146493A
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alkyl
base
alkoxyl
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段文虎
耿美玉
赵彬
丁健
艾菁
范珺
彭霞
严伟
陈奕
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Shanghai Institute of Materia Medica of CAS
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Shanghai Institute of Materia Medica of CAS
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    • C—CHEMISTRY; METALLURGY
    • C07—ORGANIC CHEMISTRY
    • C07D—HETEROCYCLIC COMPOUNDS
    • C07D231/00—Heterocyclic compounds containing 1,2-diazole or hydrogenated 1,2-diazole rings
    • C07D231/54—Heterocyclic compounds containing 1,2-diazole or hydrogenated 1,2-diazole rings condensed with carbocyclic rings or ring systems
    • C07D231/56—Benzopyrazoles; Hydrogenated benzopyrazoles
    • C—CHEMISTRY; METALLURGY
    • C07—ORGANIC CHEMISTRY
    • C07D—HETEROCYCLIC COMPOUNDS
    • C07D403/00—Heterocyclic compounds containing two or more hetero rings, having nitrogen atoms as the only ring hetero atoms, not provided for by group C07D401/00
    • C07D403/02—Heterocyclic compounds containing two or more hetero rings, having nitrogen atoms as the only ring hetero atoms, not provided for by group C07D401/00 containing two hetero rings
    • C07D403/12—Heterocyclic compounds containing two or more hetero rings, having nitrogen atoms as the only ring hetero atoms, not provided for by group C07D401/00 containing two hetero rings linked by a chain containing hetero atoms as chain links
    • C—CHEMISTRY; METALLURGY
    • C07—ORGANIC CHEMISTRY
    • C07D—HETEROCYCLIC COMPOUNDS
    • C07D471/00—Heterocyclic compounds containing nitrogen atoms as the only ring hetero atoms in the condensed system, at least one ring being a six-membered ring with one nitrogen atom, not provided for by groups C07D451/00 - C07D463/00
    • C07D471/02—Heterocyclic compounds containing nitrogen atoms as the only ring hetero atoms in the condensed system, at least one ring being a six-membered ring with one nitrogen atom, not provided for by groups C07D451/00 - C07D463/00 in which the condensed system contains two hetero rings
    • C07D471/04—Ortho-condensed systems

Abstract

The invention provides a kind of pyrazolo [3,4-b] pyridines and the preparation method of indazole compounds and purposes, specifically, the invention provides a kind of compound as shown in following formula (I), wherein, the definition of each group is as noted in the discussion.The compounds of this invention has the tyrosine-kinase enzyme inhibition activity of excellence, therefore can be used in preparing the medicine of a series for the treatment of disease relevant to tyrosine kinase activity.

Description

Pyrazolo [3,4-b] pyridines and the preparation method of indazole compounds and purposes
Technical field
The present invention relates to pyrazolo [3,4-b] pyridines of structure as shown in formula and indazole compounds, its pharmaceutical salts, before it Medicine and hydrate thereof or solvate, be directed to the preparation method of described compound, comprise the pharmaceutical composition of described compound And it is as protein tyrosine kinase inhibitor, especially as FGFR inhibitor, at relevant diseases such as treatment cancers Purposes in medicine.
Background technology
Protein kinase is a kind of by the phosphorylation of specific amino acids on protein is regulated the protein of various cell function (enzyme).Protein by the change of conformation regulate activity and with other component binding abilities.The activity of protein kinase refers to , speed that phosphate group is attached on substrate by kinases, this speed can be by being converted into product in detection certain time The amount of substrate be measured.The phosphorylation of substrate occurs on the activation site of protein kinase.
Tyrosine kinase is a kind of protease that can be catalyzed and adenosine triphosphate is transferred to protein-tyrosine residue.This A little kinases role that performer is important in somatomedin conduction trigger cell propagation, differentiation and transition process.
Fibroblast growth factor (FGF) has been identified have important regulation effect, such as device in many physiological process Official generates and angiogenesis etc..It is known that have more than 25 kinds of hypotypes, fibroblast growth factor in FGF family Receptor (FGFR) family comprises four hypotypes (FGFR1-4) altogether, and they are glycoproteins, comprises extracellular para-immunity ball egg White region, the hydrophobic region of cross-film and intracytoplasmic tyrosine kinase domain.The combination of FGF causes FGFR dimerization, enters And there is the autophosphorylation of receptor and the activation of downstream signaling pathway.Some specific components of downstream signaling pathway is raw for cell Length, metabolism and survival have very important effect.Therefore, FGFR signal path for the breeding of tumor cell, migration, Infiltration and angiogenesis have polyphenic important physiological action.
At present, it has been demonstrated that FGF signal path has with human cancer directly associates.At different types of cancerous cell In (bladder cancer, renal carcinoma and carcinoma of prostate etc.) FGF process LAN phenomenon of being all in the news out different.Therefore, FGF signal Path is a promising therapy target.
In sum, this area is in the urgent need to developing novel tyrosine kinase inhibitor.
Summary of the invention
It is an object of the present invention to provide pyrazolo [3,4-b] pyridines and the indazole class with structure shown in lower formula (I) The stereoisomer of compound, geometric isomer, tautomer, its pharmaceutical salts, its prodrug and hydrate thereof or solvate.
Further object is that above-mentioned pyrazolo [3,4-b] pyridines and the method for indazole compounds are prepared in offer.
It is yet a further object of the present invention to provide comprise therapeutically effective amount selected from above-mentioned pyrazolo [3,4-b] pyridines and Yin The pharmaceutical composition of one or more in azole compounds, its pharmaceutical salts, its prodrug and hydrate thereof or solvate.
A further object of the present invention is to provide selected from above-mentioned pyrazolo [3,4-b] pyridines and indazole compounds, it is medicinal One or more in salt, its prodrug and hydrate thereof or solvate, as protein tyrosine kinase inhibitor, are especially made For FGFR1-3 inhibitor, prevent in preparation and/or treat the medicine with FGF/FGFR signal path unconventionality expression relevant disease In purposes.
A first aspect of the present invention, it is provided that a kind of compound as shown in following formula (I) or its pharmaceutical salts, its prodrug, Its hydrate or solvate,
Wherein:
Y is selected from CH or N；
R1Selected from substituted or unsubstituted 6-10 unit aryl, substituted or unsubstituted miscellaneous former selected from N, O and S containing 1-3 The 5-10 unit heteroaryl of son；Described substituent group be one or more be selected from halogen, C1～C6 alkyl, C1～C6 alkoxyl, C1～C6 alkylthio group, halo C1～C6 alkyl, halo C1～C6 alkoxyl, cyano group, C1～C6 alkanoyl, In-NH-CO-C1～C6 alkyl, amino ,-NH (C1～C6 alkyl) ,-N (C1～C6 alkyl) (C1～C6 alkyl) and hydroxyl Substituent group；
R2Selected from hydrogen, C1～C6 alkyl, C1～C6 alkanoyl ,-CO (O)-C1～C6 alkyl；
R3Selected from substituted or unsubstituted 6-10 unit aryl, substituted or unsubstituted miscellaneous selected from N, O and S containing 1-3 The 5-10 unit heteroaryl of atom；Described substituent group be one or more be selected from halogen, C1～C6 alkyl, C1～C6 alkoxyl, C1～C6 alkylthio group, cyano group, hydroxyl, halo C1～C6 alkyl, halo C1～C6 alkoxyl, C1～C6 alkanoyl, -NH-CO-C1～C6 alkyl, amino ,-C1～C6 alkyl-NH2,-NH (C1～C6 alkyl) ,-N (C1～C6 alkane Base) (C1～C6 alkyl) ,-CO (O)-C1～C6 alkyl, C3-C8 heterocyclic radical ,-Y1-(C1～C6 alkyl)-Y2, C1～C6 Substituent group in alkyl-C3～C8 heterocyclic radical and C3～C8 cycloalkyl-C3～C8 heterocyclic radical, described Y1 is selected from O, NH and S； Described Y2 is selected from C1～C6 alkoxyl, amino ,-NH (C1～C6 alkyl) and-N (C1～C6 alkyl) (C1～C6 alkyl)； Wherein said C3-C8 heterocyclic radical contains 1-3 the hetero atom selected from N, O and S, and optionally by one or more Selected from C1～C6 alkyl, C1～C6 alkoxyl, C3～C8 cycloalkyl, C2～C6 thiazolinyl, C2～C6 alkynyl, C1～C6 Alkanoyl, C1～C6 alkoxyl substituted C1～C6 alkyl, C1～C6 alkylamino substituted C1～C6 alkyl, hydroxyl take C1～the C6 alkyl in generation ,-CO (O)-C1～C6 alkyl, hydroxyl, halogen ,-SO2Substituent group in-C1～C6 alkyl is entered One step replaces；
R4、R5It is each independently selected from hydrogen, halogen, C1～C6 alkyl, C2～C6 thiazolinyl, C2～C6 alkynyl, halo C1～C6 alkyl, C1～C6 alkoxyl, halo C1～C6 alkoxyl, C1～C6 alkylthio group, cyano group, C1～C6 alkane acyl Base, nitro, amino ,-NH (C1～C6 alkyl) ,-N (C1～C6 alkyl) (C1～C6 alkyl) ,-NH-CO-C1～C6 Alkyl and-CO (O)-C1～C6 alkyl；
And when Y is CH, R3At least there is less than one substituent group: C3-C8 heterocyclic radical；Wherein, described C3-C8 Heterocyclic radical as defined above.
In another preference, R1Selected from substituted or unsubstituted 6-10 unit aryl, substituted or unsubstituted containing 1-3 Individual selected from N, O and S heteroatomic 5-10 unit heteroaryl；Described substituent group is that one or more are selected from halogen, C1～C4 Alkyl, C1～C4 alkoxyl, C1～C4 alkylthio group, halo C1～C4 alkyl, halo C1～C4 alkoxyl, cyano group, C1～C4 alkanoyl ,-NH-CO-C1～C4 alkyl, amino ,-NH (C1～C4 alkyl) ,-N (C1～C4 alkyl) (C1～C4 Alkyl) and hydroxyl in substituent group；
R2Selected from hydrogen, C1～C4 alkyl, C1～C4 alkanoyl ,-CO (O)-C1～C4 alkyl；
R3Selected from substituted or unsubstituted 6-10 unit aryl, substituted or unsubstituted miscellaneous selected from N, O and S containing 1-3 The 5-10 unit heteroaryl of atom；Described substituent group be one or more be selected from halogen, C1～C4 alkyl, C1～C4 alkoxyl, C1～C4 alkylthio group, cyano group, hydroxyl, halo C1～C4 alkyl, halo C1～C4 alkoxyl, C1～C4 alkanoyl, -NH-CO-C1～C4 alkyl, amino ,-C1～C4 alkyl-NH2,-NH (C1～C4 alkyl) ,-N (C1～C4 alkane Base) (C1～C4 alkyl) ,-CO (O)-C1～C4 alkyl, C3-C8 heterocyclic radical ,-Y1-(C1～C4 alkyl)-Y2, C1～C4 Substituent group in alkyl-C3～C8 heterocyclic radical and C3～C8 cycloalkyl-C3～C8 heterocyclic radical, described Y1 is selected from O, NH and S； Described Y2 is selected from C1～C4 alkoxyl, amino ,-NH (C1～C4 alkyl) and-N (C1～C4 alkyl) (C1～C4 alkyl)； Wherein said C3-C8 heterocyclic radical contains 1-3 the hetero atom selected from N, O and S, and optionally by one or more Selected from C1～C4 alkyl, C1～C4 alkoxyl, C3～C8 cycloalkyl, C2～C4 thiazolinyl, C2～C4 alkynyl, C1～C4 Alkanoyl, C1～C4 alkoxyl substituted C1～C4 alkyl, C1～C4 alkylamino substituted C1～C4 alkyl, hydroxyl take C1～the C4 alkyl in generation ,-CO (O)-C1～C4 alkyl, hydroxyl, halogen ,-SO2Substituent group in-C1～C4 alkyl is entered One step replaces；
R4、R5It is each independently selected from hydrogen, halogen, C1～C4 alkyl, C2～C4 thiazolinyl, C2～C4 alkynyl, halo C1～C4 alkyl, C1～C4 alkoxyl, halo C1～C4 alkoxyl, C1～C4 alkylthio group, cyano group, C1～C4 alkane acyl Base, nitro, amino ,-NH (C1～C4 alkyl) ,-N (C1～C4 alkyl) (C1～C4 alkyl) ,-NH-CO-C1～C4 Alkyl and-CO (O)-C1～C4 alkyl；
And when Y is CH, R3At least there is less than one substituent group: C3-C8 heterocyclic radical；Wherein, described C3-C8 Heterocyclic radical as defined above.
In another preference, R1Selected from substituted or unsubstituted following aryl or heteroaryl: phenyl, naphthyl, pyrrole Piperidinyl, pyrazinyl, furyl, thienyl, pyrrole radicals, oxazolyl, thiazolyl, pyrazolyl, imidazole radicals, indole Base, benzofuranyl, benzothiazolyl and benzimidazolyl；Described substituent group is that one or more are selected from halogen, C1～C4 Substituent group in alkyl, C1～C4 alkoxyl and-N (C1～C4 alkyl) (C1～C4 alkyl)；
R3In replacement or unsubstituted following aryl or heteroaryl: phenyl, naphthyl, pyridine radicals, pyrazinyl, thiophene Base, furyl, imidazole radicals, pyrrole radicals, oxazolyl, thiazolyl, pyrazolyl, indyl, benzofuranyl, benzene Benzothiazolyl, benzimidazolyl；Described substituent group is that one or more are selected from halogen, C1～C4 alkyl, C1～C4 alkane Epoxide, amino ,-NH (C1～C4 alkyl) ,-N (C1～C4 alkyl) (C1～C4 alkyl), C3-C8 heterocyclic radical and Substituent group in-Y1-(C1～C4 alkyl)-Y2, described Y1 is selected from O, NH and S；Described Y2 selected from C1～C4 alkoxyl, Amino ,-NH (C1～C4 alkyl) and-N (C1～C4 alkyl) (C1～C4 alkyl)；Wherein said C3-C8 heterocyclic radical contains 1-3 the hetero atom selected from N, O and S, and optionally it is selected from C1～C4 alkyl, C1～C4 alcoxyl by one or more Base, C3～C8 cycloalkyl, C2～C4 thiazolinyl, C2～C4 alkynyl, C1～C4 alkanoyl, C1～C4 alkoxyl are substituted C1～C4 alkyl, hydroxyl substituted C1～C4 alkyl ,-CO (O)-C1～C4 alkyl and-SO2Replacement in-C1～C4 alkyl Base is further substituted with；
R4、R5It is each independently selected from hydrogen, halogen, C1～C4 alkyl and halo C1～C4 alkyl；
And when Y is CH, R3At least there is less than one substituent group: C3-C8 heterocyclic radical；Wherein, described C3-C8 Heterocyclic radical as defined above.
In another preference, R1Selected from substituted or unsubstituted following aryl or heteroaryl: phenyl, naphthyl, pyridine Base, furyl and benzimidazolyl (preferably phenyl or benzimidazolyl, more preferably phenyl)；Described substituent group is One or more substituent groups in F, Cl, methyl, methoxyl group and N, N-dimethylamino；
In another preference, R2Selected from lower group: hydrogen, methyl, acetyl group and tertbutyloxycarbonyl.
In another preference, when Y is N, R3Selected from substituted or unsubstituted following aryl or heteroaryl: phenyl, Naphthyl, pyridine radicals, pyrazinyl, thienyl, indyl；R3Optionally by one or more selected from fluorine, chlorine, first Base, methoxyl group, amino, dimethylamino, piperazinyl, piperidyl, morpholinyl, homopiperazine base, thio-morpholinyl, Pyrrolidinyl, tetrahydrofuran base ,-NH (CH2)2OCH3、-NH(CH2)2N(CH3)2With-O (CH2)2OCH3In take Replace for base, and piperazinyl in substituent group, piperidyl, morpholinyl, homopiperazine base, thio-morpholinyl, pyrrolidine Base and tetrahydrofuran base are selected from methyl, ethyl, isopropyl, cyclopropyl, ring by one or more the most further Butyl ,-(CH2)2OCH3, pi-allyl ,-(CH2)2Replacement in OH, acetyl group, mesyl and tertbutyloxycarbonyl Base replaces；
In another preference, when Y is C, R3In substituted following aryl or heteroaryl: phenyl, naphthyl, Pyridine radicals, pyrazinyl, thienyl, indyl；R3Can by one or more selected from fluorine, chlorine, methyl, methoxyl group, Amino, dimethylamino, piperazinyl, piperidyl, morpholinyl, homopiperazine base, thio-morpholinyl, pyrrolidinyl, four Hydrogen furyl ,-NH (CH2)2OCH3、-NH(CH2)2N(CH3)2With-O (CH2)2OCH3In substituent group replace, and At least a part of which has 1 substituent group selected from piperazinyl, piperidyl, morpholinyl, homopiperazine base, thio-morpholinyl, pyrroles In alkyl and tetrahydrofuran base, piperazinyl therein, piperidyl, morpholinyl, homopiperazine base, thio-morpholinyl, pyrrole Cough up alkyl and tetrahydrofuran base the most further by one or more selected from methyl, ethyl, isopropyl, cyclopropyl, Cyclobutyl ,-(CH2)2OCH3, pi-allyl ,-(CH2)2Taking in OH, acetyl group, mesyl and tertbutyloxycarbonyl Replace for base.
In another preference, R4、R5It is each independently selected from hydrogen, fluorine, methyl or trifluoromethyl.
In another preference, in described compound, Y, R1、R2、R3、R4、R5In any one is respectively real Execute group corresponding in particular compound described in example.
In another preference, described compound is selected from lower group:
A second aspect of the present invention, it is provided that a kind of pharmaceutical composition, it is characterised in that including: therapeutically effective amount as Compound described in first aspect present invention, or its pharmaceutical salts, its prodrug, its hydrate or solvate, and optional medicine Acceptable carrier on.
In another preference, described pharmaceutical composition is the pharmaceutical composition for treating tumor, or is used for treating cheese The pharmaceutical composition of histidine kinase (preferably FGFR, more preferably FGFR1 and FGFR2) activity related diseases.
In another preference, described pharmaceutical composition is used for treating FGF/FGFR signal path unconventionality expression and is correlated with disease Sick.
A third aspect of the present invention, it is provided that a kind of compound as described in the first aspect of the invention, or its pharmaceutical salts, its Prodrug, its hydrate or the purposes of solvate, for preparing prevention and/or the treatment medicine selected from the disease of lower group: (a) swells Tumor relevant disease, preferably cancer；B () protein tyrosine kinase activity relevant disease, preferably FGFR activity is correlated with disease Disease, more preferably FGFR1 and/or FGFR2 relevant disease.
In another preference, described tumor-related illness be selected from lower group: breast carcinoma, pulmonary carcinoma, bladder cancer, gastric cancer, Cancer of pancreas, carcinoma of prostate, colon cancer, multiple myeloma AML, hepatocarcinoma, melanoma, head and neck cancer, thyroid carcinoma, Renal cell carcinoma, glioblast cancer and carcinoma of testis；It is preferably breast carcinoma, nonsmall-cell lung cancer, bladder cancer, gastric cancer, pancreas Cancer, carcinoma of prostate, colon cancer, multiple myeloma, hepatocarcinoma, melanoma, head and neck cancer, thyroid carcinoma, renal cell carcinoma, Glioblast cancer and carcinoma of testis；More preferably nonsmall-cell lung cancer, gastric cancer, multiple myeloma.
A fourth aspect of the present invention, it is provided that a kind of protein tyrosine kinase inhibitor alive, described inhibitor contains suppression The compound as described in the first aspect of the invention of effective dose, or its pharmaceutical salts, its prodrug, its hydrate or solvate.
In another preference, described protein tyrosine kinase is FGFR, preferably FGFR1 and/or FGFR2.
A fifth aspect of the present invention, it is provided that a kind of for treating cancer or the medicine of protein tyrosine kinase activity relevant disease Compositions, described pharmaceutical composition includes the compound as described in the first aspect of the invention of therapeutically effective amount, or it is medicinal Salt, its prodrug, its hydrate or solvate are as active component.
In another preference, described cancer be selected from lower group: breast carcinoma, nonsmall-cell lung cancer, bladder cancer, gastric cancer, Cancer of pancreas, carcinoma of prostate, colon cancer, multiple myeloma, hepatocarcinoma, melanoma, head and neck cancer, thyroid carcinoma, kidney are thin Born of the same parents' cancer, glioblast cancer and carcinoma of testis.
A sixth aspect of the present invention, it is provided that the preparation method of a kind of formula (I) compound as described in the first aspect of the invention, Described method includes step (a) or (b):
A () compound (II) or its salt prepare compound (I) in the basic conditions with corresponding acetyl halide compound (III)；Or
B () compound (II) or its salt prepare compound (I) with corresponding ester (III) under lewis acid effect；
Wherein, Q is leaving group, it is therefore preferable to halogen (acyl chlorides) or C2-C6 ester group (ester)；The definition of remaining each group is such as The above.
In another preference, described reaction temperature is 30-90 DEG C.
In another preference, described reaction is carried out in non-polar solven, is preferably entering in the solvent of lower group OK: toluene, benzene and carbon tetrachloride.
In another preference, the described response time is 2-48h.
In another preference, described lewis acid is selected from lower group: trimethyl aluminium, aluminum chloride or boron trifluoride.
A seventh aspect of the present invention, it is provided that a kind for the treatment of or prophylaxis of cancer or protein tyrosine kinase activity relevant disease Method, it is characterised in that including: to treatment object of prevention administering therapeutic or prevention effective dose such as first aspect present invention Described compound, or its pharmaceutical salts, its prodrug, its hydrate or solvate, or drug regimen as described in the present invention Thing.
In should be understood that within the scope of the present invention, above-mentioned each technical characteristic of the present invention and having in below (eg embodiment) Can be combined with each other between each technical characteristic that body describes, thus constitute new or preferred technical scheme.It is limited to a piece Width, tires out the most one by one at this and states.
Accompanying drawing explanation
Fig. 1 is the I-15 growth inhibited exercising result figure to people's pulmonary carcinoma NCI-H1581 Nude Mice.
Detailed description of the invention
The present inventor, through in-depth study for a long time, has prepared a class and has had tyrosine-kinase enzyme inhibition activity Compound of formula I.Compared with tyrosine kinase inhibitor of the prior art, described compound has higher suppression Activity.Based on above-mentioned discovery, inventor completes the present invention.
Term
In this article, in place of special instruction, it is selected that term " replaces " the one or more hydrogen atoms referring on group Replace from the substituent group of lower group: C1～C10Alkyl, C3～C10Cycloalkyl, C1～C10Alkoxyl, halogen, hydroxyl, Carboxyl (-COOH), C1～C10Aldehyde radical, C2～C10Acyl group, C2～C10Ester group, amino, phenyl；Described benzene Base includes unsubstituted phenyl or has the substituted-phenyl of 1-3 substituent group, and described substituent group is selected from: halogen, C1-C10 Alkyl, cyano group, OH, nitro, C3～C10Cycloalkyl, C1～C10Alkoxyl, amino.
In place of special instruction, among all compounds of the present invention, each chiral carbon atom can be optionally R structure Type or S configuration, or R configuration and the mixture of S configuration.
Term " C1～C6 alkyl " refers to the straight or branched alkyl with 1～6 carbon atom, such as methyl, ethyl, Propyl group, isopropyl, butyl, isobutyl group, sec-butyl, the tert-butyl group or similar group.
Term " 3-8 unit heterocyclic radical " refers to that having 1-3 heteroatomic 3～8 yuan of saturated rings selected from lower group loses a hydrogen The group that atom is formed: N, S, O；Such as pyrrolidinyl, piperidyl, piperazinyl, morpholinyl or similar group.
Term " 6-10 unit aryl " refers to that 6～10 yuan of aryl lose the group that a hydrogen atom is formed；Such as phenyl, naphthalene Base, or similar group.
Term " 5-10 unit heteroaryl " refers to that having 1-3 heteroatomic 5～8 yuan of aryl selected from lower group loses a hydrogen The group that atom is formed: N, S, O, the ring-type system of the most each heteroaryl can be monocycle or multi-ring；Such as Pyrrole radicals, pyridine radicals, thienyl, furyl, imidazole radicals, pyrimidine radicals, benzothienyl, indyl, quinolyl Or similar group.
