WO2017215485A1 - 一种具有fgfr抑制活性的新型化合物及其制备和应用 - Google Patents

一种具有fgfr抑制活性的新型化合物及其制备和应用 Download PDF

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WO2017215485A1
WO2017215485A1 PCT/CN2017/087222 CN2017087222W WO2017215485A1 WO 2017215485 A1 WO2017215485 A1 WO 2017215485A1 CN 2017087222 W CN2017087222 W CN 2017087222W WO 2017215485 A1 WO2017215485 A1 WO 2017215485A1
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group
substituted
unsubstituted
pyrrole
pyrimidin
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PCT/CN2017/087222
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English (en)
French (fr)
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段文虎
耿美玉
王玉明
艾菁
范珺
戴阳
丁健
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中国科学院上海药物研究所
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Priority to EP17812596.9A priority Critical patent/EP3486244A4/en
Priority to US16/310,394 priority patent/US20190367520A1/en
Priority to CN201780037478.3A priority patent/CN109328187B/zh
Priority to JP2018566299A priority patent/JP2019521992A/ja
Publication of WO2017215485A1 publication Critical patent/WO2017215485A1/zh

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    • C—CHEMISTRY; METALLURGY
    • C07—ORGANIC CHEMISTRY
    • C07D—HETEROCYCLIC COMPOUNDS
    • C07D403/00—Heterocyclic compounds containing two or more hetero rings, having nitrogen atoms as the only ring hetero atoms, not provided for by group C07D401/00
    • C07D403/02—Heterocyclic compounds containing two or more hetero rings, having nitrogen atoms as the only ring hetero atoms, not provided for by group C07D401/00 containing two hetero rings
    • C07D403/12—Heterocyclic compounds containing two or more hetero rings, having nitrogen atoms as the only ring hetero atoms, not provided for by group C07D401/00 containing two hetero rings linked by a chain containing hetero atoms as chain links
    • C—CHEMISTRY; METALLURGY
    • C07—ORGANIC CHEMISTRY
    • C07D—HETEROCYCLIC COMPOUNDS
    • C07D487/00—Heterocyclic compounds containing nitrogen atoms as the only ring hetero atoms in the condensed system, not provided for by groups C07D451/00 - C07D477/00
    • C07D487/02—Heterocyclic compounds containing nitrogen atoms as the only ring hetero atoms in the condensed system, not provided for by groups C07D451/00 - C07D477/00 in which the condensed system contains two hetero rings
    • C07D487/04—Ortho-condensed systems
    • A—HUMAN NECESSITIES
    • A61—MEDICAL OR VETERINARY SCIENCE; HYGIENE
    • A61P—SPECIFIC THERAPEUTIC ACTIVITY OF CHEMICAL COMPOUNDS OR MEDICINAL PREPARATIONS
    • A61P35/00—Antineoplastic agents
    • C—CHEMISTRY; METALLURGY
    • C07—ORGANIC CHEMISTRY
    • C07D—HETEROCYCLIC COMPOUNDS
    • C07D209/00—Heterocyclic compounds containing five-membered rings, condensed with other rings, with one nitrogen atom as the only ring hetero atom
    • C07D209/02—Heterocyclic compounds containing five-membered rings, condensed with other rings, with one nitrogen atom as the only ring hetero atom condensed with one carbocyclic ring
    • C07D209/04—Indoles; Hydrogenated indoles
    • C—CHEMISTRY; METALLURGY
    • C07—ORGANIC CHEMISTRY
    • C07D—HETEROCYCLIC COMPOUNDS
    • C07D209/00—Heterocyclic compounds containing five-membered rings, condensed with other rings, with one nitrogen atom as the only ring hetero atom
    • C07D209/02—Heterocyclic compounds containing five-membered rings, condensed with other rings, with one nitrogen atom as the only ring hetero atom condensed with one carbocyclic ring
    • C07D209/04—Indoles; Hydrogenated indoles
    • C07D209/08—Indoles; Hydrogenated indoles with only hydrogen atoms or radicals containing only hydrogen and carbon atoms, directly attached to carbon atoms of the hetero ring
    • C—CHEMISTRY; METALLURGY
    • C07—ORGANIC CHEMISTRY
    • C07D—HETEROCYCLIC COMPOUNDS
    • C07D215/00—Heterocyclic compounds containing quinoline or hydrogenated quinoline ring systems
    • C—CHEMISTRY; METALLURGY
    • C07—ORGANIC CHEMISTRY
    • C07D—HETEROCYCLIC COMPOUNDS
    • C07D215/00—Heterocyclic compounds containing quinoline or hydrogenated quinoline ring systems
    • C07D215/58—Heterocyclic compounds containing quinoline or hydrogenated quinoline ring systems with hetero atoms directly attached to the ring nitrogen atom
    • C—CHEMISTRY; METALLURGY
    • C07—ORGANIC CHEMISTRY
    • C07D—HETEROCYCLIC COMPOUNDS
    • C07D401/00—Heterocyclic compounds containing two or more hetero rings, having nitrogen atoms as the only ring hetero atoms, at least one ring being a six-membered ring with only one nitrogen atom
    • C07D401/02—Heterocyclic compounds containing two or more hetero rings, having nitrogen atoms as the only ring hetero atoms, at least one ring being a six-membered ring with only one nitrogen atom containing two hetero rings
    • C07D401/12—Heterocyclic compounds containing two or more hetero rings, having nitrogen atoms as the only ring hetero atoms, at least one ring being a six-membered ring with only one nitrogen atom containing two hetero rings linked by a chain containing hetero atoms as chain links
    • C—CHEMISTRY; METALLURGY
    • C07—ORGANIC CHEMISTRY
    • C07D—HETEROCYCLIC COMPOUNDS
    • C07D405/00—Heterocyclic compounds containing both one or more hetero rings having oxygen atoms as the only ring hetero atoms, and one or more rings having nitrogen as the only ring hetero atom
    • C07D405/02—Heterocyclic compounds containing both one or more hetero rings having oxygen atoms as the only ring hetero atoms, and one or more rings having nitrogen as the only ring hetero atom containing two hetero rings
    • C07D405/12—Heterocyclic compounds containing both one or more hetero rings having oxygen atoms as the only ring hetero atoms, and one or more rings having nitrogen as the only ring hetero atom containing two hetero rings linked by a chain containing hetero atoms as chain links
    • Y—GENERAL TAGGING OF NEW TECHNOLOGICAL DEVELOPMENTS; GENERAL TAGGING OF CROSS-SECTIONAL TECHNOLOGIES SPANNING OVER SEVERAL SECTIONS OF THE IPC; TECHNICAL SUBJECTS COVERED BY FORMER USPC CROSS-REFERENCE ART COLLECTIONS [XRACs] AND DIGESTS
    • Y02—TECHNOLOGIES OR APPLICATIONS FOR MITIGATION OR ADAPTATION AGAINST CLIMATE CHANGE
    • Y02P—CLIMATE CHANGE MITIGATION TECHNOLOGIES IN THE PRODUCTION OR PROCESSING OF GOODS
    • Y02P20/00—Technologies relating to chemical industry
    • Y02P20/50—Improvements relating to the production of bulk chemicals
    • Y02P20/55—Design of synthesis routes, e.g. reducing the use of auxiliary or protecting groups

Definitions

  • the invention relates to the field of medicine, in particular to a novel compound having FGFR inhibitory activity and preparation and application thereof.
  • Protein kinases are proteins (enzymes) that regulate various cellular functions by phosphorylating specific amino acids on proteins. Proteins regulate activity and ability to bind to other components through changes in conformation.
  • the activity of a protein kinase refers to the rate at which a kinase binds a phosphate group to a substrate, and the rate can be determined by detecting the amount of substrate converted to a product over a period of time. Phosphorylation of the substrate occurs at the activation site of the protein kinase.
  • Tyrosine kinase is a proteinase that catalyzes the transfer of adenosine triphosphate to a protein tyrosine residue. These kinases play an important role in growth factor-mediated cell proliferation, differentiation and migration.
  • Fibroblast growth factor has been identified to have important regulatory roles in many physiological processes, such as organogenesis and angiogenesis. It is known that there are more than 25 subtypes in the FGF family, and the fibroblast growth factor receptor (FGFR) family contains four subtypes (FGFR1-4), all of which are glycoproteins, including extracellular immunity. The globulin region, the transmembrane hydrophobic region, and the tyrosine kinase region within the cytoplasm. Binding of FGF triggers FGFR dimerization, which in turn undergoes autophosphorylation of the receptor and activation of downstream signaling pathways. Certain specific components of the downstream signaling pathway play a very important role in cell growth, metabolism, and survival. Therefore, the FGFR signaling pathway plays an important physiological role in the multi-effect of tumor cell reproduction, migration, infiltration and angiogenesis.
  • FGFR signaling pathway plays an important physiological role in the multi-effect of tumor cell reproduction, migration, infiltration and angiogenesis.
  • the FGF signaling pathway is directly related to human cancer. Different FGF overexpression phenomena have been reported in different types of cancer cells (bladder cancer, kidney cancer, prostate cancer, etc.). Therefore, the FGF signaling pathway is a promising therapeutic target.
