CN103467481B

CN103467481B - Dihydropyridine compounds, a combination thereof thing, preparation method and purposes

Info

Publication number: CN103467481B
Application number: CN201210187107.2A
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: 2016-08-31
Anticipated expiration: 2032-06-07
Also published as: CN103467481A

Abstract

Dihydropyridine compounds disclosed by the invention, for having the compound of below general formula (I):Wherein, R₁Selected from hydrogen, halogen, alkyl, replacement alkyl, alkyl oxy, substituted alkyl oxy, alkyl amine group, substituted alkyl amine group, aryl, substituted aryl, heteroaryl, substituted heteroaryl, heterocyclic radical, substituted heterocyclic radical, ester group, amide groups；R₂Selected from hydrogen, cyano group, alkyl, replacement alkyl, thiazolinyl, substituted alkenyl, acyl group；Or, R₂With R₃Formed and ring；R₃Selected from alkyl, replace alkyl, aryl, substituted aryl, heteroaryl, substituted heteroaryl；Or, R₃With R₂Or R₄Formed and ring；R₄Selected from hydrogen, alkyl, replacement alkyl；Or, R⁴With R³Or R⁵Formed and ring；R₅Selected from alkyl, replace alkyl, aryl, substituted aryl, heteroaryl, substituted heteroaryl；Or, R₅With R₄Formed and ring.The invention also discloses the application in terms for the treatment of cancer drug of the compounds process for production thereof of this logical formula (I), the compositions of the compound containing this logical formula (I) and this pharmaceutical composition.

Description

Dihydropyridine compounds, a combination thereof thing, preparation method and purposes

Technical field

The present invention relates to a kind of dihydropyridine compounds, its preparation method, containing them as the medicine group of active component Compound, and it is relevant in order to treat the disease relevant to tyrosine kinase c-Met, particularly c-Met as medicine The purposes of cancer.

Background of invention

Cancer is the number one killer threatening whole world human life healthy.Although the progress of medical science makes the mankind control for cancer Treat and there are the newest means, but cancer is still considered as the most unsolved difficult medical problem at present.Cause the cause of disease of cancer very Many, in recent years, the development of the subject such as molecular weight tumor, molecular pharmacology makes the essence of tumor the most progressively illustrate, people Gradually recognize that the essence of cell carcinogenesis is that intracellular signal transduction pathway is lacked of proper care the cell infinite multiplication caused.

As the of paramount importance member of participation cellular signal transduction, protein tyrosine kinase (protein tyrosine Kinases, PTKs, be called for short tyrosine kinase) it is modal growth factor receptors, generation with tumor and develop close Cut is closed.

The hyperactivity of tyrosine kinase, causes signal pathway downstream to activate, thus causes cell transformation, propagation, right Anti-apoptotic, promotion cell survival, ultimately result in the formation of tumor.Therefore, the research and development of antitumor drug become in recent years Gesture starts to turn to the medicine for intracellular improper signal conduction from traditional cell toxicity medicament, and has related drugs to answer successively For clinic.Compared with traditional cell toxicant series antineoplastic medicament, this kind of therapeutic effects of molecular targeted agents is strong, toxic and side effects is little, It is increasingly becoming the focus of current antineoplastic medicine research and development.

Tyrosine kinase divides receptor type tyrosine kinase and non-receptor tyrosine kinase two kinds.

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 such as Src kinase families, Jak, FAK etc., include again multiple in each kinase families Hypotype.

Hepatic growth factor receptor c-Met is one (Park et al., the Proc.Natl. of receptor type tyrosine kinase Acad.Sci.USA 84:6379-83,1987；Bottaro et al., Science 2S 1:802-4,1991), Outside alpha subunit and β subunit by high glycosylation are together with extracellular domain, transmembrane segment and cytoplasmic tyrosine kinase territory Composition.Its endogenic ligand be hepatocyte growth factor (hepatocyte growth factor, HGF) (Nature, 327: 239-242(1987)；J.Cell Biol., 111:2097-2108 (1990)), part zygotic induction c-Met Dimerization, generates the activated receptor of autophosphorylation, promotes the signal transduction in downstream, mediates multiple response in tumor cell, Including epithelial cell and the increment of endotheliocyte, epithelial cell movement, cell survival and morphologic change and promotion is stimulated to invade Enter.Additionally, HGF regulates angiogenesis, for the growth of tumor with spread extremely important.C-Met and part thereof exist Process LAN in kinds of tumors (including thyroid carcinoma, ovarian cancer, cancer of pancreas etc.) also illustrate that its sending out in these tumors Effect during exhibition.At present, c-Met is as the action target spot of antitumor drug, and its advantage is progressively illustrated (Nature Reviews Cancer,2012,12,89-103)。

In the primary tumor played a key effect at c-Met receptor activation and secondary tumors transfer, targeting HGF or The biological substance (ribozyme, antibody and antisense RNA) of c-Met can suppress the generation of tumor, the selectivity of targeting c-Met Micromolecular inhibitor is the most predicted has treatment potentiality.WO2009091374, WO2009149836, WO2011003604, The patents such as WO2011042367, WO2011042368, CN200910247948.6, CN201010175273.1 are all wrapped Selective c-Met micromolecular inhibitor, Preparation Method And The Use are contained.

Dihydropyridine compounds as tyrosine kinase inhibitor involved in the present invention, as tyrosine-kinase enzyme level Agent, particularly c-Met inhibitor, be never reported.

Summary of the invention

One of the technical problem to be solved is to provide a kind of dihydropyridine compounds.

The two of the technical problem to be solved are to provide the preparation method of above-mentioned dihydropyridine compounds.

The three of the technical problem to be solved are to provide the pharmaceutical composition containing above-mentioned dihydropyridine compounds.

The four of the technical problem to be solved are to provide the purposes of above-mentioned dihydropyridine compounds.

As the dihydropyridine compounds of first aspect present invention, for having the compound of below general formula (I):

Wherein,

R₁Selected from hydrogen, halogen, alkyl, replacement alkyl, alkyl oxy, substituted alkyl oxy, alkyl amine group, take The alkyl amine group in generation, aryl, substituted aryl, heteroaryl, substituted heteroaryl, heterocyclic radical, substituted heterocyclic radical, ester group, Amide groups；

R₂Selected from hydrogen, cyano group, alkyl, replacement alkyl, thiazolinyl, substituted alkenyl, acyl group；Or, R₂With R₃Formed And ring；

R₃Selected from alkyl, replace alkyl, aryl, substituted aryl, heteroaryl, substituted heteroaryl；Or, R₃With R₂ Or R₄Formed and ring；

R₄Selected from hydrogen, alkyl, replacement alkyl；Or, R⁴With R³Or R⁵Formed and ring；

R₅Selected from alkyl, replace alkyl, aryl, substituted aryl, heteroaryl, substituted heteroaryl；Or, R₅With R₄ Formed and ring.

More specifically, the present invention lead to formula (I) compound preferably be selected from as follows (I-1) in the compound shown in (I-46) One:

Described logical formula (I) compound is any one in enantiomer, diastereomer, conformer Or arbitrarily both or the mixture of three.

Described logical formula (I) compound is pharmaceutically acceptable derivant.

Logical formula (I) compound of the present invention can exist as a pharmaceutically acceptable salt form.

Pharmaceutically acceptable salt of the present invention is the logical hydrochlorate of formula (I) compound, sulfate, phosphate, second Hydrochlorate, trifluoroacetate, mesylate, fluoroform sulphonate, tosilate, tartrate, maleate, Fumarate, succinate or malate.

As the preparation method of the logical formula (I) compound of second aspect present invention, can be (II) by corresponding structural formula Intermediate be prepared by the reaction equation of following route 1,

Route 1:

Wherein, R₁, R₂, R₃, R₄As defined above；PG is amido protection group.PG is preferably tertbutyloxycarbonyl, benzyloxy Base carbonyl, to methoxy-benzyl, 3,4-dimethoxy-benzyl.

Specifically, as R in intermediate (II)₁During for H, its preparation method as shown in Scheme 2: the 5-of structural formula 1 Methylthiophene-2-formic acid obtains the intermediate of structural formula 2 through fuming nitric aicd nitrification, the Intermediate carboxylic acids of structural formula 2 is esterified For the intermediate of structural formula 3, use reduced iron powder that the nitro of the intermediate of structural formula 3 is reduced to amino and obtain structural formula 4 Intermediate, then in the presence of acetic anhydride, nitrite, the intermediate of structural formula 4 is cyclized as in structural formula 5 Mesosome；The intermediate acetyl of structural formula 5 is removed to obtain the intermediate of structural formula 6, then by the intermediate of structural formula 6 Ester group be reduced to alcohol radical and obtain the intermediate of structural formula 7, then the intermediate oxidation of structural formula 7 is obtained structural formula for (II) Intermediate；

Route 2:

As R in intermediate (II)₁When being not H, its preparation method as shown in Scheme 3: the 5-methyl thiazolium of structural formula 1 Fen-2-formic acid obtains the intermediate of structural formula 2 through fuming nitric aicd nitrification, is esterified the Intermediate carboxylic acids of structural formula 2 for structure The intermediate of formula 3, uses reduced iron powder that the nitro of the intermediate of structural formula 3 is reduced to amino and obtains the centre of structural formula 4 Body, is then cyclized the intermediate for structural formula 5 by the intermediate of structural formula 4 in the presence of acetic anhydride, nitrite； The intermediate acetyl of structural formula 5 is removed to obtain the intermediate of structural formula 6, by intermediate 6 iodo of structural formula 6, The intermediate of structural formula 8；The pyrazoles amino of the intermediate of structural formula 8 is protected with amido protection group, obtains structural formula 9 Intermediate；Then the iodine utilizing the intermediate of structural formula 9 carries out coupling and obtains the intermediate of structural formula 10；Again by structural formula The ester group of the intermediate of 10 is reduced to alcohol radical and obtains the intermediate of structural formula 11, by the alcohol radical in the intermediate of structural formula 11 It is oxidized to aldehyde radical, obtains 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 formula (Ia), and R '=R₃=R₅Time, its preparation method is such as Shown in route 4:

Route 4:

When the compound of required preparation is the compound shown in formula (Ib), and R₃With R₅Time different, its preparation method is such as Shown in route 5:

Route 5:

When the compound of required preparation is the compound shown in formula (Ic), R₃With R₅Difference and R₄When being not H, its system Preparation Method is as shown in Scheme 6:

Route 6:

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

Route 7:

Wherein X is CH₂Or O.

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

Route 8:

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

Route 9:

Wherein X is CH₂Or O.

As the pharmaceutical composition containing dihydropyridine compounds of third aspect present invention, wherein said pharmaceutical composition bag Logical formula (I) compound containing therapeutically effective amount and pharmaceutically acceptable excipient.

As a kind of pharmaceutical composition of third aspect present invention, wherein said pharmaceutical composition comprises the logical of therapeutically effective amount The pharmaceutically acceptable derivant of formula (I) compound and pharmaceutically acceptable excipient.

As a kind of pharmaceutical composition of third aspect present invention, wherein said pharmaceutical composition comprises the logical of therapeutically effective amount The pharmaceutically acceptable salt of formula (I) compound and pharmaceutically acceptable excipient.

Described pharmaceutical composition make tablet, capsule, aqueous suspension, Oil suspensions, dispersible powder, Granule, lozenge, Emulsion, syrup, ointment, ointment, suppository or injection.

As the application of fourth aspect present invention, it is wherein that logical formula (I) compound is lived in preparation regulation protein kinase catalysis Application in property goods.

As the application of fourth aspect present invention, it is wherein that the pharmaceutically acceptable derivant of logical formula (I) compound is in system Application in standby regulation protein kinase catalysis active product.

As the application of fourth aspect present invention, it is wherein that the pharmaceutically useful salt of logical formula (I) compound is at preparation regulation egg Application in white kinase catalytic activity goods.

As the application of fourth aspect present invention, it is wherein that pharmaceutical composition treats the disease relevant with protein kinase in preparation The sick application in medicine.

Described protein kinase is c-Met receptor tyrosine kinase.

The relevant disease of described protein kinase is cancer.

Described cancer be thyroid carcinoma, colorectal carcinoma, gastric cancer, renal carcinoma, hepatocarcinoma, pulmonary carcinoma, ovarian cancer, breast carcinoma, Carcinoma of prostate, bladder cancer, head and neck cancer, cancer of pancreas, carcinoma of gallbladder, osteosarcoma, rhabdomyosarcoma, MFH/ Fibrosarcoma, glioblastoma/astrocytoma, melanoma or mesothelioma.

The dihydropyridine compounds of logical formula (I) involved in the present invention can be additionally used in grinding of biology or pharmacology's phenomenon Study carefully, tyrosine kinase participate in the research of signal transduction pathway and relatively commenting for new tyrosine kinase inhibitor Valency.

Detailed description of the invention

The present invention provides logical formula (I) compound defined above, prepares the method for these compounds, uses these to change The pharmaceutical composition of compound and the method using these compounds.

Listed below is the definition to the various terms for describing the compounds of this invention.These definition are applied to The term (unless the most defined otherwise) that each place of description uses, no matter these terms are used alone also It it is the part as more macoradical.

Unless otherwise defined, term as used herein " alkyl " (alone or as the part of another group) refers to The univalent perssad comprising 1 to 12 carbon atom that alkane is derivative.Preferably alkyl has 1 to 6 carbon atom.Alkyl For optionally substituted straight chain, side chain or cyclic saturated hydrocarbon base.Exemplary alkyl includes methyl, ethyl, propyl group, isopropyl Base, normal-butyl, the tert-butyl group, isobutyl group, amyl group, hexyl, isohesyl, heptyl, 4,4-dimethyl amyl group, octyl group, 2,2,4-tri-methyl-amyl, nonyl, decyl, undecyl, dodecyl etc..

The substituent group of " replacement alkyl " is selected from following group: alkyl, halogen (such as fluorine, chlorine, bromine, iodine), alcoxyl Base, amino/amido, haloalkyl (such as trichloromethyl, trifluoromethyl), aryl, aryloxy, alkylthio group, hydroxyl, Cyano group, nitro, carboxyl, alkoxy carbonyl, alkyl-carbonyl epoxide, carbamyl, urea or sulfydryl.

Term as used herein " aryl " (alone or as the part of another group) refer to monocyclic aromatic ring or Polycyclic aromatic ring, such as phenyl, substituted phenyl etc. and group such as naphthyl, the phenanthryl etc. condensed.Thus, aryl bag The ring having at least 6 atoms containing at least one, comprises at most five such rings (wherein comprising at most 22 atoms), And between adjacent carbon atom or suitably hetero atom, there is (conjugation) double bond alternately.Preferably aryl is in ring Comprise 6 to 14 carbon atoms.

" substituted aryl " can be optionally substituted with one or more group, described group include but not limited to halogen (such as fluorine, Chlorine, bromine), alkyl (such as methyl, ethyl, propyl group), replace alkyl (such as trifluoromethyl), cycloalkyl, alkoxyl (such as methoxy or ethoxy), hydroxyl, carboxyl, amine formyl (-C (=O) NR'R "), alkoxy carbonyl (-CO₂R)、 Amino/amido, nitro, cyano group, thiazolinyl epoxide, aryl, heteroaryl, sulfonyl (-SO₂R) etc., wherein, R, R', R are " for described alkyl.

Term as used herein " heteroaryl " (alone 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 group, these groups are extremely A few ring has at least one hetero atom (O, S or N).The each ring comprising heteroatomic heteroaryl can comprise One or two oxygen atoms or sulphur atom and/or one to four nitrogen-atoms, condition is that in each ring, heteroatomic sum is four Individual or less, and each ring has at least one carbon atom, form condensing of above-mentioned bicyclic radicals and three cyclic groups Ring can only comprise carbon atom, and can be saturated or fractional saturation.Nitrogen-atoms and sulphur atom can be oxidations, and And nitrogen-atoms can be quaternary ammoniated.The heteroaryl of dicyclo or three rings must include the ring of at least one Wholly aromatic, but its Ring or multiple ring that it condenses can be aromatics or non-aromatic.Heteroaryl can be at the most available nitrogen-atoms of any ring or carbon Connect at atom.