Term " C1～C6 alkoxyl " refers to the straight or branched alkoxyl with 1-6 carbon atom, such as methoxyl group, Ethyoxyl, propoxyl group, isopropoxy, butoxy, isobutoxy, sec-butoxy, tert-butoxy or similar group.
Term " C2-C6 ester group " refers to R-O-C (=the O)-group with 2-6 carbon atom, such as-COOCH3、 -COOC2H5、-COOC3H7、-COOC4H9, or similar group.
Term " C2-C6 thiazolinyl " refers to that the alkene with 2-6 carbon atom loses what one or two hydrogen atom was formed Group, described alkene can be monoolefine, alkadienes or alkatrienes, such as-CH=CH2、-C2H4=CH2、 -CH=C2H4, or similar group.
Term " halogen " refers to F, Cl, Br and I.
Unless stated otherwise, structural formula described in the invention be intended to include all of isomeric forms (such as enantiomerism, Diastereo-isomerism and geometric isomer (or conformer): such as contain R, S configuration of asymmetric center, (Z) of double bond, (E) isomer and (Z), the conformer of (E).Therefore the single three-dimensional chemical isomer of the compound of the present invention or its mapping The mixture of isomer, diastereomer or geometric isomer (or conformer) broadly falls into the scope of the present invention.
Term " tautomer " represents that the structural isomer with different-energy can exceed low energy barrier, thus Inversion of phases mutually.Such as, proton tautomer (i.e. prototropic change) includes being migrated by proton carrying out change, such as 1H- Indazole and 2H-indazole, 1H-benzo [d] imidazoles and 3H-benzo [d] imidazoles, valence tautomers includes by some Bonding electrons is recombinated and is carried out change.
Formula (I) compound
The present invention is by the crystal structure of FGFR and the research of other tyrosine kinase inhibitor structure activity relationship, and design is closed Become the compound of a series of novel structure, by molecule, cell and animal model, these compounds have been screened, sent out These compounds existing can substantially suppress FGFR enzymatic activity at molecular level, the various cancerous cell that FGFR is induced by cellular level Propagation also has obvious inhibiting effect, and can also significantly inhibit tumor growth in animal body.
Specifically, described compound has a structure shown in formula (I):
Wherein:
Y is selected from CH or N；
R1Selected from substituted or unsubstituted 6-10 unit aryl, substituted or unsubstituted miscellaneous former selected from N, O and S containing 1-3 The 5-10 unit heteroaryl of son；Described substituent group be one or more be selected from halogen, C1～C6 alkyl, C1～C6 alkoxyl, C1～C6 alkylthio group, halo C1～C6 alkyl, halo C1～C6 alkoxyl, cyano group, C1～C6 alkanoyl, In-NH-CO-C1～C6 alkyl, amino ,-NH (C1～C6 alkyl) ,-N (C1～C6 alkyl) (C1～C6 alkyl) and hydroxyl Substituent group；
R2Selected from hydrogen, C1～C6 alkyl, C1～C6 alkanoyl ,-CO (O)-C1～C6 alkyl；
R3Selected from substituted or unsubstituted 6-10 unit aryl, substituted or unsubstituted miscellaneous selected from N, O and S containing 1-3 The 5-10 unit heteroaryl of atom；Described substituent group be one or more be selected from halogen, C1～C6 alkyl, C1～C6 alkoxyl, C1～C6 alkylthio group, cyano group, hydroxyl, halo C1～C6 alkyl, halo C1～C6 alkoxyl, C1～C6 alkanoyl, -NH-CO-C1～C6 alkyl, amino ,-C1～C6 alkyl-NH2,-NH (C1～C6 alkyl) ,-N (C1～C6 alkane Base) (C1～C6 alkyl) ,-CO (O)-C1～C6 alkyl, C3-C8 heterocyclic radical ,-Y1-(C1～C6 alkyl)-Y2, C1～C6 Substituent group in alkyl-C3～C8 heterocyclic radical and C3～C8 cycloalkyl-C3～C8 heterocyclic radical, described Y1 is selected from O, NH and S； Described Y2 is selected from C1～C6 alkoxyl, amino ,-NH (C1～C6 alkyl) and-N (C1～C6 alkyl) (C1～C6 alkyl)； Wherein said C3-C8 heterocyclic radical contains 1-3 the hetero atom selected from N, O and S, and optionally by one or more Selected from C1～C6 alkyl, C1～C6 alkoxyl, C3～C8 cycloalkyl, C2～C6 thiazolinyl, C2～C6 alkynyl, C1～C6 Alkanoyl, C1～C6 alkoxyl substituted C1～C6 alkyl, C1～C6 alkylamino substituted C1～C6 alkyl, hydroxyl take C1～the C6 alkyl in generation ,-CO (O)-C1～C6 alkyl, hydroxyl, halogen ,-SO2Substituent group in-C1～C6 alkyl is entered One step replaces；
R4、R5It is each independently selected from hydrogen, halogen, C1～C6 alkyl, C2～C6 thiazolinyl, C2～C6 alkynyl, halo C1～C6 alkyl, C1～C6 alkoxyl, halo C1～C6 alkoxyl, C1～C6 alkylthio group, cyano group, C1～C6 alkane acyl Base, nitro, amino ,-NH (C1～C6 alkyl) ,-N (C1～C6 alkyl) (C1～C6 alkyl) ,-NH-CO-C1～C6 Alkyl and-CO (O)-C1～C6 alkyl；
And when Y is CH, R3At least there is less than one substituent group: C3-C8 heterocyclic radical；Wherein, described C3-C8 Heterocyclic radical as defined above.
In another preference, R1Selected from substituted or unsubstituted 6-10 unit aryl, substituted or unsubstituted containing 1-3 Individual selected from N, O and S heteroatomic 5-10 unit heteroaryl；Described substituent group is that one or more are selected from halogen, C1～C4 Alkyl, C1～C4 alkoxyl, C1～C4 alkylthio group, halo C1～C4 alkyl, halo C1～C4 alkoxyl, cyano group, C1～C4 alkanoyl ,-NH-CO-C1～C4 alkyl, amino ,-NH (C1～C4 alkyl) ,-N (C1～C4 alkyl) (C1～C4 Alkyl) and hydroxyl in substituent group；
R2Selected from hydrogen, C1～C4 alkyl, C1～C4 alkanoyl ,-CO (O)-C1～C4 alkyl；
R3Selected from substituted or unsubstituted 6-10 unit aryl, substituted or unsubstituted miscellaneous selected from N, O and S containing 1-3 The 5-10 unit heteroaryl of atom；Described substituent group be one or more be selected from halogen, C1～C4 alkyl, C1～C4 alkoxyl, C1～C4 alkylthio group, cyano group, hydroxyl, halo C1～C4 alkyl, halo C1～C4 alkoxyl, C1～C4 alkanoyl, -NH-CO-C1～C4 alkyl, amino ,-C1～C4 alkyl-NH2,-NH (C1～C4 alkyl) ,-N (C1～C4 alkane Base) (C1～C4 alkyl) ,-CO (O)-C1～C4 alkyl, C3-C8 heterocyclic radical ,-Y1-(C1～C4 alkyl)-Y2, C1～C4 Substituent group in alkyl-C3～C8 heterocyclic radical and C3～C8 cycloalkyl-C3～C8 heterocyclic radical, described Y1 is selected from O, NH and S； Described Y2 is selected from C1～C4 alkoxyl, amino ,-NH (C1～C4 alkyl) and-N (C1～C4 alkyl) (C1～C4 alkyl)； Wherein said C3-C8 heterocyclic radical contains 1-3 the hetero atom selected from N, O and S, and optionally by one or more Selected from C1～C4 alkyl, C1～C4 alkoxyl, C3～C8 cycloalkyl, C2～C4 thiazolinyl, C2～C4 alkynyl, C1～C4 Alkanoyl, C1～C4 alkoxyl substituted C1～C4 alkyl, C1～C4 alkylamino substituted C1～C4 alkyl, hydroxyl take C1～the C4 alkyl in generation ,-CO (O)-C1～C4 alkyl, hydroxyl, halogen ,-SO2Substituent group in-C1～C4 alkyl is entered One step replaces；
R4、R5It is each independently selected from hydrogen, halogen, C1～C4 alkyl, C2～C4 thiazolinyl, C2～C4 alkynyl, halo C1～C4 alkyl, C1～C4 alkoxyl, halo C1～C4 alkoxyl, C1～C4 alkylthio group, cyano group, C1～C4 alkane acyl Base, nitro, amino ,-NH (C1～C4 alkyl) ,-N (C1～C4 alkyl) (C1～C4 alkyl) ,-NH-CO-C1～C4 Alkyl and-CO (O)-C1～C4 alkyl；
And when Y is CH, R3At least there is less than one substituent group: C3-C8 heterocyclic radical；Wherein, described C3-C8 Heterocyclic radical as defined above.
In another preference, R1Selected from substituted or unsubstituted following aryl or heteroaryl: phenyl, naphthyl, pyridine Base, pyrazinyl, furyl, thienyl, pyrrole radicals, oxazolyl, thiazolyl, pyrazolyl, imidazole radicals, indyl, Benzofuranyl, benzothiazolyl and benzimidazolyl；Described substituent group is that one or more are selected from halogen, C1～C4 Substituent group in alkyl, C1～C4 alkoxyl and-N (C1～C4 alkyl) (C1～C4 alkyl)；
R3In replacement or unsubstituted following aryl or heteroaryl: phenyl, naphthyl, pyridine radicals, pyrazinyl, thiophene Base, furyl, imidazole radicals, pyrrole radicals, oxazolyl, thiazolyl, pyrazolyl, indyl, benzofuranyl, benzene Benzothiazolyl, benzimidazolyl；Described substituent group is that one or more are selected from halogen, C1～C4 alkyl, C1～C4 alkane Epoxide, amino ,-NH (C1～C4 alkyl) ,-N (C1～C4 alkyl) (C1～C4 alkyl), C3-C8 heterocyclic radical and Substituent group in-Y1-(C1～C4 alkyl)-Y2, described Y1 is selected from O, NH and S；Described Y2 selected from C1～C4 alkoxyl, Amino ,-NH (C1～C4 alkyl) and-N (C1～C4 alkyl) (C1～C4 alkyl)；Wherein said C3-C8 heterocyclic radical contains 1-3 the hetero atom selected from N, O and S, and optionally it is selected from C1～C4 alkyl, C1～C4 alcoxyl by one or more Base, C3～C8 cycloalkyl, C2～C4 thiazolinyl, C2～C4 alkynyl, C1～C4 alkanoyl, C1～C4 alkoxyl are substituted C1～C4 alkyl, hydroxyl substituted C1～C4 alkyl ,-CO (O)-C1～C4 alkyl and-SO2Replacement in-C1～C4 alkyl Base is further substituted with；
R4、R5It is each independently selected from hydrogen, halogen, C1～C4 alkyl and halo C1～C4 alkyl；
And when Y is CH, R3At least there is less than one substituent group: C3-C8 heterocyclic radical；Wherein, described C3-C8 Heterocyclic radical as defined above.
In another preference, R1Selected from substituted or unsubstituted following aryl or heteroaryl: phenyl, naphthyl, pyridine Base, furyl and benzimidazolyl (preferably phenyl or benzimidazolyl, more preferably phenyl)；Described substituent group is One or more substituent groups in F, Cl, methyl, methoxyl group and N, N-dimethylamino；
In another preference, R2Selected from lower group: hydrogen, methyl, acetyl group and tertbutyloxycarbonyl.
In another preference, when Y is N, R3Selected from substituted or unsubstituted following aryl or heteroaryl: phenyl, Naphthyl, pyridine radicals, pyrazinyl, thienyl, indyl；R3Optionally by one or more selected from fluorine, chlorine, first Base, methoxyl group, amino, dimethylamino, piperazinyl, piperidyl, morpholinyl, homopiperazine base, thio-morpholinyl, Pyrrolidinyl, tetrahydrofuran base ,-NH (CH2)2OCH3、-NH(CH2)2N(CH3)2With-O (CH2)2OCH3In take Replace for base, and piperazinyl in substituent group, piperidyl, morpholinyl, homopiperazine base, thio-morpholinyl, pyrrolidine Base and tetrahydrofuran base are selected from methyl, ethyl, isopropyl, cyclopropyl, ring by one or more the most further Butyl ,-(CH2)2OCH3, pi-allyl ,-(CH2)2Replacement in OH, acetyl group, mesyl and tertbutyloxycarbonyl Base replaces；
In another preference, when Y is C, R3In substituted following aryl or heteroaryl: phenyl, naphthyl, Pyridine radicals, pyrazinyl, thienyl, indyl；R3Can by one or more selected from fluorine, chlorine, methyl, methoxyl group, Amino, dimethylamino, piperazinyl, piperidyl, morpholinyl, homopiperazine base, thio-morpholinyl, pyrrolidinyl, four Hydrogen furyl ,-NH (CH2)2OCH3、-NH(CH2)2N(CH3)2With-O (CH2)2OCH3In substituent group replace, and At least a part of which has 1 substituent group selected from piperazinyl, piperidyl, morpholinyl, homopiperazine base, thio-morpholinyl, pyrroles In alkyl and tetrahydrofuran base, piperazinyl therein, piperidyl, morpholinyl, homopiperazine base, thio-morpholinyl, pyrrole Cough up alkyl and tetrahydrofuran base the most further by one or more selected from methyl, ethyl, isopropyl, cyclopropyl, Cyclobutyl ,-(CH2)2OCH3, pi-allyl ,-(CH2)2Taking in OH, acetyl group, mesyl and tertbutyloxycarbonyl Replace for base.
In another preference, R4、R5It is each independently selected from hydrogen, fluorine, methyl or trifluoromethyl.
In another preference, in described compound, Y, R1、R2、R3、R4、R5In any one is respectively real Execute group corresponding in particular compound described in example.
Preferably, the present invention leads to pyrazolo [3, the 4-b] pyridines shown in formula (I) and indazole compounds in table 1 below Compound:
Table 1
The preparation of formula (I) compound
The invention still further relates to pyrazolo [3,4-b] pyridines and the preparation method of indazole compounds with logical formula (I) structure, Specifically, the preparation method of pyrazolo [3,4-b] pyridines and indazole compounds comprises the steps:
Compound (II) or its salt react with compound (III) and prepare compound (I), and described method is as follows:
Compound (II) or its salt prepare compound (I), or compound in the basic conditions with corresponding acetyl halide compound (III) Or its salt prepares compound (I) with corresponding ester (III) under lewis acid effect (II)；
Wherein, Q is leaving group, it is therefore preferable to halogen (acyl chlorides) or C2-C6 ester group (ester)；Remaining each group is as defined above Described in literary composition.
In another preference, described reaction temperature is 30-90 DEG C.
In another preference, described reaction is carried out in non-polar solven, is preferably entering in the solvent of lower group OK: toluene, benzene and carbon tetrachloride.
In another preference, the described response time is 2-48h.
In another preference, described lewis acid is selected from lower group: trimethyl aluminium, aluminum chloride or boron trifluoride.
Wherein, described compound (II) or compound (III) can be prepared by the conventional method of this area, or by commercially available way Footpath obtains.In a preference of the present invention, described compound (II) or its salt are prepared the most by the following method:
From compound 1, first making borate, then carry out Suzuki coupling, then cyclization forms pyrazolo [3,4-b] Pyridine ring or indazole ring, finally replace and obtain compound (II) or its salt.
Specifically, described preparation method comprises the steps:
1. compound 1 reacts generation borate 2 in the basic conditions with connection pinacol borate；
2. compound 2 occurs Suzuki coupling reaction to generate compound 3 in the basic conditions with corresponding pyridine；
3. compound 3 generates compound 4 with hydrazine hydrate cyclization；
4. compound 4 and corresponding halides or anhydride reacting generating compound in the basic conditions (II) or its salt.
Or
From compound 1, first cyclization forms pyrazolo [3,4-b] pyridine ring or indazole ring, then carries out Suzuki even Connection, finally replaces and obtains compound (II) or its salt.
Specifically, described preparation method comprises the steps:
1. compound 1 generates compound 2 with hydrazine hydrate cyclization；
2. compound 2 occurs Suzuki coupling reaction to generate compound 3 in the basic conditions with corresponding borate；
3. compound 3 and corresponding halides or anhydride reacting generating compound in the basic conditions (II) or its salt.
Wherein, R1、R2、R3、R4、R5, Y as defined above and preferably, X is Br or I.
Pyrazolo [3,4-b] pyridines of the present invention and the preparation method of indazole compounds have that reaction condition is gentle, raw material is rich The advantages such as richness is easy to get, operate and post processing is simple.
Pharmaceutical composition containing formula (I) compound
The invention still further relates to a kind of pharmaceutical composition, described pharmaceutical composition comprise therapeutically effective amount selected from pyrrole shown in formula (I) One in azoles also [3,4-b] pyridines and indazole compounds, its pharmaceutical salts, its prodrug and hydrate thereof and solvate or Multiple and optionally, pharmaceutically acceptable carrier, it can be used for treating the relevant diseases such as cancer.Described drug regimen Thing can be prepared as various forms according to different way of administration.
Pyrazolo [3,4-b] pyridines shown in formula of the present invention (I) and indazole compounds, its pharmaceutical salts, its prodrug and One or more in hydrate and solvate, or above-mentioned comprise therapeutically effective amount selected from pyrazolo shown in formula (I) One or more in [3,4-b] pyridines and indazole compounds, its pharmaceutical salts, its prodrug and hydrate thereof and solvate Pharmaceutical composition especially as FGFR inhibitor, can be used for treating cancer as protein tyrosine kinase inhibitor. Described FGFR includes one or more in FGFR1, FGFR2 and FGFR3, especially FGFR1-3 inhibitor.
The preparation of the pharmaceutical salts of the compounds of this invention, can use the free alkali of compound directly to become salt with inorganic or organic acid Reaction is carried out.Inorganic or organic acid be selected from hydrochloric acid, sulphuric acid, phosphoric acid, nitric acid, Fluohydric acid., hydrobromic acid, formic acid, acetic acid, Picric acid, citric acid, maleic acid, Loprazolam, trifluoromethayl sulfonic acid, ethane sulfonic acid and p-methyl benzenesulfonic acid etc..
Owing to the compounds of this invention has the pressing down FGFR kinases (Kinase) such as FGFR1 and FGFR2 of excellence System activity, therefore the compounds of this invention and various crystal formation thereof, pharmaceutically acceptable inorganic or organic salt, hydrate or Solvate, and containing the compounds of this invention be the pharmaceutical composition of main active can be used for treating, prevent with And alleviate by the disease relevant to FGFR activity or expression, such as prevent and/or treat and FGF/FGFR signal Path unconventionality expression relevant disease.According to prior art, the compounds of this invention can be used for treating following disease: described Cancer include breast carcinoma, pulmonary carcinoma, bladder cancer, gastric cancer, cancer of pancreas, carcinoma of prostate, colon cancer, multiple bone marrow Tumor AML, hepatocarcinoma, melanoma, head and neck cancer, thyroid carcinoma, renal cell carcinoma, glioblast cancer and carcinoma of testis. More particularly, these cancers are selected from: breast carcinoma, nonsmall-cell lung cancer, bladder cancer, gastric cancer, cancer of pancreas, prostatitis Adenocarcinoma, colon cancer, multiple myeloma, hepatocarcinoma, melanoma, head and neck cancer, thyroid carcinoma, renal cell carcinoma, glue Matter blast cell cancer and carcinoma of testis.Most particularly, this cancer is nonsmall-cell lung cancer, gastric cancer or multiple myeloma.
The pharmaceutical composition of the present invention comprises the compounds of this invention in the range of safe and effective amount or it pharmacologically can accept Salt and pharmacologically acceptable excipient or carrier.Wherein " safe and effective amount " refers to: the amount foot of compound To be obviously improved the state of an illness, and it is unlikely to produce serious side effect.Generally, pharmaceutical composition contains 1-2000mg originally Invention compound/agent, more preferably, containing 5-200mg the compounds of this invention/agent.It is preferred that described " potion " It is a capsule or tablet.
" pharmaceutically acceptable carrier " refers to: one or more biocompatible solid or liquid filler or jello Matter, they are suitable for people and use and it is necessary to have enough purity and of a sufficiently low toxicity." compatibility " herein means Be in compositions each component energy and the compound of the present invention and they between mutually admix, and significantly reduce chemical combination The drug effect of thing.Pharmaceutically acceptable carrier part example have cellulose and its derivates (as sodium carboxymethyl cellulose, Ethyl cellulose sodium, cellulose ethanoate etc.), gelatin, Talcum, kollag (such as stearic acid, magnesium stearate), Calcium sulfate, vegetable oil (such as Oleum Glycines, Oleum sesami, Oleum Arachidis hypogaeae semen, olive oil etc.), polyhydric alcohol (as propylene glycol, glycerol, Mannitol, sorbitol etc.), emulsifying agent (as), wetting agent (such as sodium lauryl sulphate), coloring agent, tune Taste agent, stabilizer, antioxidant, preservative, apirogen water etc..
The method of application of the compounds of this invention or pharmaceutical composition is not particularly limited, and representational method of application includes (but being not limited to): in oral, tumor, rectum, parenteral (intravenous, intramuscular or subcutaneous) and topical.
Solid dosage forms for oral administration includes capsule, tablet, pill, powder and granule.At these solids In dosage form, reactive compound mixes with at least one conventional inert excipients (or carrier), such as sodium citrate or di(2-ethylhexyl)phosphate Calcium, or mix with following compositions: (a) filler or bulking agent, such as, starch, lactose, sucrose, glucose, sweet Dew alcohol and silicic acid；(b) binding agent, such as, hydroxymethyl cellulose, alginate, gelatin, polyvinyl pyrrolidone, Sucrose and arabic gum；(c) wetting agent, such as, glycerol；(d) disintegrating agent, such as, agar, calcium carbonate, horse Bell sweet potato starch or tapioca, alginic acid, some composition silicate and sodium carbonate；(e) retarding solvent, such as paraffin；(f) Absorb accelerator, such as, quaternary ammonium compound；(g) wetting agent, such as spermol and glyceryl monostearate；H () inhales Attached dose, such as, Kaolin；(i) lubricant, such as, the poly-second of Talcum, calcium stearate, magnesium stearate, solid Glycol, sodium lauryl sulphate, or its mixture.In capsule, tablet and pill, dosage form also can comprise buffer agent.
Solid dosage forms such as tablet, sugar pill, capsule, pill and granule can use coating and shell material to prepare, such as casing With other material well known in the art.They can comprise opacifying agent, and, in this compositions reactive compound or The release of compound can discharge in the part of certain in digestive tract in a delayed fashion.The reality of adoptable embedding component Example is polymeric material and Wax.If desired, reactive compound also can be with one or more shapes in above-mentioned excipient Become microencapsulation form.
Liquid formulation for oral administration includes pharmaceutically acceptable emulsion, solution, suspension, syrup or tincture. Except active ingredient beyond the region of objective existence, liquid dosage form can comprise in this area the conventional inert diluent used, such as water or other is molten Agent, solubilizing agent and emulsifying agent, example knows, ethanol, isopropanol, ethyl carbonate, ethyl acetate, propylene glycol, 1,3-fourth Glycol, dimethylformamide and oil, particularly Oleum Gossypii semen, Oleum Arachidis hypogaeae semen, maize embryo oil, olive oil, Oleum Ricini and Oleum sesami or the mixture etc. of these materials.
In addition to these inert diluents, compositions also can comprise auxiliary agent, such as wetting agent, emulsifying agent and suspending agent, sweet Taste agent, correctives and spice.
Except active ingredient beyond the region of objective existence, suspension can comprise suspending agent, such as, ethoxylation isooctadecane alcohol, polyoxy second Alkene sorbitol and Isosorbide Dinitrate, microcrystalline Cellulose, aluminium methoxide and agar or the mixture etc. of these materials.
Compositions for parenteral injection can comprise physiologically acceptable sterile, aqueous or anhydrous solution, dispersion liquid, Suspension or emulsion, and for being again dissolved into the sterilized powder of aseptic Injectable solution or dispersion liquid.Suitable contains Water and nonaqueous carrier, diluent, solvent or excipient include water, ethanol, polyhydric alcohol and suitable mixture thereof.