  • a compound of formula I or a stereoisomer, geometric isomer, tautomer thereof, pharmaceutically acceptable salt thereof, prodrug thereof, and hydrate or solvate thereof ,
  • R 4 is selected from H, substituted or unsubstituted C1-C6 alkyl, substituted or unsubstituted C3-C8 cycloalkyl, NR 6 R 7 , halogen, hydroxy, cyano, substituted or unsubstituted C1-C6 alkane An oxy group and a substituted or unsubstituted C1-C6 alkylthio group;
  • X 1 and X 2 may be the same or different and are each independently selected from N and CR 10 ;
  • R 10 is selected from H, substituted or unsubstituted C1-C6 alkyl, substituted or unsubstituted C3-C8 cycloalkyl, halogen a hydroxy group, a cyano group, a substituted or unsubstituted C1-C6 alkoxy group and a substituted or unsubstituted C1-C6 alkylthio group;
  • n 0, 1, 2, 3, 4 or 5;
  • Y is selected from substituted or unsubstituted 3-10 membered heterocyclic groups containing 1-3 selected from N, O and S heteroatoms, -NR 6 R 7 , substituted or unsubstituted C3-C8 cycloalkyl and L2-(substituted or unsubstituted C6-C10 aryl)-, and said substitution is independently substituted with one or more groups selected from the group consisting of C1-C6 alkyl, halogenated C1-C6 Alkyl, C3-C8 cycloalkyl, halogenated C3-C8 cycloalkyl, hydroxy, C1-C6 alkoxy, halogenated C1-C6 alkoxy, -O-(C3-C8 cycloalkyl) , -O-(C3-C8 halocycloalkyl), NR 11 R 12 , halogen, 4-10 membered heterocyclic group containing 1-3 selected from N, O
  • Z is selected from the group consisting of unsubstituted, substituted or unsubstituted C2-C6 alkenyl, substituted or unsubstituted C2-C6 alkynyl, substituted or unsubstituted C1-C6 alkyl, substituted or unsubstituted C3-C8 ring Alkyl, -(R 13 )-N(R 11 )-(R 14 )-(substituted or unsubstituted C1-C6 alkoxy);
  • R 8 and R 9 may be the same or different and are each independently selected from: H, a hydroxyl group, a substituted or unsubstituted C1-C6 alkyl group, a substituted or unsubstituted C3-C8 cycloalkyl group, a substituted or unsubstituted C1- a C6 alkoxy group, a substituted or unsubstituted C2-C4 alkenyl group, a substituted or unsubstituted C2-C4 alkynyl group, -L2-(C1-C6 alkylene)-L1 and -NR 11 R 12 ;
  • L1 is selected from -OH, C1-C4 alkoxy, -NR 11 R 12 , or a 4-7 membered heterocyclic ring having 1 or 2 N atoms;
  • L2 is selected from the group consisting of -NR 11 -, or -N (substituted or unsubstituted C3-C6 cycloalkyl);
  • R 11 and R 12 are independently selected from H, C1-C6 alkyl, C1-C6 haloalkyl, C3-C6 cycloalkyl, C3-C6 halocycloalkyl, -CO(C2-C4 alkenyl), or -CO(C2-C4 alkynyl); or R 11 and R 12 and an adjacent N form a 4-7 membered heterocyclic ring having 1-2 N atoms and 0-2 O or S atoms;
  • R 13 and R 14 are independently selected from a substituted or unsubstituted C1-C6 alkylene group, or a substituted or unsubstituted C2-C6 alkenylene group.
  • Z is a C2-C6 alkenyl group having 1, 2 or 3 substituents AA, wherein the substituent AA is selected from the group consisting of NR 11 R 12 and -(C1-C6 Alkyl)-L1, C3-C8 cycloalkyl, halogenated C3-C8 cycloalkyl, -CN, halogen.
  • Z is a C2-C4 alkenyl group having 1-2 substituents AA substituted, wherein the substituent AA is selected from the group consisting of NR 11 R 12 and -(C1-C6 alkylene)- L1, C3-C8 cycloalkyl, -CN, halogen.
  • R A , R B and R C are independently selected from the group consisting of H, the above substituent AA;
  • R A , R B and R C are not H at the same time.
  • R A or R B is a substituent AA.
  • R C is a substituent AA.
  • R 1 is a bicyclic group of the formula:
  • Ring A is a substituted or unsubstituted 5 membered heteroaryl ring; and Ring B is a substituted or unsubstituted 6-membered heteroaryl ring or a substituted or unsubstituted phenyl group.
  • R 1 is a bicyclic group of the formula:
  • the ring A is a 5-membered heteroaryl ring containing S.
  • R 1 is a bicyclic group of the formula:
  • the ring A is a 5-membered heteroaryl ring containing one or two N atoms.
  • R1 is a substituted 5-14 membered heteroaryl or substituted 6-14 membered aryl.
  • R1 is a substituted 5-14 membered heteroaryl or substituted 6-14 membered aryl, and the substitution is one or more substituents selected from the group consisting of C1-C6.
  • Alkyl, halogenated C1-C6 alkyl, C3-C8 cycloalkyl, halogenated C3-C8 cycloalkyl, hydroxy, C1-C6 alkoxy, halogenated C1-C6 alkoxy, -O -(C3-C8 cycloalkyl), -O-(C3-C8 halocycloalkyl), -NR 6 R 7 , halogen, C( O)R 8 .
  • the R 1 is a substituted 5-14 membered heteroaryl or substituted 6-14 membered aryl, and the substitution is one or more substituents selected from the group consisting of: C1-C6 alkyl, hydroxy, C1-C6 alkoxy, halogenated C1-C6 alkoxy, -NH 2, halo.
  • substitution is one having at least one substituent selected from the group consisting of C1-C6 alkoxy, halogenated C1-C6 alkoxy.
  • the substitution is a substituent having at least one selected from the group consisting of a C1-C6 alkoxy group, a halogenated C1-C6 alkoxy group, a hydroxyl group, and —NH 2 ; At least one substituent selected from the group consisting of a C1-C6 alkyl group, a C3-C8 cycloalkyl group, and a halogen.
  • R 1 is a substituted group selected from the group consisting of benzothienyl, benzofuranyl, fluorenyl, benzimidazolyl, benzoselenophene, oxazolyl, and benzene. And thiazolyl.
  • R1 is a substituted benzothienyl group.
  • R1 is a substituted benzothienyl group and has a substituent selected from the group consisting of a C1-C6 alkoxy group, a halogenated C1-C6 alkoxy group, a hydroxyl group, and -NH. 2 .
  • the compound of formula I is selected from the compounds listed in Table 1.
  • a pharmaceutical composition comprising a therapeutically effective amount of the compound of the first aspect of the present invention or a stereoisomer, geometric isomer thereof, tautomerism thereof
  • a pharmaceutically acceptable salt a pharmaceutically acceptable salt, a prodrug thereof, and a hydrate or solvate thereof, and optionally a pharmaceutically acceptable carrier.
  • the pharmaceutical composition is a pharmaceutical composition for preventing and/or treating cancer, or a pharmaceutical composition for preventing and/or treating a FGFR-related disease.
  • the FGFR is selected from the group consisting of FGFR1, FGFR2, FGFR3, FGFR4, or a combination.
  • the pharmaceutical composition is for use in preventing and/or treating a disease associated with abnormal expression of the FGF/FGFR signaling pathway.
  • the dosage form of the pharmaceutical composition is selected from the group consisting of an oral dosage form, a lyophilized preparation, and an injection.
  • a compound according to the first aspect of the present invention or a stereoisomer, a geometric isomer thereof, a tautomer thereof, a pharmaceutically acceptable salt thereof, a prodrug thereof and a hydrate thereof Or use of a solvate or a pharmaceutical composition according to the second aspect of the invention for the preparation of a medicament for preventing and/or treating a disease selected from the group consisting of:
  • the tumor-related disease is selected from the group consisting of breast cancer, lung cancer, bladder cancer, stomach cancer, pancreatic cancer, Prostate cancer, colon cancer, multiple myeloma AML, liver cancer, melanoma, head and neck cancer, thyroid cancer, renal cell carcinoma, glioblastoma, and testicular cancer.
  • the lung cancer is non-small cell lung cancer.
  • the protein tyrosine kinase activity-associated disease is selected from the group consisting of FGFR-associated diseases.
  • a FGFR inhibitor comprising an inhibitory effective amount of a compound of the first aspect of the invention, or a stereoisomer, geometric isomer thereof, tautomerism thereof One or more of a pharmaceutically acceptable salt, a prodrug thereof, and a hydrate or solvate thereof.
  • the FGFR inhibitor is a FGFR1 inhibitor and a FGFR4 inhibitor.
  • a compound according to the first aspect of the invention or a stereoisomer, a geometric isomer thereof, a tautomer thereof, a pharmaceutically acceptable salt thereof, a prodrug thereof and a hydrate thereof Or a method of preparing a solvate, the method comprising the steps of:
  • R 1 , R 2 , R 3 , R 4 , n, X 1 , X 2 , Y, E and Z have the same meanings as defined above;
  • X is selected from the group consisting of halogen and hydroxyl.
  • reaction of step (i) is carried out under basic conditions, preferably in triethylamine, pyridine, N,N-diisopropylethylamine, 4-dimethylaminopyridine, N- It is carried out under the conditions in which methylmorpholine or the like is present.
  • the acid halide compound is preferably an acid chloride compound.
  • the acid is a carboxylic acid.
  • a process for the preparation of a pharmaceutical composition according to the second aspect of the invention comprising the steps of: pharmaceutically acceptable carrier and the compound of the first aspect of the invention or a stereoisomer thereof
  • the body, geometric isomer, tautomer, pharmaceutically acceptable salt thereof, prodrug thereof, and hydrate or solvate thereof are mixed to form a pharmaceutical composition.
  • a method for non-diagnostic, non-therapeutic inhibition of FGFR activity comprising administering to a patient in need thereof an inhibitory effective amount of a compound of the first aspect of the invention or a stereoisod thereof A construct, a geometric isomer, a tautomer, a pharmaceutically acceptable salt thereof, a prodrug thereof, and a hydrate or solvate thereof, or a pharmaceutical composition according to the second aspect of the invention.
  • FGFR inhibitors prepared with the compounds of the invention achieve significant inhibition of FGFR1-4 enzymatic activity at the nM level compared to existing FGFR inhibitors, and the inhibitors are induced at the cellular level by FGFR1-4 Cancer cell proliferation also has a significant inhibitory effect, which is of great significance for the development of new anti-tumor drugs.
  • the inventors completed the present invention.