" substituted heteroaryl " ring system can comprise zero, one, two or three selected from following substituent group: halogen, alkyl, takes The alkyl in generation, thiazolinyl, block base, aryl, nitro, cyano group, hydroxyl, alkoxyl, alkylthio group ,-CO₂H ,-C (=O) H, -CO₂-alkyl ,-C (=O) alkyl, phenyl, benzyl, phenylethyl, phenyl epoxide, thiophenyl, cycloalkyl, replacement Cycloalkyl, Heterocyclylalkyl, heteroaryl ,-NR'R " ,-C (=O) NR'R " ,-CO₂NR'R " ,-C (=O) NR'R ", -NR'CO₂R " ,-NR'C (=O) R " ,-SO₂NR'R " and-NR'SO₂R ", wherein R' and R " is each independently selected from hydrogen, alkane Base, substituted alkyl and cycloalkyl, or R' with R " together with form Heterocyclylalkyl or heteroaryl ring.

The example of bicyclic heteroaryl includes pyrrole radicals, pyrazolyl, pyrazolinyl, imidazole radicals, oxazolyl, di azoly, different Oxazolyl, thiazolyl, thiadiazolyl group, isothiazolyl, furyl, thienyl, oxadiazoles base, pyridine radicals, pyrazinyl, Pyrimidine radicals, pyridazinyl, triazine radical etc..

The example of bicyclic heteroaryl include indyl, benzothiazolyl, benzodioxole base, benzoxazolyl, Benzothienyl, quinolyl, tetrahydric quinoline group, isoquinolyl, tetrahydro isoquinolyl, benzimidazolyl .alpha.-5:6-benzopyran Base, indolizine base, benzofuranyl, chromone base, coumarin base, benzofuranyl, quinoxalinyl, indazolyl, pyrroles And pyridine radicals, furopyridyl etc..

The example of tricyclic heteroaryl includes carbazyl, benzindole base, phenanthroline base, acridinyl, phenanthridinyl etc..

A carbon in term as used herein " heterocycle " (alone or as the part of another group) finger ring Atom is selected from the cycloalkanes that hetero atom replaces and at most 3 extra carbon atoms can be replaced by described hetero atom of O, S or N Base (non-aromatic).Term " heterocyclic radical " (alone or as the part of another group) used in this application refers to Comprise the stable saturated or part of 5 to 7 annular atomses (carbon atom and selected from nitrogen, sulfur and/or other atom of oxygen) Undersaturated monocyclic ring system.Heterocycle can be 5,6 or 7 yuan of monocycles, and comprises one, two or three selected from nitrogen, oxygen And/or the hetero atom of sulfur.Heterocycle can be optionally substituted, it means that heterocycle can be in one or more commutable ring position Putting replacement has one or more to be independently selected from following group: alkyl, Heterocyclylalkyl, heteroaryl, alkoxyl, nitro, list Alkyl amine group, dialkyl amino, cyano group, halogen, haloalkyl, alkanoyl, ammonia amine base carbonyl, monoalkyl amido Carbonyl, dialkyl amino carbonyl, alkylamidoalkyl, alkoxyalkyl, alkoxy carbonyl, alkyl-carbonyl epoxide and aryl, Described aryl is optionally substituted with halogen, alkyl and alkoxyl.The example of these Heterocyclylalkyls includes but not limited to: piperidines, Morpholine, high morpholine, piperazine, tetrahydro-1,4-thiazine, pyrrolidine and azetidine.

Term as used herein " alkoxyl " (alone or as the part of another group) refers to pass through oxygen atom The alkyl preferably with 1 to 6 carbon atom connected, such as-OR, wherein R is described alkyl.

Term as used herein " amino " (alone or as the part of another group) refers to-NH₂." amido " One or two substituent group (-NR'R "), wherein R' and R can be optionally substituted with " can be identical or different, such as alkyl, Aryl, aryl alkyl, thiazolinyl, alkynyl, heteroaryl, heteroaryl alkyl, Heterocyclylalkyl, alkyl, hetercycloalkylalkyl, Cycloalkyl, cycloalkyl-alkyl, haloalkyl, antelope base alkyl, alkoxyalkyl, alkylthio group, carbonyl or carboxyl.These Substituent group can be further substituted with any one in the alkyl or aryl substituent group listed by carboxylic acid or the application.At some In embodiment, amino replaces carboxyl or carbonyl, forms N-acyl group or N-carbamyl deriveding group.

Term " haloalkyl " (alone or as the part of another group) used in this application refers to by alkyl even The halogen atom connect, such as-CF₃。

Term as used herein " acyl group " (alone or as the part of another group) refers to be connected by carbonyl Alkyl or-C (=O) R, wherein R is described alkyl.

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

Term as used herein " aryl alkyl " or " aralkyl " (alone or as the part of another group) Refer to the aromatic ring (such as benzyl) connected by alkyl described above.

Term as used herein " aminoalkyl " (alone or as the part of another group) refers to pass through alkyl The amino (-NR'R ") connected.

Term as used herein " aryl-alkyl amino " (alone or as the part of another group) refers to pass through The aryl that alkyl connects, described alkyl is connected by amino.

Term " hetero atom " refers to independently selected O, S or N.It should be noted that quantivalence is ungratified the most miscellaneous Atom is considered to connection hydrogen atom, thus meets quantivalence.

Term " halogen " refers to independently selected fluorine, chlorine, bromine or iodine.

Term as used herein " cycloalkyl " (alone or as the part of another group) refers to 3 to 9 carbon The hydrocarbon ring of the fully saturated or fractional saturation of atom, preferably 3 to 7 carbon atoms.Additionally, cycloalkyl can be to replace 's." substituted cycloalkyl " refers to have one, two or three rings selected from following substituent group: halogen, alkyl, replacement Alkyl (wherein substituent group is as defined with regard to alkyl substituent above), thiazolinyl, alkynyl, nitro, cyano group, oxo (=O), Hydroxyl, alkoxyl, alkylthio group ,-CO₂H ,-C (=O) H ,-CO₂-alkyl ,-C (=O) alkyl, ketone group ,=N-OH ,=N-O- Alkyl, aryl, heteroaryl, five or hexa-atomic ketals (i.e. 1,3-dioxane or 1,3-bis-uh alkane) ,-NR ' R ", -C (=O) NR'R " ,-CO₂NR'R " ,-C (=O) NR'R " ,-NR'CO₂R " ,-NR'C (=O) R " ,-SO₂NR'R" With-NR'SO₂R ", wherein R' and R " is each independently selected from hydrogen, alkyl, substituted alkyl and cycloalkyl, or R' With R ", form Heterocyclylalkyl or heteroaryl ring together.

Term " anticarcinogen " includes the most known medicine that can be used for treating cancer, including: (1) cytotoxic drug: Chlormethine series pharmaceuticals, such as melphalan, cyclophosphamide；Platinum coordination complex, such as cisplatin, carboplatin and oxaliplatin；(2) Anti-metabolism antineoplastic agent: 5-fluorouracil, capecitabine, methotrexate, calcium folinate, Raltitrexed, purine are short of money Anti-agent (such as 6-thioguanine and Ismipur)；(3) hormones: the 17 female alcohol of alpha-acetylenes, diethylstilbestrol, testis Ketone, prednisone, fluoxymesterone, dromostanolone propionate, testolactone, megestrol acetate, methylprednisolone, methyltestosterone, Prednisolone, triamcinolone, chlorotrianisene, hydroxyprogesterone, aminoglutethimide, estramustine, medroxyprogesterone acetate, Tuo Rui Meter Fen；(4) tyrosine kinase inhibitor: EGFR inhibitor, including gefitinib (Gefitinib), Erlotinib (Erlotinib), Cetuximab (Cetuximab), Trastuzumab (Herceptin) etc.；VEGF inhibitor, all As VEGF antibody (Avastin (Avastin)) and micromolecular inhibitor such as Sunitinib, Vandetanib, Cediranib；Bcr-Abl inhibitor such as imatinib (Imatinib), Dasatinib (Dasatinib)；Src presses down Preparation, MEK inhibitors of kinases, mapk kinase inhibitor, PI3K inhibitors of kinases, c-Met inhibitor, ALK suppress Agent etc.；(5) medicine of tubulin, such as vinca medicine, taxanes medicine, epothilones medicine are acted on Thing such as ipsapirone (Ixabepilone) etc.；(6) topoisomerase I inhibitor, such as topotecan, irinotecan； (7) histon deacetylase (HDAC) (HDAC) inhibitor such as Vorinostat (Vorinostat)；(8) proteasome suppression Agent such as bortezomib (Bortezomib)；(9) anticarcinogen of other classifications such as aurora kinase (aurora kinase) presses down Preparation, biological response modifier, growth inhibitor, glu famine antagonist, angiogenesis inhibitor and anti-vascular medicine, base Matter inhibitors of metalloproteinase etc..

" mammal " includes the mankind and domestic animal, such as cat, Canis familiaris L., pig, cattle, sheep, goat, horse, rabbit etc..Preferably Ground, for the purposes of the present invention, described mammal is the mankind.

" optionally () " or " optionally () " represents that the environment event described subsequently there may be or not exist, and Described description includes the situation that described event or environment occur and situation about not occurring.Such as, " optionally substituted aryl " Represent that described aryl may be substituted or unsubstituted and described description includes substituted aryl and unsubstituted aryl.

" pharmaceutically acceptable derivant " represented when receiver is administered, it is possible to directly or indirectly provide the compound of the present invention Or the active metabolite of its inhibition or any nontoxic salt of residue, ester, the salt of ester, amide, the salt of amide or its His derivant.

" pharmaceutically acceptable excipient " includes but not limited to be ratified as available by state food and Drug Administration In the mankind or any adjuvant of domestic animal, carrier, excipient, fluidizer, sweeting agent, dispersant, diluent, preservative, Suspending agent, stabilizer, dyestuff/coloring agent, flavour enhancer, surfactant, wetting agent, isotonic agent, solvent or emulsifying Agent.

" pharmaceutically acceptable salt " includes acid-addition salts and base addition salts.

" pharmaceutically acceptable acid-addition salts " refers to such salt, and they remain biological effect and the character of free alkali, Adverse consequences will not be produced at biology or other aspects, and be with mineral acid such as but not limited to hydrochloric acid, hydrobromic acid, Sulphuric acid, nitric acid, phosphoric acid etc., and the most following acid of organic acids: formic acid, acetic acid, trifluoroacetic acid, first sulphur Acid, trifluoromethanesulfonic acid, ethyl sulfonic acid, 2-ethylenehydrinsulfonic acid, benzenesulfonic acid, p-methyl benzenesulfonic acid, 2,2-dichloroacetic acid, oneself two Acid, alginic acid, ascorbic acid, aspartic acid, benzoic acid, paraacetaminobenzoic acid, dextrocamphoric acid., Camphora-10- Sulfonic acid, capric acid, caproic acid, octanoic acid, carbonic acid, cinnamic acid, citric acid, cyclohexane sulfamic acid, lauryl sulphate acid, Ethane-1,2-disulfonic acid, Fumaric acid, galactosaccharic acid, gentisic acid, glucoheptonic acid, gluconic acid, glucuronic acid, Glutamic acid, 1,3-propanedicarboxylic acid, 2-oxo-1,3-propanedicarboxylic acid, phosphoglycerol, glycolic, hippuric acid, isopropylformic acid., lactic acid, lactose Acid, lauric acid, maleic acid, malic acid, malonic acid, mandelic acid, glactaric acid, naphthalene-2-sulfonic acid, naphthalene-1,5-disulfonic acid, 1-hydroxy-2-naphthoic acid, nicotinic acid, oleic acid, orotic acid, oxalic acid, palm fibre eleostearic acid, pamoic acid, propanoic acid, pyroglutamic acid, Acetone acid, salicylic acid, 4-ASA, decanedioic acid, stearic acid, fumaric acid, succinic acid, tartaric acid, Hydrogen thiocyanate, Undecylenic acids etc. are formed.

" pharmaceutically acceptable base addition salts " refers to such salt, and they remain biological effect and the character of free acid, Will not be improper in biology or other aspects.These salt are added to prepare on free acid by by inorganic base or organic base. The salt being derived from inorganic base includes but not limited to sodium, potassium, lithium, ammonium, calcium, magnesium, ferrum, zinc, copper, manganese, aluminium salt etc..Excellent The inorganic salt of choosing is ammonium, sodium, potassium, calcium and magnesium salt.The salt being derived from organic base includes but not limited to the salt of following substances: primary Amine, secondary amine and tertiary amine, substituted amine, including naturally occurring replacement amine, cyclammonium and deacidite, as ammonia, Methylamine, dimethylamine, trimethylamine, diethylamine, triethylamine, tripropyl amine (TPA), 2-aminopropane., diethanolamine, ethanolamine, 2- Dimethylaminoethanol, 2-diethylaminoethanol, hexanamine, lysine, arginine, histidine, caffeine, Pu Lu Caine, Hai Baming (hydrabamine), choline, glycine betaine, benethamine, ethylenediamine, glycosamine, methyl Amine, theobromine, triethanolamine, trometamol, purine, piperidines, piperazine, N-ethylpiperidine, polyamino resin etc..Excellent The organic base of choosing is 2-aminopropane., diethylamine, ethanolamine, triethylamine, hexanamine, choline and caffeine.

" pharmaceutical composition " refers to the compound of the present invention and is delivered in the mammal such as mankind by biologically active cpds The preparation that formed of the medium generally accepted.Such medium includes all of pharmaceutically acceptable carrier to this, dilute Release agent or excipient.

" therapeutically effective amount " refers to when being administered (the preferably mankind) to mammal, it is sufficient to mammal (the preferably mankind) Relevant disease or disease realize the amount of compound of the present invention for the treatment of as defined below.Constitute " therapeutically effective amount " The amount of compound of the present invention can be according to the activity of particular compound such as applied；The metabolic stability of described compound Property and effect duration；The age of patient, body weight, holistic health, sex and diet；Mode of administration and time；Excretion speed Rate；Drug combination；Specific illness or the seriousness of disease；And experience the individuality for the treatment of and change, but it can be by this Field those of ordinary skill determines routinely according to himself knowledge and the disclosure.

" treat " or " treatment " is contained the mammal with relevant disease or disease, preferably for herein time The relevant disease of the mankind or the treatment of disease, and include:

There is disease or disease in (i) prevention mammal, especially the most ill but without examining when such mammal Break when ill；

(ii) suppression disease or disease, i.e. stop it to develop；

(iii) alleviate disease or disease, i.e. cause disease or disease to disappear；

(iv) stable disease or disease.

When this paper, term " disease " and " disease " can exchange use or can be different, and reason is specific disease Sick or disease is likely not to have the inducement (thus the most not working out the cause of disease) that oneself knows, be not considered as the most disease and Being only used as improper situation or syndrome, wherein clinician have identified concrete syndrome the most more or less. The compounds of this invention shown in this article and their structure are also represented by including all isomers (such as enantiomer, non-right Reflect isomer, geometrical isomerism or conformational isomerism) form, they can be according to fixed for amino acid whose absolute stereochemical Justice be (R)-/(S)-or (D)-/(L)-or (R, R)-/(R, S)-/(S, S)-.The present invention represent include all these can The isomer of energy, and their racemic, enantiomer enrichment and optional pure form.Optically-active (+) and (-), (R)-and (S)-and (R, R)-/(R, S)-/(S, S)-or (D)-chirality synthesis, chirality can be used to tear open with (L)-isomer Divide preparation, or routine techniques can be used to split such as but not limited to the efficient liquid phase (HPLC) using chiral column.When When compound as herein described comprises thiazolinyl double bond or other geometry asymmetric centers, except as otherwise noted, described compound Including E and Z geometric isomer.Equally, all tautomeric forms are also included.

" stereoisomer " refers to be made up of with identical chemical bonding identical atom but has the change of different three dimensional structure Compound, they are non-interchangeable.The present invention contain various stereoisomer and mixture thereof and include " enantiomer " and " diastereomer ", enantiomer refers to two kinds of stereoisomers of the mirror image that its molecule can not be overlapped each other；Non-right Reflect isomer and refer to that molecule has two or more chiral centre, and the intermolecular stereoisomer for non-mirror image relationship.

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

It addition, except as otherwise noted, the compound of the present invention also includes that structure is different only in that one or more coordinatioies of existence The compound of element enriched atoms.Such as, there is the structure of the present invention, except replacing hydrogen with " deuterium " or " tritium ", or With¹⁸F-fluorine labelling (¹⁸F isotope) replace fluorine, or use¹¹C-,¹³C-, or¹⁴C-enrichment carbon (¹¹C-,¹³C-, Or¹⁴C-carbon markings；¹¹C-,¹³C-, or¹⁴C-isotope) replace the compound of carbon atom to be in the scope of the present invention In.Such compound can be used as the analytical tool in such as biological characteristis or probe, or can serve as the body of disease Interior diagnosing image tracer, or the tracer as the research of pharmacodynamics, pharmacokinetics or receptor.

