CN103467481A

CN103467481A - Dihydropyridine compounds, compositions thereof, preparation method and applications

Info

Publication number: CN103467481A
Application number: CN2012101871072A
Authority: CN
Inventors: 程建军; 秦继红; 叶斌
Original assignee: SHANGHAI HUILUN TECHNOLOGY Co Ltd
Current assignee: Shanghai Huilun Jiangsu Pharmaceutical Co ltd; Shanghai Huilun Pharmaceutical Co ltd
Priority date: 2012-06-07
Filing date: 2012-06-07
Publication date: 2013-12-25
Anticipated expiration: 2032-06-07
Also published as: CN103467481B

Abstract

Disclosed dihydropyridine compounds are the compounds possessing the following general formula (I) as shown in the specification, wherein R1 is selected from hydrogen, halogen, alkyl, substituted alkyl, alkoxy, substituted alkoxy, alkylamino, substituted alkylamino, aryl, substituted aryl, heteroaryl, substituted heteroaryl, heterocyclic groups, substituted heterocyclic groups, ester group and amide group; R2 is selected from hydrogen, cyanogroup, alkyl, substituted alky, alkenyl, substituted alkenyl and acyl; or R2 and R3 form a fused ring; R3 is selected from alkyl, substituted alkyl, aryl, substituted aryl, heteroaryl and substituted heteroaryl; or R3 and R2 or R4 form a fused ring; R4 is selected from hydrogen, alkyl and substituted alkyl; or R4 and R3 or R5 form a fused ring; R5 is selected form alkyl, substituted alky, aryl, substituted aryl, heteroaryl and substituted heteroaryl; or R5 and R4 form a fused ring. The invention also discloses a preparation method of the compounds of the general formula (I), compositions containing the compounds of the general formula (I), and applications of the compositions to medicaments for treating cancers.

Description

Dihydropyridine compounds, its composition, preparation method and purposes

Technical field

The present invention relates to a kind of dihydropyridine compounds, its preparation method, contain their pharmaceutical compositions as activeconstituents, usings and as the purposes of medicine relevant cancer in order to the treatment disease, particularly c-Met relevant to Tyrosylprotein kinase c-Met.

Background of invention

Cancer is the No.1 killer who threatens whole world human life's health.Although the progress of medical science makes the mankind have a lot of new means for the treatment of cancer, cancer still is considered to still unsolved difficult medical problem at present.Cause the cause of disease of cancer a lot, in recent years, the development of the subjects such as molecular weight tumor, molecular pharmacology is progressively being illustrated the essence of tumour, and people recognize that the essence of cell carcinogenesis is the cell infinite multiplication that the intracellular signal transduction pathway imbalance causes gradually.

As the of paramount importance member who participates in cell signaling, protein tyrosine kinase (protein tyrosine kinases, PTKs are called for short Tyrosylprotein kinase) is modal growth factor receptors, with the generation of tumour with develop closely related.

The hyperactivity of Tyrosylprotein kinase, cause its downstream signal pathway activation, thus cause cell transformation, propagation, to anti-apoptotic, promote cell survival, finally lead oncogenic formation.Therefore, the Development Trend of antitumor drug starts to turn to the medicine for abnormal signal transduction in cell from traditional cell toxicity medicament in recent years, and there have successively related drugs to be applied to be clinical.With traditional cell toxicant series antineoplastic medicament, compare, this quasi-molecule targeting medicament curative effect is strong, toxic side effect is little, becomes gradually the focus of current antitumor drug research and development.

Tyrosylprotein kinase divides two kinds of receptor type tyrosine kinase and non-receptor type Tyrosylprotein kinases.

Receptor type tyrosine kinase has EGF-R ELISA (EGFR) family, vascular endothelial growth factor receptor (VEGFR) family, platelet derived growth factor receptor (PDGFR) family, fibroblast growth factor acceptor (FGFR) family etc.

Nonreceptor tyrosine kinase is Src kinases family, Jak for example, and FAK etc. comprise again multiple hypotype in each kinases family.

Hepatic growth factor receptor c-Met is a kind of (Park et al., Proc.Natl.Acad.Sci.USA 84:6379-83,1987 of receptor type tyrosine kinase; Bottaro et al., Science 2S 1:802-4,1991), by outside alpha subunit and the β subunit of high glycosylation, together with extracellular domain, transmembrane segment and tenuigenin tyrosine kinase domain, formed.Its endogenic ligand is pHGF (hepatocyte growth factor, HGF) (Nature, 327:239-242(1987); J.Cell Biol., 111:2097-2108(1990)), ligand binding is induced the c-Met dimerization, generate the activated receptor of autophosphorylation, promote the signal transduction in downstream, multiple the replying of mediation in tumour cell, comprise that increment, stimulation epithelial cell mobility, cell survival and the form of epithelial cell and endotheliocyte changes and promote intrusion etc.In addition, HGF regulates vasculogenesis, for the growth of tumour with spread extremely important.The effect during its evolution in these tumours also has been described of excessively expressing in kinds of tumors (comprising thyroid carcinoma, ovarian cancer, carcinoma of the pancreas etc.) of c-Met and part thereof.At present, c-Met is as the action target spot of antitumor drug, its advantage progressively illustrated (Nature Reviews Cancer, 2012,12,89-103).

During the primary tumor played a key effect at the c-Met receptor activation and secondary tumors shift, the biological substance of target HGF or c-Met (ribozyme, antibody and sense-rna) can suppress the generation of tumour, and the selectivity micromolecular inhibitor of target c-Met is also predicted has treatment potentiality.WO2009091374, WO2009149836, WO2011003604, WO2011042367, WO2011042368, CN200910247948.6, all comprised optionally c-Met micromolecular inhibitor, Preparation Method And The Use in the patents such as CN201010175273.1.

Dihydropyridine compounds as tyrosine kinase inhibitor involved in the present invention, as tyrosine kinase inhibitor, particularly c-Met inhibitor, never be in the news.

Summary of the invention

One of technical problem to be solved by this invention is to provide a kind of dihydropyridine compounds.

Two of technical problem to be solved by this invention is to provide the preparation method of above-mentioned dihydropyridine compounds.

Three of technical problem to be solved by this invention is to provide the pharmaceutical composition that contains above-mentioned dihydropyridine compounds.

Four of technical problem to be solved by this invention is to provide the purposes of above-mentioned dihydropyridine compounds.

As the dihydropyridine compounds of first aspect present invention, for thering is the compound of following general formula (I):

Wherein,

R ₁be selected from alkyl oxy, alkyl amine group, the alkyl amine group of replacement, aryl, substituted aryl, heteroaryl, substituted heteroaryl, heterocyclic radical, substituted heterocyclic radical, ester group, the amide group of hydrogen, halogen, alkyl, substituted alkyl, alkyl oxy, replacement;

R ₂be selected from hydrogen, cyano group, alkyl, substituted alkyl, thiazolinyl, substituted alkenyl, acyl group; Perhaps, R ₂with R ₃form and encircle;

R ₃be selected from alkyl, substituted alkyl, aryl, substituted aryl, heteroaryl, substituted heteroaryl; Perhaps, R ₃with R ₂or R ₄form and encircle;

R ₄be selected from hydrogen, alkyl, substituted alkyl; Perhaps, R ⁴with R ³or R ⁵form and encircle;

R ₅be selected from alkyl, substituted alkyl, aryl, substituted aryl, heteroaryl, substituted heteroaryl; Perhaps, R ₅with R ₄form and encircle.

More specifically, preferred a kind of in the compound shown in following (I-1) to (I-46) of general formula of the present invention (I) compound:

Described general formula (I) compound is any one or both or three's the mixture arbitrarily in enantiomer, diastereomer, conformer.

Described general formula (I) compound is the pharmacy acceptable derivates.

General formula of the present invention (I) compound can exist with the form of pharmacy acceptable salt.

Hydrochloride, vitriol, phosphoric acid salt, acetate, trifluoroacetate, mesylate, fluoroform sulphonate, tosilate, tartrate, maleate, fumarate, succinate or malate that pharmacy acceptable salt of the present invention is general formula (I) compound.

As the preparation method of general formula (I) compound of second aspect present invention, the intermediate that can be (II) by corresponding structural formula is prepared by the reaction formula of following route 1,

route 1:

Wherein, R ₁, R ₂, R ₃, R ₄definition is with above-mentioned; PG is the amido protecting group.PG is preferably tertbutyloxycarbonyl, benzyloxycarbonyl, to methoxy-benzyl, 3, the 4-dimethoxy-benzyl.

Particularly, R in intermediate (II) ₁during for H, its preparation method is as shown in Scheme 2: the 5-thiotolene of structural formula 1-2-formic acid through nitrosonitric acid nitrated the intermediate of structural formula 2, the intermediate that is structural formula 3 by the intermediate carboxylic esterification of structural formula 2, adopting reduced iron powder is the amino intermediate that obtains structural formula 4 by the nitroreduction of the intermediate of structural formula 3, the intermediate that is then structural formula 5 by the intermediate cyclisation of structural formula 4 under the existence of acetic anhydride, nitrite; The intermediate ethanoyl of structural formula 5 is removed to obtain to the intermediate of structural formula 6, then the ester group in the intermediate of structural formula 6 is reduced to the intermediate that alcohol radical obtains structural formula 7, then the intermediate oxidation of structural formula 7 is obtained to the intermediate that structural formula is (II);

route 2:

R in intermediate (II) ₁while being not H, its preparation method is as shown in Scheme 3: the 5-thiotolene of structural formula 1-2-formic acid through nitrosonitric acid nitrated the intermediate of structural formula 2, the intermediate that is structural formula 3 by the intermediate carboxylic esterification of structural formula 2, adopting reduced iron powder is the amino intermediate that obtains structural formula 4 by the nitroreduction of the intermediate of structural formula 3, the intermediate that is then structural formula 5 by the intermediate cyclisation of structural formula 4 under the existence of acetic anhydride, nitrite; The intermediate ethanoyl of structural formula 5 is removed to obtain to the intermediate of structural formula 6, by the intermediate of structural formula 66 iodos, obtain the intermediate of structural formula 8; The pyrazoles amino of the intermediate of structural formula 8, with the protection of amido protecting group, is obtained to the intermediate of structural formula 9; Then utilize the iodine of the intermediate of structural formula 9 to carry out the intermediate that coupling obtains structural formula 10; Again the ester group of the intermediate of structural formula 10 is reduced to the intermediate that alcohol radical obtains structural formula 11, the alcohol radical in the intermediate of structural formula 11 is oxidized to aldehyde radical, obtain the intermediate that structural formula is (II);

route 3:

Described coupling is selected from Suzuki coupling, Buchwald coupling.

When the compound of required preparation is the compound shown in general formula (Ia), and R '=R ₃=R ₅the time, its preparation method as shown in Scheme 4:

route 4:

When the compound of required preparation is the compound shown in general formula (Ib), and R ₃with R ₅when different, its preparation method as shown in Scheme 5:

route 5:

When the compound of required preparation is the compound shown in general formula (Ic), R ₃with R ₅difference and R ₄while being not H, its preparation method as shown in Scheme 6:

route 6:

When the compound of required preparation is the compound shown in general formula (Id), its preparation method as shown in Scheme 7:

route 7:

Wherein X is CH ₂or O.

When the compound of required preparation is the compound shown in general formula (Ie), its preparation method as shown in Scheme 8:

route 8:

When the compound of required preparation is the compound shown in general formula (If), its preparation method as shown in Scheme 9:

route 9:

Wherein X is CH ₂or O.

As the pharmaceutical composition containing dihydropyridine compounds of third aspect present invention, wherein said pharmaceutical composition comprises general formula (I) compound and the acceptable vehicle of pharmacy for the treatment of significant quantity.

As a kind of pharmaceutical composition of third aspect present invention, pharmacy acceptable derivates and the acceptable vehicle of pharmacy that wherein said pharmaceutical composition comprises general formula (I) compound for the treatment of significant quantity.

As a kind of pharmaceutical composition of third aspect present invention, pharmacy acceptable salt and the acceptable vehicle of pharmacy that wherein said pharmaceutical composition comprises general formula (I) compound for the treatment of significant quantity.

Described pharmaceutical composition is made tablet, capsule, aqueous suspension, oiliness suspensoid, dispersible pulvis, granule, lozenge, emulsion, syrup, ointment, ointment, suppository or injection.

As the application of fourth aspect present invention, be wherein that general formula (I) compound is regulated the application in protein kinase catalytic activity goods in preparation.

As the application of fourth aspect present invention, be wherein that the pharmacy acceptable derivates of general formula (I) compound is regulated the application in protein kinase catalytic activity goods in preparation.

As the application of fourth aspect present invention, be wherein that the pharmaceutically useful salt of general formula (I) compound is regulated the application in protein kinase catalytic activity goods in preparation.

As the application of fourth aspect present invention, be wherein the application of pharmaceutical composition in the medicine for preparing the treatment disease relevant with protein kinase.

Described protein kinase is the c-Met receptor tyrosine kinase.

The relevant disease of described protein kinase is cancer.

Described cancer is thyroid carcinoma, colorectal carcinoma, cancer of the stomach, kidney, liver cancer, lung cancer, ovarian cancer, mammary cancer, prostate cancer, bladder cancer, head and neck cancer, carcinoma of the pancreas, carcinoma of gallbladder, osteosarcoma, rhabdosarcoma, MFH/ fibrosarcoma, glioblastoma/astrocytoma, melanoma or mesothelioma.

The dihydropyridine compounds of general formula involved in the present invention (I) also can be used for the research of the signal transduction pathway that the research, Tyrosylprotein kinase of biology or pharmacology phenomenon participates in and for the comparative evaluation of new tyrosine kinase inhibitor.

Embodiment

The invention provides general formula defined above (I) compound, prepare these compounds method, use the pharmaceutical composition of these compounds and use the method for these compounds.

Below listed be the definition to the various terms for describing the compounds of this invention.These definition are applied to the term (unless in specific situation, restriction separately being arranged) in each place use of specification sheets, and no matter these terms are used separately the more part of macoradical of still conduct.

Unless otherwise defined, term as used herein " alkyl " (use separately or as the part of another group) refers to the univalent perssad that comprises 1 to 12 carbon atom that alkane is derivative.Preferred alkyl has 1 to 6 carbon atom.Alkyl is optional straight chain, side chain or the cyclic saturated hydrocarbon base replaced.Exemplary alkyl comprises methyl, ethyl, propyl group, sec.-propyl, normal-butyl, the tertiary butyl, isobutyl-, amyl group, hexyl, isohexyl, heptyl, 4,4-dimethyl amyl group, octyl group, 2,2,4-tri-methyl-amyl, nonyl, decyl, undecyl, dodecyl etc.

The substituting group of " substituted alkyl " is selected from following group: alkyl, halogen (as fluorine, chlorine, bromine, iodine), alkoxyl group, amino/amido, haloalkyl (as trichloromethyl, trifluoromethyl), aryl, aryloxy, alkylthio, hydroxyl, cyano group, nitro, carboxyl, alkoxy carbonyl, alkyl-carbonyl oxygen base, carbamyl, urea or sulfydryl.

Term as used herein " aryl " (use separately or as the part of another group) refers to monocyclic aromatic ring or polycyclic aromatic ring, such as the phenyl of phenyl, replacement etc. and group such as the naphthyl condensed, phenanthryl etc.Thereby aryl comprises at least one ring with at least 6 atoms, comprise five such rings (wherein comprising 22 atoms at the most) at the most, and there are (conjugation) two keys alternately between adjacent carbon atom or suitable heteroatoms.Preferred aryl comprises 6 to 14 carbon atoms in ring.

" substituted aryl " can optionally replace has one or more group, described group to include but not limited to halogen (such as fluorine, chlorine, bromine), alkyl (such as methyl, ethyl, propyl group), substituted alkyl (as trifluoromethyl), cycloalkyl, alkoxyl group (such as methoxy or ethoxy), hydroxyl, carboxyl, amine formyl (C (=O) NR'R "), alkoxy carbonyl (CO ₂r), amino/amido, nitro, cyano group, thiazolinyl oxygen base, aryl, heteroaryl, alkylsulfonyl (SO ₂r) etc., wherein, R, R', R " are described alkyl.

Term as used herein " heteroaryl " (use separately or as the part of another group) refers to replace and unsubstituted aromatics 5 or 6 yuan of monocyclic groups, 9 or 10 yuan of bicyclic radicals and 11 to 14 yuan of three cyclic groups, and these groups have at least one heteroatoms (O, S or N) at least one encircles.Each ring that comprises heteroatomic heteroaryl can comprise one or two Sauerstoffatoms or sulphur atom and/or one to four nitrogen-atoms, condition is four or still less for heteroatomic sum in each ring, and each ring has at least one carbon atom, the ring condensed that forms above-mentioned bicyclic radicals and three cyclic groups can only comprise carbon atom, and can be saturated or fractional saturation.Nitrogen-atoms and sulphur atom can be oxidations, and nitrogen-atoms can be quaternary ammoniated.The heteroaryl of dicyclo or three rings must comprise the ring of at least one Wholly aromatic, but other ring condensed or a plurality of ring can be aromatics or non-aromatic.Heteroaryl can be in any available nitrogen-atoms or the connection of carbon atom place of ring arbitrarily.

" substituted heteroaryl " ring system can comprise zero, one, two or three be selected from following substituting group: the alkyl of halogen, alkyl, replacement, thiazolinyl, piece base, aryl, nitro, cyano group, hydroxyl, alkoxyl group, alkylthio ,-CO ₂h ,-C (=O) H ,-CO ₂-alkyl ,-cycloalkyl of C (=O) alkyl, phenyl, benzyl, phenylethyl, phenyl oxygen base, thiophenyl, cycloalkyl, replacement, Heterocyclylalkyl, heteroaryl ,-NR'R " ,-C (=O) NR'R " ,-CO ₂nR'R " ,-C (=O) NR'R " ,-NR'CO ₂r " ,-NR'C (=O) R " ,-SO ₂nR'R " and-NR'SO ₂r ", wherein R' and R " independently is selected from alkyl and the cycloalkyl of hydrogen, alkyl, replacement separately, or R' and R " together with form Heterocyclylalkyl or heteroaryl ring.

The example of bicyclic heteroaryl comprises pyrryl, pyrazolyl, pyrazolinyl, imidazolyl, oxazolyl, di azoly, isoxazolyl, thiazolyl, thiadiazolyl group, isothiazolyl, furyl, thienyl, oxadiazoles base, pyridyl, pyrazinyl, pyrimidyl, pyridazinyl, triazinyl etc.

The example of bicyclic heteroaryl comprises indyl, benzothiazolyl, benzodioxole base, benzoxazolyl, benzothienyl, quinolyl, tetrahydric quinoline group, isoquinolyl, tetrahydro isoquinolyl, benzimidazolyl-, benzopyranyl, indolizine base, benzofuryl, chromone base, tonka bean camphor base, benzofuryl, quinoxalinyl, indazolyl, pyrrolopyridinyl, furo pyridyl etc.

The example of tricyclic heteroaryl comprises carbazyl, benzindole base, phenanthroline base, acridyl, phenanthridinyl etc.

The heteroatoms that a carbon atom in term as used herein " heterocycle " (use separately or as the part of another group) finger ring is selected from O, S or N replaces and 3 cycloalkyl (non-aromatic) that extra carbon atom can be replaced by described heteroatoms at the most.The term that the application uses " heterocyclic radical " (use separately or as the part of another group) refers to comprise the undersaturated monocycle ring system of stable saturated or part of 5 to 7 annular atomses (carbon atom and be selected from other atom of nitrogen, sulphur and/or oxygen).Heterocycle can be 5,6 or 7 yuan of monocycles, and comprises one, two or three heteroatoms that is selected from nitrogen, oxygen and/or sulphur.Heterocycle can be optional the replacement; this means that heterocycle can replace and have one or more independently to be selected from following group at one or more commutable ring position: alkyl, Heterocyclylalkyl, heteroaryl, alkoxyl group, nitro, monoalkyl amido, dialkyl amino, cyano group, halogen, haloalkyl, alkyloyl, ammonia/amido carbonyl, monoalkyl amido carbonyl, dialkyl amino carbonyl, alkylamidoalkyl, alkoxyalkyl, alkoxy carbonyl, alkyl-carbonyl oxygen base and aryl, the optional replacement of described aryl has halogen, alkyl and alkoxyl group.The example of these Heterocyclylalkyls includes but not limited to: piperidines, morpholine, high morpholine, piperazine, parathiazan, tetramethyleneimine and azetidine.

Term as used herein " alkoxyl group " (use separately or as the part of another group) refers to the alkyl that preferably has 1 to 6 carbon atom connected by Sauerstoffatom, such as-OR, wherein R is described alkyl.