  • substituted means that one or more hydrogen atoms on the group are substituted with a substituent selected from the group consisting of C1-C6 alkyl, halo C1-C6 alkyl, C3-C8 cycloalkyl, halogenated C3-C8 cycloalkyl, hydroxy, C1-C6 alkoxy, halogenated C1-C6 alkoxy, -O-(C3-C8 cycloalkyl), -O -(C3-C8 halocycloalkyl), halogen, 4-10 membered heterocyclic group containing 1-3 selected from S, O, N and Se heteroatoms, amino group, phenyl group, cyano group, C2- a C6 alkenyl group, a C2-C6 alkynyl group; the phenyl group comprising an unsubstituted phenyl group or a substituted phenyl
  • each chiral carbon atom may optionally be in the R configuration or the S configuration, or a mixture of the R configuration and the S configuration.
  • 5-14 membered heteroaryl refers to a group formed by a 5- to 14-membered aryl group having from 1 to 3 heteroatoms selected from the group consisting of N, S, O, Se, each of which
  • the heteroaryl ring system may be monocyclic or polycyclic; for example, benzothienyl, benzofuranyl, fluorenyl, naphthyl, benzimidazolyl, benzoselenophene, pyridyl, furyl Phenyl, carbazolyl, thienyl, pyrrolyl, imidazolyl, pyrazolyl, pyrazinyl, oxazolyl, thiazolyl, benzothiazolyl, or the like.
  • 6-14 membered aryl refers to a group formed by the loss of a hydrogen atom of a 6-14 membered aryl group; for example, a phenyl group, a naphthyl group, or the like.
  • C1-C6 alkyl refers to a straight or branched alkyl group having from 1 to 6 carbon atoms, such as methyl, ethyl, propyl, isopropyl, butyl, isobutyl, sec-butyl, Tert-butyl, or a similar group.
  • C1-C6 alkoxy refers to a straight or branched alkoxy group having from 1 to 6 carbon atoms, such as methoxy, ethoxy, propoxy, isopropoxy, butoxy, iso Butoxy, sec-butoxy, tert-butoxy, or the like.
  • halogen refers to F, Cl, Br and I.
  • alkyl includes a saturated or unsaturated, linear, branched, cyclic all-carbon alkyl group of 1 to 10 carbon atoms or 1 to 3 carbon atoms thereof by oxygen, An alkyl group substituted with a hetero atom such as nitrogen or sulfur, and an aralkyl group bonded through one or more carbon atoms. Further, the alkyl group is unsubstituted or substituted.
  • aryl includes fused or non-fused aryl groups, usually containing from 6 to 30 carbon atoms, and representative aryl groups include phenyl, naphthyl, or oxygen, nitrogen, sulfur, etc. Aromatic group of atoms.
  • the structural formulae described herein are intended to include all isomeric forms (such as enantiomeric, diastereomeric, and geometric isomers (or conformational isomers): for example, containing asymmetric centers R, S configuration, (Z), (E) isomers of the double bond and conformational isomers of (Z), (E).
  • isomeric forms such as enantiomeric, diastereomeric, and geometric isomers (or conformational isomers): for example, containing asymmetric centers R, S configuration, (Z), (E) isomers of the double bond and conformational isomers of (Z), (E).
  • a single stereochemical isomer of the compound of the invention or its enantiomer Mixtures of isomers, diastereomers or geometric isomers (or conformational isomers) are within the scope of the invention.
  • tautomer means that structural isomers having different energies can exceed the low energy barrier and thereby transform each other.
  • proton tautomers ie, proton shifts
  • the valence tautomers include interconversion through some bonding electron recombination.
  • the present inventors designed and synthesized a series of novel FGFR inhibitor compounds, and through structural optimization, found that small molecule FGFR inhibitors with excellent activity in enzymes, cells and animals have strong inhibitory effects on FGFR1-4.
  • This series of compounds is expected to be used clinically to treat diseases caused by abnormal expression of FGF/FGFR signaling pathways, such as cancer.
  • R 1 , R 2 , R 3 , R 4 , R 6 , R 7 , R 8 , R 9 , R 10 , R 11 , R 12 , R 13 , R 14 , X 1 , X 2 , L1, L2, Y, E, Z are each a group corresponding to each specific compound in the examples.
  • the compound is preferably a compound prepared in the examples.
  • the compounds NO. 1-NO. 56 are the compounds obtained in Examples 1-56, respectively.
  • the compound is selected from the compounds listed in Table 1.
  • the compound has significant FGFR inhibitory activity.
  • the compound is substantially free of EGFR inhibitory activity.
  • the term "pharmaceutically acceptable salt” refers to a salt of the compound of the invention formed with an acid or base suitable for use as a medicament.
  • the pharmaceutically acceptable salts include inorganic salts and organic salts.
  • a preferred class of salts are the salts of the compounds of the invention with acids.
  • Suitable acids for forming salts include, but are not limited to, mineral acids such as hydrochloric acid, hydrobromic acid, hydrofluoric acid, sulfuric acid, nitric acid, phosphoric acid; formic acid, acetic acid, trifluoroacetic acid, propionic acid, oxalic acid, malonic acid, succinic acid, Fumaric acid, maleic acid, lactic acid, malic acid, tartaric acid, citric acid, picric acid, benzoic acid, methanesulfonic acid, ethanesulfonic acid, methanesulfonic acid, trifluoromethanesulfonic acid, ethanesulfonic acid, p-toluene
  • An organic acid such as acid, benzenesulfonic acid or naphthalenesulfonic acid; and an amino acid such as proline, phenylalanine, aspartic acid or glutamic acid.
  • salts of the compounds of the invention with bases such as alkali metal salts (for example sodium or potassium salts), alkaline earth metal salts (for example magnesium or calcium salts), ammonium salts (for example lower alkanolammonium).
  • bases such as alkali metal salts (for example sodium or potassium salts), alkaline earth metal salts (for example magnesium or calcium salts), ammonium salts (for example lower alkanolammonium).
  • Salts and other pharmaceutically acceptable amine salts such as methylamine, ethylamine, propylamine, dimethylamine, trimethylamine, diethylamine, triethylamine, tert-butyl
  • a base amine salt an ethylenediamine salt, a hydroxyethylamine salt, a dihydroxyethylamine salt, a trihydroxyethylamine salt, and an amine salt formed of morpholine, piperazine, and lysine, respectively.
  • solvate refers to a complex of a compound of the invention that is coordinated to a solvent molecule to form a specific ratio.
  • Hydrophilate means a complex formed by the coordination of a compound of the invention with water.
  • prodrug includes a class of compounds which, by themselves, may be biologically active or inactive, which, upon administration in an appropriate manner, are converted to a compound of formula I by metabolism or chemical reaction in the human body, or A salt or solution of a compound of I.
  • the prodrug includes, but is not limited to, a carboxylic acid ester, a carbonate, a phosphate, a nitrate, a sulfate, a sulfone ester, a sulfoxide, an amino compound, a carbamate, an azo compound of the compound. , phosphoramide, glucoside, ether, acetal and the like.
  • the preparation process of the compounds of the present invention is as follows, wherein the starting materials and reagents used are commercially available unless otherwise specified.
  • R 1 , R 2 , R 3 , R 4 , n, X 1 , X 2 , Y, E and Z have the same meanings as defined above, P is any group which can serve as an amino protecting group, and L is a nucleophilic substitution. The leaving group at the time.
  • a compound 2 is prepared by a light extension reaction or a nucleophilic substitution, and then coupled, and the target group is condensed to obtain a target product.
  • the preparation method includes the following steps:
  • Compound 1 is formed by a photo-delay reaction or nucleophilic substitution to form compound 2, or compound 13 is reacted by kinetic reaction or nucleophilic substitution to form compound 14, and compound 14 is reacted with NHR 2 R 3 or a salt thereof to form compound 2;
  • Compound 4 is condensed with the corresponding acid chloride compound or carboxylic acid to give the target product.
  • the intermediate 1 can be produced by a conventional method in the art or can be obtained by a commercially available route.
  • the compound (1a) is prepared by the following method:
  • R 2 and R 3 have the same meanings as defined above.
  • the preparation method includes the following steps:
  • the compound (1b) is produced by the following method:
  • the compound (1b) is obtained by cyclization from compound 7 to give an iodo compound.
  • the preparation method includes the following steps:
  • Compound 7 is cyclized at a degree of 180 ° in a formamide solvent to form a corresponding 8;
  • the corresponding boronic acid required in the process for preparing the structure of Formula I can be prepared by conventional methods in the art or commercially available. In a preferred embodiment of the invention, it is prepared by the following method:
  • R a is a substituent in the above R 1 and is as defined above.
  • Compound 10 is prepared by nucleophilic substitution starting from compound 9, and then compound 11 is obtained under chlorobenzene under polyphosphoric acid conditions, and then in conventional reaction conditions (n-butyllithium and triisopropylborate). Into the corresponding boric acid.
  • the preparation method includes the following steps:
  • Compound 9 is produced by nucleophilic substitution to give compound 10;
  • Compound 11 gives a boronic acid compound under the conditions of n-butyllithium and triisopropylborate.
  • the preparation method of the compound of the invention has the advantages of mild reaction conditions, abundant raw materials, easy operation and post-treatment.
  • the present invention also provides a pharmaceutical composition
  • a pharmaceutical composition comprising a therapeutically effective amount of the compound or a stereoisomer, a geometric isomer, a tautomer thereof, a pharmaceutically acceptable salt thereof, and a prodrug thereof
  • a pharmaceutically acceptable carrier One or more of a drug and a hydrate or solvate thereof, and optionally a pharmaceutically acceptable carrier.
  • the pharmaceutical composition is a pharmaceutical composition for preventing and/or treating cancer, or a pharmaceutical composition for preventing and/or treating a FGFR-related disease.
  • the FGFR is selected from the group consisting of FGFR1, FGFR2, FGFR3, FGFR4, or a combination.
  • the pharmaceutical composition is for use in preventing and/or treating a disease associated with abnormal expression of the FGF/FGFR signaling pathway.
  • the dosage form of the pharmaceutical composition is selected from the group consisting of an oral dosage form, a lyophilized preparation, and an injection.