Dosage form for the compounds of this invention of topical includes ointment, powder, patch, propellant and inhalant. Active component aseptically with physiologically acceptable carrier and any preservative, buffer agent, or if desired may The propellant needed is mixed together.
The compounds of this invention can be individually dosed, or with other pharmaceutically acceptable compound administering drug combinations.
When making pharmaceutical composition, it it is the mammal that the compounds of this invention of safe and effective amount is applicable to treatment (such as people), when wherein using, dosage is the effective dosage pharmaceutically thought, for the people of 60kg body weight, day Dosage is usually 1～2000mg, preferably 5～500mg.Certainly, concrete dosage is it is also contemplated that route of administration, disease The factors such as people's health status, within the scope of these are all skilled practitioners technical ability.
Below in conjunction with specific embodiment, the present invention is expanded on further.Should be understood that these embodiments are merely to illustrate the present invention Rather than restriction the scope of the present invention.The experimental technique of unreceipted actual conditions in the following example, generally according to conventional strip Part, or according to the condition proposed by manufacturer.Unless otherwise indicated, otherwise percentage ratio and number are calculated by weight.Except another It is defined or illustrates, the meaning that all specialties used herein are familiar with scientific words and those skilled in the art Identical.
Embodiment 1:
4-((3R, 5S)-3,5-lupetazin-1-base)-N-(6-phenyl-1H-pyrazolo [3,4-b] pyridin-3-yl) Benzoylamide (I-1)
Step 1: preparation 4,4,5,5-tetramethyl-2-phenyl-1,3,2-dioxaborinate
1 milliliter of bromobenzene is dissolved in 80 milliliters of dioxane, is sequentially added into 3.618 grams of connection pinacol borate, 2.800 grams of second Acid potassium and 0.696 gram of [1,1 '-bis-(diphenylphosphino) ferrocene] palladium chloride, be warming up to 85 DEG C under argon shield, stirs 6 Stopping heating, reacting liquid filtering, a small amount of ethyl acetate filter wash cake after hour, filtrate concentrates and is evaporated, residue column chromatography (stone Oil ether) 1.938 grams of yellow oil of isolated, yield 100%.
1H NMR(300MHz,CDCl3) δ: 7.81 (dd, J=8.0,1.4Hz, 2H), 7.50 7.43 (m, 1H), 7.41 7.33(m,2H),1.35(s,12H).
Step 2: preparation 2-chloro-6-phenyl nicotinic acid nitrile
0.865 gram of 2,6-dichloro nicotinic acid nitrile and 1.021 grams of 4,4,5,5-tetramethyl-2-phenyl-1,3,2-dioxaborinate are dissolved in 60 milliliters In glycol dimethyl ether, it is sequentially added into 20 milliliters of sodium bicarbonate aqueous solutions (1 mole every liter) and 0.366 gram of [1,1 '-bis-(diphenylphosphine Base) ferrocene] palladium chloride, under argon shield, it is warming up to 90 DEG C, stops after stirring 6 hours heating, reactant liquor separatory, Being extracted with ethyl acetate aqueous phase, organic facies is washed with saturated sodium-chloride water solution after merging, and anhydrous sodium sulfate is dried, and filters dense Contracting, residue column chromatography (petroleum ether: ethyl acetate=99:1 95:5, V/V) isolated 0.680 grammeter white solid, receive Rate 63.4%.
1H NMR(300MHz,CDCl3) δ: 8.10 8.04 (m, 2H), 8.03 (d, J=8.1Hz, 1H), 7.79 (d, J= 8.1Hz,1H),7.57–7.49(m,3H).
Step 3: preparation 6-phenyl-1H-pyrazolo [3,4-b] pyridine-3-amine
664 milligrams of 2-chloro-6-phenyl nicotinic acid nitriles are dissolved in 15 milliliters of ethanol, add 1.7 milliliters of hydrazine hydrates (85%, w/w), rise Temperature, to 70 DEG C, stops heating after stirring 20 hours, reactant liquor concentrates and is evaporated, residue column chromatography (dichloromethane: methanol=99:1 98:2, V/V) 598 milligrams of yellow solids of isolated, yield 94.8%.
1H NMR (300MHz, DMSO) δ: 11.96 (s, 1H), 8.19 (d, J=8.3Hz, 1H), 8.11 (dd, J=8.2, 1.4Hz, 2H), 7.56 (d, J=8.3Hz, 1H), 7.54 7.40 (m, 3H), 5.57 (s, 2H).
Step 4: preparation 4-((3R, 5S)-3,5-lupetazin-1-base) ethyl benzoate
0.44 milliliter of 4-ethyl fluoro benzoate is dissolved in 8 milliliters of dimethyl sulfoxides, adds 1.370 grams of (2S, 6R)-lupetazins, Under argon shield, it is warming up to 120 DEG C, stops heating after stirring 28 hours, add 40 milliliters of water dilute reaction solutions, use acetic acid Ethyl ester extracts, and organic facies is washed five times with saturated sodium-chloride water solution after merging, and anhydrous sodium sulfate is dried, filtering and concentrating, residual Excess column chromatography (dichloromethane: methanol=99:1 97:3, V/V) 0.730 gram of brown oil of isolated, yield 92.8%.
1H NMR(300MHz,CDCl3) δ: 7.91 (d, J=9.0Hz, 2H), 6.85 (d, J=9.0Hz, 2H), 4.32 (q, J=7.1Hz, 2H), 3.67 (dd, J=12.3,2.2Hz, 2H), 3.06 2.93 (m, 2H), 2.44 2.34 (m, 2H), 1.36 (t, J=7.1Hz, 3H), 1.15 (d, J=6.3Hz, 6H).
Step 5: preparation 4-((3R, 5S)-3,5-lupetazin-1-base)-N-(6-phenyl-1H-pyrazolo [3,4-b] pyridin-3-yl) Benzoylamide
By 84 milligrams of 6-phenyl-1H-pyrazolo [3,4-b] pyridine-3-amine and 126 milligrams of 4-((3R, 5S)-3,5-lupetazin-1- Base) ethyl benzoate is dissolved in 2 milliliters of toluene, is added dropwise over the toluene solution (2 moles every liter) of 0.8 milliliter of trimethyl aluminium, Being warming up to 60 DEG C under argon shield, stop heating after stirring 38 hours, methanol cancellation is reacted, and reactant liquor concentrates and is evaporated, remaining Thing column chromatography (dichloromethane: methanol=99:1 96:4, V/V) 81 milligrams of white solids of isolated, yield 47.5%.
1H NMR(300MHz,CD3OD) δ: 8.44 (d, J=8.6Hz, 1H), 8.19 8.10 (m, 2H), 7.96 (d, J =8.9Hz, 2H), 7.71 (d, J=8.6Hz, 1H), 7.57 7.42 (m, 3H), 7.05 (d, J=9.0Hz, 2H), 3.81 (dd, J=12.3,2.0Hz, 2H), 3.05 2.90 (m, 2H), 2.48 2.34 (m, 2H), 1.18 (d, J=6.4Hz, 6H).
Embodiment 2:
4-(4-methylpiperazine-1-yl)-N-(6-phenyl-1H-pyrazolo [3,4-b] pyridin-3-yl) Benzoylamide (I-2)
Step 1: preparation 4-(4-methylpiperazine-1-yl) ethyl benzoate
0.735 milliliter of 4-ethyl fluoro benzoate is dissolved in 5 milliliters of dimethyl sulfoxides, adds 0.67 milliliter of 1-methyl piperazine and 1.037 Gram potassium carbonate, is warming up to 120 DEG C under argon shield, stops heating, add 25 milliliters of water diluting reactions after stirring 23 hours Liquid, is extracted with ethyl acetate, and organic facies is washed five times with saturated sodium-chloride water solution after merging, and anhydrous sodium sulfate is dried, mistake Filter concentrates, residue column chromatography (dichloromethane: methanol=99:1 98:2, V/V) isolated 1.095 grammeter yellow solid, Yield 88.0%.
1H NMR(300MHz,CDCl3) δ 7.92 (d, J=9.0Hz, 2H), 6.87 (d, J=9.0Hz, 2H), 4.33 (q, J=7.1Hz, 2H), 3.44 3.24 (m, 4H), 2.64 2.51 (m, 4H), 2.35 (s, 3H), 1.36 (t, J=7.1Hz, 3H).
Step 2: preparation 4-(4-methylpiperazine-1-yl)-N-(6-phenyl-1H-pyrazolo [3,4-b] pyridin-3-yl) Benzoylamide
Change 4-((3R, 5S)-3,5-lupetazin-1-base) ethyl benzoate into 4-(4-methylpiperazine-1-yl) ethyl benzoate, Remaining needed raw material, reagent and preparation method, with the step 5 in embodiment 1, obtain white solid 4-(4-methyl piperazine-1- Base)-N-(6-phenyl-1H-pyrazolo [3,4-b] pyridin-3-yl) Benzoylamide.
1H NMR (300MHz, DMSO) δ 13.29 (s, 1H), 10.76 (s, 1H), 8.39 (d, J=8.5Hz, 1H), 8.17 (d, J=7.4Hz, 2H), 8.00 (d, J=8.8Hz, 2H), 7.75 (d, J=8.5Hz, 1H), 7.60 7.44 (m, 3H), 7.02 (d, J=8.6Hz, 2H), 2.48 2.39 (m, 4H), 2.24 (s, 3H).
Embodiment 3:
N-(6-(3,5-Dimethoxyphenyl)-1H-pyrazolo [3,4-b] pyridin-3-yl)-4-((3R, 5S)-3,5-lupetazin-1- Base) Benzoylamide (I-3)
Step 1: preparation 2-(3,5-Dimethoxyphenyl)-4,4,5,5-tetramethyl-1,3,2-dioxaborinate
2.171 grams of 1-bromo-3,5-dimethoxy benzene and 3.810 grams of connection pinacol borates are dissolved in 85 milliliters of dioxane, depend on 2.945 grams of potassium acetates of secondary addition and 0.732 gram of [1,1 '-bis-(diphenylphosphino) ferrocene] palladium chloride, heat up under argon shield To 85 DEG C, stopping heating, reacting liquid filtering, a small amount of ethyl acetate filter wash cake after stirring 12 hours, filtrate concentrates and is evaporated, residual Excess column chromatography (petroleum ether: ethyl acetate=99:1 98:2, V/V) 2.462 grams of white solids of isolated, yield 93.2%.
1H NMR(300MHz,CDCl3) δ 6.95 (d, J=2.4Hz, 2H), 6.57 (t, J=2.4Hz, 1H), 3.81 (s, 6H),1.34(s,12H).
Step 2: the preparation chloro-6-of 2-(3,5-Dimethoxyphenyl) nicotinic acid nitrile
By 1.384 grams of 2,6-dichloro nicotinic acid nitriles and 2.325 grams of 2-(3,5-Dimethoxyphenyl)-4,4,5,5-tetramethyl-1,3,2-dioxas Borine is dissolved in 67.2 milliliters of isopropanols, is sequentially added into 22.4 milliliters of sodium bicarbonate aqueous solutions (1 mole every liter) and 0.925 gram four (triphenylphosphine) palladium, is warming up to 90 DEG C under argon shield, stops heating, reactant liquor separatory, use acetic acid after stirring 3 hours Ethyl ester aqueous phase extracted, washs with saturated sodium-chloride water solution after organic facies merging, and anhydrous sodium sulfate is dried, and filtering and concentrating is residual Excess column chromatography (petroleum ether: ethyl acetate=98:2 96:4, V/V) isolated 1.740 grammeter yellow solid, yield 79.2%.
1H NMR(300MHz,CDCl3) δ 8.01 (d, J=8.1Hz, 1H), 7.74 (d, J=8.1Hz, 1H), 7.18 (d, J=2.2Hz, 2H), 6.61 (t, J=2.2Hz, 1H), 3.88 (s, 6H).
Step 3: preparation 6-(3,5-Dimethoxyphenyl)-1H-pyrazolo [3,4-b] pyridine-3-amine
The 1.738 grams of chloro-6-of 2-(3,5-Dimethoxyphenyl) nicotinic acid nitriles are dissolved in 30 milliliters of ethanol, add 3 milliliters of hydrazine hydrates (85%, w/w), is warming up to 70 DEG C, after stirring 22 hours stop heating, reactant liquor concentrate be evaporated, add ethyl acetate and Moisture liquid, is extracted with ethyl acetate aqueous phase, and organic facies is washed with saturated sodium-chloride water solution after merging, and anhydrous sodium sulfate is dried, Filtering and concentrating, residue is dissolved in 5 milliliters of dichloromethane, is stirred at room temperature 2 hours, filters, and dichloromethane filter wash cake, filter cake is true Empty dry, obtain 1.415 grams of yellow solids, yield 82.7%.
1H NMR (300MHz, DMSO) δ 11.98 (s, 1H), 8.17 (d, J=8.3Hz, 1H), 7.57 (d, J=8.4Hz, 1H), 7.26 (d, J=2.2Hz, 2H), 6.58 (t, J=2.2Hz, 1H), 5.58 (s, 2H), 3.83 (s, 6H).
Step 4: preparation N-(6-(3,5-Dimethoxyphenyl)-1H-pyrazolo [3,4-b] pyridin-3-yl)-4-((3R, 5S)-3,5-two Methylpiperazine-1-yl) Benzoylamide
By 54 milligrams of 6-(3,5-Dimethoxyphenyl)-1H-pyrazolo [3,4-b] pyridine-3-amine and 63 milligrams of 4-((3R, 5S)-3,5-two Methylpiperazine-1-yl) ethyl benzoate is dissolved in 2 milliliters of toluene, is added dropwise over the toluene solution (1.6 of 0.3 milliliter of trimethyl aluminium Mole every liter), under argon shield, it is warming up to 60 DEG C, after stirring 24 hours, stops heating, methanol cancellation is reacted, reactant liquor Concentration is evaporated, residue column chromatography (dichloromethane: methanol=96:4, V/V) 17 milligrams of yellow solids of isolated, yield 17.5%.
1H NMR (300MHz, DMSO) δ 13.33 (s, 1H), 10.78 (s, 1H), 8.37 (d, J=8.6Hz, 1H), 8.00 (d, J=8.9Hz, 2H), 7.76 (d, J=8.6Hz, 1H), 7.31 (d, J=2.3Hz, 2H), 7.04 (d, J=9.0Hz, 2H), 6.63 (t, J=2.2Hz, 1H), 4.01 3.73 (m, 8H), 2.98 (s, 2H), 2.47 2.30 (m, 2H), 1.12 (d, J=6.1 Hz,6H).
Embodiment 4:
N-(6-(3,5-Dimethoxyphenyl)-1H-pyrazolo [3,4-b] pyridin-3-yl)-4-(piperazine-1-base) Benzoylamide (I-4)
Step 1: preparation 4-(4-(ethoxy carbonyl) phenyl) piperazine-1-t-butyl formate
0.735 milliliter of 4-ethyl fluoro benzoate is dissolved in 5 milliliters of dimethyl sulfoxides, adds 0.933 gram of piperazine-1-t-butyl formate With 1.039 grams of potassium carbonate, under argon shield, it is warming up to 120 DEG C, stops heating after stirring 18 hours, add 25 milliliters of water dilute Releasing reactant liquor, be extracted with ethyl acetate, organic facies is washed five times with saturated sodium-chloride water solution after merging, and anhydrous sodium sulfate is done Dry, filtering and concentrating, residue column chromatography (petroleum ether: ethyl acetate=96:4 95:5, V/V) 0.871 gram of white of isolated Solid, yield 52.0%.
1H NMR(300MHz,CDCl3) δ 7.94 (d, J=8.9Hz, 2H), 6.86 (d, J=9.0Hz, 2H), 4.33 (q, J=7.1Hz, 2H), 3.70 3.54 (m, 4H), 3.37 3.21 (m, 4H), 1.48 (s, 9H), 1.37 (t, J=7.1Hz, 3H).
Step 2: preparation 4-(4-((6-(3,5-Dimethoxyphenyl)-1H-pyrazolo [3,4-b] pyridin-3-yl) carbamoyl) benzene Base) piperazine-1-t-butyl formate
Change 4-((3R, 5S)-3,5-lupetazin-1-base) ethyl benzoate into 4-(4-(ethoxy carbonyl) phenyl) piperazine-1-first Tert-butyl acrylate, remaining needed raw material, reagent and preparation method, with the step 4 in embodiment 3, obtain white solid 4-(4-((6-(3,5- Dimethoxyphenyl)-1H-pyrazolo [3,4-b] pyridin-3-yl) carbamoyl) phenyl) piperazine-1-t-butyl formate.
1H NMR(300MHz,CDCl3) δ 11.04 (s, 1H), 8.94 (s, 1H), 8.79 (d, J=8.6Hz, 1H), 7.88 (d, J=8.7Hz, 2H), 7.57 (d, J=8.6Hz, 1H), 7.22 (d, J=2.1Hz, 2H), 6.89 (d, J=8.5Hz, 2H), 6.55 (t, J=2.0Hz, 1H), 3.87 (s, 6H), 3.73 3.51 (m, 4H), 3.35 3.19 (m, 4H), 1.49 (s, 9H).
Step 3: preparation N-(6-(3,5-Dimethoxyphenyl)-1H-pyrazolo [3,4-b] pyridin-3-yl)-4-(piperazine-1-base) benzene first Amide
By 58 milligrams of 4-(4-((6-(3,5-Dimethoxyphenyl)-1H-pyrazolo [3,4-b] pyridin-3-yl) carbamoyl) phenyl) Piperazine-1-t-butyl formate is dissolved in 1 milliliter of dichloromethane, adds 0.16 milliliter of trifluoroacetic acid, stirred overnight at room temperature, reaction Liquid concentrates and is evaporated, and residue is dissolved in methanol, and dropping ammonia regulation pH value is more than 7, and concentration is evaporated, residue column chromatography (two Chloromethanes: methanol=97:3 90:10, V/V) 10 milligrams of yellow solids of isolated, yield 21.8%.
1H NMR (300MHz, DMSO) δ 13.36 (s, 1H), 10.85 (s, 1H), 8.37 (d, J=8.6Hz, 1H), 8.04 (d, J=8.7Hz, 2H), 7.77 (d, J=8.6Hz, 1H), 7.31 (d, J=1.9Hz, 2H), 7.10 (d, J=8.9Hz, 2H), 6.63(s,1H),3.85(s,6H),3.63–3.49(m,4H),3.26–3.11(m,4H).
Embodiment 5:
N-(6-(3,5-Dimethoxyphenyl)-1H-pyrazolo [3,4-b] pyridin-3-yl)-4-(4-methylpiperazine-1-yl) Benzoylamide (I-5)
Step 1: preparation N-(6-(3,5-Dimethoxyphenyl)-1H-pyrazolo [3,4-b] pyridin-3-yl)-4-(4-methyl piperazine-1- Base) Benzoylamide
Change 4-((3R, 5S)-3,5-lupetazin-1-base) ethyl benzoate into 4-(4-methylpiperazine-1-yl) ethyl benzoate, Remaining needed raw material, reagent and preparation method, with the step 4 in embodiment 3, obtain white solid N-(6-(3,5-dimethoxy benzenes Base)-1H-pyrazolo [3,4-b] pyridin-3-yl)-4-(4-methylpiperazine-1-yl) Benzoylamide.
1H NMR (400MHz, DMSO) δ 13.32 (s, 1H), 10.77 (s, 1H), 8.37 (d, J=8.6Hz, 1H), 7.99 (d, J=9.0Hz, 2H), 7.76 (d, J=8.6Hz, 1H), 7.31 (d, J=2.2Hz, 2H), 7.02 (d, J=9.1Hz, 2H), 6.62 (t, J=2.2Hz, 1H), 3.85 (s, 6H), 3.32 3.25 (m, 4H), 2.48 2.41 (m, 4H), 2.23 (s, 3H).
Embodiment 6:
N-(6-(2,6-bis-chloro-3,5-Dimethoxyphenyl)-1H-pyrazolo [3,4-b] pyridin-3-yl)-4-((3R, 5S)-3,5-diformazan Base piperazine-1-base) Benzoylamide (I-6)
Step 1: preparation 1,5-bis-chloro-2,4-dimethoxy benzene
By 5 milliliter 1,3-dimethoxy benzene is dissolved in 50 milliliters of anhydrous methylene chlorides, is added dropwise over 6.2 milliliters of sulfonic acid chlorides at 0 DEG C, Being stirred overnight under room temperature, reactant liquor concentrates and is evaporated, and residue column chromatography (petroleum ether: ethyl acetate=98:2, V/V) separates To 7.906 grams of white solids, yield 100%.
1H NMR(300MHz,CDCl3)δ7.35(s,1H),6.53(s,1H),3.91(s,6H).
Step 2: the preparation chloro-3-of 2,4-bis-iodo-1,5-dimethoxy benzene
By 7.8 milliliter 2,2,6,6-tetramethyl piperidines are dissolved in 100 milliliters of anhydrous tetrahydro furans, are added dropwise over 17.5 milliliters at 0 DEG C The hexane solution (2.4 moles every liter) of n-BuLi, stirs 20 minutes, is cooled to-78 DEG C, adds 4.141 gram 1,5-bis-chloro-2,4- Dimethoxy benzene, stirs 3 hours, is added dropwise over the tetrahydrofuran solution (0.8 mole every liter) of 50 milliliters of iodine, stirs 30 minutes, Recovering to after room temperature to stir 1 hour, add saturated sodium thiosulfate solution cancellation reaction, ethyl acetate extracts, and organic is harmonious Washing with saturated sodium-chloride water solution after and, anhydrous sodium sulfate is dried, filtering and concentrating, residue column chromatography (petroleum ether: second Acetoacetic ester=96:4 90:10, V/V) 6.352 grams of yellow solids of isolated, yield 95.4%.
1H NMR(300MHz,CDCl3)δ6.60(s,1H),3.92(s,6H).
Step 3: preparation 2-(2,6-bis-chloro-3,5-Dimethoxyphenyl)-4,4,5,5-tetramethyl-1,3,2-dioxaborinate
By molten to the 3.729 grams of chloro-3-of 2,4-bis-iodo-1,5-dimethoxy benzenes, 4.266 grams of connection pinacol borates and 3.298 grams of potassium acetates In 40 milliliters of DMFs, add 0.252 gram of palladium, under argon shield, be warming up to 90 DEG C, stir 18 Stopping heating after hour, add 200 milliliters of water dilute reaction solutions, be extracted with ethyl acetate, organic facies uses saturated chlorine after merging Changing sodium water solution to wash five times, anhydrous sodium sulfate is dried, filtering and concentrating, residue column chromatography (petroleum ether: dichloromethane=80:20 50:50, V/V) 3.041 grams of white solids of isolated, yield 80.8%.
1H NMR(300MHz,CDCl3)δ6.53(s,1H),3.88(s,6H),1.42(s,12H).
Step 4: the preparation chloro-6-of 2-(2,6-bis-chloro-3,5-Dimethoxyphenyl) nicotinic acid nitrile
By 817 milligrams of 2,6-dichloro nicotinic acid nitriles and 1.888 grams of 2-(2,6-bis-chloro-3,5-Dimethoxyphenyl)-4,4,5,5-tetramethyls -1,3,2-dioxaborinate are dissolved in 94.5 milliliters of glycol dimethyl ethers, are sequentially added into 18.9 milliliters of sodium bicarbonate aqueous solutions (1 mole Every liter) and 346 milligrams of [1,1 '-bis-(diphenylphosphino) ferrocene] palladium chlorides, under argon shield, it is warming up to 90 DEG C, stirring Stopping heating, reactant liquor separatory after 17 hours, be extracted with ethyl acetate aqueous phase, organic facies is water-soluble with saturated sodium-chloride after merging Liquid washs, and anhydrous sodium sulfate is dried, filtering and concentrating, residue column chromatography (petroleum ether: ethyl acetate=95:5 85:15, V/V) 840 milligrams of buff white solid of isolated, yield 51.8%.