Term as used herein " amino " (use separately or as the part of another group) refers to-NH ₂." amido " can optionally replace one or two substituting groups (NR'R "); wherein R' and R " can be identical or different, such as alkyl, aryl, arylalkyl, thiazolinyl, alkynyl, heteroaryl, heteroarylalkyl, Heterocyclylalkyl, alkyl, Heterocyclylalkyl alkyl, cycloalkyl, cycloalkylalkyl, haloalkyl, antelope base alkyl, alkoxyalkyl, alkylthio, carbonyl or carboxyl.These substituting groups can further replace any one having in carboxylic acid or the listed alkyl or aryl substituting group of the application.In some embodiments,, there are carboxyl or carbonyl amino the replacement, forms N-acyl group or N-carbamyl deriveding group.

The term that the application uses " haloalkyl " (use separately or as the part of another group) refers to the halogen atom connected by alkyl, such as-CF ₃.

Term as used herein " acyl group " (use separately or as the part of another group) refer to the alkyl that connects by carbonyl or-C (=O) R, wherein R is described alkyl.

The term that this text is used " alkoxy carbonyl " (use separately or as the part of another group) refers to-C (=O) OR, and wherein R is described alkyl.

Term as used herein " arylalkyl " or " aralkyl " (use separately or as the part of another group) refer to the aromatic ring (for example benzyl) connected by alkyl described above.

Term as used herein " aminoalkyl group " (use separately or as the part of another group) refers to the amino (NR'R ") connected by alkyl.

Term as used herein " aryl-alkyl amino " (use separately or as the part of another group) refers to the aryl connected by alkyl, and described alkyl connects by amino.

Term " heteroatoms " refers to independent O, S or the N selected.It should be noted, the ungratified any heteroatom of valency all is considered to be connected with hydrogen atom, thereby meets valency.

Term " halogen " refers to independent fluorine, chlorine, the bromine or iodine of selecting.

Term as used herein " cycloalkyl " (use separately or as the part of another group) refers to 3 to 9 carbon atoms, is preferably the hydrocarbon ring of the complete saturated or fractional saturation of 3 to 7 carbon atoms.In addition, cycloalkyl can replace." cycloalkyl of replacement " refer to have once, two or three be selected from following substituent ring: the alkyl of halogen, alkyl, replacement (wherein substituting group is defined as above alkyl substituent), thiazolinyl, alkynyl, nitro, cyano group, oxo (=O), hydroxyl, alkoxyl group, alkylthio ,-CO ₂h ,-C (=O) H ,-CO ₂-alkyl ,-C (=O) alkyl, ketone group ,=N-OH ,=N-O-alkyl, aryl, heteroaryl, five or hexa-atomic ketal (1,3-dioxane or 1,3-bis-uh alkane) ,-NR ' R " ,-C (=O) NR'R " ,-CO ₂nR'R " ,-C (=O) NR'R " ,-NR'CO ₂r " ,-NR'C (=O) R " ,-SO ₂nR'R " and-NR'SO ₂the alkyl and the cycloalkyl that independently are selected from separately hydrogen, alkyl, replacement in R ", wherein R' and R ", or R' and R ", form together Heterocyclylalkyl or heteroaryl ring.

Term " anticarcinogen " comprises any known medicine that can be used for treating cancer, comprising: (1) cytotoxic drug: chlormethine series pharmaceuticals, as melphalan, endoxan; Platinum coordination complex, such as cis-platinum, carboplatin and oxaliplatin; (2) anti-metabolism antitumour drug: 5 FU 5 fluorouracil, capecitabine, methotrexate, Calciumlevofolinate, Raltitrexed, purine antagonist (for example 6-thioguanine and Ismipur); (3) hormones: the female alcohol of 17 alpha-acetylenes, stilboestrol, testosterone, prednisone, Fluoxymesterone, dromostanolone propionate, testolactone, Magace, methylprednisolone, methyltestosterone, prednisolone, triamcinolone, Chlortrianisoestrol, hydroxyprogesterone, aminoglutethimide, estramustine, medroxyprogesterone acetate, toremifene; (4) tyrosine kinase inhibitor: the EGFR inhibitor comprises Gefitinib (Gefitinib), Erlotinib (Erlotinib), Cetuximab (Cetuximab), Trastuzumab (Herceptin) etc.; The VEGF inhibitor, such as VEGF antibody (Avastin (Avastin)) and micromolecular inhibitor such as Sunitinib, Vandetanib, Cediranib; The Bcr-Abl inhibitor is as imatinib (Imatinib), Dasatinib (Dasatinib); Src inhibitor, MEK kinase inhibitor, mapk kinase inhibitor, PI3K kinase inhibitor, c-Met inhibitor, ALK inhibitor etc.; (5) act on the medicine of tubulin, such as vinca medicine, taxanes medicine, epothilones medicine as ipsapirone (Ixabepilone) etc.; (6) topoisomerase I inhibitor, as topotecan, irinotecan; (7) histon deacetylase (HDAC) (HDAC) inhibitor is as Vorinostat (Vorinostat); (8) proteasome inhibitor is as Velcade (Bortezomib); (9) anticarcinogen of other classifications is as aurora kinase (aurora kinase) inhibitor, biological response modifier, growth inhibitor, glu famine antagonist, angiogenesis inhibitor and anti-vascular medicine, matrix metallo-proteinase inhibitor etc.

" Mammals " comprises the mankind and domestic animal, as cat, dog, pig, ox, sheep, goat, horse, rabbit etc.Preferably, for the purposes of the present invention, described Mammals is the mankind.

" optional () " or " optional () " mean that the environment event of describing subsequently may exist or not exist, and described description comprises the situation that described event or environment occur and situation about not occurring.For example, " optional substituted aryl " means that described aryl may be substituted or not be substituted and described description comprises aryl and the unsubstituted aryl of replacement.

When " pharmacy acceptable derivates " means to recipient's administration, can directly or indirectly provide salt, the acid amides of any nontoxic salt, ester, the ester of the active metabolite of compound of the present invention or its inhibition or resistates, salt or other derivatives of acid amides.

" the acceptable vehicle of pharmacy " includes but not limited to be can be used for by state food and Drug Administration's approval conduct any assistant agent, carrier, vehicle, glidant, sweeting agent, dispersion agent, thinner, sanitas, suspending agent, stablizer, dyestuff/tinting material, odorant, tensio-active agent, wetting agent, isotonic agent, solvent or the emulsifying agent of the mankind or domestic animal.

" pharmacologically acceptable salts " comprises acid salt and base addition salt.

" the acceptable acid salt of pharmacy " refers to such salt, and they have retained biological effect and the character of free alkali, can aspect biology or other, not produce adverse consequences, and be such as but not limited to hydrochloric acid with mineral acid, Hydrogen bromide, sulfuric acid, nitric acid, phosphoric acid etc., and organic acid is such as but not limited to following acid: formic acid, acetic acid, trifluoroacetic acid, methylsulfonic acid, trifluoromethanesulfonic acid, ethyl sulfonic acid, the 2-ethylenehydrinsulfonic acid, Phenylsulfonic acid, tosic acid, 2,2-dichloro acetic acid, hexanodioic acid, Lalgine, xitix, aspartic acid, phenylformic acid, paraacetaminobenzoic acid, dextrocamphoric acid, camphor-10-sulfonic acid, capric acid, caproic acid, sad, carbonic acid, styracin, citric acid, cyclohexane sulfamic acid, dodecyl sulphate, ethane-1, the 2-disulfonic acid, fumaric acid, tetrahydroxyadipic acid, gentisinic acid, glucoheptonic acid, glyconic acid, glucuronic acid, L-glutamic acid, pentanedioic acid, 2-oxo-pentanedioic acid, Phosphoric acid glycerol esters, oxyacetic acid, urobenzoic acid, isopropylformic acid, lactic acid, lactobionic acid, lauric acid, toxilic acid, oxysuccinic acid, propanedioic acid, amygdalic acid, glactaric acid, naphthalene-2-sulfonic acid, naphthalene-1, the 5-disulfonic acid, the 1-hydroxy-2-naphthoic acid, nicotinic acid, oleic acid, vitamin B13, oxalic acid, the palm fibre eleostearic acid, pamoic acid, propionic acid, Pyrrolidonecarboxylic acid, pyruvic acid, Whitfield's ointment, 4-ASA, sebacic acid, stearic acid, fumaric acid, succsinic acid, tartrate, thiocyanic acid, the formation such as undecylenic acid.

" the acceptable base addition salt of pharmacy " refers to such salt, and they have retained biological effect and the character of free acid, can be not improper aspect biology or other.These salt are by mineral alkali or organic bases being added on free acid and make.The salt that is derived from mineral alkali includes but not limited to sodium, potassium, lithium, ammonium, calcium, magnesium, iron, zinc, copper, manganese, aluminium salt etc.Preferred inorganic salt are ammonium, sodium, potassium, calcium and magnesium salts.The salt that is derived from organic bases includes but not limited to the salt of following substances: primary amine, secondary amine and tertiary amine, the amine replaced, comprise naturally occurring replacement amine, cyclammonium and deacidite, as ammonia, methylamine, dimethylamine, Trimethylamine 99, diethylamine, triethylamine, tripropyl amine, Isopropylamine, diethanolamine, thanomin, DMAE, 2-diethylaminoethanol, dicyclohexyl amine, Methionin, arginine, Histidine, caffeine, PROCAINE HCL, PHARMA GRADE, Hai Baming (hydrabamine), choline, trimethyl-glycine, Benethamine diacetale, quadrol, glycosamine, methylglucosamine, Theobromine, trolamine, Trometamol, purine, piperidines, piperazine, N-ethylpiperidine, versamid 900 etc.Preferred organic bases is Isopropylamine, diethylamine, thanomin, triethylamine, dicyclohexyl amine, choline and caffeine.

" pharmaceutical composition " refers to compound of the present invention and biologically active cpds is delivered to the preparation that medium formed of Mammals as the common acceptance in the mankind.Such medium comprises all pharmaceutically acceptable carriers, thinner or the vehicle to this.

" treatment significant quantity " refers to when to the Mammals administration (the preferably mankind), is enough to the amount of the compound of the present invention that relative disease or illness to Mammals (the preferably mankind) be achieved as follows civilian defined treatment.The amount that forms the compound of the present invention of " treatment significant quantity " can be according to for example activity of applied particular compound; The metabolic stability of described compound and effect duration; Patient's age, body weight, holistic health, sex and diet; Mode of administration and time; Discharge rate; Drug combination; The seriousness of specific illness or illness; And the individuality of experience treatment and changing, but it can be determined according to himself knowledge and the disclosure routinely by those of ordinary skills.

" treated " or " treatment " contained having the Mammals of relative disease or illness during for this paper, the preferably mankind's relative disease or the treatment of illness, and comprise:

(i) diseases or illness in the prevention Mammals, especially when such Mammals ill but also do not diagnose out when ill;

(ii) suppress disease or illness, stop its development;

(iii) alleviate disease or illness, cause that disease or illness disappear;

(iv) stable disease or illness.

During for this paper, term " disease " and " illness " can Alternates or can be different, reason is that specified disease or illness may not have known inducement (thereby also not working out the cause of disease), therefore also be not considered to disease and, only as improper situation or syndrome, wherein the clinician has identified concrete syndrome more or less.The compounds of this invention shown in this article and their structure also mean to comprise all isomer (for example enantiomer, diastereomer, rotamerism or conformational isomerism) form, they can according to for amino acid whose absolute stereo chemistry, be defined as (R)-/(S)-or (D)-/(L)-or (R, R)-/(R, S)-/(S, S)-.The present invention means the isomer that comprises that all these are possible, and their racemic, the enantiomorph enrichment and optional pure form.Optically-active (+) and (-), (R)-and (S)-and (R, R)-/(R, S)-/(S, S)-or (D)-and (L)-isomer can be used, and chirality is synthetic, the chiral separation preparation, or can use routine techniques to split such as but not limited to the high performance liquid phase (HPLC) that uses chiral column.When compound as herein described comprises the two keys of thiazolinyl or other how much asymmetric centers, except as otherwise noted, described compound comprises E and Z geometrical isomer.Equally, also comprise all tautomeric forms.

" steric isomer " refers to consist of with identical chemical bonding identical atom but compound with different three-dimensional structures, and they are not interchangeable.The present invention is contained various steric isomers and composition thereof and is comprised " enantiomer " and " diastereomer ", and enantiomer refers to two kinds of steric isomers of the mirror image that its molecule each other can not be overlapping; Diastereomer refers to that molecule has two or more chiral centres, and intermolecular be the steric isomer of non-mirror.

" tautomer " refers to that proton moves to another position of same a part from original position from an atom of molecule.The present invention includes the tautomer of any described compound.

In addition, except as otherwise noted, compound of the present invention also comprises that the structure difference only is to exist the compound of one or more isotopic enrichment atoms.For example, there is structure of the present invention, except with " deuterium " or " tritium ", replacing hydrogen, or use ¹⁸f-fluorine mark ( ¹⁸the F isotropic substance) replace fluorine, or use ¹¹c-, ¹³c-, or ¹⁴the carbon of C-enrichment ( ¹¹c-, ¹³c-, or ¹⁴the C-carbon markings; ¹¹c-, ¹³c-, or ¹⁴the C-isotropic substance) replace the compound of carbon atom in scope of the present invention.Such compound can be used as biological example and learns analysis tool or the probe in measuring, or can be used as the in-vivo diagnostic imaging tracer agent of disease, or the tracer agent of studying as pharmacodynamics, pharmacokinetics or acceptor.

The present invention also provides following methods: give by (I) compound of general formula as defined above and at least one other anticarcinogen combination that will treat significant quantity the patient that (while or priority) needs this treatment, via regulating the c-Met kinases, treat proliferative disease (such as cancer).In preferred embodiments, proliferative disease is cancer.

Particularly, general formula (I) compound can be used for treating kinds cancer, is specially those cancers that depend on the c-Met activation most.The c-Met activation can stimulate to regulate by gene amplification, sudden change (various mutations) and/or HGF, and wherein HGF is provided by tumour (autocrine) or host's (paracrine) tissue.Usually, compound of the present invention can be used for the treatment of to following cancer:

(A) solid tumor, comprise cancer of the stomach, lung cancer (comprising small cell lung cancer, nonsmall-cell lung cancer), colorectal carcinoma, kidney, liver cancer, mammary cancer, ovarian cancer, cervical cancer, esophagus cancer, carcinoma of gallbladder, bladder cancer, carcinoma of the pancreas, thyroid carcinoma, prostate cancer and skin carcinoma (comprising squamous cell carcinoma);

(B) the hematopoiesis tumour of lymph pedigree, comprise acute lymphoblastic leukemia (ALL), acute lymphoblast leukemia, B cell lymphoma, t cell lymphoma, Hodgkin lymphoma, non-Hodgkin′s woods bar knurl, hair cell lymphoma and Burkitt lymphoma (Burkett's lymphoma);

(C) the hematopoiesis tumour of marrow pedigree, comprise acute and chronic myelogenous leukemia, myelodysplastic syndrome and promyelocytic leukemia;

(D) tumour of mesenchyme origin, comprise fibrosarcoma and rhabdosarcoma;

(E) tumour of maincenter and peripheral nervous system, comprise astrocytoma, neuroblastoma, neurospongioma and schwannoma;

(F) other tumour, comprise melanoma, spermocytoma, teratocarcinoma, osteosarcoma, xeroderma pitmentosum, keratoacanthoma, follicular carcinoma of thyroid and Kaposi sarcoma.

General formula (I) compound also can be used for any lysis that treatment is characterized as abnormal cell proliferation, the restenosis, inflammatory bowel, graft-rejection, endotoxin shock and the fungi infestation that for example after benign prostatic hyperplasia, neurofibromatosis, atherosclerosis, pulmonary fibrosis, sacroiliitis, psoriasis, glomerulonephritis, angioplasty or vascular surgery, occur.

The adjustable ganglion cell RNA of general formula (I) compound and the synthetic level of DNA.Therefore, these materials can be used for the treatment of to virus infection (including but not limited to HIV, human papillomavirus, simplexvirus, poxvirus, Epstein-Barr virus, sindbis alphavirus and adenovirus).

General formula (I) compound can be used for the chemoprophylaxis of cancer.Chemoprophylaxis is defined as by the precellular progress that cancerates of blocking initial mutagenesis event or damaged by blocking-up carry out the development of anti-invasion cancer or suppress tumor recurrence.

General formula (I) compound can be used for suppressing tumor-blood-vessel growth and transfer.

Compound of the present invention also can be used in combination with known anticarcinogen (include but not limited to mention in above-mentioned " anticarcinogen " those) or anticancer therapy (such as radiotherapy) (together with give or successively give).

Usually can prepare according to following route 1 to route 9 by some general formula (I) compound.The tautomer of general formula (I) compound and solvate (for example hydrate, ethanol compound) are also within the scope of the invention.The preparation method of solvate is normally known in the art.Therefore, compound of the present invention can be free form or hydrate forms.In described method, the functional group of midbody compound may need to be protected by suitable protecting group hereinafter.Such functional group comprises hydroxyl, amino, sulfydryl and carboxylic acid.For the applicable protecting group of hydroxyl comprise trialkylsilkl or alkyl diaryl silyl (such as t-butyldimethylsilyl, t-butyldiphenylsilyl or trimethyl silyl), THP trtrahydropyranyl, benzyl, to methoxy-benzyl etc.For amino suitable protecting group comprise tertbutyloxycarbonyl, carbobenzoxy-(Cbz), ethanoyl, benzoyl, trifluoroacetyl group, to methoxy-benzyl etc.Suitable protecting group for carboxylic acid comprises alkyl, aryl or alkyl aryl.For heteroaryl such as the suitable protecting group of the NH functional group such as indoles or indazole ring comprise tertbutyloxycarbonyl, carbobenzoxy-(Cbz), ethanoyl, benzoyl, 2-TMS-ethoxyl methyl, to methoxy-benzyl etc.

Protecting group can according to method known to those skilled in the art, (Protective Groups in Organic Synthesis, 1999, the 3rd edition, Wiley) add with standard technique as herein described or remove for Greene, T.W..Described protecting group can be also that fluoropolymer resin is as Wang resin, Rink resin or 2-chlorine trityl chloride resin.Simultaneously, although the protected derivative of these of the compounds of this invention itself may not have pharmacological activity, they can be administered to Mammals, and then metabolism has the compounds of this invention of pharmacological activity with formation in vivo.Therefore such derivative is described to " prodrug ".All prodrugs of the compounds of this invention include within the scope of the invention.The dihydropyridine compounds of general formula of the present invention (I), can be prepared by the following method.

The preparation method of general formula (I) compound, the intermediate that can be (II) by corresponding structural formula is prepared by the reaction formula of following route 1,

route 1:

Wherein, R ₁, R ₂, R ₃, R ₄definition is with above-mentioned; PG is the amido protecting group.PG is preferably tertbutyloxycarbonyl, to methoxy-benzyl ... .(as ask the visitor in, supplement! ! )

route 2:

R in intermediate (II) ₁while being not H, its preparation method is as shown in Scheme 3: the 5-thiotolene of structural formula 1-2-formic acid through nitrosonitric acid nitrated the intermediate of structural formula 2, the intermediate that is structural formula 3 by the intermediate carboxylic esterification of structural formula 2, adopting reduced iron powder is the amino intermediate that obtains structural formula 4 by the nitroreduction of the intermediate of structural formula 3, the intermediate that is then structural formula 5 by the intermediate cyclisation of structural formula 4 under the existence of acetic anhydride, nitrite; The intermediate ethanoyl of structural formula 5 is removed to obtain to the intermediate of structural formula 6, by the intermediate of structural formula 66 iodos, obtain the intermediate of structural formula 8; The pyrazoles amino of the intermediate of structural formula 8, with the protection of amido protecting group, is obtained to the intermediate of structural formula 9; Then utilize the iodine of the intermediate of structural formula 9 to carry out the intermediate that coupling obtains structural formula 10; Again the ester group of the intermediate of structural formula 10 is reduced to the intermediate that alcohol radical obtains structural formula 11,, the alcohol radical in the intermediate of structural formula 11 is oxidized to aldehyde radical, obtain the intermediate that structural formula is (II);

route 3:

Described coupling is selected from Suzuki coupling, Buchwald coupling.

route 4:

When the compound of required preparation is the compound shown in general formula (Ib), and R ₃when different from R5, its preparation method as shown in Scheme 5:

route 5:

route 6:

route 7:

Wherein X is CH ₂or O.

route 8:

route 9:

Wherein X is CH ₂or O.

Wherein, following is the abbreviation of commonly using:

DMF:N, dinethylformamide;

DMSO: dimethyl sulfoxide (DMSO);

CDCl ₃: deuterochloroform;

¹h NMR: proton nmr spectra;

MS: mass spectrum

ESI-MS: electrospray ionization mass spectrometry;

S: unimodal;

D: bimodal;

T: triplet;

Dd: doublet of doublet;

Br: broad peak;

M: multiplet;

℃: degree centigrade;

Mol: mole;

TLC: tlc.