  • the compound of the present invention has excellent inhibitory activity against FGFR kinase (Kinase) such as FGFR1 and FGFR4, the compound of the present invention and various crystal forms thereof, pharmaceutically acceptable inorganic or organic salts, hydrates or solvates, and
  • the pharmaceutical composition containing the compound of the present invention as a main active ingredient can be used for the treatment, prevention, and alleviation of diseases associated with FGFR activity or expression, such as prevention and/or treatment of diseases associated with abnormal expression of the FGF/FGFR signaling pathway.
  • the compounds of the present invention are useful for the treatment of breast cancer, lung cancer, bladder cancer, gastric cancer, pancreatic cancer, prostate cancer, colon cancer, multiple myeloma AML, liver cancer, melanoma, head and neck. Cancer, thyroid cancer, renal cell carcinoma, glioblastoma, and testicular cancer. More specifically, these cancers are selected From: breast cancer, non-small cell lung cancer, bladder cancer, stomach cancer, pancreatic cancer, prostate cancer, colon cancer, multiple myeloma, liver cancer, melanoma, head and neck cancer, thyroid cancer, renal cell carcinoma, glioblastoma and Testicular cancer. Most particularly, the cancer is non-small cell lung cancer, gastric cancer or multiple myeloma.
  • compositions of the present invention comprise a safe or effective amount of a compound of the present invention, or a pharmaceutically acceptable salt thereof, and a pharmaceutically acceptable excipient or carrier.
  • safe and effective amount it is meant that the amount of the compound is sufficient to significantly improve the condition without causing serious side effects.
  • the pharmaceutical compositions contain from 1 to 2000 mg of the compound of the invention per agent, more preferably from 5 to 200 mg of the compound of the invention per agent.
  • the "one dose” is a capsule or tablet.
  • “Pharmaceutically acceptable carrier” means: one or more compatible solid or liquid fillers or gel materials which are suitable for human use and which must be of sufficient purity and of sufficiently low toxicity. By “compatibility” it is meant herein that the components of the composition are capable of intermingling with the compounds of the invention and with each other without significantly reducing the efficacy of the compound.
  • pharmaceutically acceptable carriers are cellulose and its derivatives (such as sodium carboxymethylcellulose, sodium ethylcellulose, cellulose acetate, etc.), gelatin, talc, solid lubricants (such as stearic acid).
  • magnesium stearate magnesium stearate
  • calcium sulfate vegetable oil (such as soybean oil, sesame oil, peanut oil, olive oil, etc.), polyol (such as propylene glycol, glycerin, mannitol, sorbitol, etc.), emulsifier (such as Tween ), a wetting agent (such as sodium lauryl sulfate), a coloring agent, a flavoring agent, a stabilizer, an antioxidant, a preservative, a pyrogen-free water, and the like.
  • vegetable oil such as soybean oil, sesame oil, peanut oil, olive oil, etc.
  • polyol such as propylene glycol, glycerin, mannitol, sorbitol, etc.
  • emulsifier such as Tween
  • a wetting agent such as sodium lauryl sulfate
  • a coloring agent such as a flavoring agent, a stabilizer, an antioxidant, a preservative
  • the mode of administration of the compound or pharmaceutical composition of the present invention is not particularly limited, and representative modes of administration include, but are not limited to, oral, intratumoral, rectal, parenteral (intravenous, intramuscular or subcutaneous) and topical administration.
  • Solid dosage forms for oral administration include capsules, tablets, pills, powders, and granules.
  • the active compound is mixed with at least one conventional inert excipient (or carrier), such as sodium citrate or dicalcium phosphate, or mixed with: (a) a filler or compatibilizer, for example, Starch, lactose, sucrose, glucose, mannitol and silicic acid; (b) binders, for example, hydroxymethylcellulose, alginates, gelatin, polyvinylpyrrolidone, sucrose and gum arabic; (c) humectants, For example, glycerin; (d) a disintegrant such as agar, calcium carbonate, potato starch or tapioca starch, alginic acid, certain complex silicates and sodium carbonate; (e) a slow solvent such as paraffin; (f) absorption Accelerators, for example, quaternary amine compounds; (g) wetting agents, such as cetyl alcohol and
  • Solid dosage forms such as tablets, dragees, capsules, pills, and granules can be prepared with coatings and shells such as enteric coatings and other materials known in the art. They may contain opacifying agents and the release of the active compound or compound in such compositions may be released in a portion of the digestive tract in a delayed manner. Examples of embedding components that can be employed are polymeric and waxy materials. If necessary, the active compound may also be in microencapsulated form with one or more of the above-mentioned excipients.
  • Liquid dosage forms for oral administration include pharmaceutically acceptable emulsions, solutions, suspensions, syrups or elixirs.
  • the liquid dosage form may contain inert diluents conventionally employed in the art, such as water or other solvents, solubilizers and emulsifiers, for example, ethanol, isopropanol, ethyl carbonate, ethyl acetate, propylene glycol, 1 , 3-butanediol, Dimethylformamide and oils, especially cottonseed oil, peanut oil, corn germ oil, olive oil, castor oil and sesame oil or a mixture of these substances.
  • compositions may contain adjuvants such as wetting agents, emulsifying and suspending agents, sweetening agents, flavoring agents and perfumes.
  • the suspension may contain suspending agents, for example, ethoxylated isostearyl alcohol, polyoxyethylene sorbitol and sorbitan esters, microcrystalline cellulose, aluminum methoxide and agar or mixtures of these and the like.
  • suspending agents for example, ethoxylated isostearyl alcohol, polyoxyethylene sorbitol and sorbitan esters, microcrystalline cellulose, aluminum methoxide and agar or mixtures of these and the like.
  • compositions for parenteral injection may comprise a physiologically acceptable sterile aqueous or nonaqueous solution, dispersion, suspension or emulsion, and a sterile powder for reconstitution into a sterile injectable solution or dispersion.
  • Suitable aqueous and nonaqueous vehicles, diluents, solvents or vehicles include water, ethanol, polyols, and suitable mixtures thereof.
  • Dosage forms for the compounds of the invention for topical administration include ointments, powders, patches, propellants and inhalants.
  • the active ingredient is admixed under sterile conditions with a physiologically acceptable carrier and any preservatives, buffers, or, if necessary, propellants.
  • the compounds of the invention may be administered alone or in combination with other therapeutic means and/or other therapeutic agents.
  • a safe and effective amount of a compound of the invention is administered to a mammal (e.g., a human) in need of treatment wherein the dosage is a pharmaceutically effective effective dosage, for a 60 kg body weight
  • the dose to be administered is usually from 1 to 2000 mg, preferably from 5 to 500 mg.
  • specific doses should also consider factors such as the route of administration, the health of the patient, etc., which are within the skill of the skilled physician.
  • the present invention provides a process for the preparation of the pharmaceutical composition comprising the steps of: pharmaceutically acceptable carrier and said compound or a stereoisomer, geometric isomer thereof, tautomer thereof A pharmaceutical salt, a prodrug thereof, and a hydrate or solvate thereof are mixed to form a pharmaceutical composition.
  • the invention also provides the use of a compound or the pharmaceutical composition for the preparation of a medicament for the prevention and/or treatment of a disease selected from the group consisting of:
  • the tumor-related disease is selected from the group consisting of breast cancer, lung cancer, bladder cancer, gastric cancer, pancreatic cancer, prostate cancer, colon cancer, multiple myeloma AML, liver cancer, melanoma, head and neck cancer, Thyroid cancer, renal cell carcinoma, glioblastoma, and testicular cancer.
  • the lung cancer is non-small cell lung cancer.
  • the protein tyrosine kinase activity-associated disease is selected from the group consisting of FGFR-associated diseases.
  • the present invention provides an FGFR inhibitor comprising an inhibitory effective amount of the compound or a stereoisomer, a geometric isomer, a tautomer thereof, a pharmaceutically acceptable salt thereof, or the like thereof One or more of a prodrug and a hydrate or solvate thereof.
  • the FGFR inhibitor is a FGFR1 inhibitor and a FGFR4 inhibitor.
  • the invention also provides a method of treating a FGFR-associated disease comprising administering to a subject in need thereof an inhibitory effective amount of said compound or a stereoisomer, geometric isomer, tautomer thereof, pharmaceutically acceptable salt thereof a prodrug thereof and a hydrate or solvate thereof or the pharmaceutical composition.
  • the invention also provides a method of non-diagnostic, non-therapeutic inhibition of FGFR activity, the method comprising administering to a patient in need thereof an inhibitory effective amount of the compound or a stereoisomer, geometric isomer thereof, An isomer, a pharmaceutically acceptable salt thereof, a prodrug thereof, and a hydrate or solvate thereof or the pharmaceutical composition.
  • the present invention has the following main advantages:
  • the inhibitor prepared by the preparation of the compound has a significant inhibitory effect on the FGFR enzyme activity
  • the compound can achieve significant inhibition of FGFR1-4 enzyme activity at the nM level;
  • the compound also has a significant inhibitory effect on FGFR1-4-induced proliferation of various cancer cells at the cellular level.
  • reaction solution was added to a hot (about 75 ° C) 11 g of potassium ethyl xanthate dissolved in 51 ml of water, stirring was continued for 1 hour, cooled to room temperature, extracted with ethyl acetate, organically separated The phase was washed with a saturated aqueous solution of sodium chloride, dried over anhydrous sodium sulfate, filtered and concentrated, then dissolved in a 1.3N aqueous solution of potassium hydroxide, and added with 3 g of glucose and refluxed for 3 hours. The reaction mixture was concentrated and cooled with 6 N sulfuric acid. The pH was adjusted to about 1, and then 5.7 g of zinc powder was added and heated and stirred at 50 ° C for 30 minutes.