The present invention also provides for following methods: by by therapeutically effective amount lead to as defined above formula (I) compound with At least one other anticancer agents gives the patient that (or successively) needs this treatment simultaneously, swashs via regulation c-Met Enzyme treats proliferative disease (such as cancer).In preferred embodiments, proliferative disease is cancer.

Specifically, logical formula (I) compound can be used for treating kinds cancer, is most specifically that depend on c-Met activation A little cancers.C-Met activation can be passed through gene amplification, sudden change (various mutations) and/or HGF stimulation and regulate, wherein HGF is provided by tumor (autocrine) or host's (paracrine) tissue.Generally, can be used for controlling by the compound of the present invention Treat following cancer:

(A) solid tumor, including gastric cancer, pulmonary carcinoma (including small cell lung cancer, nonsmall-cell lung cancer), colon cancer, renal carcinoma, Hepatocarcinoma, breast carcinoma, ovarian cancer, cervical cancer, esophageal carcinoma, carcinoma of gallbladder, bladder cancer, cancer of pancreas, thyroid carcinoma, prostatitis Adenocarcinoma and skin carcinoma (including squamous cell carcinoma)；

(B) hematopoetic tumor of lymphoid is white including acute lymphoblastic leukemia (ALL), acute lymphoblastic Disorders of blood, B cell lymphoma, t cell lymphoma, Hodgkin lymphoma, non-Hodgkin′s woods bar tumor, hairy cell lymphoma With Burkitt lymphoma (Burkett's lymphoma)；

(C) hematopoetic tumor of myeloid lineage, including acute and chronic myelogenous leukemia, myelodysplastic syndrome and morning Myelocyte leukemia；

(D) tumor of mesenchyme origin, including fibrosarcoma and rhabdomyosarcoma；

(E) maincenter and the tumor of peripheral nervous system, including astrocytoma, neuroblastoma, glioma and Schwannoma；

(F) other tumor, including melanoma, spermocytoma, teratocarcinoma, osteosarcoma, xeroderma pigmentosum, angle Change acanthoma, follicular carcinoma of thyroid and Kaposi sarcoma.

Logical formula (I) compound can be additionally used in treatment and is characterized as any lysis of abnormal cell proliferation, the most optimum before Row gland hypertrophy, neurofibromatosis, atherosclerosis, pulmonary fibrosis, arthritis, psoriasis, glomerulonephritis, After angioplasty or vascular surgery occur restenosis, inflammatory bowel, graft-rejection, endotoxin shock and Fungal infection.

Logical formula (I) compound scalable cell RNA and the level of DNA synthesis.Therefore, can be used for controlling by these materials Treat virus infection and (include but not limited to HIV, human papillomavirus, herpesvirus, poxvirus, Epstein-Barr virus, Xin Debi This virus and adenovirus).

Logical formula (I) compound can be used for the chemoprophylaxis of cancer.It is defined as chemoprophylaxis being dashed forward by the initial cause of blocking-up Change event or suffer from the progress of pre-malignant cells of damage by blocking-up and carry out the development of anti-invasion cancer or suppression is swollen Tumor recurs.

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

The compound of the present invention also can be with known anticarcinogen (including but not limited to those mentioned in above-mentioned " anticarcinogen ") Or anticancer therapy (such as radiotherapy) is applied in combination (give together or successively give).

Some logical formula (I) compound generally can be prepared according to following route 1 to route 9.Logical formula (I) compound Tautomer and solvate (such as hydrate, ethanolates) are also within the scope of the invention.The system of solvate Preparation Method is the most commonly known.Therefore, the compound of the present invention can be free form or hydrate forms. In method discussed below, the functional group of midbody compound may need to be protected by suitable protection group.Such official Group can include hydroxyl, amino, sulfydryl and carboxylic acid.The protection group being suitable for for hydroxyl includes trialkylsilkl or two Arylalkylsilyl (such as t-butyldimethylsilyl, t-butyldiphenylsilyl or trimethyl first silicon Alkyl), THP trtrahydropyranyl, benzyl, to methoxy-benzyl etc..Suitable protecting groups for amino include tertbutyloxycarbonyl, Benzyloxycarbonyl group, acetyl group, benzoyl, trifluoroacetyl group, to methoxy-benzyl etc..Suitable protecting groups for carboxylic acid Including alkyl, aryl or alkyl aryl.NH functional group suitable for heteroaryl the most such as indole or indazole ring Protection group include tertbutyloxycarbonyl, benzyloxycarbonyl group, acetyl group, benzoyl, 2-trimethylsilyl-ethoxy methyl, To methoxy-benzyl etc..

Protection group can be according to method known to those skilled in the art (Greene, T.W., Protective Groups in Organic Synthesis, 1999, the 3rd edition, Wiley) and standard technique as herein described is added or removal. Described protection group can also be fluoropolymer resin such as Wang resin, Rink resin or 2-chlorine trityl chloride resin.

Meanwhile, although these protected derivants of the compounds of this invention itself may not have pharmacological activity, but they are permissible Being administered to mammal, metabolism is to form the compounds of this invention with pharmacological activity the most in vivo.Such spread out Therefore biology is described as " prodrug ".All prodrugs of the compounds of this invention are included within the scope of the present invention.

The present invention leads to the dihydropyridine compounds of formula (I), can be prepared by the following method.

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

Route 1:

Wherein, R₁, R₂, R₃, R₄As defined above；PG is amido protection group.PG is preferably tertbutyloxycarbonyl, to first Oxy-benzyl.

Route 2:

Route 3:

Described coupling is selected from Suzuki coupling, Buchwald coupling.

Route 4:

When the compound of required preparation is the compound shown in formula (Ib), and R₃Time different from R5, its preparation method is such as Shown in route 5:

Route 5:

Route 6:

Route 7:

Wherein X is CH₂Or O.

Route 8:

Route 9:

Wherein X is CH₂Or O.

Wherein, following is conventional abbreviation:

DMF:N, dinethylformamide；

DMSO: dimethyl sulfoxide；

CDCl₃: 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；

DEG C: degree Celsius；

Mol: mole；

TLC: thin layer chromatography.

Those skilled in the art can use suitable raw material, use similar method, prepare and do not have in reaction scheme above Other compounds of the specifically disclosed present invention.

By with suitable inorganic or organic base or acid treatment, can existing with free alkali or acid form according to prepared as above All the compounds of this invention change into their pharmaceutically acceptable salt.The salt of compound prepared as above can be by mark Quasi-technical transform becomes their free alkali or acid form.

The compound of the present invention includes its all crystal formations, amorphous forms, dehydrate, hydrate, solvate and salt. Additionally, the compound of all present invention comprising ester group and amide group can oneself knows by those skilled in the art Method or change into corresponding acid by method described herein.Equally, the compounds of this invention comprising hydroxy-acid group can It is converted into corresponding ester and amide with the method that oneself knows by those skilled in the art.Those skilled in the art can also be passed through The method (such as hydrogenation, alkylation and acyl chloride reaction etc.) that oneself knows carries out other replacements on molecule and replaces.

The cyclodextrin clathrate of the present invention to be prepared, can be dissolved in the compound of the logical formula (I) defined in summary of the invention above The acceptable solvent of pharmacology such as (but not limited to) alcohol (preferred alcohol), ketone (such as acetone) or ether (such as second Ether) in, and at 20 DEG C to 80 DEG C and alpha-cyclodextrin, beta-schardinger dextrin-or the water of gamma-cyclodextrin, preferably beta-schardinger dextrin- Solution mixes；Or can be by the acid of the compound of the logical formula (I) defined in summary of the invention above with its salt (such as sodium Or potassium salt) aqueous solution form and cyclodextrin be blended, then with equivalent acid (such as HCl or H₂SO₄) solution altogether Mixed, to provide corresponding cyclodextrin clathrate.

Now or after the cooling period, corresponding cyclodextrin clathrate crystal can crystallization.Or when logical formula (I) When compound is oily and crystallization, by stirring (such as 1 hour to 14 days) the most for a long time, add cyclodextrin Aqueous solution process, it is also possible to be converted into corresponding cyclodextrin clathrate.Then by filtering and being dried, can be by inclusion Thing is separated into solid or crystal.

Cyclodextrin for the present invention is commercially available (such as from Aldrich Chemical Co.), or by this area skill Art personnel use known method to prepare.See for example Croft, A.P. et al., " Synthesis of Chemically Modified Cyclodextrins",Tetrahedron 1983,39,9,1417-1474.Suitable cyclodextrin bag Include the compound with formula listed above (I) and prepare all kinds of clathrate.

By selecting appropriate cyclodextrin and water, repeatably active substance content can be obtained according to stoichiometric composition Clathrate.Clathrate can be to be dried water suction form or form use that is aqueous but that less absorb water.Cyclodextrin and formula (I) Typical mole ratios of compound is 2:1 (cyclodextrin: compound).

Comprising logical formula (I) compound can be to be adapted for the form being administered orally, for example, as the pharmaceutical composition of active component Tablet, capsule, aqueous suspension, Oil suspensions, dispersible powder or granule, syrup etc..Orally available The compositions used can be prepared according to any means for preparing pharmaceutical composition known in the art, and these groups Compound can comprise one or more selected from sweeting agent, flavoring agent, coloring agent and the material of preservative, in order to provides pharmaceutically beautiful See and agreeable to the taste preparation.

Tablet comprises active component, and is mixed with nontoxic pharmaceutically acceptable excipient or the carrier being suitable to prepare tablet.These excipient Or carrier can be inert diluent, such as calcium carbonate, sodium carbonate, lactose, calcium phosphate or sodium phosphate；Granulating agent and disintegrate Agent, such as Celluloasun Microcrystallisatum, carmethose, corn starch or alginic acid；Binding agent, such as starch, gelatin, poly- Vinylpyrrolidone or arabic gum；And lubricant, such as magnesium stearate, stearic acid or Pulvis Talci.Tablet can be not Coating, maybe can carry out coating by known technology, thus cover unpleasant drug tastes, or prolong in the gastrointestinal tract Disintegrate late and absorption, thus provide lasting effect within the longer period.Such as, water miscible taste can be used to cover Material (such as hydroxypropyl-methylcellulose or hydroxypropyl-cellulose) or time delay material (such as ethyl cellulose, Cellulose acetate-butyrate).

Capsule comprises hard-gelatin capsules, Gelseal.Hard-gelatin capsules is dilute with inert solid by active component Release the mixing of agent such as calcium carbonate, calcium phosphate or Kaolin；Gelseal is by active component with water-solubility carrier (such as Polyethylene Glycol) or oil medium (such as Oleum Arachidis hypogaeae semen, liquid paraffin or olive oil) mixing.

Aqueous suspension comprises active substance and is suitable to prepare the excipient of aqueous suspension.These excipient are suspending agent, Such as sodium carboxymethyl cellulose, methylcellulose, hydroxypropyl methyl-cellulose, sodium alginate, polyvinylpyrrolidone and Arabic gum；Dispersant or wetting agent, the phospholipid that can be naturally-occurring (such as lecithin) or oxyalkylene and fatty acid Condensation product (such as Myrj 45) or the condensation product (such as 17 of ethylene oxide and long-chain fatty alcohol Oxygen ethylene spermol (heptadecaethylene-oxycetanol)) or ethylene oxide spread out with from fatty acid and hexitol The condensation product (such as polyoxyethylene 80 sorbitan monooleate) of raw partial ester or ethylene oxide with from fatty acid and hexose The condensation product (such as polyethylene sorbitan monoleate) of the partial ester that the liquor-saturated mixture of alcohol is derivative.Aqueous suspension Also can comprise one or more preservative (such as ethylparaben or n-propyl), one or more coloring agent, one Or multiple flavoring agent and one or more sweeting agent (such as sucrose, saccharin or aspartame).

Oil suspensions can be by being suspended in vegetable oil (such as Oleum Arachidis hypogaeae semen, olive oil, Oleum Sesami or cocos nucifera oil) by active component Or mineral oil (such as liquid paraffin) is prepared.Oil suspensions can comprise thickening agent, such as Cera Flava, hard paraffin or Spermol.Sweeting agent (such as those listed above) and flavoring agent can be added, thus agreeable to the taste oral formulations is provided. These compositionss can carry out anticorrosion by adding antioxidant (such as butylated hydroxyanisole or alpha-tocopherol).

Dispersible powder and granule comprise active component, and be mixed with dispersant or wetting agent, suspending agent and one or more Preservative.The suitably example of dispersant or wetting agent and suspending agent is above already mentioned those.Also can be comprised other Excipient, such as sweeting agent, flavoring agent and coloring agent.These compositionss can be by adding antioxidant (such as Vitamin C Acid) carry out anticorrosion.Dispersible powder and granule can prepare aqueous suspension by adding water.

Syrup can be prepared with sweeting agent (such as glycerol, propylene glycol, sorbitol or sucrose).These preparations also can wrap Containing demulcent, preservative, flavoring agent, coloring agent and antioxidant.

The pharmaceutical composition of the present invention can also be the form of oil-in-water emulsion.Oil phase can be vegetable oil (such as Fructus Canarii albi Oil or Oleum Arachidis hypogaeae semen) or mineral oil (such as liquid paraffin) or their mixture.Suitably emulsifying agent can be natural depositing Phospholipid (such as soybean lecithin), (be such as dehydrated mountain from the derivative ester of fatty acid and hexitol mixture or partial ester Pears Sorbitane monooleate) and condensation product (the such as polyoxyethylene sorbitan list oil of described partial ester and ethylene oxide Acid esters).Emulsion also can comprise sweeting agent, flavoring agent, preservative and antioxidant.

Pharmaceutical composition can be with the form being sterile injectable aqueous solution.Spendable acceptable carriers and solvent have water, woods Lattice solution (Ringer's solution), isotonic sodium chloride solution and glucose solution.

Sterile injectable preparation can also be sterile injectable water oil-packaging type micro-emulsion, wherein active component is dissolved in oil phase In.Such as, first active component is dissolved in the mixture of soybean oil and lecithin.Then, by obtained oil soluble Liquid imports in the mixture of water and glycerol and processes, thus forms microemulsion.

Injectable solution or microemulsion can be imported to by local bolus injection in the blood flow of patient, or give described in some way Solution or microemulsion, thus maintain the circulation composition of constant the compounds of this invention.In order to maintain this constant concentration, can Use the continuous intravenous administration set such as infusion pump.

Pharmaceutical composition could be for sterile injectable aqueous or the form of oil-based suspension of intramuscular or subcutaneous administration.This Plant suspension and can use above those already mentioned suitable dispersants or wetting agent and suspending agent according to known technology Configure.Sterile injectable preparation can also be nontoxic pharmaceutically acceptable diluent or the sterile injectable solution of solvent or suspendible The solution of liquid, such as 1,3 butylene glycol.It addition, sterile, fixed oils can be easily used as solvent or suspension medium.For This purpose, the gentleest fixed oil all can use, including monoglyceride or two glyceride of synthesis.It addition, Fatty acid (such as oleic acid) can use in preparing injection.

Logical formula (I) compound also can be given by the form of the suppository for rectally.These compositionss can be by mixed Composite medicine is prepared with suitable nonirritant excipient, and described excipient is solid at room temperature but is liquid at rectal temperature Body, melts the most in the rectum, thus discharges medicine.These materials include cocoa butter, glycerin gelatine, hydrogenated vegetable oil, The mixture of the Polyethylene Glycol of different molecular weight and the fatty acid ester of Polyethylene Glycol.

With regard to local use for, can prepare and use comprise the ointment of logical formula (I) compound, ointment, jelly, Solution or suspensoid etc..

The compound of the present invention can use suitable nasal carrier and doser in intranasal form to give by local, Or use those skilled in the art the form of well-known transdermal skin patches given by cutaneous routes.This The compound of invention also can by use such as following the form of suppository of substrate give: cocoa butter, glycerin gelatine, Hydrogenated vegetable oil, the mixture of Polyethylene Glycol of different molecular weight and the fatty vinegar of Polyethylene Glycol.