1H NMR(300MHz,CDCl3) δ 8.11 (d, J=7.9Hz, 1H), 7.39 (d, J=7.9Hz, 1H), 6.66 (s, 1H),3.96(s,6H)
Step 5: preparation 6-(2,6-bis-chloro-3,5-Dimethoxyphenyl)-1H-pyrazolo [3,4-b] pyridine-3-amine
837 milligrams of chloro-6-of 2-(2,6-bis-chloro-3,5-Dimethoxyphenyl) nicotinic acid nitrile is dissolved in 12 milliliters of ethanol, adds 7 milliliters of water Closing hydrazine (85%, w/w), be warming up to 70 DEG C, stop heating after stirring 13 hours, reactant liquor concentrates and is evaporated, residue post layer Analysis (dichloromethane: methanol=99:1 98:2, V/V) 463 milligrams of yellow solids of isolated, yield 55.9%.
1H NMR (300MHz, DMSO) δ 12.02 (s, 1H), 8.19 (d, J=8.1Hz, 1H), 7.01 (s, 1H), 6.86 (d, J=8.1Hz, 1H), 5.64 (s, 2H), 3.97 (s, 6H).
Step 6:N-(6-(2,6-bis-chloro-3,5-Dimethoxyphenyl)-1H-pyrazolo [3,4-b] pyridin-3-yl)-4-((3R, 5S)-3,5- Lupetazin-1-base) Benzoylamide
By 68 milligrams of 6-(2,6-bis-chloro-3,5-Dimethoxyphenyl)-1H-pyrazolo [3,4-b] pyridine-3-amine and 79 milligrams 4-((3R, 5S)-3,5-lupetazin-1-base) ethyl benzoate is dissolved in 2 milliliters of toluene, is added dropwise over 0.4 milliliter of trimethyl aluminium Toluene solution (1.6 moles every liter), under argon shield, be warming up to 60 DEG C, after stirring 35 hours stop heating, methanol cancellation Reaction, reacting liquid filtering, methylene chloride/methanol/ammonia (10:10:1, V/V) filter wash cake, filtrate concentrates and is evaporated, and residue is adopted (dichloromethane: methanol=15:1, V/V) 37 milligrams of white solids of isolated, yield 33.3% is separated with preparative thin-layer chromatography.
1H NMR(300MHz,CD3OD) δ 8.51 (d, J=8.4Hz, 1H), 8.04 (d, J=8.9Hz, 2H), 7.17 (d, J=9.0Hz, 2H), 7.13 (d, J=8.3Hz, 1H), 6.95 (s, 1H), 4.13 (dd, J=13.4,2.2Hz, 2H), 3.99 (s, 6H), 3.56 3.46 (m, 2H), 2.82 (dd, J=13.5,11.7Hz, 2H), 1.42 (d, J=6.5Hz, 6H).
Embodiment 7:
N-(6-(2,6-bis-chloro-3,5-Dimethoxyphenyl)-1H-pyrazolo [3,4-b] pyridin-3-yl)-4-(piperazine-1-base) benzoyl Amine (I-7)
Step 1: preparation 4-(4-((6-(2,6-bis-chloro-3,5-Dimethoxyphenyl)-1H-pyrazolo [3,4-b] pyridin-3-yl) amino first Acyl group) phenyl) piperazine-1-t-butyl formate
Change 4-((3R, 5S)-3,5-lupetazin-1-base) ethyl benzoate into 4-(4-(ethoxy carbonyl) phenyl) piperazine-1-first Tert-butyl acrylate, remaining needed raw material, reagent and preparation method, with the step 6 in embodiment 6, obtain yellow solid 4-(4-((6-(2,6- Two chloro-3,5-Dimethoxyphenyls)-1H-pyrazolo [3,4-b] pyridin-3-yl) carbamoyl) phenyl) the tertiary fourth of piperazine-1-formic acid Ester.
1H NMR(300MHz,CDCl3) δ 10.13 (s, 1H), 8.85 (d, J=8.4Hz, 1H), 8.74 (s, 1H), 7.91 (d, J=8.8Hz, 2H), 7.16 (d, J=8.4Hz, 1H), 6.96 (d, J=8.9Hz, 2H), 6.66 (s, 1H), 3.97 (s, 6H), 3.70–3.53(m,4H),3.45–3.25(m,4H),1.50(s,9H).
Step 2:N-(6-(2,6-bis-chloro-3,5-Dimethoxyphenyl)-1H-pyrazolo [3,4-b] pyridin-3-yl)-4-(piperazine-1-base) Benzoylamide
By 32 milligrams of 4-(4-((6-(2,6-bis-chloro-3,5-Dimethoxyphenyl)-1H-pyrazolo [3,4-b] pyridin-3-yl) carbamyl Base) phenyl) piperazine-1-t-butyl formate is dissolved in 0.5 milliliter of dichloromethane, adds 0.08 milliliter of trifluoroacetic acid, be stirred at room temperature At night, reactant liquor concentrates and is evaporated, and residue is dissolved in methanol, and dropping ammonia regulation pH value is more than 7, and concentration is evaporated, residue Preparative thin-layer chromatography is used to separate (dichloromethane: methanol=10:1, V/V) 24 milligrams of yellow solids of isolated, yield 89.2%.
1H NMR (300MHz, DMSO) δ 13.39 (s, 1H), 10.84 (s, 1H), 8.38 (d, J=8.3Hz, 1H), 8.02 (d, J=8.9Hz, 2H), 7.12 6.97 (m, 4H), 3.98 (s, 6H), 3.52 3.20 (m, 4H), 3.07 2.90 (m, 4H).
Embodiment 8:
N-(6-(2,6-bis-chloro-3,5-Dimethoxyphenyl)-1H-pyrazolo [3,4-b] pyridin-3-yl)-4-(4-methylpiperazine-1-yl) Benzoylamide (I-8)
Step 1: preparation N-(6-(2,6-bis-chloro-3,5-Dimethoxyphenyl)-1H-pyrazolo [3,4-b] pyridin-3-yl)-4-(4-methyl Piperazine-1-base) Benzoylamide
Change 4-((3R, 5S)-3,5-lupetazin-1-base) ethyl benzoate into 4-(4-methylpiperazine-1-yl) ethyl benzoate, Remaining needed raw material, reagent and preparation method, with the step 6 in embodiment 6, obtain white solid N-(6-(2,6-bis-chloro-3,5-bis- Methoxyphenyl)-1H-pyrazolo [3,4-b] pyridin-3-yl)-4-(4-methylpiperazine-1-yl) Benzoylamide.
1H NMR (300MHz, DMSO) δ 13.38 (s, 1H), 10.82 (s, 1H), 8.38 (d, J=8.4Hz, 1H), 8.00 (d, J=8.9Hz, 2H), 7.13 6.96 (m, 4H), 3.98 (s, 6H), 3.32 3.25 (m, 4H), 2.48 2.39 (m, 4H), 2.23(s,3H).
Embodiment 9:
N-(6-(2,6-bis-chloro-3,5-Dimethoxyphenyl)-1H-pyrazolo [3,4-b] pyridin-3-yl)-4-(4-ethyl piperazidine-1-base) Benzoylamide (I-9)
Step 1: preparation 4-(4-ethyl piperazidine-1-base) ethyl benzoate
0.735 milliliter of 4-ethyl fluoro benzoate is dissolved in 5 milliliters of dimethyl sulfoxides, add 0.765 milliliter of 1-ethyl piperazidine and 1.039 grams of potassium carbonate, are warming up to 120 DEG C under argon shield, stop heating after stirring 19 hours, add 25 milliliters of water dilutions Reactant liquor, is extracted with ethyl acetate, and organic facies is washed five times with saturated sodium-chloride water solution after merging, and anhydrous sodium sulfate is dried, Filtering and concentrating, residue column chromatography (dichloromethane: methanol=99:1, V/V) 1.060 grams of light orange solids of isolated, yield 80.6%.
1H NMR(300MHz,CDCl3) δ 7.92 (d, J=9.0Hz, 2H), 6.87 (d, J=9.0Hz, 2H), 4.32 (q, J=7.1Hz, 2H), 3.45 3.26 (m, 4H), 2.73 2.55 (m, 4H), 2.47 (q, J=7.2Hz, 2H), 1.36 (t, J= 7.1Hz, 3H), 1.13 (t, J=7.2Hz, 3H).
Step 2: preparation N-(6-(2,6-bis-chloro-3,5-Dimethoxyphenyl)-1H-pyrazolo [3,4-b] pyridin-3-yl)-4-(4-ethyl Piperazine-1-base) Benzoylamide
Change 4-((3R, 5S)-3,5-lupetazin-1-base) ethyl benzoate into 4-(4-ethyl piperazidine-1-base) ethyl benzoate, Remaining needed raw material, reagent and preparation method, with the step 6 in embodiment 6, obtain white solid N-(6-(2,6-bis-chloro-3,5-bis- Methoxyphenyl)-1H-pyrazolo [3,4-b] pyridin-3-yl)-4-(4-ethyl piperazidine-1-base) Benzoylamide.
1H NMR(300MHz,CD3OD) δ 8.51 (d, J=8.3Hz, 1H), 7.99 (d, J=8.9Hz, 2H), 7.12 (d, J=8.3Hz, 1H), 7.08 (d, J=9.0Hz, 2H), 6.94 (s, 1H), 3.99 (s, 6H), 3.52 3.39 (m, 4H), 2.84 2.68 (m, 4H), 2.59 (q, J=7.3Hz, 2H), 1.19 (t, J=7.2Hz, 3H).
Embodiment 10:
N-(6-(2,6-bis-chloro-3,5-Dimethoxyphenyl)-1H-pyrazolo [3,4-b] pyridin-3-yl)-4-(4-isopropyl piperazine-1- Base) Benzoylamide (I-10)
Step 1: preparation 4-(4-isopropyl piperazine-1-base) ethyl benzoate
337 milligrams of 4-ethyl fluoro benzoates are dissolved in 2 milliliters of dimethyl sulfoxides, add 0.345 milliliter of 1-isopropyl piperazine and 415 Milligram potassium carbonate, is warming up to 120 DEG C under argon shield, stops heating after stirring 18 hours, adds 10 milliliters of water dilutions anti- Answering liquid, be extracted with ethyl acetate, organic facies is washed five times with saturated sodium-chloride water solution after merging, and anhydrous sodium sulfate is dried, Filtering and concentrating, residue column chromatography (dichloromethane: methanol=99:1 98:2, V/V) 358 milligrams of light yellow solids of isolated, Yield 64.8%.
1H NMR(300MHz,CDCl3) δ 7.92 (d, J=9.0Hz, 2H), 6.86 (d, J=9.1Hz, 2H), 4.32 (q, J=7.1Hz, 2H), 3.47 3.28 (m, 4H), 2.80 2.70 (m, 1H), 2.69 2.61 (m, 4H), 1.36 (t, J=7.1 Hz, 3H), 1.09 (d, J=6.5Hz, 6H).
Step 2: preparation N-(6-(2,6-bis-chloro-3,5-Dimethoxyphenyl)-1H-pyrazolo [3,4-b] pyridin-3-yl)-4-(4-isopropyl Base piperazine-1-base) Benzoylamide
4-((3R, 5S)-3,5-lupetazin-1-base) ethyl benzoate is changed into 4-(4-isopropyl piperazine-1-base) benzoic acid second Ester, remaining needed raw material, reagent and preparation method, with the step 6 in embodiment 6, obtain white solid N-(6-(2,6-bis-chloro-3,5- Dimethoxyphenyl)-1H-pyrazolo [3,4-b] pyridin-3-yl)-4-(4-isopropyl piperazine-1-base) Benzoylamide.
1H NMR(300MHz,CDCl3) δ 9.09 (s, 1H), 8.82 (d, J=8.4Hz, 1H), 7.89 (d, J=8.4Hz, 2H), 7.14 (d, J=8.4Hz, 1H), 6.88 (d, J=8.4Hz, 2H), 6.64 (s, 1H), 3.96 (s, 6H), 3.59 3.43 (m, 4H), 3.08 2.97 (m, 1H), 2.96 2.80 (m, 4H), 1.22 (d, J=6.5Hz, 6H).
Embodiment 11:
4-(4-cyclopropylpiperazin-1-base)-N-(6-(2,6-bis-chloro-3,5-Dimethoxyphenyl)-1H-pyrazolo [3,4-b] pyridine-3- Base) Benzoylamide (I-11)
Step 1: preparation 4-(4-cyclopropylpiperazin-1-base) ethyl benzoate
337 milligrams of 4-ethyl fluoro benzoates are dissolved in 2 milliliters of dimethyl sulfoxides, add 0.29 milliliter of 1-cyclopropylpiperazin and 415 Milligram potassium carbonate, is warming up to 120 DEG C under argon shield, stops heating after stirring 23 hours, adds 10 milliliters of water dilutions anti- Answering liquid, be extracted with ethyl acetate, organic facies is washed five times with saturated sodium-chloride water solution after merging, and anhydrous sodium sulfate is dried, Filtering and concentrating, residue column chromatography (dichloromethane: methanol=99:1 98:2, V/V) 365 milligrams of buff white solid of isolated, Yield 66.5%.
1H NMR(300MHz,CDCl3) δ 7.92 (d, J=9.1Hz, 2H), 6.86 (d, J=9.0Hz, 2H), 4.32 (q, J=7.1Hz, 2H), 3.46 3.19 (m, 4H), 2.93 2.66 (m, 4H), 1.75 1.54 (m, 1H), 1.36 (t, J=7.1 Hz,3H),0.67–0.37(m,4H).
Step 2: preparation 4-(4-cyclopropylpiperazin-1-base)-N-(6-(2,6-bis-chloro-3,5-Dimethoxyphenyl)-1H-pyrazolo [3,4-b] pyridin-3-yl) Benzoylamide
Change 4-((3R, 5S)-3,5-lupetazin-1-base) ethyl benzoate into 4-(4-cyclopropylpiperazin-1-base) benzoic acid second Ester, remaining needed raw material, reagent and preparation method, with the step 6 in embodiment 6, obtain white solid 4-(4-cyclopropylpiperazin-1- Base)-N-(6-(2,6-bis-chloro-3,5-Dimethoxyphenyl)-1H-pyrazolo [3,4-b] pyridin-3-yl) Benzoylamide.
1H NMR(300MHz,CDCl3) δ 10.20 (s, 1H), 8.85 (d, J=8.4Hz, 1H), 8.73 (s, 1H), 7.89 (d, J=8.7Hz, 2H), 7.15 (d, J=8.4Hz, 1H), 6.96 (d, J=8.9Hz, 2H), 6.66 (s, 1H), 3.97 (s, 6H), 3.44 3.30 (m, 4H), 2.91 2.76 (m, 4H), 1.26 (t, J=6.9Hz, 1H), 0.54 (d, J=6.5Hz, 4H).
Embodiment 12:
4-(4-cyclobutyl piperazine-1-base)-N-(6-(2,6-bis-chloro-3,5-Dimethoxyphenyl)-1H-pyrazolo [3,4-b] pyridine-3- Base) Benzoylamide (I-12)
Step 1: preparation 4-cyclobutyl piperazine-1-t-butyl formate
1.870 grams of piperazine-1-t-butyl formates are dissolved in 75 milliliter 1, in 2-dichloroethanes, add 0.75 milliliter of cyclobutanone, 15 minutes it are stirred at room temperature under argon shield.Add 1.277 grams of sodium triacetoxy borohydrides, be stirred at room temperature 30 minutes, then add Enter 1.277 grams of sodium triacetoxy borohydrides, stirred overnight at room temperature.Add 35 milliliters of sodium hydrate aqueous solutions (1 mole every liter) Cancellation is reacted, reactant liquor separatory, is extracted with ethyl acetate aqueous phase, and organic facies uses sodium hydrate aqueous solution (1 after merging successively Mole every liter) and saturated sodium-chloride water solution washing, anhydrous sodium sulfate is dried, filtering and concentrating, residue column chromatography (dichloromethane Alkane: methanol=99:1 95:5, V/V) 1.032 grams of yellow oil of isolated, yield 42.8%.
1H NMR(300MHz,CDCl3)δ3.47–3.30(m,4H),2.77–2.64(m,1H),2.35–2.16(m, 4H),2.03–1.96(m,2H),1.95–1.78(m,2H),1.78–1.64(m,2H),1.45(s,9H).
Step 2: preparation 1-cyclobutyl piperazine
1.030 grams of 4-cyclobutyl piperazine-1-t-butyl formates are dissolved in 12 ml methanol, are added dropwise over the dioxy of 4.5 milliliters of hydrochloric acid Six ring solution (4 moles every liter), stirred overnight at room temperature.Reactant liquor concentrates and is evaporated, and residue is dissolved in 16 milliliters of ether, room Temperature stirring 30 minutes；Concentration is evaporated, and residue is dissolved in 10 milliliters of ether/methanol (10:1, V/V), is stirred at room temperature 15 minutes, Filtering, ether filter wash cake, filter cake is vacuum dried, and obtains beige solid 889 milligrams.
Being dissolved in by beige solid in 24 milliliters of methylene chloride/water (1:1, V/V), (2 rub to be added dropwise over sodium hydrate aqueous solution You have every liter) to pH value more than 9, reactant liquor separatory, use dichloromethane aqueous phase extracted, organic facies uses saturated sodium-chloride after merging Solution washing, anhydrous sodium sulfate is dried, filtering and concentrating, vacuum drying, obtains 520 milligrams of yellow oil, productivity 86.6%.
1H NMR(300MHz,CDCl3) δ 2.89 (t, J=4.9Hz, 4H), 2.77 2.62 (m, 1H), 2.29 (s, 4H), 2.09–1.96(m,2H),1.95–1.78(m,2H),1.78–1.66(m,2H).
Step 3: preparation 4-(4-cyclobutyl piperazine-1-base) ethyl benzoate
484 milligrams of 4-ethyl fluoro benzoates are dissolved in 3 milliliters of dimethyl sulfoxides, add 484 milligrams of 1-cyclobutyl piperazines and 597 Milligram potassium carbonate, is warming up to 120 DEG C under argon shield, stops heating after stirring 18 hours, adds 15 milliliters of water dilutions anti- Answering liquid, be extracted with ethyl acetate, organic facies is washed five times with saturated sodium-chloride water solution after merging, and anhydrous sodium sulfate is dried, Filtering and concentrating, residue column chromatography (dichloromethane: methanol=99:1 98:2, V/V) 533 milligrams of buff white solid of isolated, Yield 64.2%.
1H NMR(300MHz,CDCl3) δ 7.92 (d, J=9.0Hz, 2H), 6.86 (d, J=9.0Hz, 2H), 4.32 (q, J=7.1Hz, 2H), 3.41 3.28 (m, 4H), 2.84 2.71 (m, 1H), 2.56 2.38 (m, 4H), 2.13 2.01 (m, 2H), 1.99 1.83 (m, 2H), 1.79 1.67 (m, 2H), 1.36 (t, J=7.1Hz, 3H).
Step 4: preparation 4-(4-cyclobutyl piperazine-1-base)-N-(6-(2,6-bis-chloro-3,5-Dimethoxyphenyl)-1H-pyrazolo [3,4-b] pyridin-3-yl) Benzoylamide
4-((3R, 5S)-3,5-lupetazin-1-base) ethyl benzoate is changed into 4-(4-cyclobutyl piperazine-1-base) benzoic acid second Ester, remaining needed raw material, reagent and preparation method, with the step 6 in embodiment 6, obtain white solid 4-(4-cyclobutyl piperazine-1- Base)-N-(6-(2,6-bis-chloro-3,5-Dimethoxyphenyl)-1H-pyrazolo [3,4-b] pyridin-3-yl) Benzoylamide.
1H NMR(300MHz,CD3OD) δ 8.51 (d, J=8.3Hz, 1H), 8.02 (d, J=8.9Hz, 2H), 7.18 7.08(m,3H),6.95(s,1H),3.99(s,6H),3.60–3.48(m,4H),3.10–3.05(m,1H),3.04–2.89 (m,4H),2.32–2.22(m,2H),2.19–2.09(m,2H),1.92–1.81(m,2H).
Embodiment 13:
N-(6-(2,6-bis-chloro-3,5-Dimethoxyphenyl)-1H-pyrazolo [3,4-b] pyridin-3-yl)-4-(4-(2-methoxy ethyl)- Piperazine-1-base) Benzoylamide (I-13)
Step 1: preparation 4-(4-(2-methoxy ethyl) piperazine-1-base) ethyl benzoate
337 milligrams of 4-ethyl fluoro benzoates are dissolved in 2 milliliters of dimethyl sulfoxides, add 0.36 milliliter of 1-(2-methoxy ethyl) piperazine Piperazine and 415 milligrams of potassium carbonate, be warming up to 120 DEG C under argon shield, stops heating, add 10 milliliters of water after stirring 27 hours Dilute reaction solution, is extracted with ethyl acetate, and organic facies is washed five times with saturated sodium-chloride water solution after merging, anhydrous sodium sulfate It is dried, filtering and concentrating, residue column chromatography (dichloromethane: methanol=99:1 98:2, V/V) 473 milligrams of yellow of isolated Solid, yield 80.9%.
1H NMR(300MHz,CDCl3) δ 7.92 (d, J=9.1Hz, 2H), 6.86 (d, J=9.1Hz, 2H), 4.32 (q, J=7.1Hz, 2H), 3.55 (t, J=5.5Hz, 2H), 3.46 3.29 (m, 7H), 2.77 2.55 (m, 6H), 1.36 (t, J= 7.1Hz,3H).
Step 2: preparation N-(6-(2,6-bis-chloro-3,5-Dimethoxyphenyl)-1H-pyrazolo [3,4-b] pyridin-3-yl)-4-(4-(2- Methoxy ethyl)-piperazine-1-base) Benzoylamide
Change 4-((3R, 5S)-3,5-lupetazin-1-base) ethyl benzoate into 4-(4-(2-methoxy ethyl) piperazine-1-base) benzene Ethyl formate, remaining needed raw material, reagent and preparation method, with the step 6 in embodiment 6, obtain white solid N-(6-(2,6- Two chloro-3,5-Dimethoxyphenyls)-1H-pyrazolo [3,4-b] pyridin-3-yl)-4-(4-(2-methoxy ethyl)-piperazine-1-base) benzene first Amide.
1H NMR(300MHz,CDCl3) δ 10.05 (s, 1H), 8.85 (d, J=8.4Hz, 1H), 8.62 (s, 1H), 7.90 (d, J=8.9Hz, 2H), 7.16 (d, J=8.2Hz, 1H), 6.96 (d, J=8.7Hz, 2H), 6.67 (s, 1H), 3.97 (s, 6H), 3.69–3.62(m,2H),3.54–3.44(m,4H),3.39(s,3H),2.89–2.71(m,6H).
Embodiment 14:
4-(4-pi-allyl piperazine-1-base)-N-(6-(2,6-bis-chloro-3,5-Dimethoxyphenyl)-1H-pyrazolo [3,4-b] pyridine-3- Base) Benzoylamide (I-14)
Step 1: preparation 4-(piperazine-1-base) ethyl benzoate
970 milligrams of 4-(4-(ethoxy carbonyl) phenyl) piperazine-1-t-butyl formate is dissolved in 20 milliliters of dichloromethane, adds 10 milliliters of trifluoroacetic acids, are stirred at room temperature 4 hours, and reactant liquor concentrates and is evaporated, and add saturated sodium bicarbonate aqueous solution and adjust pH value big In 7, extracting with dichloromethane, organic facies is washed with saturated sodium-chloride water solution after merging, and anhydrous sodium sulfate is dried, and filters Concentrate, vacuum drying, obtain 694 milligrams of light orange solids.
1H NMR(300MHz,CDCl3) δ 7.93 (d, J=9.0Hz, 2H), 6.87 (d, J=9.1Hz, 2H), 4.33 (q, J=7.1Hz, 2H), 3.46 3.21 (m, 4H), 3.19 2.94 (m, 4H), 1.37 (t, J=7.1Hz, 3H).
Step 2: preparation 4-(4-pi-allyl piperazine-1-base) ethyl benzoate
468 milligrams of 4-(piperazine-1-base) ethyl benzoate is dissolved in 4 milliliters of anhydrous methylene chlorides, adds 0.9 milliliter of N, N-bis- After wopropyl ethyl amine, then being added dropwise over 0.2 milliliter of 3-bromopropene, stirred overnight at room temperature under argon shield, reactant liquor concentrates It is evaporated, residue column chromatography (dichloromethane: methanol=99:1 98:2, V/V) 334 milligrams of beige solid of isolated, receives Rate 60.9%.