  • Step 4 Preparation of (2,2-diethoxyethyl)(2-methoxy-4-methylphenyl) sulfide
  • Step 8 Preparation of tert-butyl 3-(4-chloro-5-iodo-7H-pyrrole[2,3-d]pyrimidin-7-yl)pyrrolidine-1-carboxylate
  • Step 9 Preparation of tert-butyl 3-(4-amino-5-iodo-7H-pyrrole[2,3-d]pyrimidin-7-yl)pyrrolidine-1-carboxylate
  • Step 10 Preparation of 3-(4-amino-5-(7-methoxy-5-methylbenzothiophen-2-yl)-7H-pyrrole[2,3-d]pyrimidin-7-yl)pyrrole Alkyl-1-carboxylic acid tert-butyl ester
  • Step 11 Preparation of 5-(7-methoxy-5-methylbenzothiophen-2-yl)-7-(pyrrolidin-3-yl)-7H-pyrrole[2,3-d]pyrimidine-4 -amino hydrochloride
  • Step 12 Preparation of 1-(3-(4-amino-5-(7-methoxy-5-methylbenzothiophen-2-yl)-7H-pyrrole[2,3-d]pyrimidin-7- Pyrrolidin-1-yl)prop-2-en-1-one
  • Step 3 Preparation of tert-butyl 3-((methylsulfonyl)oxy)pyrrolidine-1-carboxylate
  • Step 4 Preparation of tert-butyl 3-(4-amino-3-iodo-1H-pyrazolo[3,4-d]pyrimidin-1-yl)pyrrolidine-1-carboxylate
  • Step 5 Preparation of 3-(4-amino-3-(7-methoxy-5-methylbenzothiophen-2-yl)-1H-pyrazole [3,4-d]pyrimidin-1-yl) Pyrrolidine-1-carboxylic acid tert-butyl ester
  • Step 7 Preparation of 1-(3-(4-amino-3-(7-methoxy-5-methylbenzothiophen-2-yl)-1H-pyrazole[3,4-d]pyrimidine-1 -yl)pyrrolidin-1-yl)propyl-2-en-1-one
  • the propiolic acid is replaced with trans-4-(ethyl(methyl)amino) croton hydrochloride, and the remaining raw materials, reagents and preparation methods are the same as those in the third embodiment, and (E) 1-(3- (4-Amino-5-(7-methoxy-5-methylbenzothiophen-2-yl)-7H-pyrrole[2,3-d]pyrimidin-7-yl)pyrrolidin-1-yl) 4-(Ethyl(methyl)amino)but-2-en-1-one, yield 45%.
  • Step 1 Preparation of (R)-3-(4-chloro-5-iodo-7H-pyrrole[2,3-d]pyrimidin-7-yl)pyrrolidine-1-carboxylic acid tert-butyl ester
  • Step 2 Preparation of (R)-3-(4-amino-5-iodo-7H-pyrrole[2,3-d]pyrimidin-7-yl)pyrrolidine-1-carboxylic acid tert-butyl ester
  • Step 3 Preparation of (R) 3-(4-amino-5-(7-methoxy-5-methylbenzothiophen-2-yl)-7H-pyrrole[2,3-d]pyrimidin-7- Tert-butyl pyrrolidine-1-carboxylate
  • Step 5 Preparation of (R)-1-(3-(4-amino-5-(7-methoxy-5-methylbenzothiophen-2-yl)-7H-pyrrole [2,3-d] Pyrimidin-7-yl)pyrrolidin-1-yl)prop-2-en-1-one
  • Step 1 Preparation of (S)-3-(4-chloro-5-iodo-7H-pyrrole[2,3-d]pyrimidin-7-yl)pyrrolidine-1-carboxylic acid tert-butyl ester
  • Step 2 Preparation of (S)-3-(4-amino-5-iodo-7H-pyrrole[2,3-d]pyrimidin-7-yl)pyrrolidine-1-carboxylic acid tert-butyl ester
  • Step 3 Preparation of (S) 3-(4-amino-5-(7-methoxy-5-methylbenzothiophen-2-yl)-7H-pyrrole[2,3-d]pyrimidin-7- Tert-butyl pyrrolidine-1-carboxylate
  • Step 5 Preparation of (S)-1-(3-(4-amino-5-(7-methoxy-5-methylbenzothiophen-2-yl)-7H-pyrrole [2,3-d] Pyrimidin-7-yl)pyrrolidin-1-yl)prop-2-en-1-one
  • Step 1 Preparation of tert-butyl 3-(4-chloro-5-iodo-7H-pyrrole[2,3-d]pyrimidin-7-yl)azetidin-1-carboxylate
  • tert-butyl 3-hydroxypyrrolidine-1-carboxylate is replaced with the tert-butyl 3-hydroxyazetidine-1-carboxylate, and the remaining raw materials, reagents and preparation methods are the same as those in the first step of the first embodiment.
  • Step 2 Preparation of tert-butyl 3-(4-amino-5-iodo-7H-pyrrole[2,3-d]pyrimidin-7-yl)azetidin-1-carboxylate
  • Step 3 Preparation of 3-(4-amino-5-(7-methoxy-5-methylbenzothiophen-2-yl)-7H-pyrrole[2,3-d]pyrimidin-7-yl)nitrogen Tert-butyl butane-1-carboxylate
  • Step 5 Preparation of 1-(3-(4-amino-5-(7-methoxy-5-methylbenzothiophen-2-yl)-7H-pyrrole[2,3-d]pyrimidin-7- Azetidin-1-yl)prop-2-en-1-one
  • Step 1 Preparation of tert-butyl 4-(4-chloro-5-iodo-7H-pyrrole[2,3-d]pyrimidin-7-yl)piperidine-1-carboxylate
  • Step 2 Preparation of tert-butyl 4-(4-amino-5-iodo-7H-pyrrole[2,3-d]pyrimidin-7-yl)piperidine-1-carboxylate
  • Step 3 Preparation of 4-(4-amino-5-(7-methoxy-5-methylbenzothiophen-2-yl)-7H-pyrrole[2,3-d]pyrimidin-7-yl)peri Butyl-1-carboxylic acid tert-butyl ester
  • Step 4 Preparation of 5-(7-methoxy-5-methylbenzothiophen-2-yl)-7-(piperidin-4-yl)-7H-pyrrole[2,3-d]pyrimidine-4 -amino hydrochloride
  • Step 5 Preparation of 1-(4-(4-amino-5-(7-methoxy-5-methylbenzothiophen-2-yl)-7H-pyrrole[2,3-d]pyrimidin-7- Piperidin-1-yl)prop-2-en-1-one
  • Step 1 Preparation of tert-butyl 3-(4-chloro-5-iodo-7H-pyrrole[2,3-d]pyrimidin-7-yl)piperidine-1-carboxylate
  • the tert-butyl 3-hydroxypyrrolidine-1-carboxylate was replaced with the tert-butyl 3-hydroxypiperidine-1-carboxylate, and the remaining starting materials, reagents and preparation methods were the same as those in the first step of Example 1, to obtain 3-( Tert-butyl 4-chloro-5-iodo-7H-pyrrole[2,3-d]pyrimidin-7-yl)piperidine-1-carboxylate.
  • Step 2 Preparation of tert-butyl 3-(4-amino-5-iodo-7H-pyrrole[2,3-d]pyrimidin-7-yl)piperidine-1-carboxylate
  • Step 3 Preparation of 3-(4-amino-5-(7-methoxy-5-methylbenzothiophen-2-yl)-7H-pyrrole[2,3-d]pyrimidin-7-yl)piperidin Butyl-1-carboxylic acid tert-butyl ester
  • Step 4 Preparation of 5-(7-methoxy-5-methylbenzothiophen-2-yl)-7-(piperidin-3-yl)-7H-pyrrole[2,3-d]pyrimidine-4 -amino hydrochloride
  • Step 5 Preparation of 1-(3-(4-amino-5-(7-methoxy-5-methylbenzothiophen-2-yl)-7H-pyrrole[2,3-d]pyrimidin-7- Piperidin-1-yl)prop-2-en-1-one
  • acryloyl chloride is replaced by propionyl chloride, and the remaining raw materials, reagents and preparation methods are the same as step 12 in the first embodiment.
  • Step 1 Preparation of tert-butyl 3-((4-chloro-5-iodo-7H-pyrrole[2,3-d]pyrimidin-7-yl)methyl)pyrrolidine-1-carboxylate
  • tert-butyl 3-hydroxypyrrolidine-1-carboxylate is replaced with the tert-butyl 3-hydroxymethylpyrrolidine-1-carboxylate, and the remaining raw materials, reagents and preparation methods are the same as those in the first step in the first embodiment.
  • Step 2 Preparation of tert-butyl 3-((4-amino-5-iodo-7H-pyrrole[2,3-d]pyrimidin-7-yl)methyl)pyrrolidine-1-carboxylate
  • Step 3 Preparation of 3-((4-amino-5-(7-methoxy-5-methylbenzothiophen-2-yl)-7H-pyrrole[2,3-d]pyrimidin-7-yl) Methyl)pyrrolidine-1-carboxylic acid tert-butyl ester
  • Step 4 Preparation of 5-(7-methoxy-5-methylbenzothiophen-2-yl)-7-(pyrrolidin-3-methylene)-7H-pyrrole[2,3-d]pyrimidine -4-amine hydrochloride
  • Step 5 Preparation of 1-(3-((4-amino-5-(7-methoxy-5-methylbenzothiophen-2-yl)-7H-pyrrole[2,3-d]pyrimidine-7 -yl)methyl)pyrrolidin-1-yl)prop-2-en-1-one
  • Step 1 Preparation of (2,2-diethoxyethyl)(3-methoxyphenyl) sulfide
  • Step 4 Preparation of 3-(4-amino-5-(6-methoxybenzothiophen-2-yl)-7H-pyrrole[2,3-d]pyrimidin-7-yl)azetidine- 1-carboxylic acid tert-butyl ester
  • Step 6 Preparation of 1-(3-(4-amino-5-(6-methoxybenzothiophen-2-yl)-7H-pyrrole[2,3-d]pyrimidin-7-yl)azacyclocycle Butan-1-yl)prop-2-en-1-one
  • Step 1 Preparation of (2,2-diethoxyethyl)(2-methoxyphenyl) sulfide
  • Step 4 Preparation of 3-(4-amino-5-(6-methoxybenzothiophen-2-yl)-7H-pyrrole[2,3-d]pyrimidin-7-yl)nitrogen Tert-butyl butane-1-carboxylate
  • Step 6 Preparation of 1-(3-(4-amino-5-(7-methoxybenzothiophen-2-yl)-7H-pyrrole[2,3-d]pyrimidin-7-yl)azacyclocycle Butan-1-yl)prop-2-en-1-one
  • 2-methoxy-4-methylthiophenol is replaced by 2-methoxy-4-methylphenol, and the remaining raw materials, reagents and preparation methods are the same as those in step 4 of Example 1, to obtain 1-( 2,2-Diethoxyethoxy)-2-methoxy-4-methylbenzene.