When being administered in human subject body by the compound of the present invention, daily dosage is typically by the doctor of prescription Determine, and described dosage generally with age, body weight, sex and the reaction of patient and the order of severity of the symptom of patient and Change.The effective daily dose of patient typically for 70kg is about 0.001mg/kg to 100mg/kg, and preferably 0.01 Mg/kg to 50mg/kg, more preferably 1mg/kg to 25mg/kg.

If being configured to fixed dosage, then these combination products use the present invention in dosage range described above Compound and the treatment of other medical active agent in the dosage range that it is ratified.When combination preparation is improper, logical formula (I) Compound also successively can give with known anticarcinogen or cytotoxicity medicine.The present invention is not limited by order of administration；Formula (I) compound can give known anticarcinogen (multiple anticarcinogen) or cytotoxicity medicine (various kinds of cell toxicity medicine) Before or after give.

The compound of the present invention is disease or the inhibitor of disease of c-Met mediation of c-Met mediation.Term " c-Met The disease of mediation " and the disease of mediation " c-Met " represent the known c-Met effective any morbid state of tool or other Adverse conditions.Term " disease of c-Met mediation " and " disease of c-Met mediation " are also represented by by pressing down with c-Met Those diseases that preparation for treating is eased or disease.These diseases and disease include but not limited to cancer and other proliferatives Illness.

Therefore, described compound can be used for treating the following disease in such as mammal, the especially mankind or illness: stomach Cancer, pulmonary carcinoma, esophageal carcinoma, cancer of pancreas, renal carcinoma, colon cancer, thyroid carcinoma, the brain cancer, breast carcinoma, carcinoma of prostate, with And other solid tumor cancers；Atherosclerosis；Regulation blood vessel occurs；Thrombosis and pulmonary fibrosis.

Compound involved in the present invention can be additionally used in biology or the research of pharmacology's phenomenon, the letter of tyrosine kinase participation The research of number pathway and the comparative evaluation for new tyrosine kinase inhibitor.

Compound involved by Ben Wen, including, but not limited to the structure type given by above-mentioned route 1 and route 9, is known The personnel of art technology can be by suitable initiation material, the method acquisition that application is similar.

Embodiment

The concrete synthetically prepared example (for preparing the compound of the present invention) of following offer and Biological examples (are used for demonstrate,proving The detection of bright the compounds of this invention purposes) it is to aid in putting into practice the present invention, they are not considered as limiting the present invention's Scope.

Synthetically prepared example 1

Prepare 2,6-dimethyl-4-(1H-thieno [3,2-c] pyrazoles-5-the base)-1,4-dihydro pyrrole of structural formula (I-1) Pyridine-3,5-dimethoxy nitrile

Step 1 reaction equation is as follows:

Step is: acetic anhydride (14.36g, 140.7mmol) adds in three-necked bottle, is cooled to-50 DEG C, and dropping is sent out Cigarette nitric acid (5.67g, 90.0mmol), finish slowly be dividedly in some parts 5-methylthiophene 2-formic acid (4.0g, 28.13 Mmol), finish in-20 DEG C of stirrings 2 hours.Reactant liquor is poured in frozen water, stands, and filters, and dries, and obtains yellow solid Body 5-methyl-4-Nitro-thiophene-2-formic acid (4.0g, 76%).

Step 2 reaction equation is as follows:

Step is: 5-methyl-4-Nitro-thiophene-2-formic acid (4.0g, 21.37mmol) is dissolved in methanol (40mL), Add concentrated sulphuric acid (5mL), return stirring 20 hours.Being cooled to room temperature, remove methanol under reduced pressure, residue is with acetic acid second Ester (200mL) dissolves, washing, concentrates, obtains brown oil 5-methyl-4-Nitro-thiophene-2-methyl formate (2.5 G, 62%).

Step 3 reaction equation is as follows:

Step is: 5-methyl-4-Nitro-thiophene-2-methyl formate (3.34g, 16.6mmol) is dissolved in methanol (25 ML) and in the mixed solvent of water (10mL), add ammonium chloride (4.39g, 82mmol), reduced iron powder (4.2g, 75mmol), return stirring 6 hours.Being cooled to room temperature, filtration under diminished pressure, filtrate concentrates, and it is molten that residue adds ethyl acetate Solving, washing, saturated common salt is washed, and is dried, is concentrated to dryness, obtains 4-amino-5-methyl-thiophene-2-methyl formate crude product (2.1g, 74%).ESI-MS:m/z 172(M+H).

Step 4 reaction equation is as follows:

Step is: 4-amino-5-methyl-thiophene-2-methyl formate (0.4g, 2.16mmol) is dissolved in toluene (20mL) In, add KOAc (0.424g, 4.23mmol), and 18-crown-6 (57.07mg) and acetic anhydride (0.727g, 7.13 Mmol), heated and stirred to 95 DEG C, add amyl nitrite (0.607g, 5.18mmol), be heated to 100 DEG C It is stirred overnight.Room temperature in cooling, adds ethyl acetate EtOAc (100mL), and system is done with washing, anhydrous sodium sulfate Dry, concentrate, silica gel column chromatography purification obtain 1-acetyl group-1H-thieno [3,2-c] pyrazoles-5-methyl formate (0.34g, 70%).ESI-MS:m/z 225(M+H).

Step 5 reaction equation is as follows:

Step is: 1-acetyl group-1H-thieno [3,2-c] pyrazoles-5-methyl formate (10.0g, 42mmol) and anhydrous carbon Acid potassium (23.2g, 168mmol) adds to, in absolute methanol (375mL), be stirred at room temperature 20 minutes jointly, filters.Filter Liquid concentrating under reduced pressure, silica gel column chromatography purification, obtain brown solid 1H-thieno [3,2-c] pyrazoles-5-methyl formate (8.0g, 97%).

Step 6 reaction equation is as follows:

Step is: 1H-thieno [3,2-c] pyrazoles-5-methyl formate (8.0g, 33.6mmol) is dissolved in anhydrous tetrahydrochysene Furan (160mL), nitrogen is protected.It is cooled to-20 DEG C, is dividedly in some parts lithium aluminium hydride (3.8g, 100.7mmol), Stir 3 hours.Carefully drip water (3.8mL), stand a moment, filter.Filtrate is dried with anhydrous sodium sulfate, and decompression is steamed Except solvent obtains white solid 1H-thieno [3,2-c] pyrazoles-5-methanol (3.54g, 69%).

Step 7 reaction equation is as follows:

Step is: 1H-thieno [3,2-c] pyrazoles-5-methanol (1.0g, 6.5mmol) is dissolved in dichloromethane (115mL) In, add Dess-Martin oxidant (3.3g, 7.8mmol), be stirred at room temperature 3 hours.Reduce pressure after completion of the reaction Solvent, residue silica gel column chromatography purification are evaporated off, obtain 1H-thieno [3,2-c] pyrazoles-5-formaldehyde (0.90g, 91%), For white solid.

Step 8 reaction equation is as follows:

Step is: 1H-thieno [3,2-c] pyrazoles-5-formaldehyde (700mg, 4.6mmol), 3-aminochlotononitlile (831mg, 10.1mmol) jointly it is dissolved in glacial acetic acid (15mL), is heated to 90 DEG C and stirs 1 hour.Remove acetic acid under reduced pressure, remaining Thing column chromatography purification obtains target product 2,6-dimethyl-4-(1H-thieno [3,2-c] pyrazoles-5-base)-1,4-dihydropyridine -3,5-dimethoxy nitrile (820mg, 66%), for off-white color solid.MS:282(M+H)；¹H NMR(300MHz,DMSO-d₆) 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)。

Synthetically prepared example 2

Prepare 2-ethyl-6-methyl-4-(1H-thieno [3,2-c] pyrazoles-5-the base)-1,4-dihydro of structural formula (I-2) Pyridine-3,5-dimethoxy nitrile

Step 1 reaction equation is as follows:

Step is: under nitrogen protection, anhydrous tetrahydro furan (100mL) is cooled to-70 DEG C, adds n-BuLi (2.5M Hexane solution, 12.8mL, 32mmol), stirring drip anhydrous acetonitrile (1.15g, 28mmol) in a moment.Stir Mixing 3 minutes, dropping ethyl propionate (2.04g, 20mmol), during dropping and keep temperature of reaction system to be less than-66 DEG C. The system of finishing is warming up to-45 DEG C and stirs 2 hours, drips 1N hydrochloric acid (64mL) cancellation.Reactant liquor concentrates, residue Middle addition ether extracts, and extract merges, and anhydrous sodium sulfate is dried, and concentrates, residual oil thing 3-oxopentanenitrile (1.802 G, 93% thick yield) it is directly used in next step reaction.¹H NMR(300MHz,CDCl₃): 3.48 (s, 2H), 2.62(q,2H),1.11(t,3H)。

Step 2 reaction equation is as follows:

Step is: (preparation process sees conjunction to 1H-thieno [3,2-c] pyrazoles-5-formaldehyde (200mg, 1.314mmol) Become preparation example 1) and 3-oxopentanenitrile (191mg, 1.972mmol) be jointly dissolved in dichloromethane (20mL), add Enter piperidines (11mg, 0.131mmol), glacial acetic acid (12mg, 0.197mmol), reflux under nitrogen protection the most anti- Should.Reactant liquor is cooled to room temperature, adds dehydrated alcohol (20mL) dilute reaction solution, filter after stirring ten minutes, filter Liquid concentrates dry, adds ethyl acetate (5mL) and petroleum ether (10mL), filter, solid after stirring five minutes in residue It is dried to obtain 2-((1H-thieno [3,2-c] pyrazoles-5-base) methene)-3-oxopentanenitrile (190mg, 62.5%). ESI-MS:232 [M+H].

Step 3 reaction equation is as follows:

Step is: and 2-((1H-thieno [3,2-c] pyrazoles-5-base) methene)-3-oxopentanenitrile (140mg, 0.61 Mmol), 3-aminochlotononitlile (109mg, 1.33mmol) be jointly dissolved in glacial acetic acid (10mL), be heated to 100 ° C stirs 1 hour.Being cooled to room temperature, reactant liquor concentrating under reduced pressure is done, residue column chromatography (dichloromethane: methanol=80:1) Purification, obtains marking compound 2-ethyl-6-methyl-4-(1H-thieno [3,2-c] pyrazoles-5-base)-Isosorbide-5-Nitrae-dihydropyridine -3,5-dimethoxy nitrile (120mg, 67%).ESI-MS:296 [M+H]；¹H NMR(300MHz,DMSO-d₆)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).

Synthetically prepared example 3

Prepare 2-isobutyl group-6-methyl-4-(1H-thieno [3,2-c] pyrazoles-5-the base)-1,4-two of structural formula (I-3) Pyridinium hydroxide-3,5-dimethoxy nitrile

Step 1 reaction equation is as follows:

Step is: under nitrogen protection, anhydrous tetrahydro furan (100mL) is cooled to-70 DEG C, adds n-BuLi (2.5M Hexane solution, 12.8mL, 32mmol), then drip anhydrous acetonitrile (1.15g, 28mmol), finish stirring After 3 minutes, dropping ethyl isovalerate (2.6g, 20mmol), keep temperature of reaction system less than-66 ° during dropping C.Finish and be warming up to-45 DEG C of stirrings 2 hours.Dropping 1N hydrochloric acid (40mL) cancellation reactant liquor, concentrating under reduced pressure, remaining Adding ether extraction in thing, extract merges, and is dried, and concentrates, the residue grease 5-methyl own nitrile of-3-oxo (2.68 G, 100% thick yield) it is directly used in next step reaction.¹H NMR(300MHz,CDCl₃)0.93(d,6H),2.16(m, 1H),2.49(t,2H),3.43(s,2H)。

Step 2 reaction equation is as follows:

Step is: (preparation process sees conjunction to 1H-thieno [3,2-c] pyrazoles-5-formaldehyde (200mg, 1.31mmol) Become preparation example 1), the 5-own nitrile of methyl-3-oxo (329mg, 2.63mmol) be dissolved in dichloromethane (20mL), add Enter glacial acetic acid (12mg, 0.20mmol) and piperidines (11mg, 0.13mmol), be stirred at reflux under nitrogen protection overnight. Being cooled down by reactant liquor, add dehydrated alcohol (20mL) dilute reaction solution, filter after stirring ten minutes, filtrate concentrates dry, Residue column chromatography (dichloromethane: methanol=300:1) obtains 2-((1H-thieno [3,2-c] pyrazoles-5-base) methylene Base)-5-methyl own the nitrile of-3-oxo (294mg, 86%).MS(ESI+)260[M+H].

Step 3 reaction equation is as follows:

Step is: 2-((1H-thieno [3,2-c] pyrazoles-5-base) the methene)-5-methyl own nitrile of-3-oxo (100mg, 0.605mmol), 3-aminochlotononitlile (70mg, 0.848mmol) be jointly dissolved in glacial acetic acid (8mL), be heated to 100 DEG C stir 1 hour, be cooled to room temperature, concentrating under reduced pressure do, residue silica gel prepare plate purification (dichloromethane: Methanol=20:1), obtain 2-isobutyl group-6-methyl-4-(1H-thieno [3,2-c] pyrazoles-5-base)-Isosorbide-5-Nitrae-dihydropyridine -3,5-dimethoxy nitrile (73mg, 59%).MS(ESI+)324[M+H]；¹H NMR(300MHz,DMSO-d₆)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).

Synthetically prepared example 4

Prepare 2-methyl-4-(1H-thieno [3,2-c] pyrazoles-5-base)-6-(the 3,3,3-trifluoropropyl of structural formula (I-4) Base)-1,4-dihydropyridine-3,5-dimethoxy nitrile

Step 1 reaction equation is as follows:

Step is: under nitrogen protection, anhydrous tetrahydro furan (100mL) is cooled to-74 DEG C, adds n-BuLi (2.5M Hexane solution, 16mL, 40mmol), then drip anhydrous acetonitrile (1.437g, 35mmol).Stir 3 minutes After, drip 4,4,4-trifluoroacetic acid ethyl ester (4.253g, 25mmol), during keep temperature of reaction system be less than -69℃.Finish and be warming up to-45 DEG C of stirrings 2 hours.Dropping 1N hydrochloric acid (50mL) cancellation reactant liquor, concentrating under reduced pressure, Adding ether extraction in residue, extract merges, and is dried, and concentrates, and residue (3.908g, 94% thick yield) is direct React for next step¹H NMR(300MHz,CDCl₃) 2.43-2.55 (m, 2H), 2.89 (t, 2H, J=7.5 Hz),3.55(s,2H)。

Step 2 reaction equation is as follows:

Step is: (preparation process sees synthesis to 1H-thieno [3,2-c] pyrazoles-5-formaldehyde (304mg, 2.0mmol) Preparation example 1), 6,6, the 6-tri-own nitrile of fluoro-3-oxos (660mg, 4.0mmol) be dissolved in dichloromethane (20mL), add Enter glacial acetic acid (12mg, 0.20mmol) and piperidines (11mg, 0.13mmol), be stirred at reflux under nitrogen protection overnight. Being cooled down by reactant liquor, add dehydrated alcohol (20mL) dilute reaction solution, filter after stirring ten minutes, filtrate concentrates dry, Residue column chromatography (dichloromethane: methanol=200:1) obtains 2-((1H-thieno [3,2-c] pyrazoles-5-base) methylene Base)-6,6,6-three fluoro-3-own the nitrile of oxo (340mg, 57%).MS(ESI+)300[M+H].

Step 3 reaction equation is as follows:

Step is: 2-((1H-thieno [3,2-c] pyrazoles-5-base) the methene)-6,6,6-three fluoro-3-own nitrile of oxo (200 Mg, 0.67mmol), 3-aminochlotononitlile (70mg, 0.848mmol) be jointly dissolved in glacial acetic acid (8mL), Being heated to 100 DEG C to stir 1 hour, be cooled to room temperature, concentrating under reduced pressure is done, and plate purification (dichloro prepared by residue silica gel Methane: methanol=20:1), obtain 2-methyl-4-(1H-thieno [3,2-c] pyrazoles-5-base)-6-(3,3,3-trifluoropropyl Base)-1,4-dihydropyridine-3,5-dimethoxy nitrile (63mg, 26%).MS(ESI+)364[M+H]；¹H NMR(300MHz, DMSO-d₆)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)。

Synthetically prepared example 5

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

Step 1 reaction equation is as follows:

Step is: is joined by n-BuLi (31mL, 77mmol) in the THF (350ml) of-78 DEG C, then drips Add acetonitrile (2.9g, 70mmol), finish, stir 1h at low temperatures.Then 2-methoxy methyl propionate (5.9 is instilled G, 50mmol), finish, temperature is risen to-45 DEG C, and stirs 2h, at low temperatures with 2N hydrochloric acid (160mL) Cancellation, reactant liquor is slowly increased to room temperature, extracts with ether, merges organic facies, is concentrated to give 5-methoxyl group-3-oxopentanenitrile (3.5g, 55%).