Those skilled in the art can use suitable raw material, adopt similar method, prepare in reaction scheme above with no specific disclosure of other compounds of the present invention.

By inorganic or organic bases or the acid treatment with suitable, can be by the pharmacologically acceptable salts according to above preparing all the compounds of this invention that exist with free alkali or sour form and change into them.Above the salt of the compound of preparation can change into by standard technique their free alkali or sour form.

Compound of the present invention comprises its all crystal formations, amorphous forms, dehydrate, hydrate, solvate and salt.In addition, all compounds of the present invention that comprise ester group and amide group can change into corresponding acid by method known to those skilled in the art or by method described herein.Equally, the compounds of this invention that comprises hydroxy-acid group can be converted into corresponding ester and acid amides by method known to those skilled in the art.Also can carry out other replacements and replacement on molecule by method known to those skilled in the art (such as hydrogenation, alkylation, with acyl chloride reaction etc.).Prepare cyclodextrin inclusion compound of the present invention, the compound of the general formula (I) that defines in summary of the invention above can be dissolved in to the acceptable solvent of pharmacology for example for example, for example, in (but being not limited to) alcohol (preferred alcohol), ketone (acetone) or ether (ether), and in 20 ℃ to 80 ℃ and alpha-cylodextrin, beta-cyclodextrin or γ-cyclodextrin, the preferably aqueous solution of beta-cyclodextrin; Perhaps can for example, aqueous solution form and the cyclodextrin blend with its salt (sodium or sylvite) by the acid of the compound of the general formula (I) that defines in summary of the invention above, then for example, with equivalent acid (HCl or H ₂sO ₄) solution blending, so that corresponding cyclodextrin inclusion compound to be provided.

Now or after cooling, corresponding cyclodextrin inclusion compound crystal can crystallization.Perhaps, when general formula (I) compound is oily and crystallization, for example, by room temperature stirring for a long time (1 hour to 14 days), add the aqueous solution of cyclodextrin to process, also can be converted into corresponding cyclodextrin inclusion compound.Then by filtering and drying, inclusion compound can be separated into to solid or crystal.

For cyclodextrin of the present invention commercially available (for example, from Aldrich Chemical Co.), or adopt known method preparation by those skilled in the art.Referring to for example Croft, the people such as A.P., " Synthesis of Chemically Modified Cyclodextrins ", Tetrahedron 1983,39,9,1417-1474.Suitable cyclodextrin comprises all kinds that prepare inclusion compound with the compound of listed formula (I) above.

By selecting appropriate cyclodextrin and water, can obtain the repeatably inclusion compound of active substance content according to stoichiometric composition.Inclusion compound can be used for absorb water form or the moisture but form that more do not absorb water of drying.The typical mol ratio of the compound of cyclodextrin and general formula (I) is the 2:1(cyclodextrin: compound).

Comprising general formula (I) compound can be to be suitable for oral form as the pharmaceutical composition of activeconstituents, such as being tablet, capsule, aqueous suspension, oiliness suspensoid, dispersible pulvis or granule, syrup etc.Can prepare according to any means for the preparation of pharmaceutical composition known in the art by the composition that can orally use, and these compositions can comprise one or more material that is selected from sweeting agent, seasonings, tinting material and sanitas, in order to pharmaceutically attractive in appearance and agreeable to the taste preparation is provided.

Tablet comprises activeconstituents, and is mixed with nontoxic pharmaceutically acceptable vehicle or the carrier that is suitable for preparing tablet.These vehicle or carrier can be inert diluent, such as calcium carbonate, sodium carbonate, lactose, calcium phosphate or sodium phosphate; Granulating agent and disintegrating agent, for example Celluloasun Microcrystallisatum, carmethose, W-Gum or alginic acid; Tackiness agent, for example starch, gelatin, polyvinylpyrrolidone or gum arabic; And lubricant, for example Magnesium Stearate, stearic acid or talcum powder.Tablet can be dressing not, maybe can carry out dressing by known technology, thereby cover the medicine taste that makes us unhappy, or postpone disintegration and absorption in gi tract, and lasting effect is provided thus within the longer period.For example, can use water miscible taste to cover material (such as hydroxypropyl-methylcellulose gum or hydroxypropyl-Mierocrystalline cellulose) or time lag material (such as ethyl cellulose, cellulose acetate butyrate).

Capsule comprises hard-gelatin capsules, Gelseal.By activeconstituents, with inert solid diluent, for example calcium carbonate, calcium phosphate or kaolin mix hard-gelatin capsules; Gelseal for example, is mixed with water-soluble carrier (such as polyoxyethylene glycol) or oily medium (peanut oil, whiteruss or sweet oil) by activeconstituents.

Aqueous suspension comprises active substance and is suitable for preparing the vehicle of aqueous suspension.These vehicle are suspending agent, for example Xylo-Mucine, methylcellulose gum, hydroxypropylmethyl-Mierocrystalline cellulose, sodiun alginate, polyvinylpyrrolidone and gum arabic; Dispersion agent or wetting agent can be the condensation product (for example polyoxyethylene stearic acid ester) of naturally occurring phosphatide (for example Yelkin TTS) or oxyalkylene and lipid acid or the condensation product of ethylene oxide and long chain aliphatic alcohol (for example 17 oxygen ethene hexadecanols (heptadecaethylene-oxycetanol)) or ethylene oxide and from the condensation product (such as polyoxyethylene 80 sorbitan monooleate) of lipid acid and the derivative partial ester of hexitol or ethylene oxide and for example, from the condensation product (polyethylene dehydrated sorbitol mono-fatty acid ester) of lipid acid and the derivative partial ester of the liquor-saturated mixture of hexitol.Aqueous suspension also can comprise one or more sanitass (for example ethyl p-hydroxybenzoate or n-propyl), one or more tinting material, one or more seasonings and one or more sweeting agent (such as sucrose, asccharin or aspartame).

The oiliness suspensoid can for example, be prepared by activeconstituents being suspended in vegetables oil (peanut oil, sweet oil, sesame oil or cocounut oil) or mineral oil (such as whiteruss).The oiliness suspensoid can comprise thickening material, for example beeswax, paraffinum durum or hexadecanol.Can add sweeting agent (such as above listed those) and seasonings, thereby agreeable to the taste oral preparations is provided.These compositions can come anticorrosion by adding antioxidant (such as Butylated Hydroxyanisole or alpha-tocopherol).

Dispersible pulvis and granule comprise activeconstituents, and are mixed with dispersion agent or wetting agent, suspending agent and one or more sanitas.The example of suitable dispersion agent or wetting agent and suspending agent is above those that mentioned.Also can comprise other vehicle, for example sweeting agent, seasonings and tinting material.These compositions can come anticorrosion by adding antioxidant (such as xitix).Dispersible pulvis and granule can prepare aqueous suspension by adding water.

Syrup can for example, be prepared with sweeting agent (glycerine, propylene glycol, sorbyl alcohol or sucrose).These preparations also can comprise negative catalyst, sanitas, seasonings, tinting material and antioxidant.

Pharmaceutical composition of the present invention can be also the form of oil-in-water emulsion.Oil phase can be vegetables oil (for example sweet oil or peanut oil) or mineral oil (for example whiteruss) or their mixture.Suitable emulsifying agent can be naturally occurring phosphatide (for example soybean lecithin), for example, for example, from lipid acid and the derivative ester of hexitol mixture or the condensation product (Polysorbate 80) of partial ester (dehydrated sorbitol mono-fatty acid ester) and described partial ester and ethylene oxide.Emulsion also can comprise sweeting agent, seasonings, sanitas and antioxidant.

Pharmaceutical composition can be the form of the sterile injectable aqueous solution.Spendable accept carrier and solvent have water, Ringer's solution (Ringer's solution), etc. the sodium chloride solution and the glucose solution that ooze.

Sterile injectable preparation can be also the sterile injectable water oil-packaging type micro-emulsion, wherein activeconstituents is dissolved in oil phase.For example, at first activeconstituents is dissolved in the mixture of soybean oil and Yelkin TTS.Then, resulting oil solution imported in the mixture of water and glycerine and process, thereby forming micro emulsion.

Injectable solution or micro emulsion can inject to import in patient's blood flow by part, or give in some way described solution or micro emulsion, thereby maintain the circulation composition of constant the compounds of this invention.In order to maintain this constant concentration, can use the continuous intravenous administration devices such as infusion pump.

Pharmaceutical composition can be for the sterile injectable water-based of intramuscular or subcutaneous administration or the form of oil-based suspension.This suspension can configure according to those suitable dispersion agents of having been mentioned more than known utilization or wetting agent and suspending agent.Sterile injectable preparation can be also sterile injectable solution or the suspension of nontoxic pharmaceutically acceptable diluent or solvent, for example solution of 1,3 butylene glycol.In addition, aseptic fixed oil can be easily used as solvent or suspension medium.For this purpose, gentle fixed oil all can be used arbitrarily, comprises synthetic direactive glyceride or two glyceryl ester.In addition, lipid acid (such as oleic acid) can be used in preparing injection.

General formula (I) compound also can give by the form of the suppository for rectal administration.Can prepare by hybrid medicine and suitable nonirritant excipient by these compositions, described vehicle is solid at normal temperature but is liquid in rectal temperature, therefore in rectum, melts, thereby discharges medicine.These materials comprise the mixture of polyoxyethylene glycol of theobroma oil, glycogelatin, hydrogenated vegetable oil, different molecular weight and the fatty acid ester of polyoxyethylene glycol.

With regard to part is used, can prepare and ointment, ointment, jelly, solution or suspensoid etc. that use comprises general formula (I) compound.

Compound of the present invention can use carrier and doser in suitable nose to give with form in nose by part, or with those skilled in the art the well-known form through the skin skin patch by giving through the skin approach.Compound of the present invention also can be by using the form such as the suppository of following such matrix to give: the fatty vinegar of the mixture of the polyoxyethylene glycol of theobroma oil, glycogelatin, hydrogenated vegetable oil, different molecular weight and polyoxyethylene glycol.

When being administered in the human subject body by compound of the present invention, every per daily dose generally determined by the doctor of prescription, and described dosage changes with the severity of patient's age, body weight, sex and reaction and patient's symptom usually.Usually, for the effective per daily dose of the patient of 70kg, be about 0.001mg/kg to 100mg/kg, be preferably 0.01mg/kg to 50mg/kg, more preferably 1mg/kg to 25mg/kg.

If be mixed with fixed dosage, the compounds of this invention of these combined prods uses in dosage range described above and other medical active agent treatment in the dosage range of its approval so.When combination preparation is improper, general formula (I) compound also can successively give with known anticarcinogen or cytotoxicity medicine.The present invention is not subject to the restriction of order of administration; General formula (I) compound can give before or after known anticarcinogen (multiple anticarcinogen) or cytotoxicity medicine (various kinds of cell toxicity medicine) giving.

Compound of the present invention is the inhibitor of disease or the illness that c-Met mediates of c-Met mediation.Term " disease of c-Met mediation " and " illness of c-Met mediation " mean the effective any morbid state of known c-Met tool or other harmful illnesss.Term " disease of c-Met mediation " and " illness of c-Met mediation " also mean those diseases or illness by being eased with the c-Met inhibitor for treating.These diseases and illness include but not limited to cancer and other proliferative illness.

Therefore, described compound can be used for treating for example Mammals, especially following disease or the illness in the mankind: cancer of the stomach, lung cancer, esophagus cancer, carcinoma of the pancreas, kidney, colorectal carcinoma, thyroid carcinoma, the cancer of the brain, mammary cancer, prostate cancer and other solid tumor cancers; Atherosclerosis; The adjusting blood vessel occurs; Thrombosis and pulmonary fibrosis.

Compound involved in the present invention also can be used for the research of the signal transduction pathway that the research, Tyrosylprotein kinase of biology or pharmacology phenomenon participates in and for the comparative evaluation of new tyrosine kinase inhibitor.

The related compound of this paper is including, but not limited to above-mentioned route 1 and the given structure type of route 9, and the personnel that know art technology can pass through suitable starting raw material, and method obtains like application class.

Embodiment

Following concrete synthetic preparation example (for the preparation of compound of the present invention) and the biology embodiment (for proving the detection of the compounds of this invention purposes) provided is in order to help to put into practice the present invention, and they should not be considered to limit the scope of the invention.

Synthetic preparation example 1

Prepare 2 of structural formula (I-1), 6-dimethyl-4-(1H-thieno-[3,2-c] pyrazoles-5-yl)-Isosorbide-5-Nitrae-dihydropyridine-3,5-dimethoxy nitrile

Step 1 reaction formula is as follows:

Step is: diacetyl oxide (14.36g, 140.7mmol) adds in three-necked bottle, is cooled to-50 ° of C, drip nitrosonitric acid (5.67g, 90.0mmol), finish and slowly add 5-thiotolene 2-formic acid (4.0g in batches, 28.13mmol), finish in-20 ° of C and stir 2 hours.In reaction solution impouring frozen water, standing, filter, dry, obtain yellow solid 5-methyl-4-nitro-thiophene-2-carboxylic acid (4.0g, 76%).

Step 2 reaction formula is as follows:

Step is: 5-methyl-4-nitro-thiophene-2-carboxylic acid (4.0g, 21.37mmol) is dissolved in methyl alcohol (40mL), adds the vitriol oil (5mL), return stirring 20 hours.Be cooled to room temperature, remove methyl alcohol under reduced pressure, resistates dissolves with ethyl acetate (200mL), and washing is concentrated, obtains brown oil 5-methyl-4-nitro-thiophene-2-carboxylic acid methyl esters (2.5g, 62%).

Step 3 reaction formula is as follows:

Step is: 5-methyl-4-nitro-thiophene-2-carboxylic acid methyl esters (3.34g, 16.6mmol) be dissolved in the mixed solvent of methyl alcohol (25mL) and water (10mL), add ammonium chloride (4.39g, 82mmol), reduced iron powder (4.2g, 75mmol), return stirring is 6 hours.Be cooled to room temperature, filtration under diminished pressure, filtrate is concentrated, and resistates adds acetic acid ethyl dissolution, washing, the saturated common salt washing, drying, be concentrated into dryly, obtains 4-amino-5-methyl-thiophene-2-carboxylic acid methyl esters crude product (2.1g, 74%).ESI-MS:m/z?172(M+H)。

Step 4 reaction formula is as follows:

Step is: 4-amino-5-methyl-thiophene-2-carboxylic acid methyl esters (0.4g, 2.16mmol) be dissolved in toluene (20mL), add KOAc (0.424g, 4.23mmol), 18-hat-6 (57.07mg) and diacetyl oxide (0.727g, 7.13mmol), heated and stirred to 95 ° C, add Isopentyl nitrite (0.607g, 5.18mmol), be heated to 100 ° of C stirrings and spend the night.Cooling middle room temperature, add ethyl acetate EtOAc (100mL), and system is with washing, and anhydrous sodium sulfate drying, concentrate, and purification by silica gel column chromatography obtains 1-ethanoyl-1H-thieno-[3,2-c] pyrazoles-5-methyl-formiate (0.34g, 70%).ESI-MS:m/z?225(M+H)。

Step 5 reaction formula is as follows:

Step is: 1-ethanoyl-1H-thieno-[3,2-c] pyrazoles-5-methyl-formiate (10.0g, 42mmol) and Anhydrous potassium carbonate (23.2g, 168mmol) add in anhydrous methanol (375mL) jointly, and stirring at room 20 minutes is filtered.Filtrate decompression is concentrated, and purification by silica gel column chromatography, obtain brown solid 1H-thieno-[3,2-c] pyrazoles-5-methyl-formiate (8.0g, 97%).

Step 6 reaction formula is as follows:

Step is: 1H-thieno-[3,2-c] pyrazoles-5-methyl-formiate (8.0g, 33.6mmol) is dissolved in anhydrous tetrahydro furan (160mL), nitrogen protection.Be cooled to-20 ° of C, add lithium aluminium hydride (3.8g, 100.7mmol) in batches, stir 3 hours.The careful water (3.8mL) that drips, in standing a moment, filter.Filtrate, with anhydrous sodium sulfate drying, removes solvent under reduced pressure and obtains white solid 1H-thieno-[3,2-c] pyrazoles-5-methyl alcohol (3.54g, 69%).

Step 7 reaction formula is as follows:

Step is: 1H-thieno-[3,2-c] pyrazoles-5-methyl alcohol (1.0g, 6.5mmol) is dissolved in methylene dichloride (115mL), adds Dess-Martin oxygenant (3.3g, 7.8mmol), stirring at room 3 hours.Remove after completion of the reaction solvent under reduced pressure, the resistates purification by silica gel column chromatography, obtain 1H-thieno-[3,2-c] pyrazoles-5-formaldehyde (0.90g, 91%), is white solid.

Step 8 reaction formula is as follows:

Step is: 1H-thieno-[3,2-c] pyrazoles-5-formaldehyde (700mg, 4.6mmol), the amino propenyl cyanide of 3-(831mg, 10.1mmol) are dissolved in Glacial acetic acid (15mL) jointly, are heated to 90 ° of C and stir 1 hour.Remove acetic acid under reduced pressure, the resistates column chromatography purification obtains target product 2,6-dimethyl-4-(1H-thieno-[3,2-c] pyrazoles-5-yl)-Isosorbide-5-Nitrae-dihydropyridine-3, and 5-dimethoxy nitrile (820mg, 66%), be the off-white color solid.MS:282(M+H); ¹H?NMR(300MHz,DMSO-d ₆)δ13.02(s,1H),9.68(s,1H),7.75(d,1H),7.04(s,1H),4.79-4.80(d,1H),2.03(s,6H)。

Synthetic preparation example 2

Prepare the 2-ethyl of structural formula (I-2)-6-methyl-4-(1H-thieno-[3,2-c] pyrazoles-5-yl)-Isosorbide-5-Nitrae-dihydropyridine-3, the 5-dimethoxy nitrile

Step 1 reaction formula is as follows:

Step is: under nitrogen protection, anhydrous tetrahydro furan (100mL) is cooled to-70 ° of C, adds n-Butyl Lithium (hexane solution of 2.5M, 12.8mL, 32mmol), stirs and drips in a moment anhydrous acetonitrile (1.15g, 28mmol).Stir 3 minutes, drip ethyl propionate (2.04g, 20mmol), during dropping and keep temperature of reaction system lower than-66 ° of C.The system of finishing is warming up to-45 ° of C and stirs 2 hours, drips 1N hydrochloric acid (64mL) cancellation.Reaction solution is concentrated, adds extracted with diethyl ether in resistates, and extraction liquid merges, anhydrous sodium sulfate drying, and concentrated, remaining oily matter 3-oxo valeronitrile (1.802g, 93% thick yield) is directly used in next step reaction. ¹H?NMR(300MHz,CDCl ₃)：3.48(s,2H),2.62(q,2H),1.11(t,3H)。

Step 2 reaction formula is as follows:

Step is: 1H-thieno-[3; 2-c] pyrazoles-5-formaldehyde (200mg; 1.314mmol) (preparation process is referring to synthetic preparation example 1) and 3-oxo valeronitrile (191mg; 1.972mmol) jointly be dissolved in methylene dichloride (20mL); add piperidines (11mg; 0.131mmol), Glacial acetic acid (12mg, 0.197mmol), flow through next time night reaction in nitrogen protection.Reaction solution is cooled to room temperature, add dehydrated alcohol (20mL) dilute reaction solution, stir after ten minutes and filter, filtrate is concentrated dry, add ethyl acetate (5mL) and sherwood oil (10mL) in resistates, stir after five minutes and filter, solid drying obtains 2-((1H-thieno-[3,2-c] pyrazoles-5-yl) methene)-3-oxo valeronitrile (190mg, 62.5%).ESI-MS：232[M+H]。

Step 3 reaction formula is as follows:

Step is: 2-((1H-thieno-[3,2-c] pyrazoles-5-yl) methene)-3-oxo valeronitrile (140mg, 0.61mmol), the amino propenyl cyanide of 3-(109mg, 1.33mmol) is dissolved in Glacial acetic acid (10mL) jointly, be heated to 100 ° of C and stir 1 hour.Be chilled to room temperature, the reaction solution concentrating under reduced pressure is dry, resistates column chromatography (methylene dichloride: methyl alcohol=80:1) purifying, obtain marking compound 2-ethyl-6-methyl-4-(1H-thieno-[3,2-c] pyrazoles-5-yl)-Isosorbide-5-Nitrae-dihydropyridine-3,5-dimethoxy nitrile (120mg, 67%).ESI-MS：296[M+H]； ¹H?NMR(300MHz,DMSO-d ₆)δ13.00(br,1H),9.62(br,1H),7.75(s,1H),7.03(s,1H),4.79(s,1H),2.27-2.36(q,2H),2.04(s,3H),1.14(t,3H，J=7.2Hz)。