  • Step 4 Preparation of 3-(4-amino-5-(7-methoxy-5-methylbenzofuran-2-yl)-7H-pyrrole[2,3-d]pyrimidin-7-yl)nitrogen Tert-butyl butane-1-carboxylate
  • Step 6 Preparation of 1-(3-(4-amino-5-(7-methoxy-5-methylbenzofuran-2-yl)-7H-pyrrole[2,3-d]pyrimidin-7- Azetidin-1-yl)prop-2-en-1-one
  • Step 1 Preparation of 2-((4-chloro-5-iodo-7H-pyrrole[2,3-d]pyrimidin-7-yl)methyl)pyrrolidine-1-carboxylic acid tert-butyl ester
  • tert-butyl 3-hydroxypyrrolidine-1-carboxylate is replaced with the tert-butyl 2-hydroxymethylpyrrolidine-1-carboxylate, and the remaining raw materials, reagents and preparation methods are the same as those in the first step of the first embodiment.
  • Step 2 Preparation of 2-((4-amino-5-iodo-7H-pyrrole[2,3-d]pyrimidin-7-yl)methyl)pyrrolidine-1-carboxylic acid tert-butyl ester
  • Step 3 Preparation of 2-((4-amino-5-(7-methoxy-5-methylbenzothiophen-2-yl)-7H-pyrrole[2,3-d]pyrimidin-7-yl) Methyl)pyrrolidine-1-carboxylic acid tert-butyl ester
  • Step 4 Preparation of 5-(7-methoxy-5-methylbenzothiophen-2-yl)-7-(pyrrolidin-2-methylene)-7H-pyrrole[2,3-d]pyrimidine -4-amine hydrochloride
  • Step 5 Preparation of 1-(2-((4-amino-5-(7-methoxy-5-methylbenzothiophen-2-yl)-7H-pyrrole[2,3-d]pyrimidine-7 -yl)methyl)pyrrolidin-1-yl)prop-2-en-1-one
  • Step 1 Preparation of 3-(3-(4-amino-5-(7-methoxy-5-methylbenzothiophen-2-yl)-7H-pyrrole[2,3-d]pyrimidin-7- Pyrrolidin-1-yl)-3-oxopropanenitrile
  • Step 2 Preparation of 2-(3-(4-amino-5-(7-methoxy-5-methylbenzothiophen-2-yl)-7H-pyrrole[2,3-d]pyrimidin-7- Azetidinium-1-carbonyl)-3-cyclopropyl acrylonitrile
  • Step 1 Preparation of tert-butyl 3-((methylsulfonyl)oxy)azetidin-1-carboxylate
  • tert-butyl 3-hydroxypyrrolidine-1-carboxylate is replaced with the tert-butyl 3-hydroxyazetidin-1-carboxylate, and the remaining raw materials, reagents and preparation methods are the same as those in the third step in the second embodiment.
  • Step 2 Preparation of tert-butyl 3-(4-amino-3-iodo-1H-pyrazolo[3,4-d]pyrimidin-1-yl)azetidin-1-carboxylate
  • Step 3 Preparation of 3-(4-amino-3-(7-methoxy-5-methylbenzothiophen-2-yl)-1H-pyrazole [3,4-d]pyrimidin-1-yl) Azetidine azetidine-1-carboxylate
  • Step 4 Preparation of 3-(7-methoxy-5-methylbenzothiophen-2-yl)-1-(azetidin-3-yl)-1H-pyrazole [3,4-d Pyrimidine-4-amine hydrochloride
  • Step 5 Preparation of 1-(3-(4-amino-3-(7-methoxy-5-methylbenzothiophen-2-yl)-1H-pyrazole [3,4-d]pyrimidine-1 -yl)azetidin-1-yl)propyl-2-en-1-one
  • Step 3 Preparation of 2-(4-amino-7-(1-(tert-butylcarboxylate)azetidin-3-yl)-7H-pyrrole[2,3-d]pyrimidin-5-yl)- 1H-indole-1-carboxylic acid tert-butyl ester
  • Step 5 Preparation of 1-(3-(4-amino-5-(1H-indol-2-yl)-7H-pyrrole[2,3-d]pyrimidin-7-yl)azetidin-1 -yl)prop-2-en-1-one
  • Step 2 Preparation of 1-(3-(4-amino-5-(7-methoxy-5-methylbenzothiophen-2-yl)-7H-pyrrole[2,3-d]pyrimidine-7- Azetidin-1-yl)-2-(morphinolinylmethyl)prop-2-en-1-one
  • the preparation method was the same as in Example 12, and the yield was 55%.
  • the preparation method was the same as in Example 26, and the yield was 47%.
  • the preparation method was the same as in Example 26, and the yield was 39%.
  • the preparation method was the same as in Example 27, and the yield was 46%.
  • the preparation method was the same as that in Example 13, and the yield was 60%.
  • the preparation method was the same as in Example 1, and the yield was 45%.
  • the preparation method was the same as in Example 1, and the yield was 43%.
  • the preparation method was the same as in Example 31, and the yield was 45%.
  • the preparation method was the same as that of Example 31, and the yield was 47%.
  • the preparation method was the same as that of Example 31, and the yield was 49%.
  • the preparation method was the same as that in Example 31, and the yield was 43%.
  • the preparation method was the same as that of Example 31, and the yield was 44%.
  • the preparation method was the same as that in Example 31, and the yield was 49%.
  • the preparation method was the same as that of Example 31, and the yield was 46%.
  • the preparation method is the same as that of the embodiment 31
  • the preparation method is the same as that of the embodiment 31
  • the preparation method was the same as that in Example 31, and the yield was 47%.
  • the preparation method was the same as that of Example 31, and the yield was 46%.
  • the preparation method was the same as that in Example 31, and the yield was 45%.
  • the preparation method was the same as in Example 2, and the yield was 55%.
  • Step 1 The starting material 2-methoxy-4-methylthiophenol in the step 4 of Example 1 was replaced with 2,4-dimethoxythiophenol. The remaining steps were the same as in Example 1, and the title was prepared.
  • Compound 1- (3-(4-Amino-5-(5,7-dimethoxybenzothiophen-2-yl)-7H-pyrrole[2,3-d]pyrimidin-7-yl)pyrrolidine- 1-yl)prop-2-en-1-one.
  • Step 4 Substituting the starting material (2,2-diethoxyethyl)(2-methoxy-4-methylphenyl) sulfide of Step 1 of Example 1 with (2,2-diethoxy) (2-Methoxy-4-(trifluoromethyl)benzene)sulfane, the title compound was obtained according to the procedure of Example 1.
  • Step 3 Preparation of (4-chloro-2-methoxybenzene) (2,2-diethoxyethyl) sulfide
  • (4-chloro-2-methoxy) can be obtained by the method reported in Adv. Synth. Catal. 2015, 357, 2205-2212 (Benzene) (2,2-diethoxyethyl) sulfide.
  • Step 4 Substituting the starting material (2,2-diethoxyethyl)(2-methoxy-4-methylphenyl) sulfide of the first step of Example 1 with (4-chloro-2-methoxy) The title compound was obtained according to the procedure of Example 1 (2,2-diethoxyethyl) thioether.
  • Example 12 The raw material ammonia of the step 2 in Example 12 was replaced with an aqueous solution of dimethylamine, and the remaining steps were the same as those in Example 12.
  • Example 12 Prepared 1-(3-(4-(dimethylamino)-5-(7-methoxy-5-methylbenzothiophen-2-yl)-7H-pyrrole[2,3-d]pyrimidine- 7-yl)pyrrolidin-1-yl)prop-2-en-1-one. The yield was 59%.
  • Step 2 Preparation of 2-thio-1,2,3,7-tetrahydro-4H-pyrrole[2,3-d]pyrimidin-4-one.
  • Step 3 Preparation of 2-(methylthio)-3,7-dihydro-4H-pyrrole[2,3-d]pyrimidin-4-one.
  • Step 4 Preparation of 4-chloro-2-(methylthio)-7H-pyrrole [2,3-d]pyrimidine.
  • Step 5 Preparation of 4-chloro-5-iodo-2-(methylthio)-7H-pyrrole [2,3-d]pyrimidine.
  • Step 6 Preparation of tert-butyl 3-(4-chloro-5-iodo-2-(methylthio)-7H-pyrrole[2,3-d]pyrimidin-7-yl)pyrrolidine-1-carboxylate.
  • Step 7 Preparation of tert-butyl 3-(4-amino-5-iodo-2-(methylthio)-7H-pyrrole[2,3-d]pyrimidin-7-yl)pyrrolidine-1-carboxylate.
  • the preparation method was the same as that in Example 1.
  • Step 8 Preparation of tert-butyl 3-(4-amino-5-iodo-2-(methylsulfonyl)-7H-pyrrole[2,3-d]pyrimidin-7-yl)pyrrolidine-1-carboxylate.
  • Step 9 Preparation of 3-(4-amino-5-iodo-2-(2-morpholineethoxy)-7H-pyrrole[2,3-d]pyrimidin-7-yl)pyrrolidine-1-carboxylic acid Butyl ester.