Step 2 reaction equation is as follows:

Step is: by 1H-thieno [3,2-c] pyrazoles-5-formaldehyde (350mg, 2.3mmol), (preparation process sees Synthetically prepared example 1) be dissolved in dichloromethane (5mL), be subsequently adding 5-methoxyl group-3-oxopentanenitrile (292mg, 2.3 Mmol), piperidines (11mg, 0.13mmol), acetic acid (99mg, 1.643mmol) and 4A molecular sieve, finish, instead Return stirring should be heated to overnight.Being cooled down by reactant liquor, concentrate, residue is directly used in next step.

Step 3 reaction equation is as follows:

Step is: upper step gained residue (600mg, 2.3mmol) be dissolved in glacial acetic acid (6mL), adds 3-aminochlotononitlile (190mg, 2.3mmol), is heated to 100 DEG C of stirring 1h.Cooling, concentrates dry by reactant liquor, residual Excess is purified and is obtained product 25mg with preparing TLC, and 3%.MS:[M+1]=326；¹H NMR(300MHz,CDCl₃)δ 7.80(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).

Synthetically prepared example 6

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

Step 1 reaction equation is as follows:

Step is: (preparation process sees conjunction to 1H-thieno [3,2-c] pyrazoles-5-formaldehyde (150mg, 0.99mmol) Become preparation example 1) and 4-fluoro benzoyl acetonitrile (177mg, 1.08mmol) be codissolved in dichloromethane (15mL), Add acetic acid (9mg, 0.15mmol) and piperidines (8mg, 0.10mmol), reflux under nitrogen protection the most anti- Should.Being cooled down by reactant liquor, add dichloromethane (30mL) dilute reaction solution, filter, filtrate concentrates dry, residue Column chromatography (dichloromethane: methanol=100:1) obtains 2-(4-fluoro benzoyl)-3-(1H-thieno [3,2-c] pyrazoles -5-base) acrylonitrile (259mg, 88%).MS(ESI+):298[M+1]⁺。

Step 2 reaction equation is as follows:

Step is: by 2-(4-fluoro benzoyl)-3-(1H-thieno [3,2-c] pyrazoles-5-base) acrylonitrile (259 Mg, 0.87mmol) and aminochlotononitlile (157mg, 1.917mmol) be dissolved in acetic acid (15mL), be heated to 100 DEG C are stirred 1 hour.After cooling, reactant liquor is concentrated dry, preparation TLC purification (dichloromethane: methanol=20:1) Obtain 2-(4-fluorophenyl)-6-methyl-4-(1H-thieno [3,2-c] pyrazoles-5-base)-1,4-dihydropyridine-3,5-diformazan Nitrile (62mg, 20%).HPLC display purity is 97%；LC-MS(ESI+):362[M+1]⁺；¹H NMR(300MHz, CDCl₃) 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)。

Synthetically prepared example 7

Prepare 2-(4-methoxyphenyl)-6-methyl-4-(1H-thieno [3,2-c] pyrazoles-5-of structural formula (I-7) Base)-1,4-dihydropyridine-3,5-dimethoxy nitrile

Step 1 reaction equation is as follows:

Step is: methyl p-methoxybenzoate (2.0g, 12mmol) is dissolved in dry toluene (50mL), fall Temperature, to 0 DEG C, adds sodium hydrogen (1.2g, 30mmol).Acetonitrile (1.23g, 30mmol) is added after stirring 10 minutes, It is to slowly warm up to 110 DEG C, is stirred overnight.After being cooled to room temperature, filtering, filtering residue is collected after washing with toluene and is dried, Obtain 1-cyano group-2-(4-methoxyphenyl)-2-oxypropionitrile sodium salt (2.2g, 93%).

Step 2 reaction equation is as follows:

Step is: 1-cyano group-2-(4-methoxyphenyl)-2-oxypropionitrile sodium salt (500mg, 3.29mmol), 1H-thiophene Fen also [3,2-c] pyrazoles-5-formaldehyde (980mg, 4.93mmol) (preparation process sees synthetically prepared example 1) adds to two In chloromethanes (50mL), add glacial acetic acid (490mg, 4.93mmol) and piperidines (280mg, 3.29mmol), nitrogen The lower backflow of protection is stirred overnight.Reactant liquor is cooled down, adds dichloromethane (30mL) dilute reaction solution, filter, filtrate Wash with water, be dried and concentrate and do to obtain 2-(4-anisoyl)-3-(1H-thiophene [3,2-c] pyrazoles-5-base) third Alkene nitrile (1.0g), crude product is directly used in next step reaction.LC-MS(ESI+)310[M+H].

Step 3 reaction equation is as follows:

Step is: 2-(4-anisoyl)-3-(1H-thiophene [3,2-c] pyrazoles-5-base) acrylonitrile (300mg, 0.92mmol), aminochlotononitlile (152mg, 1.84mmol) be jointly dissolved in glacial acetic acid (20mL), N₂Under protection It is heated to 100 DEG C to stir 1 hour.Being cooled to room temperature, concentrated by reactant liquor dry, plate (DCM:MeOH=10:1) prepared by silica gel Purification, obtains 2-(4-methoxyphenyl)-6-methyl-4-(1H-thieno [3,2-c] pyrazoles-5-base)-Isosorbide-5-Nitrae-dihydropyridine -3,5-dimethoxy nitrile (35mg, 10%).MS(ESI+)374[M+H].¹H NMR(300MHz,CDCl₃)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)。

Synthetically prepared example 8

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

Step 1 reaction equation is as follows:

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

Step 2 reaction equation is as follows:

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

Step 3 reaction equation is as follows:

Step is: the different nicotinoyl-3-of 2-(1H-thieno [3,2-c] pyrazoles-5-yl) (550mg, 1.96mmol), amino Crotonic nitrile (400mg, 4.91mmol) is dissolved in acetic acid (20mL) jointly, is heated to 100 DEG C and stirs 1 hour. and cold But to room temperature, reactant liquor is concentrated dry, and silica gel is prepared plate purifying (DCM:MeOH=10:1) and obtained 2-methyl-6-(pyridine-4-Base)-4-(1H-thieno [3,2-c] pyrazoles-5-yl)-Isosorbide-5-Nitrae-dihydropyridine-3,5-dimethoxy nitrile (80mg, 129%) .MS (ESI+) 362[M+H];¹H NMR(300MHz,CDCl₃)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,1 H,),2.10(s,3H)。

Synthetic preparation example 9

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

Step 1 reaction equation is as follows:

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

Step 2 reaction equation is as follows:

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

Step 3 reaction equation is as follows:

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

Step 4 reaction equation is as follows:

Step is: upper step gained (piperidines-2-thiazolinyl) acetonitrile (55mg crude product) is dissolved in glacial acetic acid (16mL), Add 2-((1H-thieno [3,2-c] pyrazoles-5-yl) the methylene)-3-oxo making in synthetic preparation example 2 steps 1 Butyronitrile (65mg, 0.3mmol), is heated to 100 DEG C and stirs 1 hour. by concentrated reactant liquor dry, and residue post layer Analyse (carrene: methyl alcohol=50:1) and obtain 6-methyl-8-(1H-thieno [3,2-c] pyrazoles-5-yl)-2,3,4,8-tetra-Hydrogen-1H-quinolizine-7,9-dimethoxy nitrile (63mg, 66%) .HPLC:98.4%; LC-MS (ESI+): 322[M+1]⁺；¹H NMR(300MHz,CDCl₃)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 wherein, 2-c] pyrazoles-5-yl) methylene)-3-oxo butyronitrile, can prepare respectively Compound I-10 in following table to I-12:

Synthetic preparation example 13:

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

Step 1 reaction equation is as follows:

Step is: 3-morpholone (303mg, 3mmol) is dissolved in carrene (20mL), adds MeOTf (TFMS methyl esters) (610mg, 3.72mmol), stirring at room temperature 18 hours. add Na₂CO₃Powder (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 equation is as follows:

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

Step 3 reaction equation is as follows:

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

Step 4 reaction equation is as follows:

Step is: above walk gained 2-(morpholine-3-thiazolinyl) acetonitrile crude product (103mg, 0.83mmol) and be dissolved in ice Acetic acid (6mL), adds (the synthetic preparation of 2-((1H-thieno [3,2-c] pyrazoles-5-yl) methylene)-3-oxo butyronitrile Example 2 steps 1,102mg, 0.468mmol), be heated to 100 DEG C and stir 15 minutes. and be chilled to room temperature, decompression is steamed Desolventize, residue is prepared TLC purifying (carrene: methyl alcohol=15:1) and is obtained faint yellow solid (29mg, 19%). HPLC purity: 95%;¹H NMR(300MHz,CDCl₃)δ7.75(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, substitute 2-((1H-thieno [3,2-c] pyrazoles-5-yl) methylene)-3-oxo butyronitrile wherein with corresponding intermediate, can prepare respectively Compound I-14 to I-16:

Synthetic preparation example 17:

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

Step 1 reaction equation is as follows:

Step is: 1H-thieno [3, 2-c] pyrazoles-5-methyl formate (8.5g, 46.7mmol) (preparation process is referring to closing become preparation example 1), potash (7.75g, 56mmol) add in DMF (150mL), by p-methoxybenzyl chloride (7.3g, 46.7mmol) be slowly added dropwise to reactant liquor, stirring at normal temperature 1 hour. remove DMF under reduced pressure, residue is molten with ethyl acetate separate, wash with water, saturated common salt washing, 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 equation is as follows:

Step is: by lithium aluminium hydride reduction (1.36g, 35.76mmol) be suspended in oxolane (100mL), be cooled to 0 DEG C, drip 1-(4-methoxy-benzyl)-1H-thieno [3, 2-c] pyrazoles-5-carboxylate methyl ester (7.2g, 23.8mmol) four hydrogen furans (100mL) solution, finish stirring at room temperature 2 hours. under frozen water is cooling, slowly drip water (1.4mL) cancellation, filtering. filter residue is washed with the mixed solvent (3:1) of carrene and methyl alcohol, filter, filtrate is concentrated, column chromatography obtains (1-(4-methoxy-benzyl)-1H-thieno [3, 2-c] pyrazoles-5-yl) methyl alcohol (5.94g, 91%).

Step 3 reaction equation is as follows:

Step is: (1-(4-methoxy-benzyl)-1H-thieno [3; 2-c] pyrazoles-5-yl) methyl alcohol (5.94g; 21.7mmol) be dissolved in carrene (150mL); add Dai Si-Martin oxidant (13.8g; 32.5mmol); room temperature reaction 2 hours under nitrogen protection. by concentrated solvent dry; residue column chromatography obtains 1-(4-methoxy-benzyl)-1H-thieno [3; 2-c] pyrazoles-5-formaldehyde (3.76g, 64%).

Step 4 reaction equation 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), being heated to 100 DEG C stirs 1 hour, remove acetic acid under reduced pressure, the direct column chromatography of residue 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 equation 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) be dissolved in DMF (30mL), be cooled to 0 DEG C, by carbon acid caesium (719mg, 2.2mmol) add reactant liquor. stir 15 minutes, add iodomethane (209mg, 1.47mmol), stirring at room temperature 4 hours. in reactant liquor, add carrene dilution, water respectively, saturated common salt washing, organic phase nothing aqueous sodium persulfate is dry, concentrated, column chromatography obtains 4-(1-(4-methoxy-benzyl)-1H-thieno [3, 2-c] pyrazoles-5-base)-1, 2, 6-trimethyl-1, 4-dihydropyridine-3, 5-dimethoxy nitrile (611mg, 100%).

Step 6 reaction equation is as follows:

Step is: compound 5 (611mg, 1.47mmol) is dissolved in trifluoroacetic acid (20mL), is heated to 90 DEG C Stirring is spent the night. and by concentrated trifluoroacetic acid dry, column chromatography obtains target product 1,2,6-trimethyl-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-d₆)δ7.81(s,1H),7.02(s,1H),4.72(s,1H),3.19(s,3 H),2.20(s,6H)。

Synthetic preparation example 18:

Prepare 1-ethyl-2 of 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, replace iodomethane wherein with bromoethane, can prepare this compound .MS (ESI+) 310[M+1];¹H NMR(300MHz,DMSO-d₆)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

Prepare 1-benzyl-2 of 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, replace iodomethane wherein with cylite, can prepare this compound .MS (ESI+): 372[M+1];¹H NMR(300MHz,DMSO-d₆)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-furans is [3,4-c] quinolizine-5-formonitrile HCN also, and 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 n-amyl alcohol (5mL), return stirring 1 hour. be chilled to room temperature, add 2-(piperidines-2-thiazolinyl) acetonitrile (367mg of fresh preparation, 3.0mmol), glacial acetic acid (4mL), 100 DEG C are stirred 1.5 hours. be cooled to room temperature, reduced pressure concentration, residue is prepared TLC purifying (carrene: methyl alcohol=10:1) obtains target product, faint yellow solid (59mg, 9%) .MS (ESI+): 339[M+1].

Synthetic preparation example 21

3-oxygen-the 4-(1H-thieno [3,2-c] pyrazoles-5-yl)-1,3,4,6,8 for preparing structural formula (I-21), 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 closing become preparation example 1) and furans-2, 4 (3H, 5H)-diketone (147mg, 1.469mmol) add to n-amyl alcohol (5mL) in, return stirring 1 hour. be chilled to room temperature, add 2-(morpholine-3-thiazolinyl) acetonitrile (273mg of fresh preparation, 2.2mmol), glacial acetic acid (4mL), 100 DEG C are stirred 1.5 hours. be cooled to room temperature, reduced pressure concentration, remaining thing is prepared TLC purifying (carrene: methyl alcohol=10:1) and is obtained target product faint yellow solid (60mg, 12%) .LC-MS (ESI+): 341[M+1]⁺；¹H NMR(300MHz,CDCl₃)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-isoxazole 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)-3-oxo butyronitrile (175mg, 0.81 Mmol) (preparation process is referring to synthetic preparation example 2 steps 1) are dissolved in glacial acetic acid (10mL), add the different evil of 3-methyl Azoles-5-amine (79mg, 0.81mmol), is heated to 100 DEG C and stirs 1 hour. and be cooled to room temperature, reactant liquor is concentrated Dry, residue is prepared TLC purifying (carrene: methyl alcohol=20:1) and is obtained target compound (30mg, 13%) .HPLC Show that purity is 95.2%; LC-MS (ESI+): 298[M+1]⁺；¹H NMR(300MHz,DMSO-d₆)δ10.96 (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:

6-(4-the fluorophenyl)-3-methyl-4-(1H-thieno [3,2-c] pyrazoles-5-yl)-4 for preparing structural formula (I-23), 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) acrylonitrile (480mg; 1.614mmol) (preparation process is referring to synthetic preparation example 6) is dissolved in glacial acetic acid (30mL); add 3-methyl isoxazole-5-amine (190mg; 1.937mmol); being heated to 100 DEG C stirs 1 hour. be cooled to room temperature; by concentrated reactant liquor dry; residue silica gel is prepared plate (DCM:MeOH=20:1) purifying; obtain compound 3 (20mg, 3.3%) .MS (ESI+) 378[M+1]⁺。¹H NMR(300MHz,DMSO-d₆)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,1H₉),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-3,5-dimethoxy nitrile,

Step 1 reaction scheme is as follows:

Step is: by sodium piece (5.13g, 223mmol) add reaction in methyl alcohol (30mL) until sodium piece is complete disappearance obtains sodium methoxide solution. to 1-acetyl group-1H-thieno [3, 2-c] pyrazoles-5-methyl formate (10.0g, 44.6 mmol) in methyl alcohol (100mL) solution of (preparation process is referring to synthetic preparation example 1), add the above-mentioned sodium methoxide making solution, the mixture obtaining at room temperature stirs 30 minutes. in above-mentioned reactant liquor, drip iodine (28.3g again, 112mmol) DMF (30mL) solution. after dropwising, reactant liquor is heated to 60 DEG C of stirrings to spend the night. and reactant liquor is cooled to chamber temperature, concentrated dry, residue adds ethyl acetate and water extraction, ethyl acetate is through washing, after saturated common salt washing, dry dry, be concentrated to certain volume, filtration obtains the iodo-1H-thieno [3 of 3-, 2-c]-5-methyl formate (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 formate (5.0g, 16.23mmol), DMAP (397mg, 3.25mmol) and Et₃N (1.81g, 17.85mmol) is dissolved in carrene (100mL), then drips Boc₂O (4.25g, 19.47mmol), dropwise, under room temperature, react 30 minutes, by concentrated reactant liquor dry, residue purification by silica gel column chromatography, obtains the iodo-1H-thieno [3 of 1-tertbutyloxycarbonyl-3-, 2-c] pyrazoles-5-methyl formate (5.9g, 89%) .LC-MS (ESI+): 409[M+H]⁺。