Synthetic preparation example 3

Prepare the 2-isobutyl-of structural formula (I-3)-6-methyl-4-(1H-thieno-[3,2-c] pyrazoles-5-yl)-Isosorbide-5-Nitrae-dihydropyridine-3, the 5-dimethoxy nitrile

Step 1 reaction formula is as follows:

Step is: under nitrogen protection; anhydrous tetrahydro furan (100mL) is cooled to-70 ° of C; add n-Butyl Lithium (hexane solution of 2.5M, 12.8mL, 32mmol); then drip anhydrous acetonitrile (1.15g; 28mmol), finish and stir after 3 minutes, drip Ethylisovalerate (2.6g; 20mmol), in dropping process, keep temperature of reaction system to be no more than-66 ° of C.Finish and be warming up to-45 ° of C stirrings 2 hours.Drip 1N hydrochloric acid (40mL) cancellation reaction solution, concentrating under reduced pressure, add extracted with diethyl ether in resistates, and extraction liquid merges, drying, and concentrated, resistates oily matter 5-methyl-own nitrile of 3-oxo (2.68g, 100% thick yield) is directly used in next step reaction. ¹H?NMR(300MHz,CDCl ₃)0.93(d,6H),2.16(m,1H),2.49(t,2H),3.43(s,2H)。

Step 2 reaction formula is as follows:

Step is: 1H-thieno-[3; 2-c] pyrazoles-5-formaldehyde (200mg; 1.31mmol) (preparation process is referring to synthetic preparation example 1), the own nitrile (329mg of 5-methyl-3-oxo; 2.63mmol) be dissolved in methylene dichloride (20mL); add Glacial acetic acid (12mg; 0.20mmol) and piperidines (11mg, 0.13mmol), under nitrogen protection, stirring and refluxing is spent the night.Reaction solution is cooling, add dehydrated alcohol (20mL) dilute reaction solution, stir after ten minutes and filter, filtrate is concentrated dry, resistates column chromatography (methylene dichloride: methyl alcohol=300:1) obtain 2-((1H-thieno-[3,2-c] pyrazoles-5-yl) methene)-5-methyl-own nitrile of 3-oxo (294mg, 86%).MS(ESI+)260[M+H]。

Step 3 reaction formula is as follows:

Step is: 2-((1H-thieno-[3,2-c] pyrazoles-5-yl) methene)-the own nitrile (100mg of 5-methyl-3-oxo, 0.605mmol), the amino propenyl cyanide (70mg of 3-, 0.848mmol) jointly be dissolved in Glacial acetic acid (8mL), being heated to 100 ° of C stirs 1 hour, be cooled to room temperature, concentrating under reduced pressure is dry, resistates silica gel prepares plate purifying (methylene dichloride: methyl alcohol=20:1), obtain 2-isobutyl--6-methyl-4-(1H-thieno-[3,2-c] pyrazoles-5-yl)-Isosorbide-5-Nitrae-dihydropyridine-3,5-dimethoxy nitrile (73mg, 59%).MS(ESI+)324[M+H]； ¹H?NMR(300MHz,DMSO-d ₆)δ9.56(br,1H),7.83(s,1H),7.04(s,1H),4.83(s,1H),2.12-2.27(m,2H),2.04(s,3H),1.91-2.00(m,1H),0.92(dd,6H,J=9.9,6.0Hz)。

Synthetic preparation example 4

Prepare the 2-methyl-4-(1H-thieno-[3,2-c] pyrazoles-5-yl) of structural formula (I-4)-6-(3,3,3-trifluoro propyl)-Isosorbide-5-Nitrae-dihydropyridine-3, the 5-dimethoxy nitrile

Step 1 reaction formula is as follows:

Step is: under nitrogen protection, anhydrous tetrahydro furan (100mL) is cooled to-74 ° of C, adds n-Butyl Lithium (hexane solution of 2.5M, 16mL, 40mmol), then drips anhydrous acetonitrile (1.437g, 35mmol).Stir after 3 minutes, drip 4,4,4-trifluoroacetic acid ethyl ester (4.253g, 25mmol), in process, keep temperature of reaction system to be no more than-69 ° of C.Finish and be warming up to-45 ° of C stirrings 2 hours.Drip 1N hydrochloric acid (50mL) cancellation reaction solution, concentrating under reduced pressure, add extracted with diethyl ether in resistates, and extraction liquid merges, drying, and concentrated, resistates (3.908g, 94% thick yield) is directly used in next step reaction ¹h NMR (300MHz, CDCl ₃) 2.43-2.55 (m, 2H), 2.89 (t, 2H, J=7.5Hz), 3.55 (s, 2H).

Step 2 reaction formula is as follows:

Step is: 1H-thieno-[3; 2-c] pyrazoles-5-formaldehyde (304mg; 2.0mmol) (preparation process is referring to synthetic preparation example 1), 6; the fluoro-own nitrile of 3-oxo of 6,6-tri-(660mg, 4.0mmol) is dissolved in methylene dichloride (20mL); add Glacial acetic acid (12mg; 0.20mmol) and piperidines (11mg, 0.13mmol), under nitrogen protection, stirring and refluxing is spent the night.Reaction solution is cooling, add dehydrated alcohol (20mL) dilute reaction solution, stir after ten minutes and filter, filtrate is concentrated dry, resistates column chromatography (methylene dichloride: methyl alcohol=200:1) obtain 2-((1H-thieno-[3,2-c] pyrazoles-5-yl) methene)-6,6, the own nitrile of the fluoro-3-oxo of 6-tri-(340mg, 57%).MS(ESI+)300[M+H]。

Step 3 reaction formula is as follows:

Step is: 2-((1H-thieno-[3,2-c] pyrazoles-5-yl) methene)-6,6, own nitrile (the 200mg of the fluoro-3-oxo of 6-tri-, 0.67mmol), the amino propenyl cyanide of 3-(70mg, 0.848mmol) is dissolved in Glacial acetic acid (8mL) jointly, be heated to 100 ° of C and stir 1 hour, be cooled to room temperature, concentrating under reduced pressure is dry, and resistates silica gel prepares plate purifying (methylene dichloride: methyl alcohol=20:1), obtain 2-methyl-4-(1H-thieno-[3,2-c] pyrazoles-5-yl)-6-(3,3,3-trifluoro propyl)-Isosorbide-5-Nitrae-dihydropyridine-3,5-dimethoxy nitrile (63mg, 26%).MS(ESI+)364[M+H]； ¹H?NMR(300MHz,DMSO-d ₆)δ9.56(br,1H),7.83(s,1H),7.04(s,1H),4.83(s,1H),2.04(s,3H),1.97(m,2H),1.83(m,2H)。

Synthetic preparation example 5

Prepare the 2-(2-methoxy ethyl) of structural formula (I-5)-6-methyl-4-(1H-thieno-[3,2-c] pyrazoles-5-yl)-Isosorbide-5-Nitrae-dihydropyridine-3, the 5-dimethoxy nitrile

Step 1 reaction formula is as follows:

Step is: by n-BuLi(31mL, 77mmol) join in the THF (350ml) of-78 ° of C, then drip acetonitrile (2.9g, 70mmol), finish, stir at low temperatures 1h.Then splash into 2-methoxy methyl propionate (5.9g, 50mmol), finish, temperature is risen to-45 ° of C, and stir 2h, use at low temperatures 2N hydrochloric acid (160mL) cancellation, reaction solution slowly rises to room temperature, uses extracted with diethyl ether, merges organic phase, concentrate to obtain 5-methoxyl group-3-oxo valeronitrile (3.5g, 55%).

Step 2 reaction formula is as follows:

Step is: by 1H-thieno-[3,2-c] pyrazoles-5-formaldehyde (350mg, 2.3mmol) (preparation process is referring to synthetic preparation example 1) be dissolved in methylene dichloride (5mL), then add 5-methoxyl group-3-oxo valeronitrile (292mg, 2.3mmol), piperidines (11mg, 0.13mmol), acetic acid (99mg, 1.643mmol) and 4A molecular sieve, finish, reaction is heated to return stirring and spends the night.Reaction solution is cooling, and concentrated, resistates is directly used in next step.

Step 3 reaction formula is as follows:

Step is: upper step gained resistates (600mg, 2.3mmol) is dissolved in Glacial acetic acid (6mL), adds the amino propenyl cyanide (190mg, 2.3mmol) of 3-, be heated to 100 ° of C and stir 1h.Cooling, reaction solution is concentrated dry, and resistates is purified and is obtained product 25mg, 3% with preparation TLC.MS:[M+1]=326; ¹H?NMR(300MHz,CDCl ₃)δ7.80(s,1H),7.02(s,1H),6.40(br,1H),4.67(s,1H),3.99-3.90(m,2H),3.41(s,3H),2.93-2.88(m,2H)，2.01(s,3H)。

Synthetic preparation example 6

Prepare the 2-(4-fluorophenyl) of structural formula (I-6)-6-methyl-4-(1H-thieno-[3,2-c] pyrazoles-5-yl)-Isosorbide-5-Nitrae-dihydropyridine-3, the 5-dimethoxy nitrile

Step 1 reaction formula is as follows:

Step is: 1H-thieno-[3; 2-c] pyrazoles-5-formaldehyde (150mg; 0.99mmol) (preparation process is referring to synthetic preparation example 1) and 4-fluoro benzoyl acetonitrile (177mg; 1.08mmol) be dissolved in altogether methylene dichloride (15mL); add acetic acid (9mg; 0.15mmol) and piperidines (8mg, 0.10mmol), flow through next time the reaction at night in nitrogen protection.Reaction solution is cooling, add methylene dichloride (30mL) dilute reaction solution, filter; filtrate is concentrated dry; resistates column chromatography (methylene dichloride: methyl alcohol=100:1) obtain the 2-(4-fluoro benzoyl)-3-(1H-thieno-[3,2-c] pyrazoles-5-yl) vinyl cyanide (259mg, 88%).MS(ESI+):298[M+1] ⁺。

Step 2 reaction formula is as follows:

Step is: by the 2-(4-fluoro benzoyl)-3-(1H-thieno-[3,2-c] pyrazoles-5-yl) vinyl cyanide (259mg, 0.87mmol) and amino propenyl cyanide (157mg, 1.917mmol) be dissolved in acetic acid (15mL), be heated to 100 ° of C and stir 1 hour.After cooling, reaction solution is concentrated dry, preparation TLC purifying (methylene dichloride: methyl alcohol=20:1) obtain 2-(4-fluorophenyl)-6-methyl-4-(1H-thieno-[3,2-c] pyrazoles-5-yl)-Isosorbide-5-Nitrae-dihydropyridine-3,5-dimethoxy nitrile (62mg, 20%).HPLC shows that purity is 97%; LC-MS (ESI+): 362[M+1] ⁺; ¹h NMR (300MHz, CDCl ₃) δ 7.76 (br, 1H), 7.54-7.59 (m, 2H), 7.19 (t, 2H, J=8.7Hz), 7.04 (s, 1H), 6.36 (s, 1H), 2.24 (s, 3H).

Synthetic preparation example 7

Prepare the 2-(4-p-methoxy-phenyl) of structural formula (I-7)-6-methyl-4-(1H-thieno-[3,2-c] pyrazoles-5-yl)-Isosorbide-5-Nitrae-dihydropyridine-3, the 5-dimethoxy nitrile

Step 1 reaction formula is as follows:

Step is: methyl p-methoxybenzoate (2.0g, 12mmol) is dissolved in dry toluene (50mL), is cooled to 0 ° of C, adds sodium hydrogen (1.2g, 30mmol).Stir and add acetonitrile (1.23g, 30mmol) after 10 minutes, slowly be warming up to 110 ° of C, stirring is spent the night.After being cooled to room temperature, filter, filter residue, with after toluene wash, collecting and dry, obtains 1-cyano group-2-(4-p-methoxy-phenyl)-2-oxypropionitrile sodium salt (2.2g, 93%).

Step 2 reaction formula is as follows:

Step is: 1-cyano group-2-(4-p-methoxy-phenyl)-2-oxypropionitrile sodium salt (500mg; 3.29mmol), 1H-thieno-[3; 2-c] pyrazoles-5-formaldehyde (980mg; 4.93mmol) (preparation process is referring to synthetic preparation example 1) add in methylene dichloride (50mL); add Glacial acetic acid (490mg; 4.93mmol) and piperidines (280mg, 3.29mmol), under nitrogen protection, return stirring spends the night.Reaction solution is cooling, add methylene dichloride (30mL) dilute reaction solution, filter; the filtrate water washing; dry and concentrated 2-(4-anisoyl)-3-(1H-thiophene [3,2-c] pyrazoles-5-yl) vinyl cyanide (1.0g) of doing to obtain, crude product is directly used in next step reaction.LC-MS(ESI+)310[M+H]。

Step 3 reaction formula is as follows:

Step is: 2-(4-anisoyl)-3-(1H-thiophene [3,2-c] pyrazoles-5-yl) vinyl cyanide (300mg, 0.92mmol), amino propenyl cyanide (152mg, 1.84mmol) are dissolved in Glacial acetic acid (20mL) jointly, N ₂being heated to 100 ° of C under protection stirs 1 hour.Be cooled to room temperature, reaction solution is concentrated dry, and silica gel prepares plate (DCM:MeOH=10:1) purifying, obtain 2-(4-p-methoxy-phenyl)-6-methyl-4-(1H-thieno-[3,2-c] pyrazoles-5-yl)-Isosorbide-5-Nitrae-dihydropyridine-3,5-dimethoxy nitrile (35mg, 10%).MS(ESI+)374[M+H]。 ¹H?NMR(300MHz,CDCl ₃)δ9.85(s,1H),7.84(s,1H),7.50-7.47(m,2H),7.10-7.05(m,3H),4.92(s,1H),3.85(s,3H),2.10(s,3H)。

Synthetic preparation example 8

Prepare the 2-methyl-6-(pyridin-4-yl) of structural formula (I-8)-4-(1H-thieno-[3,2-c] pyrazoles-5-yl)-Isosorbide-5-Nitrae-dihydropyridine-3, the 5-dimethoxy nitrile

Step 1 reaction formula is as follows:

Step is: iso methyl nicotinate (1.0g, 7.30mmol) is dissolved in toluene (30mL), is cooled to 0 ° of C, after carefully adding sodium hydrogen (60%, 0.58g, 14.6mmol) to stir 10 minutes.Add acetonitrile (1.50g, 36.5mmol) in above-mentioned solution, slowly be warming up to 80 ° of C, stir 4 hours.After being cooled to room temperature, filter, filter residue is with collecting filter residue after toluene wash, dry 1-cyano group-2-oxo-2-(pyridin-4-yl) ethane sodium salt (1.2g, 98%)

Step 2 reaction formula is as follows:

Step is: 1-cyano group-2-oxo-2-(pyridin-4-yl) ethane sodium salt (300mg; 2.96mmol), 1H-thieno-[3; 2-c] pyrazoles-5-formaldehyde (300mg; 1.97mmol) (preparation process is referring to synthetic preparation example 1) add in methylene dichloride (20mL) jointly; add acetic acid (178mg; 2.96mmol) and piperidines (82mg, 1.97mmol), under nitrogen protection, return stirring spends the night.Reaction solution is cooled to room temperature, add methylene dichloride (30mL) dilute reaction solution, filter, filtrate is concentrated dry, resistates column chromatography (DCM:MeOH=100:1) obtains the different nicotinoyl-3-of 2-(1H-thieno-[3,2-c] pyrazoles-5-yl) vinyl cyanide (600mg).(DCM:MeOH=20:1),Rf=0.5.LC-MS(ESI+)281[M+H]。

Step 3 reaction formula is as follows:

Step is: the different nicotinoyl-3-of 2-(1H-thieno-[3,2-c] pyrazoles-5-yl) (550mg, 1.96mmol), amino propenyl cyanide (400mg, 4.91mmol) are dissolved in acetic acid (20mL) jointly, are heated to 100 ° of C and stir 1 hour.Be cooled to room temperature, reaction solution is concentrated dry, and silica gel prepares plate purifying (DCM:MeOH=10:1) and obtains 2-methyl-6-(pyridin-4-yl)-4-(1H-thieno-[3,2-c] pyrazoles-5-yl)-1,4-dihydropyridine-3,5-dimethoxy nitrile (80mg, 129%).MS(ESI+)362[M+H]； ¹H?NMR(300MHz,CDCl ₃)δ13.15(br,1H),10.07(s,1H),8.76-8.75(m,2H),7.84(br,1H),7.56(m,2H),7.15(s,1H),5.01(s,1H,),2.10(s,3H)。

Synthetic preparation example 9

6-methyl-the 8-(1H-thieno-[3,2-c] pyrazoles-5-yl)-2,3,4 for preparing structural formula (I-9), 8-tetrahydrochysene-1H-quinolizine-7,9-dimethoxy nitrile

Step 1 reaction formula is as follows:

Step is: in methylene dichloride (200mL) solution of 2-piperidone (4.96g, 50mmol), add MeOTf(trifluoromethanesulfonic acid methyl esters) (10.2g, 62mmol), finishing, reaction solution at room temperature reacts 18 hours.Add solid sodium carbonate (20g) and water (20mL) in reaction solution, stir 10 minutes, filter, filtrate is used anhydrous sodium sulfate drying, concentrated, remaining 6-methoxyl group-2,3,4, and 5-tetrahydropyridine crude product (4.52g, 81%), purifying is not directly used in the next step.LC-MS(ESI+)：114[M+1] ⁺。

Step 2 reaction formula is as follows:

Step is: by gained 6-methoxyl group-2,3,4 in step 1,5-tetrahydropyridine crude product (4.52g, 40mmol) is dissolved in anhydrous tetrahydro furan (150mL), adds the cyanoacetic acid tert-butyl ester (5.98g, 42.3mmol), is heated to 70 ° of C backflows and spends the night.Reaction solution is concentrated, resistates column chromatography (sherwood oil: ethyl acetate=10:1) obtain 2-cyano group-2-(piperidines-2-thiazolinyl) tert.-butyl acetate (3.2g, 36%).LC-MS(ESI+):223[M+1] ⁺。

Step 3 reaction formula is as follows:

Step is: 2-cyano group-2-(piperidines-2-thiazolinyl) tert.-butyl acetate (100mg, 0.45mmol) add in 6M hydrochloric acid (16mL), be heated to 100 ° of C and stir 15 minutes, concentrated dry, resistates (piperidines-2-thiazolinyl) acetonitrile (55mg, 100% thick yield) is directly used in the next step.

Step 4 reaction formula is as follows:

Step is: upper step gained (piperidines-2-thiazolinyl) acetonitrile (55mg crude product) is dissolved in Glacial acetic acid (16mL), add 2-((the 1H-thieno-[3 made in synthetic preparation example 2 steps 1,2-c] pyrazoles-5-yl) methylene radical)-3-oxo butyronitrile (65mg, 0.3mmol), be heated to 100 ° of C and stir 1 hour.Reaction solution is concentrated dry, resistates column chromatography (methylene dichloride: methyl alcohol=50:1) obtain 6-methyl-8-(1H-thieno-[3,2-c] pyrazoles-5-yl)-2,3,4,8-tetrahydrochysene-1H-quinolizine-7,9-dimethoxy nitrile (63mg, 66%).HPLC：98.4%；LC-MS(ESI+):322[M+1] ⁺； ¹H?NMR(300MHz,CDCl ₃)δ7.74(s,1H),6.96(s,1H),4.60(s,1H),3.53-3.60(m,2H),2.75(t,2H,J=4.7Hz),2.27(s,3H),1.79-1.93(m,4H)。

Copy the method for synthetic preparation example 9, adopt respectively corresponding intermediate to replace 2-((1H-thieno-[3,2-c] pyrazoles-5-yl) methylene radical) wherein-3-oxo butyronitrile, can prepare respectively Compound I-10 in following table to I-12:

Synthetic preparation example 13:

Prepare the 6-methyl-8-(1H-thieno-[3,2-c] pyrazoles-5-yl)-1,3,4 of structural formula (I-13), the 8-tetrahydropyridine is [2,1-c] [Isosorbide-5-Nitrae] oxazines-7 also, the 9-dimethoxy nitrile

Step 1 reaction formula is as follows:

Step is: 3-morpholone mai (303mg, 3mmol) is dissolved in methylene dichloride (20mL), adds MeOTf(trifluoromethanesulfonic acid methyl esters) (610mg, 3.72mmol), stirring at room 18 hours.Add Na ₂cO ₃powder (5g), water (1mL), stirring was filtered after half an hour, and filtrate, with anhydrous sodium sulfate drying, concentrates and to obtain 5-methoxyl group-3,6-dihydro-2H-1,4-oxazines crude product (261mg, 75%), purifying is not directly used in next step.