  • Step 10 Substituting tert-butyl 3-(4-amino-5-iodo-7H-pyrrole[2,3-d]pyrimidin-7-yl)pyrrolidine-1-carboxylate in Step 10 of Example 1 into 3 -(4-Amino-5-iodo-2-(2-morpholineethoxy)-7H-pyrrole[2,3-d]pyrimidin-7-yl)pyrrolidine-1-carboxylic acid tert-butyl ester, the remaining steps Similar to Example 1, the title compound was prepared.
  • Step 1 Preparation of tert-butyl 2-(((tert-butyldiphenylsilyl)oxy)methyl)-4-hydroxypyrrolidine-1-carboxylate
  • Step 2 Preparation of 4-(4-amino-5-(7-methoxy-5-methylbenzothiophen-2-yl)-7H-pyrrole[2,3-d]pyrimidin-7-yl)- 2-(((tert-Butyldiphenylsilyl)oxy)methylene)pyrrolidine-1-carboxylic acid tert-butyl ester.
  • Step 3 Preparation of 4-(4-amino-5-(7-methoxy-5-methylbenzothiophen-2-yl)-7H-pyrrole[2,3-d]pyrimidin-7-yl)- tert-Butyl 2-(hydroxymethyl)pyrrolidine-1-carboxylate.
  • Step 4 4-(4-Amino-5-(7-methoxy-5-methylbenzothiophen-2-yl)-7H-pyrrole[2,3-d]pyrimidine-7 obtained in Step 3
  • the title compound is obtained in the same manner as in the step 11 to the step 12 of Example 1 as the starting material.
  • Step 1 Preparation of 3-(4-chloro-5-(7-methoxy-5-methylbenzothiophen-2-yl)-7H-pyrrole[2,3-d]pyrimidin-7-yl)pyrrole Alkyl-1-carboxylic acid tert-butyl ester
  • Step 2 Preparation of 3-(4,6-dichloro-5-(7-methoxy-5-methylbenzothiophen-2-yl)-7H-pyrrole[2,3-d]pyrimidin-7- Tert-butyl pyrrolidine-1-carboxylate
  • Step 3 Preparation of 3-(4-amino-6-chloro-5-(7-methoxy-5-methylbenzothiophen-2-yl)-7H-pyrrole[2,3-d]pyrimidine-7 -yl)pyrrolidine-1-carboxylic acid tert-butyl ester
  • Step 4 According to Example 1, the starting material of Step 1 of Example 1 was 3-(4-amino-5-(7-methoxy-5-methylbenzothiophen-2-yl)-7H-pyrrole [2 , 3-d]pyrimidin-7-yl)pyrrolidine-1-carboxylic acid tert-butyl ester was replaced by 3-(4-amino-6-chloro-5-(7-methoxy-5-methylbenzothiophene- The title compound can be prepared from 2-ethyl)-7H-pyrrole[2,3-d]pyrimidin-7-yl)pyrrolidine-1-carboxylic acid tert-butyl ester.
  • Step 1 Preparation of (2-(4-chloro-5-iodo-7H-pyrrole[2,3-d]pyrimidin-7-yl)ethyl)carbamic acid tert-butyl ester
  • Example 1 the intermediate obtained in Step 1 was used as a raw material to replace 3-(4-chloro-5-iodo-7H-pyrrole[2,3-d]pyrimidin-7-yl)pyrrole in Step 9 of Example 1.
  • the title compound can be prepared from tert-butyl alkane-1-carboxylate.
  • Step 1 Preparation of 1-(tert-butyl 2-methyl(4S)-4-(4-amino-3-(7-methoxy-5-methylbenzothiophen-2-yl)-1H-pyridyl Azole [3,4-d]pyrimidin-1-yl)pyrrolidine-1,2-dicarboxylate
  • Step 2 Preparation of (4S)-4-(4-amino-3-(7-methoxy-5-methylbenzothiophen-2-yl)-1H-pyrazole [3,4-d]pyrimidine- 1-yl)-1-(tert-butyl formate) pyrrolidine-2-carboxylic acid.
  • Step 3 Preparation of (4S)-4-(4-amino-3-(7-methoxy-5-methylbenzothiophen-2-yl)-1H-pyrazole [3,4-d]pyrimidine- 1-yl)-2-((2-(dimethylamino)ethyl)(methyl)carboxamide)pyrrolidine-1-carboxylic acid tert-butyl ester
  • Step 4 (4S)-4-(4-Amino-3-(7-methoxy-5-methylbenzothiophen-2-yl)-1H-pyrazole obtained in Step 3 [3,4- d]pyrimidin-1-yl)-2-((2-(dimethylamino)ethyl)(methyl)carboxamide)pyrrolidine-1-carboxylic acid tert-butyl ester was replaced by 3-(Step 6 of Example 2) 4-amino-3-(7-methoxy-5-methylbenzothiophen-2-yl)-1H-pyrazole[3,4-d]pyrimidin-1-yl)pyrrolidine-1-carboxylic acid Butyl ester, the title compound was obtained. [M+H] + :563
  • Step 3 Replace 3-(4-chloro-5-iodo-7H-pyrrole[2,3-d]pyrimidin-7-yl)pyrrolidine-1-carboxylic acid tert-butyl ester in Step 9 of Example 1 with Step 2 4-(2-(4-Chloro-5-iodo-7H-pyrrole[2,3-d]pyrimidin-7-yl)ethyl)morpholine was obtained, and the title was obtained according to Example 9 Step 9 to Step 10. Compound.
  • Step 2 Substituting the starting material 3-(4-amino-5-iodo-7H-pyrrole[2,3-d]pyrimidin-7-yl)pyrrolidine-1-carboxylic acid tert-butyl ester in step 10 of Example 1 into a step 1-(3-Aminobenzyl)-3-iodo-1H-pyrazole[3,4-d]pyrimidin-4-amine obtained, the same procedure as in Example 1 gave the title compound N-(3- ((4-Amino-3-(7-methoxy-5-methylbenzothiophen-2-yl)-1H-pyrazolo[3,4-d]pyrimidin-1-yl)methyl)phenyl )Acrylamide.
  • Step 1 Replace the starting material (7-methoxy-5-methylbenzothiophen-2-yl)boronic acid of Step 10 of Example 1 with 3,5-dimethoxyphenylboronic acid pinacol ester, and the rest.
  • the same procedure as in Example 1 gave the title compound 1-(3-(4-amino-5-(3,5-dimethoxyphenyl)-7H-pyrrole[2,3-d]pyrimidin-7-yl) Pyrrolidin-1-yl)prop-2-en-1-one.
  • Step 1 Preparation of tert-butyl 3-(4-amino-5-(4-cyano-3-fluorophenyl)-7H-pyrrole[2,3-d]pyrimidin-7-yl)pyrrolidine-1-carboxylate
  • Step 2 Preparation of 3-(4-amino-5-(3-amino-1H-indazol-6-yl)-7H-pyrrole[2,3-d]pyrimidin-7-yl)pyrrolidine-1-carboxylic acid Tert-butyl ester
  • Step 3 3-(4-Amino-5-(7-methoxy-5-methylbenzothiophen-2-yl)-7H-pyrrole [2,3-d] in Step 11 of Example 1.
  • Tert-butyl pyrimidin-7-yl)pyrrolidine-1-carboxylate was replaced by 3-(4-amino-5-(3-amino-1H-indazol-6-yl)-7H-pyrrole obtained in the previous step [2 , 3-d]pyrimidin-7-yl)pyrrolidine-1-carboxylic acid tert-butyl ester, the title compound was obtained by the same procedure.
  • Step 1 The acryloyl chloride in the step 7 of Example 2 was replaced with chloroacetyl chloride to give the title compound.
  • Step 1 Preparation of ethyl 7-hydroxybenzothiophene-5-carboxylate
  • Ethyl 7-hydroxybenzothiophene-5-carboxylate can be obtained according to a similar procedure from J. AM. CHEM. SOC. 2007, 129, 14092-14099.
  • Step 4 Substituting 7-methoxy-5-methylbenzothiophene and n-butyllithium of step 6 of Example 1 with 7-methoxybenzothiophene-5-carboxylic acid and lithium diisopropylamide, respectively The title compound was prepared according to the same procedure as in Example 1.
  • the (7-methoxy-5-methylbenzothiophen-2-yl)boronic acid in the step 3 of Example 12 was replaced with the (7-methoxybenzothiophen-2-yl) group obtained in Example 22.
  • the title compound can be prepared from boric acid.
  • Example 1 The preparation method is referred to in Example 1.
  • the enzyme reaction substrate Poly(Glu, Tyr) 4:1 was diluted with potassium-free PBS (10 mM sodium phosphate buffer, 150 mM NaCl, pH 7.2-7.4) to 20 ⁇ g/mL, and 125 ⁇ L/well coated with the ELISA plate. The reaction was carried out at 37 ° C for 12-16 hours. After discarding the liquid in the well, the plate was washed, and the plate was washed three times with 200 ⁇ L/well of T-PBS (PBS containing 0.1% Tween-20) for 5 minutes each time. The plate was dried in an oven at 37 ° C for 1-2 hours.
  • reaction buffer 50 mM HEPES pH 7.4, 50 mM MgCl 2 , 0.5 mM MnCl 2 , 0.2 mM Na 3 VO 4 , 1 mM DTT
  • the compound was diluted with DMSO to a suitable concentration, 1 ⁇ L/well or containing the corresponding concentration of DMSO (negative control well), and then the FGFR1, FGFR4 kinase domain recombinant protein diluted with 49 ⁇ L of reaction buffer was added to initiate the reaction, and each experiment was required.
  • the reaction was carried out for 1 hour at 37 ° C on a shaker (100 rpm).
  • the plate was washed three times with T-PBS.
  • One anti-PY99 dilution was added to 100 ⁇ L/well, and the reaction was shaken at 37 ° C for 0.5 hour.
  • the plate was washed three times with T-PBS.
  • a second anti-horseradish peroxidase-labeled goat anti-mouse IgG dilution was added at 100 ⁇ L/well, and shaken at 37 ° C for 0.5 hour.
  • the plate was washed three times with T-PBS.