Step 3 reaction scheme is as follows:

Step is: iodo-1-tertbutyloxycarbonyl-3-1H-thieno [3,2-c] pyrazoles-5-methyl formate (5.5g, 13.47mmol) is dissolved in to Isosorbide-5-Nitrae-dioxane (100mL), adds Pd (dppf) Cl₂(986mg, 1.347mmol), drip zinc methide (1.15M toluene solution, 17.6mL, 20.2mmol), finish, nitrogen replacement three times, be heated to 100 DEG C stir 1 hour. be cooled to room temperature, add carrene (300mL) dilute reaction solution, drip the neutralization of 1N hydrochloric acid reactant liquor is to pH=7 left and right, organic phase is through washing, saturated common salt washing, dry, concentrated, residue column chromatography (two chloromethanes: methyl alcohol=125:1) 3-methyl isophthalic acid H-thieno [3, 2-c] pyrazoles-5-methyl formate (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 formate (1.46g, 7.45mmol) be dissolved in in DMF (30mL), add potash (2.06g, 14.9mmol), drip PMBCl (1.4g, 8.94mmol), finish, stirring at room temperature 4 hours. by concentrated reactant liquor dry, residue is dissolved in carrene (100mL), washing, full wash with salt, dry, concentrated, residue column chromatography (PE:EA=10:1), obtain 1-(4-methoxy-benzyl)-3-methyl isophthalic acid H-thieno [3, 2-c] pyrazoles-5-methyl formate (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 formate (429mg, 1.36mmol) is dissolved in to THF (20mL), adds LiBH₄(118mg, 5.43mmol), stirring at room temperature 3 hours, adds carrene (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 washing, is dried, residue 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, 1.354 mmol) be dissolved in carrene (25mL), add Dess-Martin oxidant (1.15g, 2.708mmol), room temperature reaction 3 hours under nitrogen protection, TLC (PE:EA=2:1) shows that reaction is complete, reactant liquor is concentrated, residual excess column chromatography (PE:EA=30:1) obtains 1-(4-methoxy-benzyl)-3-methyl isophthalic acid H-thieno [3, 2-c] pyrazoles-5-first aldehyde (279mg, 71%) .LC-MS (ESI+): 287[M+H]⁺。

Step 7 reaction scheme is as follows:

Step is: by 1-(4-methoxy-benzyl)-3-methyl isophthalic acid H-thieno [3, 2-c] pyrazoles-5-formaldehyde (275mg, 0.96mmol) with amino crotonic nitrile (174mg, 2.12mmol) be dissolved in glacial acetic acid (10mL), be heated to 100 DEG C reaction 1 hour. by concentrated reactant liquor, residue 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 (399mg, thick yield 100%), directly use 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-Base)-2,6-dimethyl-Isosorbide-5-Nitrae-dihydropyridine-3,5-dimethoxy nitrile crude product (399mg, 0.96mmol) is dissolved in trifluoroacetic acid (10mL) in, be heated to 85 DEG C of back flow reaction 24 hours, react completely. by concentrated reactant liquor dry, in residue Add carrene (20mL), with saturated sodium bicarbonate solution washing, then water and saturated common salt washing, dry, dense Contracting, prepares plate purifying through silica gel after residue column chromatography again, obtains target product 2,6-dimethyl-4-(3-methyl isophthalic acid H-thiophene Fen is [3,2-c] pyrazoles-5-yl also)-Isosorbide-5-Nitrae-dihydropyridine-3,5-dimethoxy nitrile (120mg, 42%) .HPLC purity: 98.3%; MS (ESI+): 296[M+H];¹H NMR(300MHz,DMSO-d₆)δ12.6(br,1H),9.68(br,1 H),6.97(s,1H),4.77(s,1H),2.31(s,3H),2.06(s,6H)。

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

Synthetic preparation example 29

Prepare the 4-(3-ethyl-1H-thieno [3,2-c] pyrazoles-5-yl)-2 of structural formula (I-29), 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 1-tertbutyloxycarbonyl-3-, 2-c] pyrazoles-5-methyl formate (8.2g, 20mmol) (preparation process is referring to synthetic preparation example 24) is dissolved in Isosorbide-5-Nitrae-dioxane (200mL), adds Pd (dppf) Cl₂(1.46g, 2mmol), drip diethyl zinc (1M toluene solution, 30mL, 30mmol), finish nitrogen replacement three times, being heated to 100 DEG C stirs 1 hour. and add carrene (200mL) dilute reaction solution, drip 1N hydrochloric acid neutralization reaction liquid to pH 7 left and right, organic phase is through washing, saturated common salt washing, dry, concentrated, residue column chromatography (carrene: methyl alcohol=400:1) obtains 3-ethyl-1H-thieno [3,2-c] pyrazoles-5-methyl formate 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 formate crude product (9.73g, 46.28mmol) is dissolved in anhydrous tetrahydro furan (250mL), is cooled to-15 DEG C, adds LiAlH₄(5.27g, 138.83mmol), nitrogen protection is stirred 2 hours, adds LiAlH₄(1.32g, 34.72mmol), rising to room temperature continues to stir 1 hour. the careful water (10mL) that drips, stir 10 minutes, add again methyl alcohol (100mL) and carrene (100mL), stir mix after ten minutes and filter, in filter residue, add methyl alcohol (50mL) and carrene (50mL), stir after ten minutes and filter, filtrate merges, through washing, saturated common salt washing, dry, concentrated, residue column chromatography (carrene: 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 carrene (200mL); add Dess-Martin oxidant (6.36g; 15mmol); room temperature reaction 2 hours under nitrogen protection; reactant liquor is concentrated; residue column chromatography (carrene: methyl alcohol=300:1) obtains 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 crotons Nitrile (397mg, 4.84mmol) is dissolved in glacial acetic acid (20mL), is heated to 100 DEG C of reactions 1 hour. will react Liquid is concentrated, and residue column chromatography (carrene: methyl alcohol=80:1) obtains 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 is aobvious Show that purity is 98.2%; LC-MS (ESI+): 310[M+1]⁺；¹H NMR(300MHz,CDCl₃)δ8.99(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).

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

Synthetic preparation example 34

Prepare the 4-(3-methoxyl group-1H-thieno [3,2-c] pyrazoles-5-yl)-2 of 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 formate (10.0g, 32.4mmol) (preparation step suddenly referring to synthetic preparation example 1) be dissolved in DMF (100ml), under frozen water is cooling, add potash (9.0g, 64.9 mmol), drip methoxyl group benzyl chloride (6.9g, 38.9mmol), stirring at room temperature 2 hours. in system, add water, second acetoacetic ester extraction, merge ester layer, 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 formate (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 formate (0.86g, 2 mmol), sodium methoxide (0.44g, 8mmol), 3, 4, 7, luxuriant and rich with fragrance quinoline (the 0.24g of 8-tetramethyl, 1mmol), iodine change cuprous (0.38g, 2mmol) be mixed in single neck bottle, add absolute methanol (50mL), under nitrogen protection, 80 DEG C are stirred 16 hours. be cooled to room temperature, in system, add water, 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] pyrrole azoles-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), is cooled to-10 DEG C, adds LiAlH₄(500mg), stir 2 hours. carefully drip the shrend reaction of going out, leave standstill, filter. filter residue is washed with carrene, filtrate is with dichloromethane extraction, carrene 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-Base) methanol (140mg, 0.46mmol) is dissolved in dry dichloromethane (20mL), is cooled to-10 DEG C, add Dess-Martin reagent (293mg, 0.69mmol), stirs 2 hours. rise to room temperature, stir 2 hours .Dropping Saturated sodium bicarbonate aqueous solution, separates dichloromethane phase, and aqueous phase extracts with dichloromethane, and dichloromethane merges, and is dried, Concentrate, column chromatography purification (ethyl acetate/petroleum ether=1/4), obtain 3-methoxyl group-1-(4-methoxy-benzyl)-1H-thiophene And [3, 2-c] pyrazoles-5-formaldehyde (115mg, yield 83%), yellow solid.¹H NMR(300MHz,CDCl₃):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) it is dissolved in glacial acetic acid (7mL), addition aminochlotononitlile (142mg, 1.73mmol), 90 DEG C Stir 1 hour.Removing acetic acid under reduced pressure, residue is 4-(3-methoxyl group-1-(4-methoxy-benzyl)-1H-thieno [3,2-c] pyrazoles-5-base)-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), return stirring 16 hours in above-mentioned brown oil.Remove three under reduced pressure Fluorine acetic acid, residue frozen water is cooling, saturated sodium bicarbonate aqueous solution alkalization, dichloromethane extraction .Extract merges, nothing Aqueous sodium persulfate is dried, and concentrates, and preparation TLC purification (methylene chloride/methanol=20:1) obtains 4-(3-methoxyl group-1H-thiophene Fen also [3,2-c] pyrazoles-5-base)-2,6-dimethyl-Isosorbide-5-Nitrae-dihydropyridine-3,5-dimethoxy nitrile, faint yellow solid 90mg, Yield 37%.MS:(312, M+H);¹H NMR(DMSO-d₆,300MHz):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, use with front State compound I-9, the preparation method that I-13, I-6 are similar, can prepare following compound respectively:

Synthetically prepared example 38

Prepare 4-(3-amino-1H-thieno [3,2-c] pyrazoles-5-the base)-2,6-dimethyl-1,4-of structural formula (I-38) Dihydropyridine-3,5-dimethoxy nitrile

Step 1 reaction scheme is as follows:

Step is: the 1-tert-butyl group, 5-methyl-3-iodo-1H-thieno [3,2-c] pyrazoles-1, and 5-dicarboxylic acid esters (0.82g, 2 mmol)、Pd₂(dba)₃(183mg, 0.2mmol), Xantphos (347mg, 0.6mmol), cesium carbonate (980mg, 3mmol) jointly it is dissolved in Isosorbide-5-Nitrae-dioxane (20mL), adds benzophenone imine (540mg, 3mmol), Nitrogen is protected, and is heated to 100 DEG C and stirs 6 hours.It is cooled to room temperature, removes solvent under reduced pressure, residue adds water and second Acetoacetic ester, separates organic facies, and anhydrous sodium sulfate is dried, and concentrates.Residue column chromatography purification, obtains faint yellow solid 1- The tert-butyl group, 5-methyl-3-((diphenylmethyl thiazolinyl) amino)-1H-thieno [3,2-c] pyrazoles-1,5-dicarboxylic acid esters (0.68 G, 74%).

Step 2 reaction scheme is as follows:

Step is: the 1-tert-butyl group, 5-methyl-3-((diphenylmethyl thiazolinyl) amino)-1H-thieno [3,2-c] pyrazoles-1,5- Dicarboxylic acid esters (0.68g, 1.47mmol) is dissolved in methanol (10mL), addition oxammonium hydrochloride. (0.112g, 1.62mmol), 6 hours it are stirred at room temperature.Removing solvent under reduced pressure, residue dissolves with dichloromethane, saturated sodium bicarbonate Aqueous solution is washed, and anhydrous sodium sulfate is dried, and column chromatography obtains the 1-tert-butyl group, 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) It is dissolved in acetonitrile (20mL), adds 4-dimethylamino pyridine (159mg, 1.30mmol) and two dimethyl dicarbonates Butyl ester (Boc₂O) (284mg, 1.30mmol), return stirring 2 hours.It is cooled to room temperature, removes solvent under reduced pressure, residual Excess column chromatography purification obtains the white solid 1-tert-butyl group, 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-tert-butyl group, 5-methyl 3-((tertbutyloxycarbonyl) amino)-1H-thieno [3,2-c] pyrazoles-1,5- Dicarboxylic acid esters (0.32g, 0.805mmol) is dissolved in anhydrous tetrahydro furan (10mL), add lithium borohydride (70mg, 3.2mmol), nitrogen is protected, and is stirred at room temperature 5 hours.React rear frozen water to lower the temperature, and dropping dilute hydrochloric acid (1N, 0.5 ML), stirring 0.5 hour is finished.Removing solvent under reduced pressure, residue adds acetic acid ethyl dissolution, washing, is dried after concentrating Column chromatography.Obtain 3-((tertbutyloxycarbonyl) amino)-5-(methylol)-1H-thieno [3,2-c] the pyrazoles tertiary fourth of-1-formic acid Ester (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) is dissolved in dichloromethane (10mL), add Dess-Martin oxidant (509mg, 1.2mmol), it is stirred at room temperature 2 hours under nitrogen protection.Reactant liquor concentrates, 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), 3-aminochlotononitlile (61mg, 0.738mmol) add to glacial acetic acid (5mL) jointly In, 95 DEG C are stirred 15 minutes.Acetic acid is divided exactly in decompression, and residue prepares TLC purification, obtains (5-(3,5-dicyano-2,6- Dimethyl-Isosorbide-5-Nitrae-dihydropyridine-4-base)-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-base)-1H-thieno [3,2-c] pyrazoles-3- Base) t-butyl carbamate (67mg, 0.169mmol) is dissolved in dichloromethane (5mL), adds trifluoroacetic acid (1mL), It is stirred at room temperature 5 hours.Reactant liquor dilutes with ethyl acetate (10mL), adds solid sodium carbonate (3g), stirs 0.5 Hour, to filter, filtrate concentrates, preparation TLC purification (dichloromethane: methanol=10:1), obtains 4-(3-amino-1H- Thieno [3,2-c] pyrazoles-5-base)-2,6-dimethyl-1,4-dihydropyridine-3,5-dimethoxy nitrile (35mg, 70%). ESI-MS:297(M+H)；¹H NMR(300MHz,DMSO-d₆)δ11.46(s,1H),9.69(s,1H),6.78(s, 1H),5.08(s,2H),4.69(s,1H),2.03(s,6H)。

Synthetically prepared example 39

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

Step 1 reaction scheme is as follows:

Step is: and 1-tertbutyloxycarbonyl-3-iodo-1H-thieno [3,2-c] pyrazoles-5-methyl formate (200mg, 0.49 Mmol) (preparation process sees synthetically prepared example 24), pyridine-3-boric acid (90mg, 0.74mmol), Pd(dppf)Cl₂-CH₂Cl₂(42mg, 0.049mmol) and Cs₂CO₃(639mg, 1.96mmol) adds to 1,4- In the mixed solvent of dioxane (10mL) and water (4mL), nitrogen is replaced three times, is heated to 120 DEG C of stirrings 30 Minute.Being cooled to room temperature, concentrated by reactant liquor, residue column chromatography (dichloromethane: methanol=75:1) obtains 3-(pyridine -3-base)-1H-thieno [3,2-c] pyrazoles-5-methyl formate (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 formate (487mg, 1.878mmol) Be dissolved in oxolane (20mL), be cooled to-20 DEG C, add under nitrogen protection lithium aluminium hydride (214mg, 5.635 Mmol), finish, be heated to room temperature reaction 3 hours.Reactant liquor is dripped under frozen water cools down water (0.4mL) cancellation Reaction, filters, and filtering residue washs three times through methanol, and filtrate concentrates, residue column chromatography (dichloromethane: methanol=20:1) Obtain (3-(pyridin-3-yl)-1H-thieno [3,2-c] pyrazoles-5-base) methanol (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-base) methanol (270mg, 1.167 Mmol) it is dissolved in dichloromethane (30mL), adds Dess-Martin oxidant (990mg, 2.335mmol), Room temperature reaction 3 hours under nitrogen protection, concentrate reactant liquor, and residue column chromatography obtains 3-(pyridin-3-yl)-1H-thiophene Fen also [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 Aminochlotononitlile (69mg, 0.884mmol) is dissolved in glacial acetic acid (5mL), is heated to 95 DEG C and reacts 2 hours. Being cooled to room temperature, reactant liquor concentrates, and residue column chromatography obtains 2,6-dimethyl-4-(3-(pyridin-3-yl)-1H-thieno [3,2-c] pyrazoles-5-base)-Isosorbide-5-Nitrae-dihydropyridine-3,5-dimethoxy nitrile (52mg, 38%).HPLC display purity is 97%；¹H NMR(300MHz,DMSO)δ13.43(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]⁺。