Step 2 reaction formula is as follows:

Step is: above walk gained 5-methoxyl group-3, and 6-dihydro-2H-1,4-oxazines crude product (261mg, 75%) is dissolved in anhydrous tetrahydro furan (10mL) altogether with the cyanoacetic acid tert-butyl ester (461mg, 2.81mmo), and return stirring spends the night.Be cooled to room temperature, concentrated, resistates column chromatography (sherwood oil: ethyl acetate=10:1), obtain white solid 2-cyano group-2-(morpholine-3-thiazolinyl) tert.-butyl acetate (106mg, 21%).

Step 3 reaction formula is as follows:

Step is: 2-cyano group-2-(morpholine-3-thiazolinyl) tert.-butyl acetate (189mg, 0.842mmol) joins in 6M hydrochloric acid (6mL), is heated to 100 ° of C and stirs 15 minutes.Be cooled to room temperature, concentrating under reduced pressure obtains faint yellow solid 2-(morpholine-3-thiazolinyl) acetonitrile (103mg, 99% thick yield), and purifying is not directly used in next step.

Step 4 reaction formula is as follows:

Step is: above walk gained 2-(morpholine-3-thiazolinyl) acetonitrile crude product (103mg, 0.83mmol) be dissolved in Glacial acetic acid (6mL), add 2-((1H-thieno-[3,2-c] pyrazoles-5-yl) methylene radical)-3-oxo butyronitrile (synthetic preparation example 2 steps 1,102mg, 0.468mmol), be heated to 100 ° of C and stir 15 minutes.Be chilled to room temperature, remove solvent under reduced pressure, resistates prepares TLC purifying (methylene dichloride: methyl alcohol=15:1) obtain faint yellow solid (29mg, 19%).HPLC purity: 95%; ¹h NMR (300MHz, CDCl ₃) δ 7.75 (s, 1H), 6.98 (s, 1H), 4.53-4.75 (m, 3H), 3.89-4.09 (m, 2H), 3.47-3.67 (m, 2H), 2.30 (s, 3H).

Copy the method for synthetic preparation example 13, with corresponding intermediate, substitute 2-((1H-thieno-[3,2-c] pyrazoles-5-yl) methylene radical) wherein-3-oxo butyronitrile, can prepare respectively Compound I-14 to I-16:

Synthetic preparation example 17:

Prepare 1,2 of structural formula (I-17), 6-trimethylammonium-4-(1H-thieno-[3,2-c] pyrazoles-5-yl)-Isosorbide-5-Nitrae-dihydropyridine-3,5-dimethoxy nitrile

Step 1 reaction formula is as follows:

Step is: 1H-thieno-[3,2-c] pyrazoles-5-methyl-formiate (8.5g, 46.7mmol) (preparation process is referring to synthetic preparation example 1), salt of wormwood (7.75g, 56mmol) add to DMF(150mL) in, by p-methoxybenzyl chloride (7.3g, 46.7mmol) slowly be added dropwise to reaction solution, stirring at normal temperature 1 hour.Remove DMF under reduced pressure, resistates, with acetic acid ethyl dissolution, washes with water, the saturated common salt washing, and anhydrous sodium sulfate drying, concentrated, column chromatography purification obtains 1-(4-methoxy-benzyl)-1H-thieno-[3,2-c] pyrazoles-5-carboxylate methyl ester (13.8g, 98%).

Step 2 reaction formula is as follows:

Step is: by lithium aluminum hydride (1.36g, 35.76mmol) be suspended in tetrahydrofuran (THF) (100mL), be cooled to 0 ℃, drip 1-(4-methoxy-benzyl)-1H-thieno-[3,2-c] pyrazoles-5-carboxylate methyl ester (7.2g, 23.8mmol) tetrahydrofuran (THF) (100mL) solution, finish stirring at room 2 hours.Under frozen water is cooling, slowly drip water (1.4mL) cancellation, filter.Filter residue is washed with the mixed solvent (3:1) of methylene dichloride and methyl alcohol, filters, and filtrate is concentrated, and column chromatography obtains (1-(4-methoxy-benzyl)-1H-thieno-[3,2-c] pyrazoles-5-yl) methyl alcohol (5.94g, 91%).

Step 3 reaction formula is as follows:

Step is: (1-(4-methoxy-benzyl)-1H-thieno-[3; 2-c] pyrazoles-5-yl) methyl alcohol (5.94g, 21.7mmol) is dissolved in methylene dichloride (150mL), adds Dai Si-Martin's oxygenant (13.8g; 32.5mmol), under nitrogen protection, room temperature reaction is 2 hours.Solvent is concentrated dry, and the resistates column chromatography obtains 1-(4-methoxy-benzyl)-1H-thieno-[3,2-c] pyrazoles-5-formaldehyde (3.76g, 64%).

Step 4 reaction formula is as follows:

Step is: 1-(4-methoxy-benzyl)-1H-thieno-[3,2-c] pyrazoles-5-formaldehyde (500mg, 1.84mmol), amino crotons cyanogen (332mg, 4.04mmol) be dissolved in Glacial acetic acid (50mL), be heated to 100 ° of C and stir 1 hour, remove acetic acid under reduced pressure, the direct column chromatography of resistates obtains 4-(1-(4-methoxy-benzyl)-1H-thieno-[3,2-c] pyrazoles-5-yl)-2,6-dimethyl-Isosorbide-5-Nitrae-dihydropyridine-3,5-dimethoxy nitrile (590mg, 80%)

Step 5 reaction formula is as follows:

Step is: by 4-(1-(4-methoxy-benzyl)-1H-thieno-[3,2-c] pyrazoles-5-yl)-2,6-dimethyl-1,4-dihydropyridine-3,5-dimethoxy nitrile (590mg, 1.47mmol) is dissolved in DMF(30mL) in, be cooled to 0 ℃, cesium carbonate (719mg, 2.2mmol) is added to reaction solution.Stir 15 minutes, add methyl iodide (209mg, 1.47mmol), stirring at room 4 hours.Add the methylene dichloride dilution in reaction solution, the difference water, saturated common salt washing, organic phase anhydrous sodium sulfate drying, concentrated, column chromatography obtains 4-(1-(4-methoxy-benzyl)-1H-thieno-[3,2-c] pyrazoles-5-yl)-1,2,6-trimethylammonium-1,4-dihydropyridine-3,5-dimethoxy nitrile (611mg, 100%).

Step 6 reaction formula is as follows:

Step is: compound 5(611mg, 1.47mmol) be dissolved in trifluoroacetic acid (20mL), be heated to 90 ℃ of stirrings and spend the night.Trifluoroacetic acid is concentrated dry, and column chromatography obtains target product 1,2,6-trimethylammonium-4-(1H-thieno-[3,2-c] pyrazoles-5-yl)-Isosorbide-5-Nitrae-dihydropyridine-3,5-dimethoxy nitrile (43mg, 10%).MS(ESI+)：296[M+1]； ¹H?NMR(300MHz,DMSO-d ₆)δ7.81(s,1H),7.02(s,1H),4.72(s,1H),3.19(s,3H),2.20(s,6H)。

Synthetic preparation example 18:

The 1-ethyl-2 for preparing structural formula (I-18), 6-dimethyl-4-(1H-thieno-[3,2-c] pyrazoles-5-yl)-Isosorbide-5-Nitrae-dihydropyridine-3,5-dimethoxy nitrile

Copy the method for preparation example 17, with monobromethane, replace methyl iodide wherein, can prepare this compound.MS(ESI+)310[M+1]； ¹H?NMR(300MHz,DMSO-d ₆)δ7.81(br,1H),7.03(s,1H),4.74(s,1H),3.66(q,2H),2.25(s,6H),1.14(t,3H)。

Synthetic preparation example 19

The 1-benzyl-2 for preparing structural formula (I-19), 6-dimethyl-4-(1H-thiophene [3,2-c] pyrazoles-5-yl)-Isosorbide-5-Nitrae-dihydropyridine-3,5-dimethoxy nitrile

Copy the method for preparation example 17, with cylite, replace methyl iodide wherein, can prepare this compound.MS(ESI+)：372[M+1]； ¹H?NMR(300MHz,DMSO-d ₆)δ13.21(br,1H)，7.87(s,1H),7.42(m,3H),7.33(m，1H)，7.19(m，2H)，7.09(s，1H)，4.97(s,2H),4.89(s，1H)，2.16(s,6H)。

Synthetic preparation example 20

3-oxygen-the 4-(1H-thieno-[3,2-c] pyrazoles-5-yl)-3,4,6,7,8 for preparing structural formula (I-20), 9-six hydrogen-1H-furo [3,4-c] quinolizine-5-formonitrile HCN, concrete reaction scheme is as follows:

Step is: 1H-thieno-[3,2-c] pyrazoles-5-formaldehyde (304mg, 2.0mmol) (preparation process is referring to synthetic preparation example 1) and furans-2,4 (3H, 5H)-diketone (200mg, 2.0mmol) add in Pentyl alcohol (5mL), return stirring 1 hour.Be chilled to room temperature, add 2-(piperidines-2-thiazolinyl) acetonitrile (367mg, 3.0mmol), the Glacial acetic acid (4mL) of fresh preparation, 100 ℃ are stirred 1.5 hours.Be cooled to room temperature, concentrating under reduced pressure, resistates prepares TLC purifying (methylene dichloride: methyl alcohol=10:1) obtain target product, faint yellow solid (59mg, 9%).MS(ESI+):339[M+1]。

Synthetic preparation example 21

Prepare the 3-oxygen-4-(1H-thieno-[3,2-c] pyrazoles-5-yl)-1,3,4,6,8 of structural formula (I-21), the 9-hexahydro furyl is [3', 4':5,6] pyridos [2,1-c] [Isosorbide-5-Nitrae] oxazines-5-formonitrile HCN also, and concrete reaction scheme is as follows:

Step is: 1H-thieno-[3,2-c] pyrazoles-5-formaldehyde (223mg, 1.469mmol) (preparation process is referring to synthetic preparation example 1) and furans-2,4 (3H, 5H)-diketone (147mg, 1.469mmol) adds in Pentyl alcohol (5mL), return stirring 1 hour.Be chilled to room temperature, add 2-(morpholine-3-thiazolinyl) acetonitrile (273mg, 2.2mmol), the Glacial acetic acid (4mL) of fresh preparation, 100 ° of C stir 1.5 hours.Be cooled to room temperature, concentrating under reduced pressure, resistates prepares TLC purifying (methylene dichloride: methyl alcohol=10:1) obtain target product faint yellow solid (60mg, 12%).LC-MS(ESI+):341[M+1] ⁺； ¹H?NMR(300MHz,CDCl ₃)δ13.04(s,1H),δ7.76(d,1H),7.07(s,1H),5.09-5.10(d,2H),4.94(s,1H),4.52-4.65(m,2H),3.93-3.99(m,2H),3.50-3.57(m,2H)。

Synthetic preparation example 22

Prepare 3 of structural formula (I-22), 6-dimethyl-4-(1H-thieno-[3,2-c] pyrazoles-5-yl)-4,7-dihydro-isoxzzole is [5,4-b] pyridine-5-carbonitriles also, and concrete reaction scheme is as follows:

Step is: by 2-((1H-thieno-[3,2-c] pyrazoles-5-yl) methylene radical)-3-oxo butyronitrile (175mg, 0.81mmol) (preparation process is dissolved in Glacial acetic acid (10mL) referring to synthetic preparation example 2 step 1), add 3-methyl isoxzzole-5-amine (79mg, 0.81mmol), be heated to 100 ° of C and stir 1 hour.Be cooled to room temperature, reaction solution is concentrated dry, and resistates prepares TLC purifying (methylene dichloride: methyl alcohol=20:1) obtain target compound (30mg, 13%).HPLC shows that purity is 95.2%; LC-MS (ESI+): 298[M+1] ⁺; ¹h NMR (300MHz, DMSO-d ₆) δ 10.96 (br, 1H), 7.79 (s, 1H), 7.10 (s, 1H), 5.27 (s, 1H), 2.12 (s, 3H), 1.87 (s, 3H).

Synthetic preparation example 23:

Prepare the 6-(4-fluorophenyl) of structural formula (I-23)-3-methyl-4-(1H-thieno-[3,2-c] pyrazoles-5-yl)-4, the 7-dihydro-isoxazole is [5,4-b] pyridine-5-carbonitriles also, and concrete reaction scheme is as follows:

Step is: by 2-(4-fluoro benzoyl)-3-(1H-thieno-[3; 2-c] pyrazoles-5-yl) vinyl cyanide (480mg; 1.614mmol) (is preparation process referring to synthetic preparation example?) be dissolved in Glacial acetic acid (30mL); add 3-methyl isoxzzole-5-amine (190mg; 1.937mmol), be heated to 100 ° of C and stir 1 hour.Be cooled to room temperature, reaction solution is concentrated dry, and resistates silica gel prepares plate (DCM:MeOH=20:1) purifying, obtains compound 3 (20mg, 3.3%).MS(ESI+)378[M+1] ⁺。 ¹H?NMR(300MHz,DMSO-d ₆)δ13.02(br,1H),11.23(br,1H),7.65(t,2H,J=4.4Hz),7.62(s,1H),7.37(t,2H,J=7.2Hz),7.18(s,1H),5.44(s,1?H9),1.94(s,3H)。

Synthetic preparation example 24

Prepare 2 of structural formula (I-24), 6-dimethyl-4-(3-methyl isophthalic acid H-thieno-[3,2-c] pyrazoles-5-yl)-Isosorbide-5-Nitrae-dihydropyridine-3, the 5-dimethoxy nitrile,

Step 1 reaction scheme is as follows:

Step is: sodium piece (5.13g, 223mmol) is added to reaction in methyl alcohol (30mL) until the completely dissolve of sodium piece obtains sodium methoxide solution.To 1-ethanoyl-1H-thieno-[3; 2-c] pyrazoles-5-methyl-formiate (10.0g; 44.6mmol) adding the above-mentioned sodium methoxide solution made in methyl alcohol (100mL) solution of (preparation process referring to synthetic preparation example 1), the mixture obtained at room temperature stirs 30 minutes.Again to the DMF(30mL that drips iodine (28.3g, 112mmol) in above-mentioned reaction solution) solution.After dropwising, reaction solution is heated to 60 ° of C stirrings and spends the night.Reaction solution is cooled to room temperature, concentrated dry, and resistates adds ethyl acetate and water extraction, and ethyl acetate is through washing, and after the saturated common salt washing, drying, be concentrated to certain volume, filters and obtain the iodo-1H-thieno-of 3-[3,2-c]-5-methyl-formiate (10.3g, 73%).LC-MS(ESI+)：309[M+H]。

Step 2 reaction scheme is as follows:

Step is: the iodo-1H-thieno-of 3-[3,2-c]-5-methyl-formiate (5.0g, 16.23mmol), DMAP(397mg, 3.25mmol) and Et ₃n(1.81g, 17.85mmol) be dissolved in methylene dichloride (100mL), then drip Boc ₂o(4.25g, 19.47mmol), dropwise, under room temperature, reaction is 30 minutes, and reaction solution is concentrated dry, and the resistates purification by silica gel column chromatography, obtain the iodo-1H-thieno-of 1-tertbutyloxycarbonyl-3-[3,2-c] pyrazoles-5-methyl-formiate (5.9g, 89%).LC-MS(ESI+)：409[M+H] ⁺。

Step 3 reaction scheme is as follows:

Step is: the iodo-1H-thieno-of 1-tertbutyloxycarbonyl-3-[3,2-c] pyrazoles-5-methyl-formiate (5.5g, 13.47mmol) is dissolved in to Isosorbide-5-Nitrae-dioxane (100mL), adds Pd (dppf) Cl ₂(986mg, 1.347mmol), drip zinc methide (1.15M toluene solution, 17.6mL, 20.2mmol), finishes, and nitrogen replacement three times, be heated to 100 ° of C and stir 1 hour.Be cooled to room temperature, add methylene dichloride (300mL) dilute reaction solution, drip 1N hydrochloric acid neutralization reaction liquid to the pH=7 left and right, organic phase is through washing, and saturated common salt is washed, drying, concentrated, resistates column chromatography (methylene dichloride: methyl alcohol=125:1) obtain 3-methyl isophthalic acid H-thieno-[3,2-c] pyrazoles-5-methyl-formiate (1.96g, 74%).LC-MS(ESI+)：197[M+H] ⁺。

Step 4 reaction scheme is as follows:

Step is: 3-methyl isophthalic acid H-thieno-[3,2-c] pyrazoles-5-methyl-formiate (1.46g, 7.45mmol) is dissolved in DMF(30mL) in, add salt of wormwood (2.06g, 14.9mmol), drip PMBCl(1.4g, 8.94mmol), finish stirring at room 4 hours.Reaction solution is concentrated dry, and resistates is dissolved in methylene dichloride (100mL), washing, saturated common salt washing, drying, concentrated, resistates column chromatography (PE:EA=10:1), obtain 1-(4-methoxy-benzyl)-3-methyl isophthalic acid H-thieno-[3,2-c] pyrazoles-5-methyl-formiate (0.96g, 41%).LC-MS(ESI+)：317[M+H] ⁺。

Step 5 reaction scheme is as follows:

Step is: 1-(4-methoxy-benzyl)-3-methyl isophthalic acid H-thieno-[3,2-c] pyrazoles-5-methyl-formiate (429mg, 1.36mmol) is dissolved in to THF(20mL), add LiBH ₄(118mg, 5.43mmol), stirring at room 3 hours, add methylene dichloride (30mL) dilute reaction solution, drip 1N hydrochloric acid and be adjusted to pH 7 left and right, organic phase is through washing, and saturated common salt is washed, drying, resistates column chromatography (PE:EA=10:1), obtain 1-(4-methoxy-benzyl)-3-methyl isophthalic acid H-thieno-[3,2-c] pyrazoles-5-methyl alcohol (390mg, thick yield 100%).LC-MS(ESI+)：289[M+H] ⁺。

Step 6 reaction scheme is as follows:

Step is: 1-(4-methoxy-benzyl)-3-methyl isophthalic acid H-thieno-[3; 2-c] pyrazoles-5-methyl alcohol (390mg; 1.354mmol) be dissolved in methylene dichloride (25mL); add Dess-Martin oxygenant (1.15g; 2.708mmol); under nitrogen protection, room temperature reaction is 3 hours; TLC(PE:EA=2:1) show that reaction is complete; reaction solution is concentrated; resistates column chromatography (PE:EA=30:1) obtains 1-(4-methoxy-benzyl)-3-methyl isophthalic acid H-thieno-[3; 2-c] pyrazoles-5-formaldehyde (279mg, 71%).LC-MS(ESI+)：287[M+H] ⁺。

Step 7 reaction scheme is as follows:

Step is: 1-(4-methoxy-benzyl)-3-methyl isophthalic acid H-thieno-[3,2-c] pyrazoles-5-formaldehyde (275mg, 0.96mmol) and amino propenyl cyanide (174mg, 2.12mmol) are dissolved in glacial acetic acid (10mL), are heated to 100 ° of C reactions 1 hour.Reaction solution is concentrated, resistates 4-(1-(4-methoxy-benzyl)-3-methyl isophthalic acid H-thieno-[3,2-c] pyrazoles-5-yl)-2,6-dimethyl-Isosorbide-5-Nitrae-dihydropyridine-3,5-dimethoxy nitrile (399mg, thick yield 100%), be directly used in next step reaction.LC-MS(ESI+)：416[M+H] ⁺。

Step 8 reaction scheme is as follows:

Step is: above walk gained 4-(1-(4-methoxy-benzyl)-3-methyl isophthalic acid H-thieno-[3,2-c] pyrazoles-5-yl)-2,6-dimethyl-1,4-dihydropyridine-3,5-dimethoxy nitrile crude product (399mg, 0.96mmol) be dissolved in trifluoroacetic acid (10mL), be heated to 85 ° of C back flow reaction 24 hours, react completely.Reaction solution is concentrated dry, add methylene dichloride (20mL) in resistates, with saturated sodium bicarbonate solution, wash, water and saturated common salt washing again, drying, concentrated, prepare the plate purifying through silica gel again after the resistates column chromatography, obtain target product 2,6-dimethyl-4-(3-methyl isophthalic acid H-thieno-[3,2-c] pyrazoles-5-yl)-Isosorbide-5-Nitrae-dihydropyridine-3,5-dimethoxy nitrile (120mg, 42%).HPLC purity: 98.3%; MS (ESI+): 296[M+H]; ¹h NMR (300MHz, DMSO-d ₆) δ 12.6 (br, 1H), 9.68 (br, 1H), 6.97 (s, 1H), 4.77 (s, 1H), 2.31 (s, 3H), 2.06 (s, 6H).

Intermediate 1-(4-methoxy-benzyl) in the synthetic preparation example 24 of utilization-3-methyl isophthalic acid H-thieno-[3,2-c] pyrazoles-5-formaldehyde, copy above-mentioned preparation Compound I-9, I-13, and I-6, the method for I-22 can prepare respectively Compound I-25 to I-28.