1H NMR(300MHz,CDCl3) δ 7.92 (d, J=9.1Hz, 2H), 6.86 (d, J=9.1Hz, 2H), 5.89 (ddt, J=16.8,10.2,6.6Hz, 1H), 5.27 5.16 (m, 2H), 4.32 (q, J=7.1Hz, 2H), 3.40 3.30 (m, 4H), 3.06 (d, J=6.6Hz, 2H), 2.65 2.55 (m, 4H), 1.36 (t, J=7.1Hz, 3H).
Step 3: preparation 4-(4-pi-allyl piperazine-1-base)-N-(6-(2,6-bis-chloro-3,5-Dimethoxyphenyl)-1H-pyrazolo [3,4-b] pyridin-3-yl) Benzoylamide
4-((3R, 5S)-3,5-lupetazin-1-base) ethyl benzoate is changed into 4-(4-pi-allyl piperazine-1-base) benzoic acid second Ester, remaining needed raw material, reagent and preparation method, with the step 6 in embodiment 6, obtain white solid 4-(4-pi-allyl piperazine-1- Base)-N-(6-(2,6-bis-chloro-3,5-Dimethoxyphenyl)-1H-pyrazolo [3,4-b] pyridin-3-yl) Benzoylamide.
1H NMR(300MHz,CD3OD) δ 8.51 (d, J=8.3Hz, 1H), 7.98 (d, J=8.9Hz, 2H), 7.12 (d, J=8.3Hz, 1H), 7.07 (d, J=9.0Hz, 2H), 6.95 (s, 1H), 5.94 (ddt, J=16.8,10.1,6.6Hz, 1H), 5.35 5.22 (m, 2H), 3.99 (s, 6H), 3.49 3.38 (m, 4H), 3.12 (d, J=6.7Hz, 2H), 2.76 2.60 (m, 4H).
Embodiment 15:
N-(6-(2,6-bis-chloro-3,5-Dimethoxyphenyl)-1H-pyrazolo [3,4-b] pyridin-3-yl)-4-(4-(2-hydroxyethyl)-piperazine Piperazine-1-base) Benzoylamide (I-15)
Step 1: preparation 4-(4-(2-hydroxyethyl)-piperazine-1-base) ethyl benzoate
585 milligrams of 4-(piperazine-1-base) ethyl benzoate is dissolved in 10 milliliters of acetonitriles, adds 180 microlitre ethylene bromohyrins and 415 Milligram potassium carbonate, is warming up to backflow under argon shield, stops heating, reacting liquid filtering, dichloromethane after stirring 10 hours Filter wash cake, filtrate concentrates and is evaporated, residue column chromatography (dichloromethane: methanol=99:1 98:2, V/V) isolated 480 Milligram pale pink solid, yield 69.0%.
1H NMR(300MHz,CDCl3) δ 7.93 (d, J=8.9Hz, 2H), 6.87 (d, J=8.9Hz, 2H), 4.33 (q, J=7.2Hz, 2H), 3.67 (t, J=5.3Hz, 2H), 3.37 3.29 (m, 4H), 2.70 2.64 (m, 4H), 2.64 2.56 (m, 2H), 1.37 (t, J=7.1Hz, 3H).
Step 2: preparation 4-(4-(2-((triisopropylsilyl) epoxide) ethyl) piperazine-1-base) ethyl benzoate
418 milligrams of 4-(4-(2-hydroxyethyl)-piperazine-1-base) ethyl benzoate and 133 milligrams of imidazoles are dissolved in 3 milliliters anhydrous two In chloromethanes, being added dropwise over 386 microlitre tri isopropyl chlorosilanes at 0 DEG C, recover to room temperature, stir 13 hours, add water cancellation Reaction, extracts with dichloromethane, and organic facies is washed with saturated sodium-chloride water solution after merging, and anhydrous sodium sulfate is dried, and filters Concentrate, residue column chromatography (dichloromethane: methanol=99:1, V/V) 624 milligrams of light orange grease of isolated, yield 95.7%.
1H NMR(300MHz,CDCl3) δ 7.92 (d, J=8.9Hz, 2H), 6.86 (d, J=9.0Hz, 2H), 4.32 (q, J=7.1Hz, 2H), 3.88 (t, J=6.2Hz, 2H), 3.38 3.27 (m, 4H), 2.74 2.66 (m, 4H), 2.62 (t, J= 6.2Hz, 2H), 1.36 (t, J=7.1Hz, 3H), 1.18 0.92 (m, 21H).
Step 3: preparation N-(6-(2,6-bis-chloro-3,5-Dimethoxyphenyl)-1H-pyrazolo [3,4-b] pyridine-3- Base)-4-(4-(2-((triisopropylsilyl) epoxide) ethyl) piperazine-1-base) Benzoylamide
4-((3R, 5S)-3,5-lupetazin-1-base) ethyl benzoate is changed into 4-(4-(2-((triisopropylsilyl) epoxide) second Base) piperazine-1-base) ethyl benzoate, remaining needed raw material, reagent and preparation method, with the step 6 in embodiment 6, obtain yellow Solid N-(6-(2,6-bis-chloro-3,5-Dimethoxyphenyl)-1H-pyrazolo [3,4-b] pyridin-3-yl)-4-(4-(2-((triisopropyl silicon Base) epoxide) ethyl) piperazine-1-base) Benzoylamide.
1H NMR(300MHz,CDCl3) δ 10.50 (s, 1H), 8.88 (s, 1H), 8.85 (d, J=8.4Hz, 1H), 7.89 (d, J=8.8Hz, 2H), 7.15 (d, J=8.4Hz, 1H), 6.94 (d, J=9.0Hz, 2H), 6.65 (s, 1H), 3.96 (s, 6H), 3.90 (t, J=6.2Hz, 2H), 3.40 3.29 (m, 4H), 2.78 2.68 (m, 4H), 2.64 (t, J=6.2Hz, 2H), 1.15 –0.99(m,21H).
Step 4: preparation N-(6-(2,6-bis-chloro-3,5-Dimethoxyphenyl)-1H-pyrazolo [3,4-b] pyridin-3-yl)-4-(4-(2- Hydroxyethyl)-piperazine-1-base) Benzoylamide
By 73 milligrams of N-(6-(2,6-bis-chloro-3,5-Dimethoxyphenyl)-1H-pyrazolo [3,4-b] pyridin-3-yl)-4-(4-(2-((three Isopropyl is silica-based) epoxide) ethyl) piperazine-1-base) Benzoylamide is dissolved in 1 milliliter of oxolane, is added dropwise over 0.11 milli at 0 DEG C Rising the tetrahydrofuran solution (1 mole every liter) of tetra-n-butyl ammonium fluoride, recover to being stirred at room temperature 17 hours, reactant liquor concentrates and steams Dry, residue column chromatography (dichloromethane: methanol=99:1 96:4, V/V) 25 milligrams of white solids of isolated, yield 43.7%.
Embodiment 16:
N-(6-(2,6-bis-chloro-3,5-Dimethoxyphenyl)-1H-pyrazolo [3,4-b] pyridin-3-yl)-4-((3R, 5S)-3,4,5-front three Base piperazine-1-base) Benzoylamide (I-16)
Step 1: preparation 4-((3R, 5S)-3,4,5-tri methyl piperazine-1-base) ethyl benzoate
262 milligrams of 4-((3R, 5S)-3,5-lupetazin-1-base) ethyl benzoate is dissolved in 5 milliliters of dichloromethane, successively Add 165 milligrams of formaldehyde, 115 microlitre acetic acid and 530 milligrams of sodium triacetoxy borohydrides, be stirred at room temperature 5 hours, add full Reacting with sodium bicarbonate aqueous solution cancellation, extract with dichloromethane, organic facies is washed with saturated sodium-chloride water solution after merging, Anhydrous sodium sulfate is dried, filtering and concentrating, residue column chromatography (dichloromethane: methanol=99:1 98:2, V/V) isolated 265 milligrams of yellow orange solid, yield 95.9%.
1H NMR(300MHz,CDCl3) δ 7.91 (d, J=9.0Hz, 2H), 6.84 (d, J=9.0Hz, 2H), 4.32 (q, J=7.1Hz, 2H), 3.66 3.58 (m, 2H), 2.68 (t, J=11.6Hz, 2H), 2.41 2.32 (m, 2H), 2.31 (s, 3H), 1.37 (t, J=7.1Hz, 3H), 1.18 (d, J=6.2Hz, 6H).
Step 2: preparation N-(6-(2,6-bis-chloro-3,5-Dimethoxyphenyl)-1H-pyrazolo [3,4-b] pyridine-3- Base)-4-((3R, 5S)-3,4,5-tri methyl piperazine-1-base) Benzoylamide
Change 4-((3R, 5S)-3,5-lupetazin-1-base) ethyl benzoate into 4-((3R, 5S)-3,4,5-tri methyl piperazine-1-base) Ethyl benzoate, remaining needed raw material, reagent and preparation method, with the step 6 in embodiment 6, obtain white solid N-(6-(2,6- Two chloro-3,5-Dimethoxyphenyls)-1H-pyrazolo [3,4-b] pyridin-3-yl)-4-((3R, 5S)-3,4,5-tri methyl piperazine-1-base) benzene Methanamide.
1H NMR(300MHz,CD3OD) δ 8.51 (d, J=8.3Hz, 1H), 8.02 (d, J=8.9Hz, 2H), 7.18 7.10 (m, 3H), 6.95 (s, 1H), 4.04 (d, J=13.7Hz, 2H), 3.99 (s, 6H), 2.99 2.88 (m, 2H), 2.81 (s, 3H), 1.44 (d, J=6.4Hz, 6H).
Embodiment 17:
4-(4-Acetylpiperazine-1-base)-N-(6-(2,6-bis-chloro-3,5-Dimethoxyphenyl)-1H-pyrazolo [3,4-b] pyridine-3- Base) Benzoylamide (I-17)
Step 1: preparation 4-(4-Acetylpiperazine-1-base) ethyl benzoate
337 milligrams of 4-ethyl fluoro benzoates are dissolved in 2 milliliters of dimethyl sulfoxides, add 310 milligrams of 1-(piperazine-1-base) ethyl ketone and 415 milligrams of potassium carbonate, are warming up to 120 DEG C under argon shield, stop heating after stirring 20 hours, add 10 milliliters of water dilutions Reactant liquor, is extracted with ethyl acetate, and organic facies is washed five times with saturated sodium-chloride water solution after merging, and anhydrous sodium sulfate is dried, Filtering and concentrating, residue column chromatography (dichloromethane: methanol=99:1 98:2, V/V) 350 milligrams of white solids of isolated, Yield 63.3%.
1H NMR(300MHz,CDCl3) δ 7.95 (d, J=9.1Hz, 2H), 6.87 (d, J=9.1Hz, 2H), 4.33 (q, J=7.1Hz, 2H), 3.81 3.73 (m, 2H), 3.67 3.58 (m, 2H), 3.39 3.28 (m, 4H), 2.15 (s, 3H), 1.37 (t, J=7.1Hz, 3H).
Step 2: preparation 4-(4-Acetylpiperazine-1-base)-N-(6-(2,6-bis-chloro-3,5-Dimethoxyphenyl)-1H-pyrazolo [3,4-b] pyridin-3-yl) Benzoylamide
Change 4-((3R, 5S)-3,5-lupetazin-1-base) ethyl benzoate into 4-(4-Acetylpiperazine-1-base) benzoic acid second Ester, remaining needed raw material, reagent and preparation method, with the step 6 in embodiment 6, obtain white solid 4-(4-Acetylpiperazine-1- Base)-N-(6-(2,6-bis-chloro-3,5-Dimethoxyphenyl)-1H-pyrazolo [3,4-b] pyridin-3-yl) Benzoylamide.
1H NMR (300MHz, DMSO) δ 13.37 (s, 1H), 10.82 (s, 1H), 8.38 (d, J=8.3Hz, 1H), 8.02 (d, J=8.7Hz, 2H), 7.13 6.99 (m, 4H), 3.98 (s, 6H), 3.70 3.53 (m, 4H), 3.43 3.36 (m, 2H), 2.05(s,3H).
Embodiment 18:
N-(6-(2,6-bis-chloro-3,5-Dimethoxyphenyl)-1H-pyrazolo [3,4-b] pyridin-3-yl)-4-(4-(methyl sulphonyl) piperazine Piperazine-1-base) Benzoylamide (I-18)
Step 1: preparation 4-(4-(methyl sulphonyl) piperazine-1-base) ethyl benzoate
234 milligrams of 4-(piperazine-1-base) ethyl benzoate is dissolved in 3 milliliters of anhydrous methylene chlorides, is added dropwise over 102 microlitre first Sulfonic acid chloride and 350 microlitre triethylamines, be stirred at room temperature 23 hours under argon shield, and the cancellation that adds water is reacted, and extracts with dichloromethane Taking, organic facies is washed with saturated sodium-chloride water solution after merging, and anhydrous sodium sulfate is dried, filtering and concentrating, and residue is dissolved in 3 Milliliter methylene chloride/methanol (10:1, V/V), is stirred at room temperature 2 hours, filters, dichloromethane filter wash cake, and filter cake is vacuum dried, Obtain 261 milligrams of white solids, productivity 83.6%.
1H NMR (300MHz, DMSO) δ 7.81 (d, J=9.0Hz, 2H), 7.03 (d, J=9.0Hz, 2H), 4.24 (q, J=7.1Hz, 2H), 3.55 3.40 (m, 4H), 3.29 3.17 (m, 4H), 2.92 (s, 3H), 1.29 (t, J=7.1Hz, 3H).
Step 2: preparation N-(6-(2,6-bis-chloro-3,5-Dimethoxyphenyl)-1H-pyrazolo [3,4-b] pyridin-3-yl)-4-(4-(first Base sulfonyl) piperazine-1-base) Benzoylamide
Change 4-((3R, 5S)-3,5-lupetazin-1-base) ethyl benzoate into 4-(4-(methyl sulphonyl) piperazine-1-base) benzene first Acetoacetic ester, remaining needed raw material, reagent and preparation method, with the step 6 in embodiment 6, obtain white solid N-(6-(2,6-bis- Chloro-3,5-Dimethoxyphenyl)-1H-pyrazolo [3,4-b] pyridin-3-yl)-4-(4-(methyl sulphonyl) piperazine-1-base) Benzoylamide.
1H NMR (400MHz, DMSO) δ 13.39 (s, 1H), 10.86 (s, 1H), 8.38 (d, J=8.3Hz, 1H), 8.03 (d, J=8.7Hz, 2H), 7.12 7.06 (m, 3H), 7.05 (s, 1H), 3.98 (s, 6H), 3.51 3.40 (m, 4H), 3.29 3.21(m,4H),2.93(s,3H).
Embodiment 19:
4-(4-((6-(2,6-bis-chloro-3,5-Dimethoxyphenyl)-1H-pyrazolo [3,4-b] pyridin-3-yl) carbamoyl) benzene Base)-1,4-Diazesuberane-1-carboxylic acid tert-butyl ester (I-19)
Step 1: preparation 4-(4-(ethoxy carbonyl) phenyl)-1,4-Diazesuberane-1-t-butyl formate
504 milligrams of 4-ethyl fluoro benzoates are dissolved in 3 milliliters of dimethyl sulfoxides, add 710 microlitre Isosorbide-5-Nitraes-Diazesuberane-1- T-butyl formate and 622 milligrams of potassium carbonate, be warming up to 120 DEG C under argon shield, stops heating, add after stirring 37 hours 15 milliliters of water dilute reaction solutions, are extracted with ethyl acetate, and organic facies is washed five times with saturated sodium-chloride water solution after merging, nothing Aqueous sodium persulfate is dried, filtering and concentrating, residue column chromatography (petroleum ether: ethyl acetate=95:5 90:10, V/V) isolated 353 milligrams of colorless oil, yield 33.8%.
1H NMR(300MHz,CDCl3) δ 7.90 (d, J=9.0Hz, 2H), 6.67 (d, J=8.6Hz, 2H), 4.31 (q, J=7.1Hz, 2H), 3.71 3.53 (m, 6H), 3.31 (t, J=6.1Hz, 1H), 3.20 (t, J=6.1Hz, 1H), 2.03 1.91(m,2H),1.50–1.31(m,12H).
Step 2: preparation 4-(4-((6-(2,6-bis-chloro-3,5-Dimethoxyphenyl)-1H-pyrazolo [3,4-b] pyridin-3-yl) amino first Acyl group) phenyl)-1,4-Diazesuberane-1-carboxylic acid tert-butyl ester
Change 4-((3R, 5S)-3,5-lupetazin-1-base) ethyl benzoate into 4-(4-(ethoxy carbonyl) phenyl)-1,4-phenodiazine Trioxepane-1-t-butyl formate, remaining needed raw material, reagent and preparation method, with the step 6 in embodiment 6, obtain white Solid 4-(4-((6-(2,6-bis-chloro-3,5-Dimethoxyphenyl)-1H-pyrazolo [3,4-b] pyridin-3-yl) carbamoyl) benzene Base)-1,4-Diazesuberane-1-carboxylic acid tert-butyl ester.
1H NMR(300MHz,CDCl3) δ 8.86 (d, J=8.4Hz, 1H), 8.71 (d, J=10.1Hz, 1H), 7.88 (d, J=8.8Hz, 2H), 7.15 (d, J=8.4Hz, 1H), 6.76 (d, J=8.8Hz, 2H), 6.66 (s, 1H), 3.97 (s, 6H), 3.71 3.55 (m, 6H), 3.29 (dt, J=31.1,5.9Hz, 2H), 2.08 1.94 (m, 2H), 1.41 (d, J=16.3Hz, 9H).
Embodiment 20:
4-(1,4-Diazesuberane-1-base)-N-(6-(2,6-bis-chloro-3,5-Dimethoxyphenyl)-1H-pyrazolo [3,4-b] pyridine -3-base) Benzoylamide (I-20)
Step 1: preparation 4-(1,4-Diazesuberane-1-base)-N-(6-(2,6-bis-chloro-3,5-Dimethoxyphenyl)-1H-pyrazolo [3,4-b] pyridin-3-yl) Benzoylamide
By 25 milligrams of 4-(4-((6-(2,6-bis-chloro-3,5-Dimethoxyphenyl)-1H-pyrazolo [3,4-b] pyridin-3-yl) carbamyl Base) phenyl)-Isosorbide-5-Nitrae-Diazesuberane-1-carboxylic acid tert-butyl ester is dissolved in 0.5 milliliter of dichloromethane, adds 0.06 milliliter of trifluoro second Acid, stirred overnight at room temperature, reactant liquor concentrates and is evaporated, and residue is dissolved in methanol, and dropping ammonia regulation pH value is more than 7, dense Contracting is evaporated, and residue uses preparative thin-layer chromatography to separate (dichloromethane: methanol=10:1, V/V) 18 milligrams of rice of isolated White solid, yield 85.2%.
1H NMR(300MHz,CD3OD) δ 8.50 (d, J=8.3Hz, 1H), 8.01 (d, J=8.8Hz, 2H), 7.13 (d, J=8.3Hz, 1H), 6.99 6.91 (m, 3H), 3.99 (s, 6H), 3.92 3.86 (m, 2H), 3.73 (t, J=6.1Hz, 2H), 3.44–3.37(m,2H),3.28–3.21(m,2H),2.25–2.15(m,2H).
Embodiment 21:
N-(6-(2,6-bis-chloro-3,5-Dimethoxyphenyl)-1H-pyrazolo [3,4-b] pyridin-3-yl)-4-(4-methyl isophthalic acid, 4-diaza Cycloheptane-1-base) Benzoylamide (I-21)
Step 1: preparation 4-(4-methyl isophthalic acid, 4-Diazesuberane-1-base) ethyl benzoate
337 milligrams of 4-ethyl fluoro benzoates are dissolved in 2 milliliters of dimethyl sulfoxides, add 300 microlitre 1-methyl isophthalic acids, 4-diazacyclo Heptane and 415 milligrams of potassium carbonate, be warming up to 120 DEG C under argon shield, stops heating, add 10 milliliters after stirring 25 hours Water dilute reaction solution, is extracted with ethyl acetate, and organic facies is washed five times with saturated sodium-chloride water solution after merging, anhydrous slufuric acid Sodium is dried, filtering and concentrating, residue column chromatography (dichloromethane: methanol=99:1 98:2, V/V) 305 milligrams of Huangs of isolated Color solid, yield 58.1%.
1H NMR(300MHz,CDCl3) δ 7.89 (d, J=9.1Hz, 2H), 6.65 (d, J=9.1Hz, 2H), 4.31 (q, J=7.1Hz, 2H), 3.67 3.59 (m, 2H), 3.54 (t, J=6.3Hz, 2H), 2.76 2.67 (m, 2H), 2.59 2.51 (m, 2H), 2.38 (s, 3H), 2.08 1.96 (m, 2H), 1.36 (t, J=7.1Hz, 3H).
Step 2: preparation N-(6-(2,6-bis-chloro-3,5-Dimethoxyphenyl)-1H-pyrazolo [3,4-b] pyridin-3-yl)-4-(4-methyl -1,4-Diazesuberane-1-base) Benzoylamide
4-((3R, 5S)-3,5-lupetazin-1-base) ethyl benzoate is changed into 4-(4-methyl isophthalic acid, 4-Diazesuberane-1-base) Ethyl benzoate, remaining needed raw material, reagent and preparation method, with the step 6 in embodiment 6, obtain beige solid N-(6-(2,6-bis-chloro-3,5-Dimethoxyphenyl)-1H-pyrazolo [3,4-b] pyridin-3-yl)-4-(4-methyl isophthalic acid, 4-diaza cycloheptyl Alkane-1-base) Benzoylamide.
1H NMR(300MHz,CD3OD) δ 8.50 (d, J=8.3Hz, 1H), 8.00 (d, J=9.0Hz, 2H), 7.12 (d, J=8.3Hz, 1H), 6.95 (s, 1H), 6.92 (d, J=9.1Hz, 2H), 3.99 (s, 6H), 3.89 3.79 (m, 2H), 3.66 (t, J=6.3Hz, 2H), 3.28 3.20 (m, 2H), 3.17 3.09 (m, 2H), 2.75 (s, 3H), 2.28 2.17 (m, 2H).
Embodiment 22:
N-(6-(2,6-bis-chloro-3,5-Dimethoxyphenyl)-1H-pyrazolo [3,4-b] pyridin-3-yl)-4-(piperidin-1-yl) benzoyl Amine (I-22)
Step 1: preparation 4-(piperidin-1-yl) ethyl benzoate
337 milligrams of 4-ethyl fluoro benzoates are dissolved in 2 milliliters of dimethyl sulfoxides, add 0.24 milliliter of piperidines and 415 milligrams of carbonic acid Potassium, is warming up to 100 DEG C under argon shield, stops heating, adds 10 milliliters of water dilute reaction solutions, use after stirring 18 hours Ethyl acetate extracts, and organic facies is washed five times with saturated sodium-chloride water solution after merging, and anhydrous sodium sulfate is dried, filtering and concentrating, Residue column chromatography (petroleum ether: ethyl acetate=99:1 98:2, V/V) 400 milligrams of white solids of isolated, yield 85.7%.
1H NMR(300MHz,CDCl3) δ 7.90 (d, J=9.1Hz, 2H), 6.85 (d, J=9.1Hz, 2H), 4.32 (q, J=7.1Hz, 2H), 3.36 3.29 (m, 4H), 1.73 1.60 (m, 6H), 1.36 (t, J=7.1Hz, 3H).
Step 2: preparation N-(6-(2,6-bis-chloro-3,5-Dimethoxyphenyl)-1H-pyrazolo [3,4-b] pyridin-3-yl)-4-(piperidines -1-base) Benzoylamide
Change 4-((3R, 5S)-3,5-lupetazin-1-base) ethyl benzoate into 4-(piperidin-1-yl) ethyl benzoate, remaining institute Need raw material, reagent and preparation method with the step 6 in embodiment 6, obtain yellow solid N-(6-(and 2,6-bis-chloro-3,5-dimethoxy Phenyl)-1H-pyrazolo [3,4-b] pyridin-3-yl)-4-(piperidin-1-yl) Benzoylamide.
1H NMR(300MHz,CDCl3) δ 10.30 (s, 1H), 8.86 (d, J=8.4Hz, 1H), 8.81 (s, 1H), 7.87 (d, J=8.6Hz, 2H), 7.15 (d, J=8.4Hz, 1H), 6.94 (d, J=8.7Hz, 2H), 6.66 (s, 1H), 3.97 (s, 6H), 3.44–3.24(m,4H),1.93–1.65(m,6H).