Synthetically prepared example 40

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

Step is: the method copying example 39, replaces the pyridine-3-boric acid in step 1 with pyridine-4-boric acid, can prepare This target compound.LC-MS (ESI+): 359 [M+H]⁺；¹H NMR(300MHz,DMSO)δ13.58(s,1 H), 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)。

Synthetically prepared example 41

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

Step 1 reaction scheme is as follows:

Step is 1-tertbutyloxycarbonyl-3-iodo-1H-thieno [3,2-c] pyrazoles-5-methyl formate (2.5g, 6.12mmol) (preparation process sees synthetically prepared example 24), 1-methyl-4-(4,4,5,5-tetramethyl-1,3,2-dioxaborolan-2- Base)-1H-pyrazoles (3.82g, 18.37mmol), Pd (dppf) Cl₂(896mg, 1.22mmol) and Cs₂CO₃(3.99 G, 12.24mmol) add Isosorbide-5-Nitrae-dioxane (125mL) and water (50mL) mixed solvent in, nitrogen is put Change three times, be heated to 100 DEG C and react 1 hour.Being cooled to room temperature, reactant liquor concentrates, and residue is dissolved in dichloromethane (100 ML) and in water (30mL), separating organic facies, washing, saturated common salt is washed, and is dried, and concentrates, residue post layer Analysis (methylene chloride/methanol=200/1) obtains compound 3-(1-methyl isophthalic acid H-pyrazoles-4-base)-1H-thieno [3,2-c] Pyrazoles-5-methyl formate (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-base)-1H-thieno [3,2-c] pyrazoles-5-methyl formate (888mg, 3.386mmol) it is dissolved in DMF (25mL), adds K₂CO₃(936mg, 6.772mmol), drips PMBCl (636 Mg, 4.063mmol), finish, stir 4 hours.Being concentrated by reactant liquor dry, residue is dissolved in dichloromethane (50mL) With in water (20mL), organic facies is washed through washing, saturated common salt, is dried, and concentrates, residue column chromatography (PE:EA =10:1), obtain compound 1-(4-methoxy-benzyl)-3-(1-methyl isophthalic acid H-pyrazoles-4-base)-1H-thieno [3,2-c] Pyrazoles-5-methyl formate (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-base)-1H-thieno [3,2-c] pyrazoles-5- Methyl formate (722mg, 1.89mmol) is dissolved in anhydrous THF (20mL), adds LiBH₄(165mg, mmol), Stir 3 hours.Adding dichloromethane (30mL) dilute reaction solution, dropping 1N hydrochloric acid adjusts about pH=7, organic facies Through washing, saturated common salt is washed, and is dried, and residue column chromatography (PE:EA=6:1) obtains compound (1-(4-methoxybenzyl Base)-3-(1-methyl isophthalic acid H-pyrazoles-4-base)-1H-thieno [3,2-c] pyrazoles-5-base) methanol (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-base)-1H-thieno [3,2-c] pyrazoles-5-base) methanol (403mg, 1.138mmol) is dissolved in dichloromethane (20mL), adds Dess-Martin reagent (996mg, 2.276mmol), under nitrogen protection room temperature reaction 3 hours, TLC (PE:EA =2:1) show that reaction is complete, reactant liquor is concentrated, residue column chromatography (dichloromethane) obtains compound 1-(4-first Oxy-benzyl)-3-(1-methyl isophthalic acid H-pyrazoles-4-base)-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-base)-1H-thieno [3,2-c] pyrazoles -5-formaldehyde (391mg, 1.11mmol) and aminochlotononitlile (200mg, 2.44mmol) are dissolved in glacial acetic acid (10mL) In, it is heated to 100 DEG C and stirs 1 hour.Reactant liquor is concentrated, residue 4-(1-(4-methoxy-benzyl)-3-(1-first Base-1H-pyrazoles-4-base)-1H-thieno [3,2-c] pyrazoles-5-base)-2,6-dimethyl-1,4-dihydropyridine-3,5-two Formonitrile HCN (533mg, 99% thick yield), the most purified be directly used in next step reaction.LC-MS(ESI+):482[M+1]⁺。

Step 6 reaction scheme is as follows:

Step is: above step gained residue (533mg, 1.107mmol) is dissolved in trifluoroacetic acid (10mL), adds Heat stirs 24 hours to 85 DEG C.Being cooled to room temperature, reactant liquor concentrates dry, adds dichloromethane (30mL) in residue, Wash with saturated sodium bicarbonate solution, then wash with water and saturated common salt, be dried, concentrate, residue column chromatography (dichloro Methane: methanol=50:1) obtain target compound 2,6-dimethyl-4-(3-(1-methyl isophthalic acid H-pyrazoles-4-base)-1H-thieno [3,2-c] pyrazoles-5-base)-Isosorbide-5-Nitrae-dihydropyridine-3,5-dimethoxy nitrile (100mg, 25%).LC-MS(ESI+):362 [M+1]⁺；¹H NMR(300MHz,CD₃OD)δ7.90(s,1H),7.82(s,1H),6.97(s,1H),4.72 (s,1H),3.95(s,3H),2.12(s,6H)。

Synthetically prepared example 42

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

Step 1 reaction scheme is as follows:

Step is: by 1-tertbutyloxycarbonyl-3-iodo-1H-thieno [3,2-c] pyrazoles-5-methyl formate (2.0g, 4.9 Mmol), thiophene-2-boric acid (940mg, 7.35mmol), Pd (dppf) Cl₂(717mg, 0.98mmol) and Cs₂CO₃ (4.79g, 14.7mmol) adds in Isosorbide-5-Nitrae-dioxane (50mL) and water (20mL), and nitrogen is replaced three times, It is heated to 100 DEG C to stir 1 hour.Being cooled to room temperature, reactant liquor concentrates, and residue is dissolved in dichloromethane (100mL) In, washing, saturated common salt is washed, and is dried, and concentrates, and residue column chromatography (dichloromethane: methanol=200:1) obtains 3-(thiophene Fen-2-base)-1H-thieno [3,2-c] pyrazoles-5-methyl formate (761mg, 59%).LC-MS(ESI+):265 [M+1]⁺。

Step 2 reaction scheme is as follows:

Step is: by 3-(thiophene-2-base)-1H-thieno [3,2-c] pyrazoles-5-methyl formate (468mg, 1.77mmol) It is dissolved in DMF (15mL), adds K₂CO₃(489mg, 3.54mmol), dropping PMBCl (333mg, 2.12mmol), Finish and be stirred at room temperature 4 hours.Reactant liquor concentrating under reduced pressure, residue is dissolved in dichloromethane, washing, and saturated common salt is washed, It is dried, concentrates, residue column chromatography (PE:EA=10:1), obtain 1-(4-methoxy-benzyl)-3-(thiophene-2-base)-1H- Thieno [3,2-c] pyrazoles-5-methyl formate (680mg, 100%).LC-MS(ESI+):385[M+1]⁺。

Step 3 reaction scheme is as follows:

Step is: by 1-(4-methoxy-benzyl)-3-(thiophene-2-base)-1H-thieno [3,2-c] pyrazoles-5-first Acid methyl ester (680mg, 1.77mmol) is dissolved in anhydrous tetrahydro furan (20mL), adds LiBH₄(154mg, 1.08mmol), 3 hours it are stirred at room temperature.Adding dichloromethane (30mL) dilute reaction solution, dropping 1N hydrochloric acid is adjusted About pH=7, organic facies is washed through washing, saturated common salt, dry, and residue column chromatography (PE:EA=10:1) obtains (1-(4- Methoxy-benzyl)-3-(thiophene-2-base)-1H-thieno [3,2-c] pyrazoles-5-base) methanol (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-base)-1H-thieno [3,2-c] pyrazoles-5-base) methanol (208mg, 0.49mmol) is dissolved in dichloromethane (10mL), addition Dess-Martin reagent (312mg, 0.74mmol), 3 hours it are stirred at room temperature under nitrogen protection.Reactant liquor concentrates, and residue column chromatography (wash by dichloromethane De-) (1-(4-methoxy-benzyl)-3-(thiophene-2-base)-1H-thieno [3,2-c] pyrazoles-5-base) 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-base)-1H-thieno [3,2-c] pyrazoles-5-base) formaldehyde (173mg, 0.49mmol) and aminochlotononitlile (88mg, 1.08mmol) are dissolved in glacial acetic acid (5mL), It is heated to 100 DEG C to stir 1 hour.It is cooled to room temperature, reactant liquor concentrating under reduced pressure, residue column chromatography (PE:EA=6:1) Obtain 4-(1-(4-methoxy-benzyl)-3-(thiophene-2-yl)-1H-thieno [3,2-c] pyrazoles-5-base)-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- Base)-2,6-dimethyl-Isosorbide-5-Nitrae-dihydropyridine-3,5-dimethoxy nitrile (215mg, 0.445mmol) is dissolved in trifluoroacetic acid (7.5 ML), in, it is heated to 70 DEG C and stirs 8 hours.Reactant liquor concentrates dry, and residue adds dichloromethane (30mL) and dissolves, Saturated sodium bicarbonate solution is washed, and saturated common salt is washed, and is dried, and concentrates, residue column chromatography (dichloromethane: methanol=50:1) Obtain 2,6-dimethyl-4-(3-(thiophene-2-base)-1H-thieno [3,2-c] pyrazoles-5-base)-1,4-dihydropyridine-3,5- Dimethoxy nitrile (32mg, 20%).HPLC display purity is 95.54%；LC-MS(ESI+):364[M+1]⁺；¹H NMR(300 MHz,CDCl₃) 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)。

Synthetically prepared example 43

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

Step 1 reaction scheme is as follows:

Step is: and the iodo-1-of 3-(4-methoxy-benzyl)-1H-thieno [3,2-c] pyrazoles-5-methyl formate (0.86g, 2 Mmol) (preparation process sees synthetically prepared example 34), morpholine (0.87g, 10mmol), potassium carbonate (1.38g, 10mmol), L-PROLINE (68mg, 0.5mmol), Hydro-Giene (Water Science). (0.38g, 2mmol) are mixed in single neck bottle In, adding anhydrous DMSO (5mL), under nitrogen protection, 80 DEG C are stirred 20 hours.It is cooled to room temperature, in system Adding water, ethyl acetate extracts, and extract merges, and anhydrous sodium sulfate is dried, and concentrates, and column chromatography for separation obtains 1-(4-first Oxy-benzyl)-3-morpholine-1H-thieno [3,2-c] pyrazoles-5-methyl formate, greenish yellow 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-first Acid methyl ester (0.19g, 0.49mmol) is dissolved in anhydrous tetrahydro furan (20mL), is cooled to-20 DEG C, adds LiAlH₄(34mg, 0.89mmol), stirs 2 hours.Careful dropping shrend is gone out reaction, dichloromethane extraction, and two Chloromethanes is dried with anhydrous sodium sulfate, concentrates, and obtains (1-(4-methoxy-benzyl)-3-morpholine-1H-thieno [3,2-c] Pyrazoles-5-base) methanol crude product 0.19g, thick 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- Base) methanol (0.19g, 0.49mmol) is dissolved in dry dichloromethane (10mL), is cooled to-5 DEG C, add Dess-Martin reagent (285mg, 0.67mmol), stirs 2 hours.Dropping saturated sodium bicarbonate aqueous solution, point Go out dichloromethane phase, be dried, concentrate, column chromatography purification (EA/PE=1/4), obtain 1-(4-methoxy-benzyl)-3-morphine Quinoline 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) it is dissolved in glacial acetic acid (5mL), adds aminochlotononitlile (180mg, 2.2mmol), be heated to 90 DEG C are stirred 1 hour.Removing acetic acid under reduced pressure, residual brownish oily thing is 4-(1-(4-methoxy-benzyl)-3-morpholine -1H-thieno [3,2-c] pyrazoles-5-base)-2,6-dimethyl-Isosorbide-5-Nitrae-dihydropyridine-3,5-dimethoxy nitrile crude product, purification is not straight Tap into and go next step.

Step 5 reaction scheme is as follows:

Step is: above walk 4-(1-(4-methoxy-benzyl)-3-morpholine-1H-thieno [3,2-c] pyrazoles-5-of gained Base)-2,6-dimethyl-Isosorbide-5-Nitrae-dihydropyridine-3,5-dimethoxy nitrile crude product adds trifluoracetic acid (10mL), 80 DEG C of stirrings Overnight.Being cooled to room temperature, remove solvent under reduced pressure, residue cools down with frozen water, dropping saturated sodium bicarbonate aqueous solution alkalization, Dichloromethane extracts, and ethyl acetate extracts, and extract merges, and anhydrous sodium sulfate is dried, and concentrates, column chromatography (dichloromethane Alkane/methanol, 50/1～20/1) 2,6-dimethyl-4-(3-morpholine-1H-thieno [3,2-c] pyrazoles-5-base)-Isosorbide-5-Nitrae- Dihydropyridine-3,5-dimethoxy nitrile, yellow solid 180mg, yield 50%.MS:367[M+H]；¹H NMR(DMSO-d₆, 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)。

Synthetically prepared example 44

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

With 1-(4-methoxy-benzyl)-3-morpholine base-1H-thieno [3,2-c] pyrazoles-5-in synthetically prepared example 43 Formaldehyde is raw material, copies aforementioned similar method, can prepare this target compound.MS:407[M+H]；¹H NMR (DMSO-d₆,300MHz)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)。

Synthetically prepared example 45

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

Step 1 reaction scheme is as follows:

Step is: and the iodo-1-of 3-(4-methoxy-benzyl)-1H-thieno [3,2-c] pyrazoles-5-methyl formate (1.29g, 3 Mmol) (preparation process sees synthetically prepared example 34), N methyl piperazine (0.905g, 9mmol), potassium carbonate (2.08 G, 15mmol), L-PROLINE (104mg, 0.9mmol), Hydro-Giene (Water Science). (0.29g, 1.5mmol) be mixed in In single neck bottle, adding under the protection of DMSO (5mL) nitrogen, 80 DEG C are stirred overnight.It is cooled to room temperature, adds in system Water, ethyl acetate extracts, and extract merges, and anhydrous sodium sulfate is dried, and evaporating column chromatographs, and obtains 1-(4-methoxybenzyl Base)-3-(4-methylpiperazine-1-yl)-1H-thieno [3,2-c] pyrazoles-5-methyl formate, greenish yellow 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 formate (250mg, 0.624mmol) is dissolved in anhydrous tetrahydro furan (20mL), is cooled to-20 DEG C, add LiAlH₄, stir 2 hours.Careful dropping shrend is gone out reaction, and dichloromethane extraction, dichloromethane is with nothing Aqueous sodium persulfate is dried, and concentrates, and obtains (1-(4-methoxy-benzyl)-3-(4-methylpiperazine-1-yl)-1H-thieno [3,2-c] Pyrazoles-5-base) methanol 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-base) methanol (200mg, 0.537mmol) is dissolved in dry dichloromethane (10mL), is cooled to-5 DEG C, Add Dess-Martin reagent (312mg, 0.736mmol), stir 2 hours.Dropping saturated sodium bicarbonate water is molten Liquid, separates organic facies, is dried, and concentrates, column chromatography purification (methylene chloride/methanol=50/1), obtains (1-(4-methoxyl group Benzyl)-3-(4-methylpiperazine-1-yl)-1H-thieno [3,2-c] pyrazoles-5-base) 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-base) In dichloromethane (15mL) solution of methanol (250mg, 0.675mmol) add cyano propanone sodium salt (78mg, 0.742mmol), piperidines (6mg, 0.07mmol), acetic acid (51mg, 0.843mmol) and 4A molecular sieve (0.2 G), finish, be heated to return stirring overnight.Reactant liquor is cooled down, filters, solid saturated sodium bicarbonate solution and water Washing, is dissolved in ethanol by solid, and insoluble matter filters, and filtrate concentrates dry, and residue column chromatography obtains (E)-2-((1-(4-first Oxy-benzyl)-3-(4-methylpiperazine-1-yl)-1H-thieno [3,2-c] pyrazoles-5-base) methylene)-3-oxygen butyronitrile (275mg, 94%).