Synthetic preparation example 29

The 4-(3-ethyl-1H-thieno-[3,2-c] pyrazoles-5-yl)-2 for preparing structural formula (I-29), 6-dimethyl-Isosorbide-5-Nitrae-dihydropyridine-3,5-dimethoxy nitrile

Step 1 reaction scheme is as follows:

Step is: (is preparation process referring to synthetic preparation example for the iodo-1H-thieno-of 1-tertbutyloxycarbonyl-5-methyl 3-[3,2-c] pyrazoles-5-methyl-formiate (8.2g, 20mmol)?) be dissolved in Isosorbide-5-Nitrae-dioxane (200mL), add Pd (dppf) Cl ₂(1.46g, 2mmol), drip zinc ethyl (1M toluene solution, 30mL, 30mmol), finishes nitrogen replacement three times, is heated to 100 ° of C and stirs 1 hour.Add methylene dichloride (200mL) dilute reaction solution, drip 1N hydrochloric acid neutralization reaction liquid to pH 7 left and right, organic phase is through washing, the saturated common salt washing, drying, concentrated, resistates column chromatography (methylene dichloride: methyl alcohol=400:1) obtain 3-ethyl-1H-thieno-[3,2-c] pyrazoles-5-methyl-formiate crude product (4.46g, 106% thick yield).LC-MS(ESI+):211[M+1] ⁺。

Step 2 reaction scheme is as follows:

Step is: 3-ethyl-1H-thieno-[3,2-c] pyrazoles-5-methyl-formiate crude product (9.73g, 46.28mmol) is dissolved in anhydrous tetrahydro furan (250mL), is cooled to-15 ° of C, adds LiAlH ₄(5.27g, 138.83mmol), nitrogen protection is stirred 2 hours, adds LiAlH ₄(1.32g, 34.72mmol), rise to room temperature and continue to stir 1 hour.The careful water (10mL) that drips, stir 10 minutes, then add methyl alcohol (100mL) and methylene dichloride (100mL), stir after ten minutes and filter, add methyl alcohol (50mL) and methylene dichloride (50mL) in filter residue, stir after ten minutes and filter, filtrate merges, through washing, and the saturated common salt washing, dry, concentrated, resistates column chromatography (methylene dichloride: methyl alcohol=100:1), obtain 3-ethyl-1H-thieno-[3,2-c] pyrazoles-5-methyl alcohol (3.85g, 46%).LC-MS(ESI+):183[M+1] ⁺。

Step 3 reaction scheme is as follows:

Step is: 3-ethyl-1H-thieno-[3; 2-c] pyrazoles-5-methyl alcohol (1.82g; 10mmol) be dissolved in methylene dichloride (200mL); add Dess-Martin oxygenant (6.36g, 15mmol), under nitrogen protection, room temperature reaction is 2 hours; reaction solution is concentrated; resistates column chromatography (methylene dichloride: methyl alcohol=300:1) obtain 3-ethyl-1H-thieno-[3,2-c] pyrazoles-5-formaldehyde (1.75g, 97%).LC-MS(ESI+)：181[M+1] ⁺。

Step 4 reaction scheme is as follows:

Step is: 3-ethyl-1H-thieno-[3,2-c] pyrazoles-5-formaldehyde (400mg, 2.2mmol) and amino propenyl cyanide (397mg, 4.84mmol) are dissolved in glacial acetic acid (20mL), are heated to 100 ° of C reactions 1 hour.Reaction solution is concentrated, resistates column chromatography (methylene dichloride: methyl alcohol=80:1) obtain target compound 4-(3-ethyl-1H-thieno-[3,2-c] pyrazoles-5-yl)-2,6-dimethyl-Isosorbide-5-Nitrae-dihydropyridine-3,5-dimethoxy nitrile (136mg, 20%).HPLC shows that purity is 98.2%; LC-MS (ESI+): 310[M+1] ⁺; ¹h NMR (300MHz, CDCl ₃) δ 8.99 (br, 1H), 6.66 (s, 1H), 4.36 (s, 1H), 2.55-2.65 (m, 2H), 1.87 (s, 6H), 1.11 (t, 3H, J=7.5Hz).

Intermediate 3-ethyl in the synthetic preparation example 29 of utilization-1H-thieno-[3,2-c] pyrazoles-5-formaldehyde, copy aforementioned preparation Compound I-9, I-13, and I-6, the method for I-22 can prepare respectively Compound I-30 to I-33.

Synthetic preparation example 34

The 4-(3-methoxyl group-1H-thieno-[3,2-c] pyrazoles-5-yl)-2 for preparing structural formula (I-34), 6-dimethyl-Isosorbide-5-Nitrae-dihydropyridine-3,5-dimethoxy nitrile

Step 1 reaction scheme is as follows:

Step is: the iodo-1H-thieno-[3 of 3-, 2-c] pyrazoles-5-methyl-formiate (10.0g, 32.4mmol) (preparation process is referring to synthetic preparation example 1) be dissolved in DMF (100ml), under frozen water is cooling, add salt of wormwood (9.0g, 64.9mmol), drip methoxyl group benzyl chloride (6.9g, 38.9mmol), stirring at room 2 hours.Add water in system, the ethyl acetate extraction, merge the ester layer, the saturated common salt washing, anhydrous sodium sulfate drying, concentrated, column chromatography purification (EA/PE=1/8), obtain the iodo-1-of near-white solid 3-(4-methoxy-benzyl)-1H-thieno-[3,2-c] pyrazoles-5-methyl-formiate (13.9g, yield 100%).

Step 2 reaction scheme is as follows:

Step is: the iodo-1-of 3-(4-methoxy-benzyl)-1H-thieno-[3; 2-c] pyrazoles-5-methyl-formiate (0.86g, 2mmol), sodium methylate (0.44g, 8mmol), 3; 4; 7,8-tetramethyl-phenanthroline (0.24g, 1mmol), cuprous iodide (0.38g; 2mmol) be mixed in single neck bottle; add anhydrous methanol (50mL), under nitrogen protection, 80 ° of C stir 16 hours.Be cooled to room temperature, in system, add water, the 1N hcl acidifying, dichloromethane extraction, extraction liquid merges, anhydrous sodium sulfate drying, concentrate to obtain 3-methoxyl group-1-(4-methoxy-benzyl)-1H-thieno-[3,2-c] pyrazoles-5-formic acid crude product (1.1g), blackish green solid.LC-MS display-object product (M+1=319) content is about 30%.

Step 3 reaction scheme is as follows:

Step is: under nitrogen protection, upper step gained 3-methoxyl group-1-(4-methoxy-benzyl)-1H-thieno-[3,2-c] pyrazoles-5-formic acid crude product (1.1g) is dissolved in THF(100mL) in, be cooled to-10 ° of C, add LiAlH ₄(500mg), stir 2 hours.Careful drip the shrend reaction of going out, standing, filtration.Filter residue is washed with methylene dichloride, filtrate is with dichloromethane extraction, methylene dichloride merges mutually, with anhydrous sodium sulfate drying, concentrated, column chromatography purification obtains (3-methoxyl group-1-(4-methoxy-benzyl)-1H-thieno-[3,2-c] pyrazoles-5-yl) methyl alcohol (210mg, two step yields 35%), pistac product.LC-MS:M+1=305。

Step 4 reaction scheme is as follows:

Step is: under nitrogen protection; (3-methoxyl group-1-(4-methoxy-benzyl)-1H-thieno-[3; 2-c] pyrazoles-5-yl) methyl alcohol (140mg; 0.46mmol) be dissolved in dry methylene dichloride (20mL); be cooled to-10 ° of C; add Dess-Martin reagent (293mg, 0.69mmol), stir 2 hours.Rise to room temperature, stir 2 hours.Drip saturated sodium bicarbonate aqueous solution, separate the methylene dichloride phase, water is with dichloromethane extraction, and methylene dichloride merges, drying, concentrated, column chromatography purification (ethyl acetate/petroleum ether=1/4), obtain 3-methoxyl group-1-(4-methoxy-benzyl)-1H-thieno-[3,2-c] pyrazoles-5-formaldehyde (115mg, yield 83%), yellow solid. ¹H?NMR(300MHz,CDCl ₃):9.80(s,1H),7.23(d,2H,J=8.4Hz),7.02(s,1H),6.88(d,2H,J=8.4Hz),5.24(s,2H),4.04(s,3H),3.81(s,3H)。

Step 5 reaction scheme is as follows:

Step is: 3-methoxyl group-1-(4-methoxy-benzyl)-1H-thieno-[3,2-c] pyrazoles-5-formaldehyde (237mg, 0.78mmol) is dissolved in Glacial acetic acid (7mL), adds amino propenyl cyanide (142mg, 1.73mmol), and 90 ° of C stir 1 hour.Remove acetic acid under reduced pressure, resistates is 4-(3-methoxyl group-1-(4-methoxy-benzyl)-1H-thieno-[3,2-c] pyrazoles-5-yl)-2,6-dimethyl-Isosorbide-5-Nitrae-dihydropyridine-3,5-dimethoxy nitrile crude product, brown oil.LC-MS:432[M+1]。

Step 6 reaction scheme is as follows:

Step is: add trifluoracetic acid (10mL) in above-mentioned brown oil, return stirring 16 hours.Remove trifluoracetic acid under reduced pressure, the resistates frozen water is cooling, saturated sodium bicarbonate aqueous solution alkalization, dichloromethane extraction.Extraction liquid merges, and anhydrous sodium sulfate drying is concentrated, preparation TLC purifying (methylene chloride/methanol=20:1), obtain 4-(3-methoxyl group-1H-thieno-[3,2-c] pyrazoles-5-yl)-2,6-dimethyl-Isosorbide-5-Nitrae-dihydropyridine-3, the 5-dimethoxy nitrile, faint yellow solid 90mg, yield 37%.MS:(312, M+H); ¹h NMR (DMSO-d ₆, 300MHz): 12.01 (s, 1H), 9.69 (s, 1H), 6.94 (s, 1H), 4.78 (s, 1H), 3.89 (s, 3H), 2.03 (s, 6H); HPLC: purity 96.8%.

With 3-methoxyl group-1-(4-methoxy-benzyl)-1H-thieno-[3,2-c] pyrazoles-5-formaldehyde, for raw material, adopt and aforesaid compound I-9, I-13, the similar preparation method of I-6 can prepare respectively following compound:

Synthetic preparation example 38

The 4-(3-amino-1H-thieno-[3,2-c] pyrazoles-5-yl)-2 for preparing structural formula (I-38), 6-dimethyl-Isosorbide-5-Nitrae-dihydropyridine-3,5-dimethoxy nitrile

Step 1 reaction scheme is as follows:

Step is: the 1-tertiary butyl, the iodo-1H-thieno-of 5-methyl-3-[3,2-c] pyrazoles-1,5-dicarboxylic acid esters (0.82g, 2mmol), Pd ₂(dba) ₃(183mg, 0.2mmol), Xantphos (347mg, 0.6mmol), cesium carbonate (980mg; 3mmol) jointly be dissolved in Isosorbide-5-Nitrae-dioxane (20mL), add benzophenone imine (540mg; 3mmol), nitrogen protection, be heated to 100 ℃ and stir 6 hours.Be cooled to room temperature, remove solvent under reduced pressure, add water and ethyl acetate in resistates, separate organic phase, anhydrous sodium sulfate drying, concentrated.The resistates column chromatography purification, obtain the faint yellow solid 1-tertiary butyl, 5-methyl-3-((diphenylmethyl thiazolinyl) amino)-1H-thieno-[3,2-c] pyrazoles-1,5-dicarboxylic acid esters (0.68g, 74%).

Step 2 reaction scheme is as follows:

Step is: the 1-tertiary butyl, 5-methyl-3-((diphenylmethyl thiazolinyl) amino)-1H-thieno-[3,2-c] pyrazoles-1,5-dicarboxylic acid esters (0.68g, 1.47mmol) is dissolved in methyl alcohol (10mL), adds oxammonium hydrochloride (0.112g, 1.62mmol), stirring at room 6 hours.Remove solvent under reduced pressure, resistates dissolves with methylene dichloride, and saturated sodium bicarbonate aqueous solution is washed, anhydrous sodium sulfate drying, and column chromatography obtains the 1-tertiary butyl, 5-methyl-3-amino-1H-thieno-[3,2-c] pyrazoles-1,5-dicarboxylic acid esters (0.35g, 80%).

Step 3 reaction scheme is as follows:

Step is: 5-methyl-3-amino-1H-thieno-[3,2-c] pyrazoles-1,5-dicarboxylic acid esters (0.35g, 1.18mmol) be dissolved in acetonitrile (20mL), add 4-dimethylamino pyridine (159mg, 1.30mmol) and tert-Butyl dicarbonate (Boc2O) (284mg, 1.30mmol), return stirring 2 hours.Be chilled to room temperature, remove solvent under reduced pressure, the resistates column chromatography purification obtains the white solid 1-tertiary butyl, 5-methyl 3-((tertbutyloxycarbonyl) amino)-1H-thieno-[3,2-c] pyrazoles-1,5-dicarboxylic acid esters (0.32g, 68%).

Step 4 reaction scheme is as follows:

Step is: the 1-tertiary butyl; 5-methyl 3-((tertbutyloxycarbonyl) amino)-1H-thieno-[3; 2-c] pyrazoles-1; 5-dicarboxylic acid esters (0.32g; 0.805mmol) be dissolved in anhydrous tetrahydro furan (10mL), add lithium borohydride (70mg, 3.2mmol); nitrogen protection, stirring at room 5 hours.Reacted rear frozen water cooling, dripped dilute hydrochloric acid (1N, 0.5mL), finished and stir 0.5 hour.Remove solvent under reduced pressure, resistates adds acetic acid ethyl dissolution, washing, dry concentrated rear column chromatography.Obtain the 3-((tertbutyloxycarbonyl) amino)-5-(methylol)-1H-thieno-[3,2-c] pyrazoles-1-t-butyl formate (0.22g, 74%).

Step 5 reaction scheme is as follows:

Step is: ((tertbutyloxycarbonyl) amino)-5-(methylol)-1H-thieno-[3; 2-c] pyrazoles-1-t-butyl formate (0.22g; 0.6mmol) be dissolved in methylene dichloride (10mL); add Dess-Martin oxygenant (509mg; 1.2mmol), under nitrogen protection, stirring at room is 2 hours.Reaction solution is concentrated, and column chromatography purification obtains 3-((tertbutyloxycarbonyl) amino)-5-aldehyde radical-1H-thieno-[3,2-c] pyrazoles-1-t-butyl formate (0.17g, 77%).

Step 6 reaction scheme is as follows:

Step is: 3-((tertbutyloxycarbonyl) amino)-5-aldehyde radical-1H-thieno-[3,2-c] pyrazoles-1-t-butyl carbamate (124mg, 0.336mmol), the amino propenyl cyanide of 3-(61mg, 0.738mmol) adds in Glacial acetic acid (5mL) jointly, 95 ℃ are stirred 15 minutes.Acetic acid is divided exactly in decompression, and resistates prepares the TLC purifying, obtains (5-(3,5-dicyano-2,6-dimethyl-Isosorbide-5-Nitrae-dihydropyridine-4-yl)-1H-thieno-[3,2-c] pyrazole-3-yl) t-butyl carbamate (49mg, 36%).

Step 7 reaction scheme is as follows:

Step is: (5-(3,5-dicyano-2,6-dimethyl-1,4-dihydropyridine-4-yl)-1H-thieno-[3,2-c] pyrazole-3-yl) t-butyl carbamate (67mg, 0.169mmol) be dissolved in methylene dichloride (5mL), add trifluoroacetic acid (1mL), stirring at room 5 hours.Reaction solution dilutes with ethyl acetate (10mL), add solid sodium carbonate (3g), stir 0.5 hour, filter, filtrate is concentrated, preparation TLC purifying (methylene dichloride: methyl alcohol=10:1), obtain 4-(3-amino-1H-thieno-[3,2-c] pyrazoles-5-yl)-2,6-dimethyl-1,4-dihydropyridine-3,5-dimethoxy nitrile (35mg, 70%).ESI-MS:297（M+H）; ¹H?NMR(300MHz,DMSO-d ₆)δ11.46(s,1H),9.69(s,1H),6.78(s,1H),5.08(s,2H),4.69(s,1H),2.03(s,6H)。

Synthetic preparation example 39

Prepare 2 of structural formula (I-39), 6-dimethyl-4-(3-(pyridin-3-yl)-1H-thieno-[3,2-c] pyrazoles-5-yl)-Isosorbide-5-Nitrae-dihydropyridine-3,5-dimethoxy nitrile

Step 1 reaction scheme is as follows:

Step is: (is preparation process referring to synthetic preparation example for the iodo-1H-thieno-of 1-tertbutyloxycarbonyl-5-methyl 3-[3,2-c] pyrazoles-5-methyl-formiate (200mg, 0.49mmol)?), pyridine-3-boric acid (90mg, 0.74mmol), Pd (dppf) Cl ₂-CH ₂cl ₂(42mg, 0.049mmol) and Cs ₂cO ₃(639mg, 1.96mmol) is added in the mixed solvent of Isosorbide-5-Nitrae-dioxane (10mL) and water (4mL), and nitrogen replacement three times is heated to 120 ° of C and stirs 30 minutes.Be cooled to room temperature, reaction solution is concentrated, resistates column chromatography (methylene dichloride: methyl alcohol=75:1) obtain 3-(pyridin-3-yl)-1H-thieno-[3,2-c] pyrazoles-5-methyl-formiate (65mg, 51%).LC-MS(ESI+):260[M+1] ⁺。

Step 2 reaction scheme is as follows:

Step is: 3-(pyridin-3-yl)-1H-thieno-[3; 2-c] pyrazoles-5-methyl-formiate (487mg; 1.878mmol) be dissolved in tetrahydrofuran (THF) (20mL); be cooled to-20 ° of C; add lithium aluminium hydride (214mg under nitrogen protection; 5.635mmol), finish, be heated to room temperature reaction 3 hours.Reaction solution is reacted in the cooling lower dropping water of frozen water (0.4mL) cancellation, filter, filter residue is through methanol wash three times, filtrate is concentrated, resistates column chromatography (methylene dichloride: methyl alcohol=20:1) obtain (3-(pyridin-3-yl)-1H-thieno-[3,2-c] pyrazoles-5-yl) methyl alcohol (276mg, 64%).LC-MS(ESI+)：232[M+H] ⁺。

Step 3 reaction scheme is as follows:

Step is: by (3-(pyridin-3-yl)-1H-thieno-[3; 2-c] pyrazoles-5-yl) methyl alcohol (270mg; 1.167mmol) be dissolved in methylene dichloride (30mL); add Dess-Martin oxygenant (990mg, 2.335mmol), under nitrogen protection, room temperature reaction is 3 hours; reaction solution is concentrated; the resistates column chromatography obtains 3-(pyridin-3-yl)-1H-thieno-[3,2-c] pyrazoles-5-formaldehyde (88mg, 33%).LC-MS(ESI+)：230[M+H] ⁺。

Step 4 reaction scheme is as follows:

Step is: 3-(pyridin-3-yl)-1H-thieno-[3,2-c] pyrazoles-5-formaldehyde (88mg, 0.384mmol) and amino propenyl cyanide (69mg, 0.884mmol) are dissolved in glacial acetic acid (5mL), are heated to 95 ° of C reactions 2 hours.Be cooled to room temperature, reaction solution is concentrated, and the resistates column chromatography obtains 2,6-dimethyl-4-(3-(pyridin-3-yl)-1H-thieno-[3,2-c] pyrazoles-5-yl)-Isosorbide-5-Nitrae-dihydropyridine-3,5-dimethoxy nitrile (52mg, 38%).HPLC shows that purity is 97%; ¹h NMR (300MHz, DMSO) δ 13.43 (s, 1H), 9.77 (s, 1H), 8.99 (s, 1H), 8.54-8.56 (m, 1H), 8.14 (t, 1H, J=3.8Hz), 7.49-7.53 (m, 1H), (7.18 s, 1H), 4.91 (s, 1H), 2.06 (s, 6H); LC-MS (ESI+): 359[M+H] ⁺.