Embodiment 23:
N-(6-(2,6-bis-chloro-3,5-Dimethoxyphenyl)-1H-pyrazolo [3,4-b] pyridin-3-yl)-4-morpholinyl Benzoylamide (I-23)
Step 1: preparation 4-morpholinyl benzoic acid ethyl ester
Being dissolved in 2 milliliters of morpholines by 337 milligrams of 4-ethyl fluoro benzoates, be warming up to 120 DEG C under argon shield, stirring 25 is little Stopping heating time after, reactant liquor concentrates and is evaporated, residue column chromatography (petroleum ether: ethyl acetate=95:5 90:10, V/V) 330 milligrams of white solids of isolated, yield 70.1%.
1H NMR(300MHz,CDCl3) δ 7.94 (d, J=8.9Hz, 2H), 6.86 (d, J=8.9Hz, 2H), 4.33 (q, J=7.1Hz, 2H), 3.89 3.82 (m, 4H), 3.31 3.24 (m, 4H), 1.37 (t, J=7.1Hz, 3H).
Step 2: preparation N-(6-(2,6-bis-chloro-3,5-Dimethoxyphenyl)-1H-pyrazolo [3,4-b] pyridin-3-yl)-4-morpholinyl Benzoylamide
Change 4-((3R, 5S)-3,5-lupetazin-1-base) ethyl benzoate into 4-morpholinyl benzoic acid ethyl ester, former needed for remaining Material, reagent and preparation method with the step 6 in embodiment 6, obtain yellow solid N-(6-(and 2,6-bis-chloro-3,5-dimethoxy benzene Base)-1H-pyrazolo [3,4-b] pyridin-3-yl)-4-morpholinyl Benzoylamide.
1H NMR (300MHz, DMSO) δ 13.38 (s, 1H), 10.83 (s, 1H), 8.39 (d, J=8.3Hz, 1H), 8.02 (d, J=8.8Hz, 2H), 7.16 6.95 (m, 4H), 3.98 (s, 6H), 3.85 3.68 (m, 4H), 3.31 3.18 (m, 4H).
Embodiment 24:
N-(6-(2,6-bis-chloro-3,5-Dimethoxyphenyl)-1H-pyrazolo [3,4-b] pyridin-3-yl)-4-(1-methyl piperidine-4-base) Benzoylamide (I-24)
Step 1: preparation 1-methyl 4-phenyl piperidines
645 milligrams of 4-Phenylpiperidines are dissolved in 13 milliliters of oxolanes, add 1.7 milliliters of formaldehyde and 1.700 grams of triacetyl oxygen Base sodium borohydride, is stirred at room temperature 19 hours, and reactant liquor concentrates and is evaporated, and residue is dissolved in dichloromethane and saturated sodium bicarbonate water In solution, separatory, extract with dichloromethane, organic facies is washed with saturated sodium-chloride water solution after merging, and anhydrous sodium sulfate is done Dry, filtering and concentrating, residue column chromatography (dichloromethane: methanol=95:5, V/V) 380 milligrams of colorless oil of isolated, Yield 54.2%.
1H NMR(300MHz,CDCl3) δ 7.34 7.16 (m, 5H), 2.97 (d, J=11.6Hz, 2H), 2.55 2.41 (m,1H),2.32(s,3H),2.10–1.99(m,2H),1.91–1.76(m,4H).
Step 2: preparation 4-(1-methyl piperidine-4-base) essence of Niobe
380 milligrams of 1-methyl 4-phenyl piperidines are dissolved in 3.8 milliliters of anhydrous methylene chlorides, are cooled under argon shield -30 DEG C, add 0.75 milliliter of oxalyl chloride and 1.182 grams of aluminum chlorides, stir 1 hour, recover to room temperature to stir 2 hours； It is cooled to 0 DEG C, is slowly added dropwise absolute methanol and clarifies to reactant liquor, recover to room temperature to stir 18 hours；Drip saturated at 0 DEG C Aqueous sodium carbonate cancellation is reacted, diluted ethyl acetate reactant liquor, filters, ethyl acetate filter wash cake, filtrate separatory, organic The most successively with water, saturated sodium-chloride water solution washing, anhydrous sodium sulfate is dried, filtering and concentrating, residue column chromatography (dichloro Methane: methanol=98:2 90:10, V/V) 437 milligrams of yellow oil of isolated, yield 86.3%.
1H NMR(300MHz,CDCl3) δ 7.96 (d, J=8.2Hz, 2H), 7.29 (d, J=8.2Hz, 2H), 3.89 (s, 3H), 3.04 (d, J=11.3Hz, 2H), 2.62 2.49 (m, 1H), 2.36 (s, 3H), 2.19 2.06 (m, 2H), 1.92 1.77(m,4H).
Step 3: preparation N-(6-(2,6-bis-chloro-3,5-Dimethoxyphenyl)-1H-pyrazolo [3,4-b] pyridin-3-yl)-4-(1-methyl Piperidin-4-yl) Benzoylamide
Change 4-((3R, 5S)-3,5-lupetazin-1-base) ethyl benzoate into 4-(1-methyl piperidine-4-base) benzoic acid first Ester, remaining needed raw material, reagent and preparation method, with the step 6 in embodiment 6, obtain yellow solid N-(6-(2,6-bis-chloro-3,5- Dimethoxyphenyl)-1H-pyrazolo [3,4-b] pyridin-3-yl)-4-(1-methyl piperidine-4-base) Benzoylamide.
1H NMR(300MHz,CD3OD) δ 8.53 (d, J=8.3Hz, 1H), 8.06 (d, J=8.4Hz, 2H), 7.49 (d, J=8.3Hz, 2H), 7.14 (d, J=8.3Hz, 1H), 6.95 (s, 1H), 3.99 (s, 6H), 3.56 (d, J=12.2Hz, 2H), 3.07 (dd, J=13.1,11.4Hz, 2H), 3.01 2.93 (m, 1H), 2.87 (s, 3H), 2.21 1.92 (m, 4H).
Embodiment 25:
N-(6-(3,5-Dimethoxyphenyl)-1H-indazole-3-base)-4-(piperazine-1-base) Benzoylamide (I-25)
Step 1: preparation 6-bromo-1H-indazole-3-amine
400 milligrams of 4-bromo-2-fluorobenzonitriles are dissolved in 6 milliliters of n-butyl alcohol, add 0.46 milliliter of hydrazine hydrate (85%, w/w), rise Temperature, to 100 DEG C, stops heating after stirring 14 hours, reactant liquor is concentrated into 2 milliliters, filters, water and a small amount of ethyl acetate filter wash Cake, filter cake is vacuum dried, obtains 336 milligrams of light orange solids, yield 79.2%.
1H NMR (300MHz, DMSO) δ 11.50 (s, 1H), 7.63 (d, J=8.5Hz, 1H), 7.41 (d, J=1.0Hz, 1H), 7.02 (dd, J=8.5,1.5Hz, 1H), 5.46 (s, 2H).
Step 2: preparation 6-(3,5-Dimethoxyphenyl)-1H-indazole-3-amine
By 212 milligrams of 6-bromo-1H-indazole-3-amine and 265 milligrams of 2-(3,5-Dimethoxyphenyl)-4,4,5,5-tetramethyl-1,3,2-two Oxa-borine is dissolved in 10 milliliters of dioxane, be sequentially added into 4 milliliters of aqueous sodium carbonates (1 mole every liter) and 74 milligrams [1,1 '- Double (diphenylphosphino) ferrocene] palladium chloride, under argon shield, it is warming up to 100 DEG C, after stirring 11 hours, stops heating, Reacting liquid filtering, ethyl acetate filter wash cake, filtrate concentrates and is evaporated, and residue is dissolved in ethyl acetate, uses saturated sodium-chloride water Solution washs, and anhydrous sodium sulfate is dried, filtering and concentrating, residue column chromatography (dichloromethane: methanol=99:1 98:2, V/V) 209 milligrams of pink solid of isolated, yield 77.6%.
1H NMR (300MHz, DMSO) δ 11.42 (s, 1H), 7.73 (d, J=8.3Hz, 1H), 7.42 (s, 1H), 7.19 (dd, J=8.4,1.4Hz, 1H), 6.79 (d, J=2.3Hz, 2H), 6.50 (t, J=2.2Hz, 1H), 5.37 (s, 2H), 3.81 (s, 6H).
Step 3: preparation 4-(4-((6-(3,5-Dimethoxyphenyl)-1H-indazole-3-base) carbamoyl) phenyl) piperazine-1-formic acid The tert-butyl ester
Change 6-(3,5-Dimethoxyphenyl)-1H-pyrazolo [3,4-b] pyridine-3-amine into 6-(3,5-Dimethoxyphenyl)-1H-Yin Azoles-3-amine, remaining needed raw material, reagent and preparation method, with the step 2 in embodiment 4, obtain buff white solid 4-(4-((6-(3,5- Dimethoxyphenyl)-1H-indazole-3-base) carbamoyl) phenyl) piperazine-1-t-butyl formate.
1H NMR(300MHz,CDCl3) δ 8.66 (s, 1H), 8.15 (d, J=8.6Hz, 1H), 7.90 (d, J=8.7Hz, 2H), 7.51 (s, 1H), 7.37 (d, J=8.6Hz, 1H), 6.92 (d, J=8.8Hz, 2H), 6.76 (d, J=2.1Hz, 2H), 6.49 (t, J=2.1Hz, 1H), 3.85 (s, 6H), 3.71 3.52 (m, 4H), 3.45 3.22 (m, 4H), 1.50 (s, 9H).
Step 4: preparation N-(6-(3,5-Dimethoxyphenyl)-1H-indazole-3-base)-4-(piperazine-1-base) Benzoylamide
By 48 milligrams of 4-(4-((6-(3,5-Dimethoxyphenyl)-1H-indazole-3-base) carbamoyl) phenyl) piperazine-1-formic acid uncle Butyl ester is dissolved in 0.8 milliliter of dichloromethane, adds 0.13 milliliter of trifluoroacetic acid, stirred overnight at room temperature, and reactant liquor concentrates and is evaporated, Residue is dissolved in methanol, and dropping ammonia regulation pH value is more than 7, and concentration is evaporated, residue column chromatography (dichloromethane: first Alcohol=98:2 96:4, V/V) 6 milligrams of yellow solids of isolated, yield 15.2%.
1H NMR (300MHz, DMSO) δ 12.82 (s, 1H), 10.60 (s, 1H), 8.03 (d, J=8.8Hz, 2H), 7.76 (d, J=8.3Hz, 1H), 7.67 (s, 1H), 7.37 (dd, J=8.4,1.0Hz, 1H), 7.09 (d, J=8.8Hz, 2H), 6.84 (d, J=2.2Hz, 2H), 6.54 (t, J=1.9Hz, 1H), 3.83 (s, 6H), 3.58 3.47 (m, 4H), 3.20 (s, 4H).
Embodiment 26:
N-(6-(2,6-bis-chloro-3,5-Dimethoxyphenyl)-1H-indazole-3-base)-4-((3R, 5S)-3,5-lupetazin-1-base) benzene Methanamide (I-26)
Step 1: preparation 6-(2,6-bis-chloro-3,5-Dimethoxyphenyl)-1H-indazole-3-amine
By 848 milligrams of 6-bromo-1H-indazole-3-amine and 2.664 grams of 2-(2,6-bis-chloro-3,5-Dimethoxyphenyl)-4,4,5,5-tetramethyls -1,3,2-dioxaborinate are dissolved in 40 milliliters of dioxane, be sequentially added into 16 milliliters of aqueous sodium carbonates (1 mole every liter) and 300 milligrams of [1,1 '-bis-(diphenylphosphino) ferrocene] palladium chlorides, are warming up to 100 DEG C under argon shield, stir 18 hours Rear stopping is heated, reacting liquid filtering, ethyl acetate filter wash cake, and filtrate separatory is extracted with ethyl acetate, after organic facies merges Washing with saturated sodium-chloride water solution, anhydrous sodium sulfate is dried, filtering and concentrating, residue column chromatography (dichloromethane: methanol =99:1 98:2, V/V) 1.308 grams of yellow solids of isolated, yield 96.7%.
1H NMR (300MHz, DMSO) δ 11.45 (s, 1H), 7.73 (d, J=8.4Hz, 1H), 6.99 (s, 1H), 6.97 (s, 1H), 6.66 (d, J=8.2Hz, 1H), 5.40 (s, 2H), 3.96 (s, 6H).
Step 2: preparation N-(6-(2,6-bis-chloro-3,5-Dimethoxyphenyl)-1H-indazole-3-base)-4-((3R, 5S)-3,5-dimethyl Piperazine-1-base) Benzoylamide
By 203 milligrams of 6-(2,6-bis-chloro-3,5-Dimethoxyphenyl)-1H-indazole-3-amine and 236 milligrams of 4-((3R, 5S)-3,5-two Methylpiperazine-1-yl) ethyl benzoate is dissolved in 3 milliliters of toluene, and (2 rub to be added dropwise over the toluene solution of 1.2 milliliters of trimethyl aluminiums You have every liter), under argon shield, it is warming up to 60 DEG C, after stirring 65 hours, stops heating, methanol cancellation is reacted, and reactant liquor is dense Contracting is evaporated, residue column chromatography (dichloromethane: methanol=99:1 90:10, V/V) 5 milligrams of white solids of isolated, receives Rate 1.5%.
1H NMR (400MHz, MeOD) δ 7.97 (d, J=8.9Hz, 2H), 7.84 (d, J=8.5Hz, 1H), 7.28 (s, 1H), 7.06 (d, J=9.0Hz, 2H), 6.92 (dd, J=8.5,1.1Hz, 1H), 6.88 (s, 1H), 3.98 (s, 6H), 3.81 (dd, J=12.4,2.4Hz, 2H), 3.03 2.93 (m, 2H), 2.46 2.36 (m, 2H), 1.18 (d, J=6.4Hz, 6H).
Embodiment 27:
N-(6-(2,6-bis-chloro-3,5-Dimethoxyphenyl)-1H-indazole-3-base)-4-(4-methylpiperazine-1-yl) Benzoylamide (I-27)
Step 1: preparation N-(6-(2,6-bis-chloro-3,5-Dimethoxyphenyl)-1H-indazole-3-base)-4-(4-methylpiperazine-1-yl) benzene Methanamide
Change 4-((3R, 5S)-3,5-lupetazin-1-base) ethyl benzoate into 4-(4-methylpiperazine-1-yl) ethyl benzoate, Remaining needed raw material, reagent and preparation method, with the step 2 in embodiment 26, obtain white solid N-(6-(2,6-bis-chloro-3,5-bis- Methoxyphenyl)-1H-indazole-3-base)-4-(4-methylpiperazine-1-yl) Benzoylamide.
1H NMR(300MHz,CD3OD) δ 7.98 (d, J=8.8Hz, 2H), 7.85 (d, J=8.4Hz, 1H), 7.28 (s, 1H), 7.06 (d, J=8.9Hz, 2H), 6.92 (dd, J=8.4,0.9Hz, 1H), 6.88 (s, 1H), 3.97 (s, 6H), 3.46 3.35(m,4H),2.73–2.53(m,4H),2.37(s,3H).
Embodiment 28:
N-(6-(2,6-bis-chloro-3,5-Dimethoxyphenyl)-1H-indazole-3-base)-4-(4-methyl isophthalic acid, 4-Diazesuberane-1-base) Benzoylamide (I-28)
Step 1: preparation N-(6-(2,6-bis-chloro-3,5-Dimethoxyphenyl)-1H-indazole-3-base)-4-(4-methyl isophthalic acid, 4-diazacyclo Heptane-1-base) Benzoylamide
4-((3R, 5S)-3,5-lupetazin-1-base) ethyl benzoate is changed into 4-(4-methyl isophthalic acid, 4-Diazesuberane-1-base) Ethyl benzoate, remaining needed raw material, reagent and preparation method, with the step 2 in embodiment 26, obtain Fructus Citri tangerinae color solid N-(6-(2,6- Two chloro-3,5-Dimethoxyphenyls)-1H-indazole-3-base)-4-(4-methyl isophthalic acid, 4-Diazesuberane-1-base) Benzoylamide.
1H NMR (400MHz, MeOD) δ 7.96 (d, J=8.9Hz, 2H), 7.84 (d, J=8.5Hz, 1H), 7.28 (s, 1H), 6.92 (d, J=8.6Hz, 1H), 6.89 (s, 1H), 6.85 (d, J=9.0Hz, 2H), 3.98 (s, 6H), 3.74 3.66 (m, 2H), 3.62 (t, J=6.2Hz, 2H), 2.83 2.76 (m, 2H), 2.66 2.60 (m, 2H), 2.40 (s, 3H), 2.11 2.03(m,2H).
Embodiment 29:
N-(6-(2,6-bis-chloro-3,5-Dimethoxyphenyl)-1-methyl isophthalic acid H-pyrazolo [3,4-b] pyridin-3-yl)-4-(4-methyl piperazine Piperazine-1-base) Benzoylamide (I-29)
Step 1: preparation 6-(2,6-bis-chloro-3,5-Dimethoxyphenyl)-1-methyl isophthalic acid H-pyrazolo [3,4-b] pyridine-3-amine
339 milligrams of 6-(2,6-bis-chloro-3,5-Dimethoxyphenyl)-1H-pyrazolo [3,4-b] pyridine-3-amine is dissolved in 1.5 milliliters of nothings In water DMF, add 40 milligrams of sodium hydrides at 0 DEG C, stir 30 minutes under argon shield；Add 64 micro- Rise potassium iodide, recover to stirred overnight at room temperature.The cancellation that adds water is reacted, and is extracted with ethyl acetate, with saturated after organic facies merging Sodium-chloride water solution washs five times, and anhydrous sodium sulfate is dried, filtering and concentrating, residue column chromatography (dichloromethane: methanol=99:1 98:2, V/V) 281 milligrams of yellow solids of isolated, yield 79.6%.
1H NMR (400MHz, DMSO) δ 8.20 (d, J=8.0Hz, 1H), 7.02 (s, 1H), 6.87 (d, J=8.0Hz, 1H),5.76(s,2H),3.97(s,6H),3.72(s,3H).
Step 2: preparation N-(6-(2,6-bis-chloro-3,5-Dimethoxyphenyl)-1-methyl isophthalic acid H-pyrazolo [3,4-b] pyridine-3- Base)-4-(4-methylpiperazine-1-yl) Benzoylamide
Change 6-(2,6-bis-chloro-3,5-Dimethoxyphenyl)-1H-pyrazolo [3,4-b] pyridine-3-amine into 6-(the chloro-3,5-of 2,6-bis-bis- Methoxyphenyl)-1-methyl isophthalic acid H-pyrazolo [3,4-b] pyridine-3-amine, remaining needed raw material, reagent and preparation method are with implementing Step 1 in example 8, obtains white solid N-(6-(2,6-bis-chloro-3,5-Dimethoxyphenyl)-1-methyl isophthalic acid H-pyrazolo [3,4-b] pyrrole Pyridine-3-base)-4-(4-methylpiperazine-1-yl) Benzoylamide.
1H NMR (300MHz, DMSO) δ 10.94 (s, 1H), 8.43 (d, J=8.2Hz, 1H), 8.04 (d, J=8.2Hz, 2H),7.22–6.98(m,4H),4.07(s,3H),3.99(s,8H),3.10(s,4H),2.67(s,3H).
Embodiment 30:
N-(6-(2,6-bis-chloro-3,5-Dimethoxyphenyl)-1-methyl isophthalic acid H-pyrazolo [3,4-b] pyridin-3-yl)-4-(4-methyl isophthalic acid, 4- Diazesuberane-1-base) Benzoylamide (I-30)
Step 1: preparation N-(6-(2,6-bis-chloro-3,5-Dimethoxyphenyl)-1-methyl isophthalic acid H-pyrazolo [3,4-b] pyridine-3- Base)-4-(4-methyl isophthalic acid, 4-Diazesuberane-1-base) Benzoylamide
Change 6-(2,6-bis-chloro-3,5-Dimethoxyphenyl)-1H-pyrazolo [3,4-b] pyridine-3-amine into 6-(the chloro-3,5-of 2,6-bis-bis- Methoxyphenyl)-1-methyl isophthalic acid H-pyrazolo [3,4-b] pyridine-3-amine, remaining needed raw material, reagent and preparation method are with implementing Step 2 in example 21, obtains white solid N-(6-(2,6-bis-chloro-3,5-Dimethoxyphenyl)-1-methyl isophthalic acid H-pyrazolo [3,4-b] Pyridin-3-yl)-4-(4-methylpiperazine-1-yl) Benzoylamide.
1H NMR (300MHz, DMSO) δ 10.85 (s, 1H), 8.43 (d, J=8.3Hz, 1H), 8.03 (d, J=8.8Hz, 2H), 7.10 (d, J=8.3Hz, 1H), 7.06 (s, 1H), 6.88 (d, J=8.9Hz, 2H), 3.99 (s, 9H), 3.93 3.71 (m, 2H), 3.55 (t, J=6.1Hz, 2H), 3.25 2.98 (m, 2H), 2.78 (s, 3H), 2.43 2.09 (m, 2H).
Embodiment 31:
N-(6-(2,6-bis-chloro-3,5-Dimethoxyphenyl)-1H-pyrazolo [3,4-b] pyridin-3-yl)-3-(4-methylpiperazine-1-yl) Benzoylamide (I-31)
Step 1: preparation N-(6-(2,6-bis-chloro-3,5-Dimethoxyphenyl)-1H-pyrazolo [3,4-b] pyridin-3-yl)-3-(4-methyl Piperazine-1-base) Benzoylamide
Change 4-((3R, 5S)-3,5-lupetazin-1-base) ethyl benzoate into 3-(4-methylpiperazine-1-yl) essence of Niobe, Remaining needed raw material, reagent and preparation method, with the step 6 in embodiment 6, obtain white solid N-(6-(2,6-bis-chloro-3,5-bis- Methoxyphenyl)-1H-pyrazolo [3,4-b] pyridin-3-yl)-3-(4-methylpiperazine-1-yl) Benzoylamide.
1H NMR (300MHz, DMSO) δ 13.46 (s, 1H), 11.10 (s, 1H), 8.41 (d, J=8.4Hz, 1H), 7.68 (s, 1H), 7.53 (d, J=7.3Hz, 1H), 7.40 (t, J=7.8Hz, 1H), 7.22 (dd, J=7.9,1.8Hz, 1H), 7.09 (d, J=8.3Hz, 1H), 7.05 (s, 1H), 3.99 (s, 6H), 3.38 (s, 4H), 2.85 (s, 4H).
Embodiment 32:
The chloro-N-of 2-(6-(2,6-bis-chloro-3,5-Dimethoxyphenyl)-1H-pyrazolo [3,4-b] pyridin-3-yl)-4-(4-methyl piperazine -1-base) Benzoylamide (I-32)
Step 1: the preparation chloro-4-of 2-(4-methylpiperazine-1-yl) essence of Niobe
0.72 milliliter of 2-chloro-4-fluorophenyl carbamate is dissolved in 5 milliliters of dimethyl sulfoxides, add 0.67 milliliter of 1-methyl piperazine and 1.041 grams of potassium carbonate, are warming up to 120 DEG C under argon shield, stop heating after stirring 18 hours, add 25 milliliters of water dilutions Reactant liquor, is extracted with ethyl acetate, and organic facies is washed five times with saturated sodium-chloride water solution after merging, and anhydrous sodium sulfate is dried, Filtering and concentrating, residue column chromatography (dichloromethane: methanol=99:1 98:2, V/V) 379 milligrams of yellow oil of isolated, Yield 28.1%.
1H NMR(300MHz,CDCl3) δ 7.84 (d, J=8.9Hz, 1H), 6.87 (d, J=2.6Hz, 1H), 6.73 (dd, J=9.0,2.6Hz, 1H), 3.87 (s, 3H), 3.38 3.28 (m, 4H), 2.58 2.48 (m, 4H), 2.34 (s, 3H).