Step 5 reaction scheme is as follows:

Step is: nitrogen is protected, 2-((1-(4-methoxy-benzyl)-3-(4-methylpiperazine-1-yl)-1H-thieno [3,2-c] pyrazoles-5-base) methylene)-3-oxygen butyronitrile (500mg, 1.15mmol) and 2-(morpholine-3-thiazolinyl) second Nitrile (210mg, 1.70mmol) is dissolved in glacial acetic acid (10mL) jointly, and 90 DEG C are stirred 1 hour.Decompression is steamed Except acetic acid, residual brownish oily thing is 8-(1-(4-methoxy-benzyl)-3-(4-methylpiperazine-1-yl)-1H-thieno [3,2-c] pyrazoles-5-base)-6-methyl isophthalic acid, 3,4,8-tetrahydropyridines also [2,1-c] [Isosorbide-5-Nitrae] oxazines-7,9-dimethoxy nitrile crude product, The most purified directly carry out next step reaction.

Step 6 reaction scheme is as follows:

Step is: above walk 8-(1-(4-methoxy-benzyl)-3-(4-the methylpiperazine-1-yl)-1H-thieno of gained [3,2-c] pyrazoles-5-base)-6-methyl isophthalic acid, 3,4,8-tetrahydropyridine also [2,1-c] [1,4] oxazines-7,9-dimethoxy nitrile crude product Middle addition trifluoroacetic acid (10mL), return stirring is overnight.Being cooled to room temperature, remove solvent under reduced pressure, residue is cold with frozen water But, dropping saturated sodium bicarbonate aqueous solution alkalization, dichloromethane extracts, and ethyl acetate extracts, and extract merges, anhydrous Sodium sulfate is dried, and concentrates, and preparation TLC divide to obtain target compound, yellow solid 83mg, two step yields 17%.MS: 422[M+H]；¹H NMR(CDCl₃,300MHz)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)。

Synthetically prepared example 46:

Prepare the 4-(3-(2-hydroxyl-oxethyl)-1H-thieno [3,2-c] pyrazoles-5-base) of structural formula (I-46) -2,6-dimethyl-1,4-dihydropyridine-3,5-dimethoxy nitrile

Step 1 reaction scheme is as follows:

Step is: the iodo-1-of 3-(4-methoxy-benzyl)-1H-thieno [3,2-c] pyrazoles-5-methyl formate (1.29g, 3mmol) (preparation process sees synthetically prepared example 34), 3,4,7,8-tetramethyl-1,10-phenanthrene quinoline (472mg, 2mmol), Cesium carbonate (4.89g, 15mmol), ethylene glycol mono-tert-butyl ether (106mg, 0.9mmol), Hydro-Giene (Water Science). (0.29 G, 1.5mmol) it is mixed in single neck bottle, add toluene (5mL), under nitrogen protection, 100 DEG C are stirred overnight.Cold But to room temperature, adding water in system, ethyl acetate extracts, and extract merges, and anhydrous sodium sulfate is dried, and is concentrated to give 3-(2- Tert-butoxyethoxy)-1-(4-methoxy-benzyl)-1H-thieno [3,2-c] pyrazoles-5-formic acid crude product, greenish yellow solid 3.4g。

Step 2 reaction scheme is as follows:

Step is: above step gained 3-(2-tert-butoxyethoxy)-1-(4-methoxy-benzyl)-1H-thieno [3,2-c] Pyrazoles-5-formic acid dissolving crude product, in anhydrous tetrahydro furan (60mL), is cooled to-20 DEG C, adds LiAlH₄(1.05g, 27.66mmol), stirring 2 hours.Careful dropping shrend is gone out reaction, and dichloromethane extraction, dichloromethane is with anhydrous Sodium sulfate is dried, and concentrates, and silica gel column chromatography obtains (3-(2-tert-butoxyethoxy)-1-(4-methoxy-benzyl)-1H-thiophene And [3,2-c] pyrazoles-5-base) methanol, yellow solid 120mg, two step yields 10%.

Step 3 reaction scheme is as follows:

Step is: under nitrogen protection, (3-(2-tert-butoxyethoxy)-1-(4-methoxy-benzyl)-1H-thieno [3,2-c] pyrazoles-5-base) methanol (120mg, 0.307mmol) is dissolved in dry dichloromethane (10mL), fall Temperature, to-5 DEG C, adds Dess-Martin reagent (179mg, 0.42mmol), stirs 2 hours.Dropping unsaturated carbonate Hydrogen sodium water solution, separates dichloromethane phase, is dried, and concentrates, column chromatography purification (methylene chloride/methanol=50/1), 3-(2-tert-butoxyethoxy)-1-(4-methoxy-benzyl)-1H-thieno [3,2-c] pyrazoles-5-base) formaldehyde, yellow is solid Body 100mg, yield 83%.

Step 4 reaction scheme is as follows:

Step is: under nitrogen protection, 3-(2-tert-butoxyethoxy)-1-(4-methoxy-benzyl)-1H-thieno [3,2-c] Pyrazoles-5-base) formaldehyde (100mg, 0.257mmol) is dissolved in glacial acetic acid (5mL), adds aminochlotononitlile (47 Mg, 0.566mmol), finish and be heated to 90 DEG C of stirrings 1.5 hours.Removing acetic acid under reduced pressure, residual oil thing is 4-(3-(2- Tert-butoxyethoxy)-1-(4-methoxy-benzyl)-1H-thieno [3,2-c] pyrazoles-5-base)-2,6-dimethyl-1,4- Dihydropyridine-3,5-dimethoxy nitrile crude product, the most purified directly carry out next step.

Step 5 reaction scheme is as follows:

Step is: above walk 4-(3-(2-tert-butoxyethoxy)-1-(4-the methoxy-benzyl)-1H-thieno of gained [3,2-c] pyrazoles-5-base)-2,6-dimethyl-1,4-dihydropyridine-3,5-dimethoxy nitrile crude product adds trifluoroacetic acid (10 ML), return stirring is overnight.Being cooled to room temperature, solvent is steamed in decompression, and residue cools down with frozen water, drips saturated sodium bicarbonate Aqueous solution alkalizes, and dichloromethane extracts, and ethyl acetate extracts, and extract merges, and anhydrous sodium sulfate is dried, and concentrates, system Standby TLC separates to obtain white solid 4-(3-(2-hydroxyl-oxethyl)-1H-thieno [3,2-c] pyrazoles-5-base)-2,6- Dimethyl-Isosorbide-5-Nitrae-dihydropyridine-3,5-dimethoxy nitrile (28mg, two step yields 32%).MS:342[M+H]；¹H NMR(CDCl₃ 300MHz): 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)。

Biological test embodiment 1: the compound In-vitro Inhibitory Effect to c-Met enzyme

The method utilizing Mobility Shift Assay, measures the compound inhibitory action in vitro to c-Met enzyme.

Experimental technique

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

1.1 containing MnCl₂1.25 times of kinase buffer liquid 62.5mM HEPES, pH 7.5;

0.001875％Brij-35；

12.5mM MgCl₂；

2.5mM DTT；

1.2 containing MnCl₂1.25 times of kinase buffer liquid.

62.5mM HEPES,pH 7.5；

0.001875％Brij-35；

12.5mM MgCl₂；

12.5mM MnCl₂；

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

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

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

2.2 transferase 45s times compound is to reaction plate

Get 10 μ L to another piece 96 orifice plates from each hole of above-mentioned 96 orifice plates, add 90 μ L ultra-pure waters. therefore In the second hole to the 11-holes, being the compound being dissolved in 10%DMSO, is 10%DMSO. in the first hole and the 12 hole From above-mentioned 96 orifice plates, take out 5 μ L to 384 hole reaction plates. therefore, in 384 hole reaction plates, just have 5 μ L's In 5 times of compounds that 10%DMSO dissolves and the 10%DMSO. negative control hole of 5 μ L, add 5 μ L 250mM's EDTA.

3. kinase reaction

2.5 times of enzyme solutions of 3.1 preparations

Kinases is added to 1.25 times of kinase buffer liquid, forms 2.5 times of enzyme solutions.

The substrate solution of 3.2 2.5 times of preparations

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 solutions in 384 orifice plates

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

In 384 hole reaction plates, add 2.5 times of enzyme solutions of 10 μ L.

Under room temperature, hatch 10 minutes.

3.4 add substrate solution in 384 orifice plates

In 384 hole reaction plates, add 2.5 times of substrate solutions of 10 μ L.

3.5 kinase reactions and termination

At 28 DEG C, hatch 1 hour. add 25 μ L stop buffer cessation reactions.

4.Caliper reading out data

The upper reading and converting rate of Caliper data.

5. inhibiting rate calculates

Copy conversion data from Caliper. conversion ratio is changed into inhibiting rate data. wherein max refers to the conversion ratio of DMSO contrast, and min refers to the conversion ratio without enzyme contrast alive.

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

Biological test embodiment 1: experimental result

Remarks: "+" represents 500nM < IC₅₀< 5uM; " ++ " represents 50nM < IC₅₀< 500nM; " +++ " representative IC₅₀<50nM。

Claims

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

Wherein,

R₁Be selected from hydrogen, methyl, ethyl, methoxyl group, amino, hydroxy ethoxy, pyridine radicals, thienyl, morpholinyl,

R₂Be selected from cyano group;

R₃Be selected from C₁-C₆Alkyl, methoxyethyl, can be by the phenyl of fluorine, methoxy substitution, pyridine; Or, R₂With R₃And the carbon atom connecting ring formation

R₄Be selected from hydrogen, methyl, ethyl, benzyl; Or, R₄With R₃And ring forms piperidine ring, morpholine ring;

R₅Be selected from methyl or to fluorophenyl; Or, R₅With R₄And ring forms morpholine ring, piperidine ring.

2. dihydropyridine compounds as claimed in claim 1, is characterized in that, described general formula (I) compound is selected from the one 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 exists with the form of pharmaceutically acceptable salt.

4. dihydropyridine compounds as claimed in claim 3, it is characterized in that, described pharmaceutically acceptable salt is hydrochloride, sulfate, phosphate, acetate, trifluoroacetate, mesylate, fluoroform sulphonate, tosilate, tartrate, maleate, fumarate, succinate or the malate of general formula (I) compound.

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

Route 1:

Wherein, R₁，R₂，R₃，R₄，R₅Definition is with described in claim 1; PG is amido protecting group.

6. preparation method as claimed in claim 5, is characterized in that, PG is tertbutyloxycarbonyl, benzyloxycarbonyl, to methoxy-benzyl, 3,4-dimethoxy-benzyl.

7. preparation method as claimed in claim 5, is characterized in that, as R in intermediate (II)₁During for H, its system Preparation Method is as shown in Scheme 2: 5-methylthiophene-2-formic acid of structural formula 1 through fuming nitric aicd nitrated structural formula 2 in Mesosome, is the intermediate of structural formula 3 by the intermediate carboxylic esterification of structural formula 2, adopts reduced iron powder by structural formula 3 The nitroreduction of intermediate is the amino intermediate that obtains structural formula 4, then under the existence of acetic anhydride, nitrite, will tie The intermediate cyclisation of structure formula 4 is the intermediate of structural formula 5; The intermediate acetyl group of structural formula 5 is removed to obtain to structural formula 6 Intermediate, then the ester group in the intermediate of structural formula 6 is reduced to alcohol radical and obtains the intermediate of structural formula 7, then by structure The intermediate of formula 7 is oxidized to such an extent that structural formula is the intermediate of (II);

Route 2:

8. preparation method as claimed in claim 5, is characterized in that, as R in intermediate (II)₁While being not H, its Preparation method is as shown in Scheme 3: 5-methylthiophene-2-formic acid of structural formula 1 through fuming nitric aicd nitrated structural formula 2 Intermediate, is the intermediate of structural formula 3 by the intermediate carboxylic esterification of structural formula 2, adopts reduced iron powder by structural formula 3 The nitroreduction of intermediate be the intermediate of amino structural formula 4, then will under the existence of acetic anhydride, nitrite The intermediate cyclisation of structural formula 4 is the intermediate of structural formula 5; The intermediate acetyl group of structural formula 5 is removed to obtain to structural formula 6 intermediate, by the intermediate of structural formula 66 iodos, obtains the intermediate of structural formula 8; By the intermediate of structural formula 8 Pyrazoles amino with amido protecting group protection, obtain the intermediate of structural formula 9; Then utilize the iodine of the intermediate of structural formula 9 Carry out coupling and obtain the intermediate of structural formula 10; Again the ester group of the intermediate of structural formula 10 is reduced to alcohol radical and obtains structural formula 11 intermediate, is oxidized to aldehyde radical by the alcohol radical in the intermediate of structural formula 11, obtains the intermediate that structural formula is (II);

Route 3:

9. preparation method as claimed in claim 8, is characterized in that, described coupling is selected from Suzuki coupling, Buchwald coupling.

10. preparation method as claimed in claim 5, is characterized in that, when the compound of required preparation is the compound shown in general formula (Ia), and R '=R₃＝R₅Time, its preparation method as shown in Scheme 4:

Route 4:

11. preparation methods as claimed in claim 5, is characterized in that, when the compound of required preparation is the compound shown in general formula (Ib), and R₃With R₅When different, its preparation method as shown in Scheme 5:

Route 5:

12. preparation methods as claimed in claim 5, is characterized in that, when the compound of required preparation is the compound shown in general formula (Ic), and R₃With R₅Difference and R₄While being not H, its preparation method as shown in Scheme 6:

Route 6:

13. preparation methods as claimed in claim 5, is characterized in that, in the time that 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 CH₂Or O.

14. preparation methods as claimed in claim 5, is characterized in that, in the time that the compound of required preparation is the compound shown in general formula (Ie), its preparation method as shown in Scheme 8:

Route 8:

15. preparation methods as claimed in claim 5, is characterized in that, in the time that 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 CH₂Or O.

16. 1 kinds of pharmaceutical compositions containing the dihydropyridine compounds described in claim 1, wherein said pharmaceutical composition comprises general formula (I) compound and the acceptable excipient of pharmacy for the treatment of effective dose.

17. as the pharmaceutical composition of claim 16, and wherein said pharmaceutical composition is made tablet, capsule, aqueous suspension, oiliness supensoid agent, dispersible pulvis, granule, lozenge, emulsion, syrup, cream, ointment, suppository or injection.

18. 1 kinds of pharmaceutical compositions, pharmaceutically acceptable salt and the acceptable excipient of pharmacy that wherein said pharmaceutical composition comprises general formula claimed in claim 1 (I) compound for the treatment of effective dose.

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

20. 1 kinds of dihydropyridine compounds claimed in claim 1 regulate in protein kinase catalytic activity goods in preparation Application; Described protein kinase is c-Met receptor tyrosine kinase.

The pharmaceutically useful salt of 21. 1 kinds of dihydropyridine compounds claimed in claim 1 regulates protein kinase to urge in preparation Change the application in active product; Described protein kinase is c-Met receptor tyrosine kinase.

Pharmaceutical composition described in 22. 1 kinds of claims 16 is at the medicine of the preparation treatment disease relevant with protein kinase Application in thing; Described protein kinase is c-Met receptor tyrosine kinase.

23. application as claimed in claim 22, the relevant disease of wherein said protein kinase is cancer.

24. application as claimed in claim 23, wherein said cancer is thyroid cancer, colorectal cancer, cancer of the stomach, kidney, liver cancer, lung cancer, oophoroma, breast cancer, prostate cancer, carcinoma of urinary bladder, head and neck cancer, cancer of pancreas, carcinoma of gallbladder, osteosarcoma, rhabdomyosarcoma, MFH, fibrosarcoma, spongioblastoma, astrocytoma, melanoma or celiothelioma.

The application of the pharmaceutical composition described in 25. 1 kinds of claims 18 is wherein pharmaceutical composition preparation treatment with Application in the medicine of the relevant disease of protein kinase; Described protein kinase is c-Met receptor tyrosine kinase.

26. application as claimed in claim 25, the relevant disease of wherein said protein kinase is cancer.

27. application as claimed in claim 26, wherein said cancer is thyroid cancer, colorectal cancer, cancer of the stomach, kidney, liver cancer, lung cancer, oophoroma, breast cancer, prostate cancer, carcinoma of urinary bladder, head and neck cancer, cancer of pancreas, carcinoma of gallbladder, osteosarcoma, rhabdomyosarcoma, MFH, fibrosarcoma, spongioblastoma, astrocytoma, melanoma or celiothelioma.