Synthetic preparation example 40

Prepare 2 of structural formula (I-40), 6-dimethyl-4-(3-(pyridin-4-yl)-1H-thieno-[3,2-c] pyrazoles-5-yl)-Isosorbide-5-Nitrae-dihydropyridine-3,5-dimethoxy nitrile

Step is: copy the method for example 39, with the pyridine in pyridine-4-boric acid replacement step 1-3-boric acid, can prepare this target compound.LC-MS(ESI+)：359[M+H] ⁺； ¹H?NMR(300MHz,DMSO)δ13.58(s,1H),9.76(s,1H),8.65(d,2H,J=4.8Hz),7.72(d,2H,J=5.4Hz),7.20(s,1H),4.92(s,1H),2.06(s,6H)。

Synthetic preparation example 41

Prepare 2 of structural formula (I-41), 6-dimethyl-4-(3-(1-methyl isophthalic acid H-pyrazoles-4-yl)-1H-thieno-[3,2-c] pyrazoles-5-yl)-Isosorbide-5-Nitrae-dihydropyridine-3,5-dimethoxy nitrile

Step 1 reaction scheme is as follows:

Step is the iodo-1H-thieno-[3 of 1-tertbutyloxycarbonyl-5-methyl 3-, 2-c] pyrazoles-5-methyl-formiate 2.5g, 6.12mmol) (is preparation process referring to synthetic preparation example?), 1-methyl-4-(4,4,5,5-tetramethyl--1,3,2-dioxa pentaborane-2-yl)-1H-pyrazoles (3.82g, 18.37mmol), Pd (dppf) Cl ₂(896mg, 1.22mmol) and Cs ₂cO ₃(3.99g, 12.24mmol) adds in the mixed solvent of Isosorbide-5-Nitrae-dioxane (125mL) and water (50mL), and nitrogen replacement three times is heated to 100 ° of C reactions 1 hour.Be cooled to room temperature, reaction solution is concentrated, resistates is dissolved in methylene dichloride (100mL) and water (30mL), separates organic phase, washing, the saturated common salt washing, drying, concentrated, resistates column chromatography (methylene chloride/methanol=200/1) obtains compound 3-(1-methyl isophthalic acid H-pyrazoles-4-yl)-1H-thieno-[3,2-c] pyrazoles-5-methyl-formiate (888mg, 55.3%).LC-MS(ESI+)：263[M+1] ⁺。

Step 2 reaction scheme is as follows:

Step is: 3-(1-methyl isophthalic acid H-pyrazoles-4-yl)-1H-thieno-[3,2-c] pyrazoles-5-methyl-formiate (888mg, 3.386mmol) is dissolved in DMF(25mL) in, add K ₂cO ₃(936mg, 6.772mmol), drip PMBCl(636mg, 4.063mmol), finish, stir 4 hours.Reaction solution is concentrated dry, resistates is dissolved in methylene dichloride (50mL) and water (20mL), organic phase is through washing, the saturated common salt washing, drying, concentrated, resistates column chromatography (PE:EA=10:1), obtain compound 1-(4-methoxy-benzyl)-3-(1-methyl isophthalic acid H-pyrazoles-4-yl)-1H-thieno-[3,2-c] pyrazoles-5-methyl-formiate (722mg, 55.8%).LC-MS(ESI+):383[M+1] ⁺.

Step 3 reaction scheme is as follows:

Step is: 1-(4-methoxy-benzyl)-3-(1-methyl isophthalic acid H-pyrazoles-4-yl)-1H-thieno-[3,2-c] pyrazoles-5-methyl-formiate (722mg, 1.89mmol) is dissolved in anhydrous THF(20mL), add LiBH ₄(165mg, mmol), stir 3 hours.Add methylene dichloride (30mL) dilute reaction solution, drip 1N hydrochloric acid and adjust the pH=7 left and right, organic phase is through washing, the saturated common salt washing, dry, resistates column chromatography (PE:EA=6:1) obtains compound (1-(4-methoxy-benzyl)-3-(1-methyl isophthalic acid H-pyrazoles-4-yl)-1H-thieno-[3,2-c] pyrazoles-5-yl) methyl alcohol (403mg, 60%).LC-MS(ESI+):355[M+1] ⁺。

Step 4 reaction scheme is as follows:

Step is: above walk gained (1-(4-methoxy-benzyl)-3-(1-methyl isophthalic acid H-pyrazoles-4-yl)-1H-thieno-[3, 2-c] pyrazoles-5-yl) methyl alcohol (403mg, 1.138mmol) be dissolved in methylene dichloride (20mL), add Dess-Martin reagent (996mg, 2.276mmol), under nitrogen protection, room temperature reaction is 3 hours, TLC(PE:EA=2:1) show that reaction is complete, reaction solution is concentrated, resistates column chromatography (methylene dichloride) obtains compound 1-(4-methoxy-benzyl)-3-(1-methyl isophthalic acid H-pyrazoles-4-yl)-1H-thieno-[3, 2-c] pyrazoles-5-formaldehyde (391mg, 97%).LC-MS(ESI+):353[M+1] ⁺。

Step 5 reaction scheme is as follows:

Step is: by 1-(4-methoxy-benzyl)-3-(1-methyl isophthalic acid H-pyrazoles-4-yl)-1H-thieno-[3,2-c] pyrazoles-5-formaldehyde (391mg, 1.11mmol) and amino propenyl cyanide (200mg, 2.44mmol) be dissolved in glacial acetic acid (10mL), be heated to 100 ° of C and stir 1 hour.Reaction solution is concentrated, resistates 4-(1-(4-methoxy-benzyl)-3-(1-methyl isophthalic acid H-pyrazoles-4-yl)-1H-thieno-[3,2-c] pyrazoles-5-yl)-2,6-dimethyl-1,4-dihydropyridine-3,5-dimethoxy nitrile (533mg, 99% thick yield), not purifiedly be directly used in next step reaction.LC-MS(ESI+):482[M+1] ⁺。

Step 6 reaction scheme is as follows:

Step is: above walk gained resistates (533mg, 1.107mmol) and be dissolved in trifluoroacetic acid (10mL), be heated to 85 ° of C and stir 24 hours.Be cooled to room temperature, reaction solution is concentrated dry, adds methylene dichloride (30mL) in resistates, with saturated sodium bicarbonate solution, wash, water and saturated common salt washing again, drying, concentrated, resistates column chromatography (methylene dichloride: methyl alcohol=50:1) obtain target compound 2,6-dimethyl-4-(3-(1-methyl isophthalic acid H-pyrazoles-4-yl)-1H-thieno-[3,2-c] pyrazoles-5-yl)-Isosorbide-5-Nitrae-dihydropyridine-3,5-dimethoxy nitrile (100mg, 25%).LC-MS(ESI+):362[M+1] ⁺； ¹H?NMR(300MHz,CD ₃OD)δ7.90(s,1H),7.82(s,1H),6.97(s,1H),4.72(s,1H),3.95(s,3H),2.12(s,6H)。

Synthetic preparation example 42

Prepare 2 of structural formula (I-42), 6-dimethyl-4-(3-(thiophene-2-yl)-1H-thieno-[3,2-c] pyrazoles-5-yl)-Isosorbide-5-Nitrae-dihydropyridine-3,5-dimethoxy nitrile

Step 1 reaction scheme is as follows:

Step is: by the iodo-1H-thieno-of 1-tertbutyloxycarbonyl-3-[3,2-c] pyrazoles-5-methyl-formiate (2.0g, 4.9mmol), and thiophene-2-boric acid (940mg, 7.35mmol), Pd (dppf) Cl ₂(717mg, 0.98mmol) and Cs ₂cO ₃(4.79g, 14.7mmol) adds in Isosorbide-5-Nitrae-dioxane (50mL) and water (20mL), and nitrogen replacement three times is heated to 100 ° of C and stirs 1 hour.Be cooled to room temperature, reaction solution is concentrated, resistates is dissolved in methylene dichloride (100mL), washing, saturated common salt washing, drying, concentrated, resistates column chromatography (methylene dichloride: methyl alcohol=200:1) obtain 3-(thiophene-2-yl)-1H-thieno-[3,2-c] pyrazoles-5-methyl-formiate (761mg, 59%).LC-MS(ESI+):265[M+1] ⁺。

Step 2 reaction scheme is as follows:

Step is: 3-(thiophene-2-yl)-1H-thieno-[3,2-c] pyrazoles-5-methyl-formiate (468mg, 1.77mmol) is dissolved in to DMF(15mL) in, add K ₂cO ₃(489mg, 3.54mmol), drip PMBCl (333mg, 2.12mmol), finishes stirring at room 4 hours.The reaction solution concentrating under reduced pressure, resistates is dissolved in methylene dichloride, washing, the saturated common salt washing, drying, concentrated, resistates column chromatography (PE:EA=10:1), obtain 1-(4-methoxy-benzyl)-3-(thiophene-2-yl)-1H-thieno-[3,2-c] pyrazoles-5-methyl-formiate (680mg, 100%).LC-MS(ESI+):385[M+1] ⁺。

Step 3 reaction scheme is as follows:

Step is: by the 1-(4-methoxy-benzyl)-3-(thiophene-2-yl)-1H-thieno-[3,2-c] pyrazoles-5-methyl-formiate (680mg, 1.77mmol) is dissolved in anhydrous tetrahydro furan (20mL), adds LiBH ₄(154mg, 1.08mmol), stirring at room 3 hours.Add methylene dichloride (30mL) dilute reaction solution, drip 1N hydrochloric acid and adjust the pH=7 left and right, organic phase is through washing, the saturated common salt washing, dry, resistates column chromatography (PE:EA=10:1) obtains (1-(4-methoxy-benzyl)-3-(thiophene-2-yl)-1H-thieno-[3,2-c] pyrazoles-5-yl) methyl alcohol (208mg, yield 33%).LC-MS(ESI+):357[M+1] ⁺。

Step 4 reaction scheme is as follows:

Step is: by (1-(4-methoxy-benzyl)-3-(thiophene-2-yl)-1H-thieno-[3; 2-c] pyrazoles-5-yl) methyl alcohol (208mg; 0.49mmol) be dissolved in methylene dichloride (10mL); add Dess-Martin reagent (312mg; 0.74mmol), under nitrogen protection, stirring at room is 3 hours.Reaction solution is concentrated, and resistates column chromatography (methylene dichloride wash-out) obtains (1-(4-methoxy-benzyl)-3-(thiophene-2-yl)-1H-thieno-[3,2-c] pyrazoles-5-yl) formaldehyde (173mg, 84%).LC-MS(ESI+):355[M+1] ⁺。

Step 5 reaction scheme is as follows:

Step is: (1-(4-methoxy-benzyl)-3-(thiophene-2-yl)-1H-thieno-[3,2-c] pyrazoles-5-yl) formaldehyde (173mg, 0.49mmol) and amino propenyl cyanide (88mg, 1.08mmol) be dissolved in glacial acetic acid (5mL), be heated to 100 ° of C and stir 1 hour.Be cooled to room temperature, the reaction solution concentrating under reduced pressure, resistates column chromatography (PE:EA=6:1) obtains 4-(1-(4-methoxy-benzyl)-3-(thiophene-2-yl)-1H-thieno-[3,2-c] pyrazoles-5-yl)-2,6-dimethyl-Isosorbide-5-Nitrae-dihydropyridine-3,5-dimethoxy nitrile (225mg, 95%).LC-MS(ESI+):484[M+1] ⁺。

Step 6 reaction scheme is as follows:

Step is: 4-(1-(4-methoxy-benzyl)-3-(thiophene-2-yl)-1H-thieno-[3,2-c] pyrazoles-5-yl)-2,6-dimethyl-1,4-dihydropyridine-3,5-dimethoxy nitrile (215mg, 0.445mmol) be dissolved in trifluoroacetic acid (7.5mL), be heated to 70 ° of C and stir 8 hours.Reaction solution is concentrated dry, resistates adds methylene dichloride (30mL) to dissolve, and saturated sodium bicarbonate solution is washed, the saturated common salt washing, dry, concentrated, resistates column chromatography (methylene dichloride: methyl alcohol=50:1) obtain 2,6-dimethyl-4-(3-(thiophene-2-yl)-1H-thieno-[3,2-c] pyrazoles-5-yl)-1,4-dihydropyridine-3,5-dimethoxy nitrile (32mg, 20%).HPLC shows that purity is 95.54%; LC-MS (ESI+): 364[M+1] ⁺; ¹h NMR (300MHz, CDCl ₃) δ 13.17 (br, 1H), 9.74 (br, 1H), 7.53 (d, 1H, J=4.5), 7.31 (s, 1H), 7.13-7.18 (m, 2H), 4.89 (s, 1H), 2.06 (s, 6H).

Synthetic preparation example 43

Prepare 2 of structural formula (I-43), 6-dimethyl-4-(3-morpholine-1H-thieno-[3,2-c] pyrazoles-5-yl)-Isosorbide-5-Nitrae-dihydropyridine-3,5-dimethoxy nitrile

Step 1 reaction scheme is as follows:

Step is: the iodo-1-(4-methoxy-benzyl of 3-)-1H-thieno-[3; 2-c] pyrazoles-5-methyl-formiate (0.86g; 2mmol) (preparation process is referring to synthetic preparation example 34), morpholine (0.87g; 10mmol), salt of wormwood (1.38g, 10mmol), L-PROLINE (68mg, 0.5mmol), cuprous iodide (0.38g; 2mmol) be mixed in single neck bottle; add anhydrous DMSO(5mL), under nitrogen protection, 80 ° of C stir 20 hours.Be cooled to room temperature, in system, add water, the ethyl acetate extraction, extraction liquid merges, anhydrous sodium sulfate drying, concentrated, column chromatography for separation obtains 1-(4-methoxy-benzyl)-3-morpholine-1H-thieno-[3,2-c] pyrazoles-5-methyl-formiate, yellow-green colour solid 0.20g, yield 23%.

Step 2 reaction scheme is as follows:

Step is: under nitrogen protection, 1-(4-methoxy-benzyl)-3-morpholine-1H-thieno-[3,2-c] pyrazoles-5-methyl-formiate (0.19g, 0.49mmol) is dissolved in anhydrous tetrahydro furan (20mL), is cooled to-20 ° of C, adds LiAlH ₄(34mg, 0.89mmol), stir 2 hours.Careful drip the shrend reaction of going out, dichloromethane extraction, methylene dichloride is with anhydrous sodium sulfate drying, concentrated, obtains (1-(4-methoxy-benzyl)-3-morpholine-1H-thieno-[3,2-c] pyrazoles-5-yl) methyl alcohol crude product 0.19g, slightly yield 100%.

Step 3 reaction scheme is as follows:

Step is: under nitrogen protection; (1-(4-methoxy-benzyl)-3-morpholine-1H-thieno-[3; 2-c] pyrazoles-5-yl) methyl alcohol (0.19g; 0.49mmol) be dissolved in dry methylene dichloride (10mL); be cooled to-5 ° of C; add Dess-Martin reagent (285mg, 0.67mmol), stir 2 hours.Drip saturated sodium bicarbonate aqueous solution, separate the methylene dichloride phase, drying, concentrated, column chromatography purification (EA/PE=1/4), obtain 1-(4-methoxy-benzyl)-3-morpholine base-1H-thieno-[3,2-c] pyrazoles-5-formaldehyde, yellow solid 110mg, yield 63%.

Step 4 reaction scheme is as follows:

Step is: 1-(4-methoxy-benzyl)-3-morpholine base-1H-thieno-[3,2-c] pyrazoles-5-formaldehyde (357mg, 1.0mmol) is dissolved in Glacial acetic acid (5mL), adds amino propenyl cyanide (180mg, 2.2mmol), be heated to 90 ° of C and stir 1 hour.Remove acetic acid under reduced pressure, remaining brown oil is 4-(1-(4-methoxy-benzyl)-3-morpholine-1H-thieno-[3,2-c] pyrazoles-5-yl)-2,6-dimethyl-Isosorbide-5-Nitrae-dihydropyridine-3, and 5-dimethoxy nitrile crude product, purifying does not directly carry out next step.

Step 5 reaction scheme is as follows:

Step is: 4-(1-(4-the methoxy-benzyl)-3-morpholine-1H-thieno-[3 that above walks gained, 2-c] pyrazoles-5-yl)-2,6-dimethyl-Isosorbide-5-Nitrae-dihydropyridine-3, add trifluoracetic acid (10mL) in 5-dimethoxy nitrile crude product, 80 ° of C stir and spend the night.Be chilled to room temperature, remove solvent under reduced pressure, resistates is cooling with frozen water, the alkalization of dropping saturated sodium bicarbonate aqueous solution, dichloromethane extraction, ethyl acetate extraction, extraction liquid merges, and anhydrous sodium sulfate drying is concentrated, column chromatography (methylene chloride/methanol, 50/1～20/1) obtains 2,6-dimethyl-4-(3-morpholine-1H-thieno-[3,2-c] pyrazoles-5-yl)-Isosorbide-5-Nitrae-dihydropyridine-3, the 5-dimethoxy nitrile, yellow solid 180mg, yield 50%.MS:367[M+H]； ¹H?NMR(DMSO-d ₆,300MHz)11.97(s,1H),9.67(s,1H),6.93(s,1H),4.78(s,1H),3.71(m,4H),3.16(m,4H),2.03(s,6H)。

Synthetic preparation example 44

6-methyl-the 8-(3-morpholine-1H-thieno-[3,2-c] pyrazoles-5-yl)-2,3,4 for preparing structural formula (I-44), 8-tetrahydrochysene-1H-quinolizine-7,9-dimethoxy nitrile

To synthesize 1-(4-methoxy-benzyl) in preparation example 43-3-morpholine base-1H-thieno-[3,2-c] pyrazoles-5-formaldehyde for raw material, copy aforementioned similar method, can prepare this target compound.MS:407[M+H]； ¹H?NMR(DMSO-d ₆,300MHz)δ12.01(br,1H),6.93(s,1H),4.74(s,1H),3.72(m,4H),3.57(m,2H),3.18(m,4H),2.62(m,2H),1.75-1.63(m,4H)。

Synthetic preparation example 45

Prepare the 6-methyl-8-(3-(4-methylpiperazine-1-yl)-1H-thieno-[3,2-c] pyrazoles-5-yl)-1,3,4 of structural formula (I-45), the 8-tetrahydropyridine is [2,1-c] [Isosorbide-5-Nitrae] oxazines-7 also, the 9-dimethoxy nitrile

Step 1 reaction scheme is as follows:

Step is: the iodo-1-(4-methoxy-benzyl of 3-)-1H-thieno-[3; 2-c] pyrazoles-5-methyl-formiate (1.29g; 3mmol) (is preparation process referring to synthetic preparation example?), N methyl piperazine (0.905g; 9mmol), salt of wormwood (2.08g; 15mmol), L-PROLINE (104mg, 0.9mmol), cuprous iodide (0.29g, 1.5mmol) are mixed in single neck bottle; adding DMSO(5mL) under nitrogen protection, 80 ° of C stir and spend the night.Be cooled to room temperature, add water in system, the ethyl acetate extraction, extraction liquid merges, anhydrous sodium sulfate drying, evaporating column chromatography, obtain 1-(4-methoxy-benzyl)-3-(4-methylpiperazine-1-yl)-1H-thieno-[3,2-c] pyrazoles-5-methyl-formiate, yellow-green colour solid 0.42g, yield 35%.

Step 2 reaction scheme is as follows:

Step is: under nitrogen protection; 1-(4-methoxy-benzyl)-3-(4-methylpiperazine-1-yl)-1H-thieno-[3,2-c] pyrazoles-5-methyl-formiate (250mg, 0.624mmol) is dissolved in anhydrous tetrahydro furan (20mL); be cooled to-20 ° of C, add LiAlH ₄, stir 2 hours.Careful drip the shrend reaction of going out, dichloromethane extraction, methylene dichloride is with anhydrous sodium sulfate drying, concentrated, obtain (1-(4-methoxy-benzyl)-3-(4-methylpiperazine-1-yl)-1H-thieno-[3,2-c] pyrazoles-5-yl) methyl alcohol crude product 200mg, thick yield 74%.

Step 3 reaction scheme is as follows:

Step is: under nitrogen protection; (1-(4-methoxy-benzyl)-3-(4-methylpiperazine-1-yl)-1H-thieno-[3; 2-c] pyrazoles-5-yl) methyl alcohol (200mg; 0.537mmol) be dissolved in dry methylene dichloride (10mL); be cooled to-5 ° of C; add Dess-Martin reagent (312mg, 0.736mmol), stir 2 hours.Drip saturated sodium bicarbonate aqueous solution, separate organic phase, dry, concentrated, column chromatography purification (methylene chloride/methanol=50/1), obtain (1-(4-methoxy-benzyl)-3-(4-methylpiperazine-1-yl)-1H-thieno-[3,2-c] pyrazoles-5-yl) formaldehyde, yellow solid 144mg, yield 72%.

Step 4 reaction scheme is as follows:

Step is: (1-(4-methoxy-benzyl)-3-(4-methylpiperazine-1-yl)-1H-thieno-[3,2-c] pyrazoles-5-yl) methyl alcohol (250mg, 0.675mmol) methylene dichloride (15mL) solution in add cyano propanone sodium salt (78mg, 0.742mmol), piperidines (6mg, 0.07mmol), acetic acid (51mg, 0.843mmol) and 4A molecular sieve (0.2g), finish, be heated to return stirring and spend the night.Reaction solution is cooling, filter, saturated sodium bicarbonate solution and water washing for solid, solid is dissolved in to ethanol, insolubles filters, and filtrate is concentrated dry, and the resistates column chromatography obtains (E)-2-((1-(4-methoxy-benzyl)-3-(4-methylpiperazine-1-yl)-1H-thieno-[3,2-c] pyrazoles-5-yl) methylene radical)-3-oxygen butyronitrile (275mg, 94%).