Step 2: the preparation chloro-N-of 2-(6-(2,6-bis-chloro-3,5-Dimethoxyphenyl)-1H-pyrazolo [3,4-b] pyridin-3-yl)-4-(4- Methylpiperazine-1-yl) Benzoylamide
Change 4-((3R, 5S)-3,5-lupetazin-1-base) ethyl benzoate into the chloro-4-of 2-(4-methylpiperazine-1-yl) benzoic acid Methyl ester, remaining needed raw material, reagent and preparation method, with the step 6 in embodiment 6, obtain white solid 2-chloro-N-(6-(2,6- Two chloro-3,5-Dimethoxyphenyls)-1H-pyrazolo [3,4-b] pyridin-3-yl)-4-(4-methylpiperazine-1-yl) Benzoylamide.
1H NMR (300MHz, DMSO) δ 13.40 (s, 1H), 10.95 (s, 1H), 8.44 (d, J=8.5Hz, 1H), 7.54 (d, J=8.9Hz, 1H), 7.10 (d, J=8.2Hz, 1H), 7.05 (s, 1H), 7.03 (s, 1H), 6.98 (d, J=8.9Hz, 1H), 3.98(s,6H),3.31–3.20(m,4H),2.48–2.39(m,4H),2.23(s,3H).
Embodiment 33:
The chloro-N-of 3-(6-(2,6-bis-chloro-3,5-Dimethoxyphenyl)-1H-pyrazolo [3,4-b] pyridin-3-yl)-4-(4-methyl piperazine -1-base) Benzoylamide (I-33)
Step 1: the preparation chloro-4-of 3-(4-methylpiperazine-1-yl) essence of Niobe
0.36 milliliter of 3-chloro-4-fluorophenyl carbamate is dissolved in 2.5 milliliters of dimethyl sulfoxides, adds 0.334 milliliter of 1-methyl piperazine With 520 milligrams of potassium carbonate, under argon shield, it is warming up to 120 DEG C, stops heating after stirring 14 hours, add 13 milliliters of water dilute Releasing reactant liquor, be extracted with ethyl acetate, organic facies is washed five times with saturated sodium-chloride water solution after merging, and anhydrous sodium sulfate is done Dry, filtering and concentrating, residue column chromatography (dichloromethane: methanol=99:1 97:3, V/V) 325 milligrams of yellow oils of isolated Shape thing, yield 48.2%.
1H NMR(300MHz,CDCl3) δ 8.02 (d, J=2.0Hz, 1H), 7.88 (dd, J=8.4,2.0Hz, 1H), 7.03 (d, J=8.5Hz, 1H), 3.89 (s, 3H), 3.28 3.09 (m, 4H), 2.72 2.53 (m, 4H), 2.37 (s, 3H).
Step 2: the preparation chloro-N-of 3-(6-(2,6-bis-chloro-3,5-Dimethoxyphenyl)-1H-pyrazolo [3,4-b] pyridin-3-yl)-4-(4- Methylpiperazine-1-yl) Benzoylamide
Change 4-((3R, 5S)-3,5-lupetazin-1-base) ethyl benzoate into the chloro-4-of 3-(4-methylpiperazine-1-yl) benzoic acid Methyl ester, remaining needed raw material, reagent and preparation method, with the step 6 in embodiment 6, obtain white solid 3-chloro-N-(6-(2,6- Two chloro-3,5-Dimethoxyphenyls)-1H-pyrazolo [3,4-b] pyridin-3-yl)-4-(4-methylpiperazine-1-yl) Benzoylamide.
1H NMR (300MHz, DMSO) δ 13.46 (s, 1H), 11.12 (s, 1H), 8.39 (d, J=8.3Hz, 1H), 8.17 (d, J=2.1Hz, 1H), 8.06 (dd, J=8.5,2.0Hz, 1H), 7.29 (d, J=8.5Hz, 1H), 7.09 (d, J=8.3Hz, 1H),7.05(s,1H),3.98(s,6H),3.23–3.08(m,4H),2.81–2.57(m,4H),2.36(s,3H).
Embodiment 34:
N-(6-(2,6-bis-chloro-3,5-Dimethoxyphenyl)-1H-pyrazolo [3,4-b] pyridin-3-yl)-2-methoxyl group-4-(4-methyl Piperazine-1-base) Benzoylamide (I-34)
Step 1: preparation 4-fluoro-2-methoxybenzoic acid methyl ester
1.702 grams of fluoro-2-methoxybenzoic acid of 4-are dissolved in 14 ml methanol, add 0.5 milliliter of sulphuric acid, be warming up to backflow, Stopping heating after stirring 23 hours, reactant liquor concentrates and is evaporated, and residue is dissolved in ethyl acetate, molten with saturated sodium bicarbonate water Liquid is washed till bubble-free and produces, then washs with saturated sodium-chloride water solution, and anhydrous sodium sulfate is dried, filtering and concentrating, residue post Chromatography (dichloromethane: methanol=97:3, V/V) 1.805 grams of colorless oil of isolated, yield 98.0%.
1H NMR(300MHz,CDCl3)δ7.91–7.81(m,1H),6.74–6.62(m,2H),3.90(s,3H), 3.87(s,3H).
Step 2: preparation 2-methoxyl group-4-(4-methylpiperazine-1-yl) essence of Niobe
921 milligrams of 4-fluoro-2-methoxybenzoic acid methyl ester are dissolved in 5 milliliters of dimethyl sulfoxides, add 0.67 milliliter of 1-methyl piperazine With 1.040 grams of potassium carbonate, under argon shield, it is warming up to 120 DEG C, stops heating after stirring 18 hours, add 25 milliliters of water dilute Releasing reactant liquor, be extracted with ethyl acetate, organic facies is washed five times with saturated sodium-chloride water solution after merging, and anhydrous sodium sulfate is done Dry, filtering and concentrating, residue column chromatography (dichloromethane: methanol=99:1 98:2, V/V) 783 milligrams of yellow oils of isolated Shape thing, yield 59.2%.
1H NMR(300MHz,CDCl3) δ 7.80 (d, J=8.8Hz, 1H), 6.45 (dd, J=8.9,2.0Hz, 1H), 6.37 (d, J=1.8Hz, 1H), 3.89 (s, 3H), 3.83 (s, 3H), 3.38 3.28 (m, 4H), 2.59 2.50 (m, 4H), 2.35(s,3H).
Step 3: preparation N-(6-(2,6-bis-chloro-3,5-Dimethoxyphenyl)-1H-pyrazolo [3,4-b] pyridin-3-yl)-2-methoxyl group -4-(4-methylpiperazine-1-yl) Benzoylamide
Change 4-((3R, 5S)-3,5-lupetazin-1-base) ethyl benzoate into 2-methoxyl group-4-(4-methylpiperazine-1-yl) benzene Methyl formate, remaining needed raw material, reagent and preparation method, with the step 6 in embodiment 6, obtain white solid N-(6-(2,6- Two chloro-3,5-Dimethoxyphenyls)-1H-pyrazolo [3,4-b] pyridin-3-yl)-2-methoxyl group-4-(4-methylpiperazine-1-yl) benzoyl Amine.
1H NMR (300MHz, DMSO) δ 13.36 (s, 1H), 10.28 (s, 1H), 8.58 (d, J=8.4Hz, 1H), 7.90 (d, J=8.8Hz, 1H), 7.08 (d, J=8.4Hz, 1H), 7.05 (s, 1H), 6.69 (dd, J=9.1,1.7Hz, 1H), 6.64 (s, 1H),4.05(s,3H),3.98(s,6H),3.48–3.34(m,4H),2.27(s,3H).
Embodiment 35:
N-(6-(2,6-bis-chloro-3,5-Dimethoxyphenyl)-1H-pyrazolo [3,4-b] pyridin-3-yl)-3-methoxyl group-4-(4-methyl Piperazine-1-base) Benzoylamide (I-35)
Step 1: preparation 4-fluoro-3-methoxyl methyl benzoate
1.702 grams of 4-fluoro-3-methoxybenzoic acids are dissolved in 14 ml methanol, add 0.15 milliliter of sulphuric acid, be warming up to backflow, Stopping heating after stirring 17 hours, reactant liquor concentrates and is evaporated, and residue is dissolved in ethyl acetate, molten with saturated sodium bicarbonate water Liquid is washed till bubble-free and produces, then washs with saturated sodium-chloride water solution, and anhydrous sodium sulfate is dried, and filtering and concentrating is evaporated, vacuum It is dried, obtains 1.772 grams of yellow oil, yield 96.2%.
1H NMR(300MHz,CDCl3)δ7.71–7.60(m,2H),7.17–7.07(m,1H),3.94(s,3H), 3.91(s,3H).
Step 2: preparation 3-methoxyl group-4-(4-methylpiperazine-1-yl) essence of Niobe
921 milligrams of 4-fluoro-3-methoxyl methyl benzoates are dissolved in 5 milliliters of dimethyl sulfoxides, add 0.67 milliliter of 1-methyl piperazine With 1.037 grams of potassium carbonate, under argon shield, it is warming up to 120 DEG C, stops heating after stirring 22 hours, add 25 milliliters of water dilute Releasing reactant liquor, be extracted with ethyl acetate, organic facies is washed five times with saturated sodium-chloride water solution after merging, and anhydrous sodium sulfate is done Dry, filtering and concentrating, residue column chromatography (dichloromethane: methanol=99:1 97:3, V/V) 253 milligrams of yellow oils of isolated Shape thing, yield 19.1%.
1H NMR(300MHz,CDCl3) δ 7.63 (dd, J=8.2,1.7Hz, 1H), 7.51 (d, J=1.6Hz, 1H), 6.91 (d, J=8.2Hz, 1H), 3.91 (s, 3H), 3.88 (s, 3H), 3.28 3.08 (m, 4H), 2.69 2.55 (m, 4H), 2.36(s,3H).
Step 3: preparation N-(6-(2,6-bis-chloro-3,5-Dimethoxyphenyl)-1H-pyrazolo [3,4-b] pyridin-3-yl)-3-methoxyl group -4-(4-methylpiperazine-1-yl) Benzoylamide
Change 4-((3R, 5S)-3,5-lupetazin-1-base) ethyl benzoate into 3-methoxyl group-4-(4-methylpiperazine-1-yl) benzene Methyl formate, remaining needed raw material, reagent and preparation method, with the step 6 in embodiment 6, obtain white solid N-(6-(2,6- Two chloro-3,5-Dimethoxyphenyls)-1H-pyrazolo [3,4-b] pyridin-3-yl)-3-methoxyl group-4-(4-methylpiperazine-1-yl) benzoyl Amine.
1H NMR (300MHz, DMSO) δ 13.44 (s, 1H), 11.04 (s, 1H), 8.40 (d, J=8.2Hz, 1H), 7.79 –7.65(m,2H),7.15–6.97(m,3H),3.99(s,6H),3.90(s,3H),3.28–3.06(m,4H),3.00–2.68 (m,4H).
Embodiment 36:
N-(6-(2,6-bis-chloro-3,5-Dimethoxyphenyl)-1H-pyrazolo [3,4-b] pyridin-3-yl)-6-(4-methylpiperazine-1-yl) Nicotiamide (I-36)
Step 1: preparation 6-(4-methylpiperazine-1-yl) methyl nicotinate
858 milligrams of 6-chlorine apellagrin methyl ester are dissolved in 13 milliliters of DMFs, add 0.67 milliliter of 1-methyl piperazine With 5 milliliters of DIPEAs, under argon shield, it is warming up to 80 DEG C, stops heating after stirring 9 hours, add 65 Milliliter water dilute reaction solution, is extracted with ethyl acetate, and organic facies is washed five times with saturated sodium-chloride water solution after merging, anhydrous Sodium sulfate is dried, filtering and concentrating, residue column chromatography (dichloromethane: methanol=99:1 97:3, V/V) isolated 1.063 Gram crocus solid, yield 90.4%.
1H NMR(300MHz,CDCl3) δ 8.79 (d, J=1.7Hz, 1H), 8.01 (dd, J=9.0,1.6Hz, 1H), 6.59 (d, J=9.0Hz, 1H), 3.86 (s, 3H), 3.74 3.66 (m, 4H), 2.54 2.45 (m, 4H), 2.34 (s, 3H).
Step 2: preparation N-(6-(2,6-bis-chloro-3,5-Dimethoxyphenyl)-1H-pyrazolo [3,4-b] pyridin-3-yl)-6-(4-methyl Piperazine-1-base) nicotiamide
Change 4-((3R, 5S)-3,5-lupetazin-1-base) ethyl benzoate into 6-(4-methylpiperazine-1-yl) methyl nicotinate, its Remaining needed raw material, reagent and preparation method, with the step 6 in embodiment 6, obtain buff white solid N-(6-(2,6-bis-chloro-3,5-bis- Methoxyphenyl)-1H-pyrazolo [3,4-b] pyridin-3-yl)-6-(4-methylpiperazine-1-yl) nicotiamide.
1H NMR (300MHz, DMSO) δ 13.42 (s, 1H), 10.98 (s, 1H), 8.88 (d, J=2.4Hz, 1H), 8.40 (d, J=8.3Hz, 1H), 8.25 (dd, J=9.0,2.4Hz, 1H), 7.08 (d, J=8.3Hz, 1H), 7.05 (s, 1H), 7.02 (d, J=9.1Hz, 1H), 3.98 (s, 6H), 3.84 (s, 4H), 2.94 (s, 4H), 2.58 (s, 3H).
Embodiment 37:
N-(6-(2,6-bis-chloro-3,5-Dimethoxyphenyl)-1H-pyrazolo [3,4-b] pyridin-3-yl)-5-(4-methylpiperazine-1-yl) Ascorbyl palmitate (I-37)
Step 1: preparation 5-(4-methylpiperazine-1-yl) pyridine carbonitrile
281 milligrams of sodium hydrides are dissolved in 14 milliliters of anhydrous DMFs, are added dropwise over 0.72 milliliter of 1-methyl piperazine Piperazine, is stirred at room temperature 30 minutes under argon shield；It is added dropwise over the N,N-dimethylformamide solution (611 of 5-fluorine pyridine carbonitrile Milligram 5-fluorine pyridine carbonitrile is dissolved in 4.5 milliliters of DMFs), it is stirred at room temperature 24 hours.The cancellation that adds water is reacted, and uses Ethyl acetate extracts, and organic facies is washed five times with saturated sodium-chloride water solution after merging, and anhydrous sodium sulfate is dried, filtering and concentrating, Residue column chromatography (dichloromethane: methanol=99:1 97:3, V/V) 589 milligrams of light yellow solids of isolated, yield 58.2%.
1H NMR(300MHz,CDCl3) δ 8.31 (d, J=2.9Hz, 1H), 7.51 (dd, J=8.8,0.5Hz, 1H), 7.08 (dd, J=8.8,3.0Hz, 1H), 3.43 3.33 (m, 4H), 2.60 2.51 (m, 4H), 2.36 (s, 3H).
Step 2: preparation N-(6-(2,6-bis-chloro-3,5-Dimethoxyphenyl)-1H-pyrazolo [3,4-b] pyridin-3-yl)-5-(4-methyl Piperazine-1-base) ascorbyl palmitate
Change 4-((3R, 5S)-3,5-lupetazin-1-base) ethyl benzoate into 5-(4-methylpiperazine-1-yl) pyridine carbonitrile, its Remaining needed raw material, reagent and preparation method with the step 6 in embodiment 6, obtain white solid N-(6-(and 2,6-bis-chloro-3,5-diformazan Phenyl)-1H-pyrazolo [3,4-b] pyridin-3-yl)-5-(4-methylpiperazine-1-yl) ascorbyl palmitate.
1H NMR (300MHz, DMSO) δ 13.48 (s, 1H), 10.64 (s, 1H), 8.56 (d, J=8.3Hz, 1H), 8.46 (d, J=1.7Hz, 1H), 8.04 (d, J=8.8Hz, 1H), 7.55 (dd, J=8.7,2.3Hz, 1H), 7.11 (d, J=8.3Hz, 1H),7.05(s,1H),3.98(s,6H),3.56(s,4H),2.87(s,4H).
Embodiment 38:
N-(6-(2,6-bis-chloro-3,5-Dimethoxyphenyl)-1H-pyrazolo [3,4-b] pyridin-3-yl)-5-(4-methylpiperazine-1-yl) Pyrazine-2-Methanamide (I-38)
Step 1: preparation 5-(4-methylpiperazine-1-yl) pyrazine-2-methyl formate
863 milligrams of 5-chloropyrazine-2-methyl formates are dissolved in 13 milliliters of DMFs, add 0.67 milliliter of 1-first Base piperazine and 5 milliliters of DIPEAs, be warming up to 80 DEG C under argon shield, stops heating after stirring 2 hours, Adding 65 milliliters of water dilute reaction solutions, be extracted with ethyl acetate, organic facies is washed five times with saturated sodium-chloride water solution after merging, Anhydrous sodium sulfate is dried, filtering and concentrating, residue column chromatography (dichloromethane: methanol=99:1 97:3, V/V) isolated 1.024 grammeter yellow solids, yield 86.7%.
1H NMR(300MHz,CDCl3) δ 8.78 (d, J=1.3Hz, 1H), 8.12 (d, J=1.3Hz, 1H), 3.95 (s, 3H),3.81–3.72(m,4H),2.56–2.47(m,4H),2.35(s,3H).
Step 2: preparation N-(6-(2,6-bis-chloro-3,5-Dimethoxyphenyl)-1H-pyrazolo [3,4-b] pyridin-3-yl)-5-(4-methyl Piperazine-1-base) pyrazine-2-Methanamide
Change 4-((3R, 5S)-3,5-lupetazin-1-base) ethyl benzoate into 5-(4-methylpiperazine-1-yl) pyrazine-2-formic acid Methyl ester, remaining needed raw material, reagent and preparation method, with the step 6 in embodiment 6, obtain white solid N-(6-(2,6-dichloros -3,5-Dimethoxyphenyl)-1H-pyrazolo [3,4-b] pyridin-3-yl)-5-(4-methylpiperazine-1-yl) pyrazine-2-Methanamide.
1H NMR (300MHz, DMSO) δ 13.47 (s, 1H), 10.56 (s, 1H), 8.79 (d, J=1.1Hz, 1H), 8.51 (d, J=8.3Hz, 1H), 8.42 (d, J=0.7Hz, 1H), 7.10 (d, J=8.3Hz, 1H), 7.05 (s, 1H), 3.98 (s, 6H), 3.78(s,4H),2.29(s,3H).
Embodiment 39:
N-(6-(2,6-bis-chloro-3,5-Dimethoxyphenyl)-1H-pyrazolo [3,4-b] pyridin-3-yl)-5-(4-methylpiperazine-1-yl) Thiophene-2-carboxamide derivatives (I-39)
Step 1: preparation 5-(4-methylpiperazine-1-yl) thiophene-2-carboxylic acid ethyl ester
0.75 milliliter of 5-bromothiophene-2-Ethyl formate is dissolved in 50 milliliters of dry toluenes, is sequentially added into 0.67 milliliter of 1-methyl piperazine Piperazine, 312 milligrams (±)-2,2 '-bis-(diphenylphosphine)-1,1 '-dinaphthalene, 2.281 grams of cesium carbonates and 113 milligrams of palladium, argon protect Protecting down and be warming up to 110 DEG C, stop heating, reacting liquid filtering after stirring 18 hours, with ethyl acetate filter wash cake, filtrate concentrates It is evaporated, residue column chromatography (dichloromethane: methanol=99:1 98:2, V/V) 710 milligrams of yellow-brown solid of isolated, receives Rate 55.7%.
1H NMR(300MHz,CDCl3) δ 7.54 (d, J=4.3Hz, 1H), 6.01 (d, J=4.3Hz, 1H), 4.27 (q, J=7.1Hz, 2H), 3.34 3.18 (m, 4H), 2.61 2.49 (m, 4H), 2.34 (s, 3H), 1.32 (t, J=7.1Hz, 3H).
Step 2: preparation N-(6-(2,6-bis-chloro-3,5-Dimethoxyphenyl)-1H-pyrazolo [3,4-b] pyridin-3-yl)-5-(4-methyl Piperazine-1-base) thiophene-2-carboxamide derivatives
Change 4-((3R, 5S)-3,5-lupetazin-1-base) ethyl benzoate into 5-(4-methylpiperazine-1-yl) thiophene-2-carboxylic acid Ethyl ester, remaining needed raw material, reagent and preparation method, with the step 6 in embodiment 6, obtain yellow solid N-(6-(2,6-dichloros -3,5-Dimethoxyphenyl)-1H-pyrazolo [3,4-b] pyridin-3-yl)-5-(4-methylpiperazine-1-yl) thiophene-2-carboxamide derivatives.
1H NMR (300MHz, DMSO) δ 13.39 (s, 1H), 10.92 (s, 1H), 8.39 (d, J=8.2Hz, 1H), 7.96 (d, J=4.0Hz, 1H), 7.06 (d, J=9.4Hz, 2H), 6.36 (d, J=3.8Hz, 1H), 3.98 (s, 6H), 3.49 (s, 4H), 3.20(s,4H),2.72(s,3H).
Embodiment 40:
N-(6-(2,6-bis-fluoro-3,5-Dimethoxyphenyl)-1H-pyrazolo [3,4-b] pyridin-3-yl)-4-(4-methylpiperazine-1-yl) Benzoylamide (I-40)
Step 1: the preparation chloro-6-of 2-(2,6-bis-fluoro-3,5-Dimethoxyphenyl) nicotinic acid nitrile
The 550 milligrams of chloro-6-of 2-(3,5-Dimethoxyphenyl) nicotinic acid nitriles are dissolved in 20 milliliters of anhydrous acetonitriles, add 1.420 grams of 1-chlorine Methyl-4-fluoro-Isosorbide-5-Nitrae-diazabicyclo [2.2.2] octane two (Tetrafluoroboric acid) salt, is stirred at room temperature 24 hours under argon shield, instead Answer liquid to concentrate to be evaporated, residue column chromatography (petroleum ether: ethyl acetate=93:7 90:10, V/V) 91 milligrams of yellow of isolated Solid, yield 14.6%.
1H NMR(300MHz,CDCl3) δ 8.09 (d, J=8.0Hz, 1H), 7.56 (dt, J=7.9,1.3Hz, 1H), 6.77 (t, J=8.0Hz, 1H), 3.92 (s, 6H).
Step 2: preparation 6-(2,6-bis-fluoro-3,5-Dimethoxyphenyl)-1H-pyrazolo [3,4-b] pyridine-3-amine
91 milligrams of chloro-6-of 2-(2,6-bis-fluoro-3,5-Dimethoxyphenyl) nicotinic acid nitrile is dissolved in 3 milliliters of ethanol, adds 335 microliters of water Closing hydrazine (85%, w/w), be warming up to 70 DEG C, stop heating after stirring 18 hours, reactant liquor concentrates and is evaporated, residue post layer Analysis (dichloromethane: methanol=99:1 97:3, V/V) 46 milligrams of yellow-brown solid of isolated, yield 51.8%.
1H NMR (300MHz, DMSO) δ 12.09 (s, 1H), 8.22 (d, J=8.0Hz, 1H), 7.16 7.03 (m, 2H),5.67(s,2H),3.91(s,6H).
Step 3: preparation N-(6-(2,6-bis-fluoro-3,5-Dimethoxyphenyl)-1H-pyrazolo [3,4-b] pyridin-3-yl)-4-(4-methyl Piperazine-1-base) Benzoylamide
Change 6-(2,6-bis-chloro-3,5-Dimethoxyphenyl)-1H-pyrazolo [3,4-b] pyridine-3-amine into 6-(the fluoro-3,5-of 2,6-bis-bis- Methoxyphenyl)-1H-pyrazolo [3,4-b] pyridine-3-amine, remaining needed raw material, reagent and preparation method are with in embodiment 8 Step 1, obtains beige solid N-(6-(2,6-bis-fluoro-3,5-Dimethoxyphenyl)-1H-pyrazolo [3,4-b] pyridin-3-yl)-4-(4- Methylpiperazine-1-yl) Benzoylamide.
1H NMR (300MHz, DMSO) δ 13.49 (s, 1H), 10.92 (s, 1H), 8.41 (d, J=8.3Hz, 1H), 8.06 (d, J=8.6Hz, 2H), 7.29 (d, J=8.4Hz, 1H), 7.22 7.05 (m, 3H), 4.16 3.99 (m, 2H), 3.93 (s, 6H),3.64–3.41(m,2H),3.29–2.99(m,4H),2.81(s,3H).
Embodiment 41:
N-(6-(2,6-bis-chloro-3,5-Dimethoxyphenyl)-1H-pyrazolo [3,4-b] pyridin-3-yl)-2-methyl-4-(4-methyl piperazine -1-base) Benzoylamide (I-41)
Step 1: preparation 2-methyl-4-(4-methylpiperazine-1-yl) essence of Niobe
0.44 milliliter of 4-fluoro-2-methylbenzene methyl formate is dissolved in 1.5 milliliters of dimethyl sulfoxides, adds 0.40 milliliter of 1-methyl Piperazine and 495 milligrams of potassium carbonate, be warming up to 100 DEG C under argon shield, stops heating, add 7.5 after stirring 23 hours Milliliter water dilute reaction solution, is extracted with ethyl acetate, and organic facies is washed five times with saturated sodium-chloride water solution after merging, anhydrous Sodium sulfate is dried, filtering and concentrating, residue column chromatography (dichloromethane: methanol=99:1 97:3, V/V) isolated 342 milligrams of yellow oil, yield 46.2%.
1H NMR(300MHz,CDCl3) δ 7.88 (d, J=8.5Hz, 1H), 6.70 (d, J=9.2Hz, 2H), 3.83 (s, 3H),3.39–3.26(m,4H),2.63–2.50(m,7H),2.35(s,3H).
Step 2: preparation N-(6-(2,6-bis-chloro-3,5-Dimethoxyphenyl)-1H-pyrazolo [3,4-b] pyridin-3-yl)-2-methyl -4-(4-methylpiperazine-1-yl) Benzoylamide
Change 4-((3R, 5S)-3,5-lupetazin-1-base) ethyl benzoate into 2-methyl-4-(4-methylpiperazine-1-yl) benzene first Acid methyl ester, remaining needed raw material, reagent and preparation method, with the step 6 in embodiment 6, obtain white solid N-(6-(2,6- Two chloro-3,5-Dimethoxyphenyls)-1H-pyrazolo [3,4-b] pyridin-3-yl)-2-methyl-4-(4-methylpiperazine-1-yl) benzoyl Amine.
1H NMR (300MHz, DMSO) δ 13.35 (s, 1H), 10.75 (s, 1H), 8.42 (d, J=8.3Hz, 1H), 7.58 (d, J=8.2Hz, 1H), 7.09 (d, J=8.4Hz, 1H), 7.05 (s, 1H), 6.88 (d, J=9.7Hz, 2H), 3.98 (s, 6H), 3.41(s,4H),2.89(s,4H),2.53(s,3H),2.47(s,3H).
Embodiment 42:
The impact that FGFR1, FGFR2 enzyme is lived by compound molecule level
1, test method
By enzyme reaction substrate Poly (Glu, Tyr) 4:1 with without potassium ion PBS (10mM sodium phosphate buffer, 150mM NaCl, PH 7.2-7.4) it is diluted to 20 μ g/mL, 125 μ L/ hole coated elisa plates, put 37 DEG C and react 12-16 hour.Discard liquid in hole Wash plate after body, wash plate three times with the T-PBS (PBS containing 0.1%Tween-20) in 200 μ L/ holes, each 5 minutes.In 37 DEG C Baking oven is dried ELISA Plate 1-2 hour.
Every hole adds with reaction buffer (50mM HEPES pH 7.4,50mM MgCl2,0.5mM MnCl2,0.2mM Na3VO4, 1mM DTT) and the ATP solution 50 μ L that dilutes, final concentration 5 μMs.Compound DMSO is diluted to suitably Concentration, 1 μ L/ hole or or containing the DMSO (negative control hole) of respective concentration, add and dilute with 49 μ L reaction buffers FGFR1, FGFR2 kinases territory recombiant protein start reaction, experiment need to set without ATP control wells holes every time.Put 37 DEG C to shake Bed (100rpm) reacts 1 hour.T-PBS washes plate three times.Add an anti-PY99 diluent 100 μ L/ hole, 37 DEG C of shaking table reactions 0.5 hour.T-PBS washes plate three times.Add the IgG diluent 100 μ L/ hole of two anti-horseradish peroxidase-labeled sheep anti mouses, 37 DEG C of shaking tables react 0.5 hour.T-PBS washes plate three times.Add the OPD nitrite ion 100 μ L/ hole of 2mg/ml (with containing 0.03% H2O20.1M citric acid-sodium citrate buffer (pH=5.4) dilution), 25 DEG C of lucifuges are reacted 1-10 minute.(OPD dissolves Time need to ultrasonic, nitrite ion need now with the current).Add 2M H2SO450 μ L/ hole stopped reactions, use wavelengthtunable type micro-hole Plate microplate reader SPECTRA MAX 190 reading, wavelength is 490nm.
The suppression ratio of sample is tried to achieve by following equation:
IC50Value uses the random bundled software of microplate reader to return with four parametric methods and tries to achieve.
2, experimental result
Table 2. compound inhibitory activity alive to FGFR1 and FGFR2 enzyme
Compound in embodiment FGFR1 FGFR2
I-1 C B
I-2 C N/A
I-3 A B
I-4 N/A B
I-5 C B
I-6 A N/A
I-7 N/A A
I-8 A N/A
I-9 A N/A
I-10 A N/A
I-11 A N/A
I-12 A N/A
I-13 A N/A
I-14 A N/A
I-15 A A
I-16 A A
I-17 A A
I-18 A A
I-19 B N/A
I-20 A N/A
I-21 A N/A
I-22 B N/A
I-23 A N/A
I-24 A N/A
I-25 N/A A
I-26 A A
I-27 A N/A
I-28 A A
I-29 C C
I-30 C C
I-31 A A
I-32 A A
I-33 A N/A
I-34 B B
I-35 A A
I-36 A A
I-37 A A
I-38 A A
I-39 A A
I-40 A A
Wherein: A represents IC50Less than 10nM
B represents IC50Less than 100nM
C represents IC50More than 100nM
Embodiment 43:
The compound impact on BaF3/TEL-FGFR1 cell proliferation
1, test method
Compound to BaF3/TEL-FGFR1 cell, (in cell, stably express at endochylema by TEL-FGFR1 kinases district fusion protein In, FGFR1 sustained activation, for FGFR1 dependent cell strain) growth inhibited detection CCK-8 Cell counting Kit (Dojindo) detection.Specifically comprise the following steps that the BaF3/TEL-FGFR1 cell being in exponential phase is inoculated by proper density To 96 well culture plates, every hole 90 μ L, after overnight incubation, add compound effects 72hr of variable concentrations, and set molten Agent matched group (negative control).After compound effects cell 72h, the impact of compound on intracellular propagation uses CCK-8 thin Born of the same parents' counting reagent kit (Dojindo) detects, and every hole adds 10 μ L CCK-8 reagent, is placed in 37 DEG C of incubators placement 2-4 hour After, declining orifice plate microplate reader SpectraMax 190 reading by all-wave length, mensuration wavelength is 450nm.Use with following equation The computerized compound suppression ratio (%) to growth of tumour cell: suppression ratio (%)=(OD negative control hole-OD dosing holes)/OD Negative control hole × 100%.IC50 value uses the random bundled software of microplate reader to return with four parametric methods and tries to achieve.
2, experimental result
Table 3. compound is to BaF3/TEL-FGFR1 cell inhibitory effect activity
Compound in embodiment Activity
I-4 D
I-7 A
I-25 D
Wherein: A represents IC50Less than 8nM
B represents IC50Less than 40nM
C represents IC50Less than 200nM
D represents IC50More than 200nM
Embodiment 44:
The compound impact on KG1 cell proliferation
1, test method
Compound is to acute marrow source leukemia cell line KG1 cell (FGFR1OP2-FGFR1 kinases district fusion protein in cell Stable expressing in endochylema, FGFR1 sustained activation, for FGFR1 dependent cell strain) growth inhibited detection CCK-8 thin Born of the same parents' counting reagent kit (Dojindo) detects.Specifically comprise the following steps that the KG1 cell being in exponential phase is inoculated by proper density To 96 well culture plates, every hole 90 μ L, after overnight incubation, add compound effects 72hr of variable concentrations, and set molten Agent matched group (negative control).After compound effects cell 72h, the impact of compound on intracellular propagation uses CCK-8 thin Born of the same parents' counting reagent kit (Dojindo) detects, and every hole adds 10 μ L CCK-8 reagent, is placed in 37 DEG C of incubators placement 2-4 hour After, declining orifice plate microplate reader SpectraMax 190 reading by all-wave length, mensuration wavelength is 450nm.Use with following equation The computerized compound suppression ratio (%) to growth of tumour cell: suppression ratio (%)=(OD negative control hole-OD dosing holes)/OD Negative control hole × 100%.IC50 value uses the random bundled software of microplate reader to return with four parametric methods and tries to achieve.
2, experimental result
Table 4. compound is to KG1 cell inhibitory effect activity
Compound in embodiment Activity
I-6 A
I-8 A
I-9 A
I-10 A
I-11 B
I-12 A
I-13 A
I-14 A
I-15 A
I-16 A
I-17 B
I-18 C
I-19 D
I-20 B
I-21 A
I-22 D
I-23 B
I-24 A
I-27 C
Wherein: A represents IC50Less than 8nM
B represents IC50Less than 40nM
C represents IC50Less than 200nM
D represents IC50More than 200nM
Embodiment 45:
The impact that non-small cell lung cancer cell NCI-H1581 Nude Mice is grown by compound
The tumor tissue taking growth animated period cuts into 1.5mm3Left and right, aseptically, is inoculated on the right side of nude mouse Axillary fossa is subcutaneous.Nude mouse subcutaneous transplantation tumor vernier caliper measurement transplanted tumor diameter, treats that tumor growth to average external volume is 120mm3Left and right is by animal random packet.I-15 100mg/kg and 12.5mg/kg group, positive control medicine AZD4547 12.5mg/kg group, equal once a day oral administration, successive administration three weeks.Solvent control group then gives normal saline. In whole experimentation, measure transplanted tumor diameter 2 times a week, weigh Mouse Weight simultaneously.Gross tumor volume growth inhibition Rate TGI%, computing formula is as follows: TGI%=[1-(TVt-TV0)/(CVt-CV0)] × 100%, TVt is that treatment group is each The tumor volume measured；TV0Gained tumor volume when being administered for therapeutic component cage；CVt is the tumor body that matched group is measured every time Long-pending；CV0For gained tumor volume during matched group sub-cage administration.Experimental result (t student ' s test vs solvent as shown in Figure 1 Matched group, * * p < 0.01, * * * p < 0.001).Result shows, I-15 100mg/kg group tumor growth inhibition rate (TGI) It is 83.5%, and 12.5mg/kg group tumor growth inhibition rate (TGI) is 67.7%.
The all documents mentioned in the present invention are incorporated as reference the most in this application, just as each document coverlet Solely it is incorporated as with reference to like that.In addition, it is to be understood that after the above-mentioned teachings having read the present invention, this area skill The present invention can be made various changes or modifications by art personnel, and these equivalent form of values fall within the application claims equally Book limited range.

Claims (10)

1. the compound as shown in following formula (I) or its pharmaceutical salts, its prodrug, its hydrate or a solvate,
Wherein:
Y is selected from CH or N；
R1Selected from substituted or unsubstituted 6-10 unit aryl, substituted or unsubstituted miscellaneous former selected from N, O and S containing 1-3 The 5-10 unit heteroaryl of son；Described substituent group be one or more be selected from halogen, C1～C6 alkyl, C1～C6 alkoxyl, C1～C6 alkylthio group, halo C1～C6 alkyl, halo C1～C6 alkoxyl, cyano group, C1～C6 alkanoyl, In-NH-CO-C1～C6 alkyl, amino ,-NH (C1～C6 alkyl) ,-N (C1～C6 alkyl) (C1～C6 alkyl) and hydroxyl Substituent group；
R2Selected from hydrogen, C1～C6 alkyl, C1～C6 alkanoyl ,-CO (O)-C1～C6 alkyl；
R3Selected from substituted or unsubstituted 6-10 unit aryl, substituted or unsubstituted miscellaneous selected from N, O and S containing 1-3 The 5-10 unit heteroaryl of atom；Described substituent group be one or more be selected from halogen, C1～C6 alkyl, C1～C6 alkoxyl, C1～C6 alkylthio group, cyano group, hydroxyl, halo C1～C6 alkyl, halo C1～C6 alkoxyl, C1～C6 alkanoyl, -NH-CO-C1～C6 alkyl, amino ,-C1～C6 alkyl-NH2,-NH (C1～C6 alkyl) ,-N (C1～C6 alkane Base) (C1～C6 alkyl) ,-CO (O)-C1～C6 alkyl, C3-C8 heterocyclic radical ,-Y1-(C1～C6 alkyl)-Y2, C1～C6 Substituent group in alkyl-C3～C8 heterocyclic radical and C3～C8 cycloalkyl-C3～C8 heterocyclic radical, described Y1 is selected from O, NH and S； Described Y2 is selected from C1～C6 alkoxyl, amino ,-NH (C1～C6 alkyl) and-N (C1～C6 alkyl) (C1～C6 alkyl)； Wherein said C3-C8 heterocyclic radical contains 1-3 the hetero atom selected from N, O and S, and optionally by one or more Selected from C1～C6 alkyl, C1～C6 alkoxyl, C3～C8 cycloalkyl, C2～C6 thiazolinyl, C2～C6 alkynyl, C1～C6 Alkanoyl, C1～C6 alkoxyl substituted C1～C6 alkyl, C1～C6 alkylamino substituted C1～C6 alkyl, hydroxyl take C1～the C6 alkyl in generation ,-CO (O)-C1～C6 alkyl, hydroxyl, halogen ,-SO2Substituent group in-C1～C6 alkyl is entered One step replaces；
R4、R5It is each independently selected from hydrogen, halogen, C1～C6 alkyl, C2～C6 thiazolinyl, C2～C6 alkynyl, halo C1～C6 alkyl, C1～C6 alkoxyl, halo C1～C6 alkoxyl, C1～C6 alkylthio group, cyano group, C1～C6 alkane acyl Base, nitro, amino ,-NH (C1～C6 alkyl) ,-N (C1～C6 alkyl) (C1～C6 alkyl) ,-NH-CO-C1～C6 Alkyl and-CO (O)-C1～C6 alkyl；
And when Y is CH, R3At least there is less than one substituent group: C3-C8 heterocyclic radical；Wherein, described C3-C8 Heterocyclic radical as defined above.
2. formula (I) compound as claimed in claim 1 or its pharmaceutical salts, its prodrug, its hydrate or solvate, It is characterized in that,
R1Selected from substituted or unsubstituted 6-10 unit aryl, substituted or unsubstituted miscellaneous former selected from N, O and S containing 1-3 The 5-10 unit heteroaryl of son；Described substituent group be one or more be selected from halogen, C1～C4 alkyl, C1～C4 alkoxyl, C1～C4 alkylthio group, halo C1～C4 alkyl, halo C1～C4 alkoxyl, cyano group, C1～C4 alkanoyl, In-NH-CO-C1～C4 alkyl, amino ,-NH (C1～C4 alkyl) ,-N (C1～C4 alkyl) (C1～C4 alkyl) and hydroxyl Substituent group；
R2Selected from hydrogen, C1～C4 alkyl, C1～C4 alkanoyl ,-CO (O)-C1～C4 alkyl；
R3Selected from substituted or unsubstituted 6-10 unit aryl, substituted or unsubstituted miscellaneous selected from N, O and S containing 1-3 The 5-10 unit heteroaryl of atom；Described substituent group be one or more be selected from halogen, C1～C4 alkyl, C1～C4 alkoxyl, C1～C4 alkylthio group, cyano group, hydroxyl, halo C1～C4 alkyl, halo C1～C4 alkoxyl, C1～C4 alkanoyl, -NH-CO-C1～C4 alkyl, amino ,-C1～C4 alkyl-NH2,-NH (C1～C4 alkyl) ,-N (C1～C4 alkane Base) (C1～C4 alkyl) ,-CO (O)-C1～C4 alkyl, C3-C8 heterocyclic radical ,-Y1-(C1～C4 alkyl)-Y2, C1～C4 Substituent group in alkyl-C3～C8 heterocyclic radical and C3～C8 cycloalkyl-C3～C8 heterocyclic radical, described Y1 is selected from O, NH and S； Described Y2 is selected from C1～C4 alkoxyl, amino ,-NH (C1～C4 alkyl) and-N (C1～C4 alkyl) (C1～C4 alkyl)； Wherein said C3-C8 heterocyclic radical contains 1-3 the hetero atom selected from N, O and S, and optionally by one or more Selected from C1～C4 alkyl, C1～C4 alkoxyl, C3～C8 cycloalkyl, C2～C4 thiazolinyl, C2～C4 alkynyl, C1～C4 Alkanoyl, C1～C4 alkoxyl substituted C1～C4 alkyl, C1～C4 alkylamino substituted C1～C4 alkyl, hydroxyl take C1～the C4 alkyl in generation ,-CO (O)-C1～C4 alkyl, hydroxyl, halogen ,-SO2Substituent group in-C1～C4 alkyl is entered One step replaces；
R4、R5It is each independently selected from hydrogen, halogen, C1～C4 alkyl, C2～C4 thiazolinyl, C2～C4 alkynyl, halo C1～C4 alkyl, C1～C4 alkoxyl, halo C1～C4 alkoxyl, C1～C4 alkylthio group, cyano group, C1～C4 alkane acyl Base, nitro, amino ,-NH (C1～C4 alkyl) ,-N (C1～C4 alkyl) (C1～C4 alkyl) ,-NH-CO-C1～C4 Alkyl and-CO (O)-C1～C4 alkyl；
And when Y is CH, R3At least there is less than one substituent group: C3-C8 heterocyclic radical；Wherein, described C3-C8 Heterocyclic radical as defined above.
3. formula (I) compound as claimed in claim 2 or its pharmaceutical salts, its prodrug, its hydrate or solvate, It is characterized in that,
R1Selected from substituted or unsubstituted following aryl or heteroaryl: phenyl, naphthyl, pyridine radicals, pyrazinyl, furan Mutter base, thienyl, pyrrole radicals, oxazolyl, thiazolyl, pyrazolyl, imidazole radicals, indyl, benzofuranyl, Benzothiazolyl and benzimidazolyl；Described substituent group is that one or more are selected from halogen, C1～C4 alkyl, C1～C4 Substituent group in alkoxyl and-N (C1～C4 alkyl) (C1～C4 alkyl)；
R3In replacement or unsubstituted following aryl or heteroaryl: phenyl, naphthyl, pyridine radicals, pyrazinyl, thiophene Base, furyl, imidazole radicals, pyrrole radicals, oxazolyl, thiazolyl, pyrazolyl, indyl, benzofuranyl, benzene Benzothiazolyl, benzimidazolyl；Described substituent group is that one or more are selected from halogen, C1～C4 alkyl, C1～C4 alkane Epoxide, amino ,-NH (C1～C4 alkyl) ,-N (C1～C4 alkyl) (C1～C4 alkyl), C3-C8 heterocyclic radical and Substituent group in-Y1-(C1～C4 alkyl)-Y2, described Y1 is selected from O, NH and S；Described Y2 selected from C1～C4 alkoxyl, Amino ,-NH (C1～C4 alkyl) and-N (C1～C4 alkyl) (C1～C4 alkyl)；Wherein said C3-C8 heterocyclic radical contains 1-3 the hetero atom selected from N, O and S, and optionally it is selected from C1～C4 alkyl, C1～C4 alcoxyl by one or more Base, C3～C8 cycloalkyl, C2～C4 thiazolinyl, C2～C4 alkynyl, C1～C4 alkanoyl, C1～C4 alkoxyl are substituted C1～C4 alkyl, hydroxyl substituted C1～C4 alkyl ,-CO (O)-C1～C4 alkyl and-SO2Replacement in-C1～C4 alkyl Base is further substituted with；
R4、R5It is each independently selected from hydrogen, halogen, C1～C4 alkyl and halo C1～C4 alkyl；
And when Y is CH, R3At least there is less than one substituent group: C3-C8 heterocyclic radical；Wherein, described C3-C8 Heterocyclic radical as defined above.
4. according to formula (I) compound according to any one of claim 1-3 or its pharmaceutical salts, its prodrug, its hydrate or Solvate, it is characterised in that described formula (I) compound is selected from lower group:
5. a pharmaceutical composition, it is characterised in that including: the institute as any one of claim 1-4 of therapeutically effective amount The compound stated, or its pharmaceutical salts, its prodrug, its hydrate or solvate, and optional pharmaceutically acceptable carrier.
6. the compound as according to any one of claim 1-4, or its pharmaceutical salts, its prodrug, its hydrate or solvent The purposes of compound, it is characterised in that for preparing prevention and/or the treatment medicine selected from the disease of lower group: (a) tumor is correlated with Disease, preferably cancer；(b) protein tyrosine kinase activity relevant disease, preferably FGFR activity related diseases, more It is preferably FGFR1 and/or FGFR2 relevant disease.
7. purposes as claimed in claim 6, it is characterised in that described tumor-related illness is selected from lower group: breast carcinoma, Pulmonary carcinoma, bladder cancer, gastric cancer, cancer of pancreas, carcinoma of prostate, colon cancer, multiple myeloma AML, hepatocarcinoma, melanin Tumor, head and neck cancer, thyroid carcinoma, renal cell carcinoma, glioblast cancer and carcinoma of testis；It is preferably breast carcinoma, non-small cell lung Cancer, bladder cancer, gastric cancer, cancer of pancreas, carcinoma of prostate, colon cancer, multiple myeloma, hepatocarcinoma, melanoma, head and neck Cancer, thyroid carcinoma, renal cell carcinoma, glioblast cancer and carcinoma of testis；More preferably nonsmall-cell lung cancer, gastric cancer, multiple Property myeloma.
8. a protein tyrosine kinase is lived inhibitor, it is characterised in that containing suppression effective dose such as claim 1-4 Described compound, or its pharmaceutical salts, its prodrug, its hydrate or solvate.
9. one kind is used for treating cancer or the pharmaceutical composition of protein tyrosine kinase activity relevant disease, it is characterised in that Described pharmaceutical composition includes the compound as described in arbitrary in claim 1-4 of therapeutically effective amount, or its pharmaceutical salts, its Prodrug, its hydrate or solvate are as active component.
10. the preparation method of formula (I) compound as claimed in claim 1, it is characterised in that include step (a) or (b):
A () compound (II) or its salt prepare compound (I) in the basic conditions with corresponding acetyl halide compound (III)；Or
B () compound (II) or its salt prepare compound (I) with corresponding ester (III) under lewis acid effect；
Wherein, Q is leaving group, it is therefore preferable to halogen (acyl chlorides) or C2-C6 ester group (ester)；The definition of remaining each group is such as The above.
CN201510192164.3A 2015-04-21 2015-04-21 Pyrazolo [3,4-b] pyridines and the preparation method of indazole compounds and purposes Pending CN106146493A (en)

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Cited By (4)

* Cited by examiner, † Cited by third party
Publication number Priority date Publication date Assignee Title
CN112521336A (en) * 2020-02-21 2021-03-19 温州医科大学 Indazole and pyrrolopyridine compounds and application thereof
WO2022015975A1 (en) * 2020-07-15 2022-01-20 Ifm Due, Inc. Compounds and compositions for treating conditions associated with sting activity
CN114008026A (en) * 2019-04-17 2022-02-01 罗达制药生物技术有限责任公司 Benzopyrazoles and pyridopyrazoles as protein kinase inhibitors
WO2022162034A1 (en) 2021-01-29 2022-08-04 F. Hoffmann-La Roche Ag Pyrazoleamide derivatives

Cited By (4)

* Cited by examiner, † Cited by third party
Publication number Priority date Publication date Assignee Title
CN114008026A (en) * 2019-04-17 2022-02-01 罗达制药生物技术有限责任公司 Benzopyrazoles and pyridopyrazoles as protein kinase inhibitors
CN112521336A (en) * 2020-02-21 2021-03-19 温州医科大学 Indazole and pyrrolopyridine compounds and application thereof
WO2022015975A1 (en) * 2020-07-15 2022-01-20 Ifm Due, Inc. Compounds and compositions for treating conditions associated with sting activity
WO2022162034A1 (en) 2021-01-29 2022-08-04 F. Hoffmann-La Roche Ag Pyrazoleamide derivatives

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Application publication date: 20161123