Step 5 reaction scheme is as follows:

Step is: nitrogen protection; 2-((1-(4-methoxy-benzyl)-3-(4-methylpiperazine-1-yl)-1H-thieno-[3; 2-c] pyrazoles-5-yl) methylene radical)-3-oxygen butyronitrile (500mg; 1.15mmol) and 2-(morpholine-3-thiazolinyl) acetonitrile (210mg; 1.70mmol) jointly be dissolved in Glacial acetic acid (10mL), 90 ° of C stir 1 hour.Remove acetic acid under reduced pressure, remaining brown oil is 8-(1-(4-methoxy-benzyl)-3-(4-methylpiperazine-1-yl)-1H-thieno-[3,2-c] pyrazoles-5-yl)-the 6-methyl isophthalic acid, 3,4,8-tetrahydropyridine also [2,1-c] [1,4] oxazines-7,9-dimethoxy nitrile crude product, not purifiedly directly carry out next step reaction.

Step 6 reaction scheme is as follows:

Step is: 8-(1-(4-methoxy-benzyl)-3-(4-the methylpiperazine-1-yl)-1H-thieno-[3 that above walks gained, 2-c] pyrazoles-5-yl)-the 6-methyl isophthalic acid, 3,4, the 8-tetrahydropyridine is [2,1-c] [Isosorbide-5-Nitrae] oxazines-7 also, add trifluoroacetic acid (10mL) in 9-dimethoxy nitrile crude product, return stirring spends the night.Be chilled to room temperature, remove solvent under reduced pressure, resistates is cooling with frozen water, drips the saturated sodium bicarbonate aqueous solution alkalization, dichloromethane extraction, the ethyl acetate extraction, extraction liquid merges, anhydrous sodium sulfate drying, concentrated, preparation TLC divide to obtain target compound, yellow solid 83mg, two step yields 17%.MS:422[M+H]； ¹H?NMR(CDCl ₃，300MHz)6.79(s,1H),4.57(m,3H),4.00(d,2H),3.52(d,2H),3.37(t,4H),2.57(t,4H),2.44(s,3H),2.27(s,3H)。

Synthetic preparation example 46:

The 4-(3-(2-hydroxyl-oxethyl for preparing structural formula (I-46))-1H-thieno-[3,2-c] pyrazoles-5-yl)-2,6-dimethyl-Isosorbide-5-Nitrae-dihydropyridine-3,5-dimethoxy nitrile

Step 1 reaction scheme is as follows:

Step is: the iodo-1-(4-methoxy-benzyl of 3-)-1H-thieno-[3,2-c] pyrazoles-5-methyl-formiate (1.29g, 3mmol) (preparation process is referring to synthetic preparation example 34), 3; 4,7,8-tetramethyl--1; 10-phenanthroline (472mg; 2mmol), cesium carbonate (4.89g, 15mmol), glycol monomethyl tertbutyl ether (106mg, 0.9mmol), cuprous iodide (0.29g; 1.5mmol) be mixed in single neck bottle; add toluene (5mL), under nitrogen protection, 100 ° of C stir and spend the night.Be cooled to room temperature, in system, add water, the ethyl acetate extraction, extraction liquid merges, and anhydrous sodium sulfate drying concentrates to obtain 3-(2-tert.-butoxy oxyethyl group)-1-(4-methoxy-benzyl)-1H-thieno-[3,2-c] pyrazoles-5-formic acid crude product, yellow-green colour solid 3.4g.

Step 2 reaction scheme is as follows:

Step is: above walk gained 3-(2-tert.-butoxy oxyethyl group)-1-(4-methoxy-benzyl)-1H-thieno-[3,2-c] pyrazoles-5-formic acid dissolving crude product in anhydrous tetrahydro furan (60mL), be cooled to-20 ° of C, add LiAlH ₄(1.05g, 27.66mmol), stir 2 hours.Careful drip the shrend reaction of going out, dichloromethane extraction, methylene dichloride is with anhydrous sodium sulfate drying, concentrated, silica gel column chromatography obtains (3-(2-tert.-butoxy oxyethyl group)-1-(4-methoxy-benzyl)-1H-thieno-[3,2-c] pyrazoles-5-yl) methyl alcohol, yellow solid 120mg, two step yields 10%.

Step 3 reaction scheme is as follows:

Step is: under nitrogen protection; (3-(2-tert.-butoxy oxyethyl group)-1-(4-methoxy-benzyl)-1H-thieno-[3; 2-c] pyrazoles-5-yl) methyl alcohol (120mg; 0.307mmol) be dissolved in dry methylene dichloride (10mL); be cooled to-5 ° of C; add Dess-Martin reagent (179mg, 0.42mmol), stir 2 hours.Drip saturated sodium bicarbonate aqueous solution, separate the methylene dichloride phase, dry, concentrated, column chromatography purification (methylene chloride/methanol=50/1), obtain 3-(2-tert.-butoxy oxyethyl group)-1-(4-methoxy-benzyl)-1H-thieno-[3,2-c] pyrazoles-5-yl) formaldehyde, yellow solid 100mg, yield 83%.

Step 4 reaction scheme is as follows:

Step is: under nitrogen protection; 3-(2-tert.-butoxy oxyethyl group)-1-(4-methoxy-benzyl)-1H-thieno-[3; 2-c] pyrazoles-5-yl) formaldehyde (100mg; 0.257mmol) be dissolved in Glacial acetic acid (5mL); add amino propenyl cyanide (47mg; 0.566mmol), finish and be heated to 90 ° of C stirrings 1.5 hours.Remove acetic acid under reduced pressure, remaining oily matter is 4-(3-(2-tert.-butoxy oxyethyl group)-1-(4-methoxy-benzyl)-1H-thieno-[3,2-c] pyrazoles-5-yl)-2,6-dimethyl-1,4-dihydropyridine-3,5-dimethoxy nitrile crude product, not purifiedly directly carry out next step.

Step 5 reaction scheme is as follows:

Step is: 4-(3-(2-tert.-butoxy oxyethyl group)-1-(4-the methoxy-benzyl)-1H-thieno-[3 that above walks gained, 2-c] pyrazoles-5-yl)-2,6-dimethyl-Isosorbide-5-Nitrae-dihydropyridine-3, add trifluoroacetic acid (10mL) in 5-dimethoxy nitrile crude product, return stirring spends the night.Be chilled to room temperature, solvent is steamed in decompression, and resistates is cooling with frozen water, the alkalization of dropping saturated sodium bicarbonate aqueous solution, dichloromethane extraction, ethyl acetate extraction, extraction liquid merges, and anhydrous sodium sulfate drying is concentrated, preparation TLC separates to obtain white solid 4-(3-(2-hydroxyl-oxethyl)-1H-thieno-[3,2-c] pyrazoles-5-yl)-2,6-dimethyl-Isosorbide-5-Nitrae-dihydropyridine-3,5-dimethoxy nitrile (28mg, two step yields 32%).MS:342[M+H]； ¹H?NMR(CDCl ₃300MHz)：12.07(s,1H),9.69(s,1H),6.93(s,1H),4.77(s,1H),4.18(t,2H),3.68(t,2H),2.03(s,6H)。

Bioassay embodiment 1: the In-vitro Inhibitory Effect of compound to the c-Met enzyme

Utilize the method for Mobility Shift Assay, measure compound in vitro to the restraining effect of c-Met enzyme.

Experimental technique

1. prepare kinase buffer liquid and the stop buffer of 1.25x

1.1 not containing MnCl ₂1.25 times of kinase buffer liquid 62.5mM HEPES, pH 7.5;

0.001875%?Brij-35；

12.5mM?MgCl ₂；

2.5mM?DTT；

1.2 containing MnCl ₂1.25 times of kinase buffer liquid.

62.5mM?HEPES,pH?7.5；

0.001875%?Brij-35；

12.5mM?MgCl ₂；

12.5mM?MnCl ₂；

2.5mM?DTT；

1.3 stop buffer

100mM?HEPES,pH?7.5；

0.015%?Brij-35；

0.2%?Coating?Reagent?#3；

50mM?EDTA。

2. compound solution preparation

2.1 diluted chemical compound

The 50mM compound that adds 20 μ L in the EP pipe, add 100% DMSO of 80 μ L, is made into the 10mM compound of 100 μ L.Add the 10mM compound of 30 μ L in another EP pipe, add 100% DMSO of 70 μ L, be made into the 3mM compound of 100 μ L.

Add 100% DMSO of 95 μ L and the 3mM compound of 5uL in second hole on 96 orifice plates, other holes add 100% DMSO of 60 μ L.Get 30 μ L compounds and add in the 3rd hole from the 2nd hole, down do successively 3 times of dilutions, dilute altogether 10 concentration.The compound concentration scope is 150uM to 7.6nM.

2.2 transferase 45 times compound is to Sptting plate

Get 10 μ L to another piece 96 orifice plates from each hole of above-mentioned 96 orifice plates, add 90 μ L ultrapure waters.Therefore in the second hole to the 11-holes, being the compound be dissolved in 10% DMSO, is 10% DMSO in the first hole and the 12 hole.Take out 5 μ L to 384 hole Sptting plates from above-mentioned 96 orifice plates.Therefore, 5 times of compounds that just have 10% DMSO of 5 μ L to dissolve in 384 hole Sptting plates and 10% DMSO of 5 μ L.The EDTA that adds 5 μ L 250mM in negative control hole.

3. kinase reaction

3.1 prepare 2.5 times of enzyme solution

Kinases is added to 1.25 times of kinase buffer liquid, form 2.5 times of enzyme solution.

3.2 prepare the substrate solution of 2.5 times

The polypeptide of FAM mark and ATP are added to 1.25 times of kinase buffer liquid, form 2.5 times of substrate solutions.

3.3 add enzyme solution in 384 orifice plates

5 times of compounds that in 384 hole Sptting plates, 10% DMSO of existing 5 μ L dissolves.

2.5 times of enzyme solution that add 10 μ L in 384 hole Sptting plates.

Under room temperature, hatch 10 minutes.

3.4 add substrate solution in 384 orifice plates

2.5 times of substrate solutions that add 10 μ L in 384 hole Sptting plates.

3.5 kinase reaction and termination

Under 28 ℃, hatch 1 hour.Add 25 μ L stop buffer termination reactions.

4.Caliper reading of data

The upper reading and converting rate of Caliper data.

5. inhibiting rate calculates

Copy conversion data from Caliper.Transformation efficiency is changed into to the inhibiting rate data.Wherein max refers to the transformation efficiency of DMSO contrast, and min refers to the transformation efficiency of without enzyme, living and contrasting.

Inhibiting rate=(max-conversion)/(max-min) * 100

Bioassay embodiment 1: experimental result

Remarks: "+" represents 500nM<IC ₅₀<5uM; " ++ " represents 50nM<IC ₅₀<500nM; " +++" represents IC ₅₀<50nM.

Claims

1. dihydropyridine compounds, is characterized in that, for thering is the compound of following general formula (I):

Wherein,

2. dihydropyridine compounds as claimed in claim 1, is characterized in that, described general formula (I) compound is selected from a kind of in the compound shown in (I-1) to (I-46) as follows:

3. dihydropyridine compounds as claimed in claim 1, is characterized in that, described general formula (I) compound is any one or both or three's the mixture arbitrarily in enantiomer, diastereomer, conformer.

4. dihydropyridine compounds as claimed in claim 1, is characterized in that, described general formula (I) compound is the pharmacy acceptable derivates.

5. dihydropyridine compounds as claimed in claim 1, is characterized in that, described general formula (I) compound exists with the form of pharmacy acceptable salt.

6. dihydropyridine compounds as claimed in claim 5, it is characterized in that hydrochloride, vitriol, phosphoric acid salt, acetate, trifluoroacetate, mesylate, fluoroform sulphonate, tosilate, tartrate, maleate, fumarate, succinate or malate that described pharmacy acceptable salt is general formula (I) compound.

7. the preparation method of a dihydropyridine compounds claimed in claim 1, the intermediate that wherein said general formula (I) compound can be (II) by corresponding structural formula is prepared by the reaction formula of following route 1,

route 1:

Wherein, R ₁, R ₂, R ₃, R ₄definition is described with claim 1; PG is the amido protecting group.

8. preparation method as claimed in claim 7, is characterized in that, PG is tertbutyloxycarbonyl, benzyloxycarbonyl, to methoxy-benzyl, 3, the 4-dimethoxy-benzyl.

9. preparation method as claimed in claim 7, is characterized in that, R in intermediate (II) ₁during for H, its preparation method is as shown in Scheme 2: the 5-thiotolene of structural formula 1-2-formic acid through nitrosonitric acid nitrated the intermediate of structural formula 2, the intermediate that is structural formula 3 by the intermediate carboxylic esterification of structural formula 2, adopting reduced iron powder is the amino intermediate that obtains structural formula 4 by the nitroreduction of the intermediate of structural formula 3, the intermediate that is then structural formula 5 by the intermediate cyclisation of structural formula 4 under the existence of acetic anhydride, nitrite; The intermediate ethanoyl of structural formula 5 is removed to obtain to the intermediate of structural formula 6, then the ester group in the intermediate of structural formula 6 is reduced to the intermediate that alcohol radical obtains structural formula 7, then the intermediate oxidation of structural formula 7 is obtained to the intermediate that structural formula is (II);

route 2:

10. preparation method as claimed in claim 7, is characterized in that, R in intermediate (II) ₁while being not H, its preparation method is as shown in Scheme 3: the 5-thiotolene of structural formula 1-2-formic acid through nitrosonitric acid nitrated the intermediate of structural formula 2, the intermediate that is structural formula 3 by the intermediate carboxylic esterification of structural formula 2, adopting reduced iron powder is the amino intermediate that obtains structural formula 4 by the nitroreduction of the intermediate of structural formula 3, the intermediate that is then structural formula 5 by the intermediate cyclisation of structural formula 4 under the existence of acetic anhydride, nitrite; The intermediate ethanoyl of structural formula 5 is removed to obtain to the intermediate of structural formula 6, by the intermediate of structural formula 66 iodos, obtain the intermediate of structural formula 8; The pyrazoles amino of the intermediate of structural formula 8, with the protection of amido protecting group, is obtained to the intermediate of structural formula 9; Then utilize the iodine of the intermediate of structural formula 9 to carry out the intermediate that coupling obtains structural formula 10; Again the ester group of the intermediate of structural formula 10 is reduced to the intermediate that alcohol radical obtains structural formula 11,, the alcohol radical in the intermediate of structural formula 11 is oxidized to aldehyde radical, obtain the intermediate that structural formula is (II);

route 3:

10. preparation method as claimed in claim 9, is characterized in that, described coupling is selected from Suzuki coupling, Buchwald coupling.

11. preparation method as claimed in claim 7, is characterized in that, when the compound of required preparation is the compound shown in general formula (Ia), and R '=R ₃=R ₅the time, its preparation method as shown in Scheme 4:

route 4:

12. preparation method as claimed in claim 7, is characterized in that, when the compound of required preparation is the compound shown in general formula (Ib), and R ₃when different from R5, its preparation method as shown in Scheme 5:

route 5:

13. preparation method as claimed in claim 7, is characterized in that, when the compound of required preparation is the compound shown in general formula (Ic), and R ₃with R ₅difference and R ₄while being not H, its preparation method as shown in Scheme 6:

route 6:

14. preparation method as claimed in claim 7, is characterized in that, when the compound of required preparation is the compound shown in general formula (Id), its preparation method as shown in Scheme 7:

route 7:

Wherein X is CH ₂or O.

15. preparation method as claimed in claim 7, is characterized in that, when the compound of required preparation is the compound shown in general formula (Ie), its preparation method as shown in Scheme 8:

route 8:

16. preparation method as claimed in claim 7, is characterized in that, when the compound of required preparation is the compound shown in general formula (If), its preparation method as shown in Scheme 9:

route 9:

Wherein X is CH ₂or O.

17. the pharmaceutical composition containing dihydropyridine compounds claimed in claim 1, wherein said pharmaceutical composition comprises general formula (I) compound and the acceptable vehicle of pharmacy for the treatment of significant quantity.

18., as the pharmaceutical composition of claim 17, wherein said pharmaceutical composition is made tablet, capsule, aqueous suspension, oiliness suspensoid, dispersible pulvis, granule, lozenge, emulsion, syrup, ointment, ointment, suppository or injection.

19. a pharmaceutical composition, pharmacy acceptable derivates and the acceptable vehicle of pharmacy that wherein said pharmaceutical composition comprises general formula (I) compound for the treatment of significant quantity.

20., as the pharmaceutical composition of claim 19, wherein said pharmaceutical composition is made tablet, capsule, aqueous suspension, oiliness suspensoid, dispersible pulvis, granule, lozenge, emulsion, syrup, ointment, ointment, suppository or injection.

21. a pharmaceutical composition, pharmacy acceptable salt and the acceptable vehicle of pharmacy that wherein said pharmaceutical composition comprises general formula (I) compound for the treatment of significant quantity.

22., as the pharmaceutical composition of claim 21, wherein said pharmaceutical composition is made tablet, capsule, aqueous suspension, oiliness suspensoid, dispersible pulvis, granule, lozenge, emulsion, syrup, ointment, ointment, suppository or injection.

23. the application of a dihydropyridine compounds claimed in claim 1 is wherein that general formula (I) compound is regulated the application in protein kinase catalytic activity goods in preparation.

24. application as claimed in claim 23, wherein said protein kinase is the c-Met receptor tyrosine kinase.

25. the application of a dihydropyridine compounds claimed in claim 1 is wherein that the pharmacy acceptable derivates of general formula (I) compound is regulated the application in protein kinase catalytic activity goods in preparation.

26. application as claimed in claim 25, wherein said protein kinase is the c-Met receptor tyrosine kinase.

27. the application of a dihydropyridine compounds claimed in claim 1 is wherein that the pharmaceutically useful salt of general formula (I) compound is regulated the application in protein kinase catalytic activity goods in preparation.

28. application as claimed in claim 27, wherein said protein kinase is the c-Met receptor tyrosine kinase.

29. the application of the described pharmaceutical composition of claim 17 is wherein the application of pharmaceutical composition in the medicine for preparing the treatment disease relevant with protein kinase.

30. application as claimed in claim 29, wherein said protein kinase is the c-Met receptor tyrosine kinase.

31. application as claimed in claim 29, the relevant disease of wherein said protein kinase is cancer.

32. application as claimed in claim 31, wherein said cancer is thyroid carcinoma, colorectal carcinoma, cancer of the stomach, kidney, liver cancer, lung cancer, ovarian cancer, mammary cancer, prostate cancer, bladder cancer, head and neck cancer, carcinoma of the pancreas, carcinoma of gallbladder, osteosarcoma, rhabdosarcoma, MFH/ fibrosarcoma, glioblastoma/astrocytoma, melanoma or mesothelioma.

33. the application of the described pharmaceutical composition of claim 19 is wherein the application of pharmaceutical composition in the medicine for preparing the treatment disease relevant with protein kinase.

34. application as claimed in claim 33, wherein said protein kinase is the c-Met receptor tyrosine kinase.

35. application as claimed in claim 33, the relevant disease of wherein said protein kinase is cancer.

36. application as claimed in claim 35, wherein said cancer is thyroid carcinoma, colorectal carcinoma, cancer of the stomach, kidney, liver cancer, lung cancer, ovarian cancer, mammary cancer, prostate cancer, bladder cancer, head and neck cancer, carcinoma of the pancreas, carcinoma of gallbladder, osteosarcoma, rhabdosarcoma, MFH/ fibrosarcoma, glioblastoma/astrocytoma, melanoma or mesothelioma.

37. the application of the described pharmaceutical composition of claim 21 is wherein the application of pharmaceutical composition in the medicine for preparing the treatment disease relevant with protein kinase.

38. application as claimed in claim 37, wherein said protein kinase is the c-Met receptor tyrosine kinase.

39. application as claimed in claim 37, the relevant disease of wherein said protein kinase is cancer.

40. application as claimed in claim 39, wherein said cancer is thyroid carcinoma, colorectal carcinoma, cancer of the stomach, kidney, liver cancer, lung cancer, ovarian cancer, mammary cancer, prostate cancer, bladder cancer, head and neck cancer, carcinoma of the pancreas, carcinoma of gallbladder, osteosarcoma, rhabdosarcoma, MFH/ fibrosarcoma, glioblastoma/astrocytoma, melanoma or mesothelioma.