  • the inhibition rate of the sample is obtained by the following formula:
  • IC 50 values were obtained by four-parameter regression using software attached to the microplate reader.
  • A indicates that the IC 50 is less than ( ⁇ ) 10nM
  • C means IC 50 is greater than (>) 100nM
  • the compound NO. 12 which is a representative compound of the present invention, has substantially no EGFR inhibitory activity.
  • the inhibitory effect of the compound on proliferation of SNU16 cells was detected by CCK-8 Cell Counting Kit (Dojindo).
  • the specific steps are as follows: SNU16 cells in logarithmic growth phase were inoculated into 96-well culture plates at a suitable density, 90 ⁇ L per well, and after incubation overnight, different concentrations of compounds were added for 72 hr, and a solvent control group (negative control) was set. After the compound was treated for 72h, the effect of the compound on cell proliferation was detected by CCK-8 cell counting kit (Dojindo). Add 10 ⁇ L of CCK-8 reagent to each well and place in a 37 ° C incubator for 2-4 hours.
  • the full-wavelength microplate reader SpectraMax 190 reads at a wavelength of 450 nm.
  • the IC50 value was obtained by four-parameter regression using the software attached to the microplate reader.
  • A indicates that the IC 50 is less than ( ⁇ ) 10nM
  • C means IC 50 is greater than (>) 100nM
  • the compounds of the present invention have a significant inhibitory effect on the reproductive activity of FGFR-dependent cell lines.
  • the tumor tissue in the vigorous growth period was cut into 1.5 mm 3 and inoculated subcutaneously in the right axilla of nude mice under aseptic conditions.
  • Subcutaneous xenografts in nude mice tumor diameter measurements with calipers the mean volume of the tumors grew to approximately 90mm after approximately 3 groups of animals randomized.
  • the compound No. 12 obtained in Example 12 was orally administered once a day at a dose of 100 mg/kg and 20 mg/kg, respectively, for 14 days, and the compound NO.12 was measured for subcutaneous transplantation of human lung cancer NCI-H1581 nude mice. Growth inhibition.
  • the solvent control group was given the same amount of physiological saline.
  • the diameter of the transplanted tumor was measured twice a week during the entire experiment, and the body weight of the mice was weighed.
  • Calculate the relative tumor volume (RTV) according to the measured result, and calculate the formula: RTV V t /V 0 , where V 0 is the tumor volume measured when the cage is administered (ie, d 0 ), V t The tumor volume for each measurement.

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Abstract

本发明涉及一种具有FGFR抑制活性的新型化合物及其制备和应用。具体地，本发明所述化合物具有式I所示结构，其中各基团和取代基如说明书中所定义。本发明还公开了所述化合物的制备方法及其在制备治疗和/或预防肿瘤相关疾病和/或FGFR相关疾病的药物中的用途。

Description

一种具有FGFR抑制活性的新型化合物及其制备和应用 技术领域
本发明涉及药物领域，具体地涉及一种具有FGFR抑制活性的新型化合物及其制备和应用。
背景技术
蛋白激酶是一种通过对蛋白质上特定氨基酸的磷酸化来调节各种细胞功能的蛋白质(酶)。蛋白质通过构象的改变来调节活性以及与其他组分结合能力。蛋白质激酶的活性指的是，激酶将磷酸基团结合到底物上的速率，该速率可以通过检测一定时间内转化为产物的底物的量来进行测定。底物的磷酸化发生在蛋白质激酶的活化位点上。
酪氨酸激酶是一种可以催化将三磷酸腺苷转移到蛋白质酪氨酸残基的蛋白质酶。这些激酶在生长因子传导引发细胞增殖、分化和迁移过程中扮演着重要的角色。
成纤维细胞生长因子(FGF)已经被确认在许多生理过程中有重要的调节作用，例如器官生成和血管生成等等。目前已知，在FGF家族中有超过25种亚型，成纤维细胞生长因子受体(FGFR)家族共包含四个亚型(FGFR1-4)，他们都是糖蛋白，包含细胞外的类免疫球蛋白区域、跨膜的疏水区域和细胞质内的酪氨酸激酶区域。FGF的结合引发FGFR二聚，进而发生受体的自磷酸化和下游信号通路的活化。下游信号通路的某些特定组分对于细胞生长、代谢和存活有非常重要的作用。因此，FGFR信号通路对于肿瘤细胞的繁殖、迁移、浸润和血管生成有多效性的重要生理作用。
目前，已有证据表明FGF信号通路与人类癌症有直接的关联。在不同种类的癌细胞中(膀胱癌、肾癌和前列腺癌等等)都被报道出有不同的FGF过表达现象。因此，FGF信号通路是一个有前景的治疗靶点。
综上所述，本领域急需开发新型的FGFR抑制剂。
发明内容
本发明的目的在于提供一种具有FGFR抑制活性的新型化合物及其制备和应用。
本发明的第一方面，提供了一种式I所示化合物或其立体异构体、几何异构体、互变异构体、其药用盐、其前药及其水合物或溶剂合物，
Figure PCTCN2017087222-appb-000001
式中，R1选自下组：取代或未取代的含有1-3个选自S、O、N和Se杂原子的5-14元杂芳基和取代或未取代的6-14元芳基，并且所述取代指被选自下组的1个或多个基团取代：C1-C6烷基、卤代的C1-C6烷基、C3-C8环烷基、卤代的C3-C8环烷基、羟基、C1-C6烷氧基、卤代的C1-C6烷氧基、-O-(C3-C8环烷基)、-O-(C3-C8的卤代环烷基)、-NR6R7、卤素、-(C1-C6亚烷基)-L1、C(＝O)R8；并且当R1为含N的5-14元杂芳基时，R1不是选自下组的基团：未取代的喹啉基、未取代的异喹啉基；
R2和R3可相同或不同，分别独立地选自：H、取代或未取代的C1-C6烷基、取代或未取代的C3-C8环烷基、C(＝O)R8和S(＝O)2R9；
R4选自H、取代或未取代的C1-C6烷基、取代或未取代的C3-C8环烷基、NR6R7、卤素、羟基、氰基、取代或未取代的C1-C6烷氧基和取代或未取代的C1-C6烷硫基；
X1和X2可相同或不同，分别独立地选自N和CR10；R10选自H、取代或未取代的C1-C6烷基、取代或未取代的C3-C8环烷基、卤素、羟基、氰基、取代或未取代的C1-C6烷氧基和取代或未取代的C1-C6烷硫基；
n为0、1、2、3、4或5；
Y选自取代或未取代的含有1-3个选自N、O和S杂原子的3-10元杂环基、-NR6R7、取代或未取代的C3-C8环烷基和-L2-(取代或未取代的C6-C10芳基)-，并且所述取代指被选自下组的1个或多个基团独立地取代：C1-C6烷基、卤代的C1-C6烷基、C3-C8环烷基、卤代的C3-C8环烷基、羟基、C1-C6烷氧基、卤代的C1-C6烷氧基、-O-(C3-C8环烷基)、-O-(C3-C8的卤代环烷基)、NR11R12、卤素、含有1-3个选自N、O和S杂原子的4-10元杂环基、-(C1-C6亚烷基)-L1、C(＝O)R8和-Boc,
E选自下组：无、C(＝O)、S(＝O)2、C(＝S)和S(＝O)；
Z选自下组：无、取代或未取代的C2-C6烯基、取代或未取代的C2-C6炔基、取代或未取代的C1-C6烷基、取代或未取代的C3-C8环烷基、-(R13)-N(R11)-(R14)-(取代或未取代的C1-C6烷氧基)；
R6和R7可相同或不同，分别独立地选自：H、取代或未取代的C1-C6烷基、取代或未取代的C3-C8环烷基、C(＝O)R8、-(C1-C6亚烷基)-L1、-(C1-C6亚烷基)-(取代或未取代的含1-3个选自N、O和S杂原子的4-8元杂环基)、-CN、卤素、-OH、-(C1-C6亚烷基)-(取代或未取代的C4-C8环烷基)、或-(C1-C6亚烷基)-L2-(C1-C6亚烷基)-(取代或未取代的C1-C6烷氧基)；
R8和R9可相同或不同，分别独立地选自：H、羟基、取代或未取代的C1-C6烷基、取代或未取代的C3-C8环烷基、取代或未取代的C1-C6烷氧基、取代或未取代的C2-C4烯基、取代或未取代的C2-C4炔基、-L2-(C1-C6亚烷基)-L1和-NR11R12；
L1选自-OH、C1-C4烷氧基、-NR11R12、或含1或2个N原子的4-7元杂环；和
L2选自下组：-NR11-、或-N(取代或未取代的C3-C6环烷基)；
对于R2至R10、X1、X2、E、Z和L2，所述取代指被选自下组的1个或多个基团独立地取代： C1-C6烷基、卤代的C1-C6烷基、C3-C8环烷基、卤代的C3-C8环烷基、羟基、C1-C6烷氧基、卤代的C1-C6烷氧基、-O-(C3-C8环烷基)、-O-(C3-C8的卤代环烷基)、NR11R12、卤素、-CN、含有1-3个选自N、O和S杂原子的4-10元杂环基、-(C1-C6亚烷基)-L1和C(＝O)R8；其中，所述的4-10元杂环基可任选地具有1-3个选自下组的取代基：C1-C3烷基、卤素、-OH、C1-C3卤代烷基、C3-C4环烷基；
R11和R12独立地选自H、C1-C6烷基、C1-C6卤代烷基、C3-C6环烷基、C3-C6卤代环烷基、-CO(C2-C4烯基)、或-CO(C2-C4炔基)；或R11和R12与相邻的N构成含1-2个N原子和0-2个O或S原子的4-7元杂环；
R13和R14独立地选自取代或未取代的C1-C6亚烷基、或取代或未取代的C2-C6亚烯基。
在另一优选例中，当Z为具有1、2或3个取代基AA的C2-C6烯基，其中所述的取代基AA选自下组：NR11R12、-(C1-C6亚烷基)-L1、C3-C8环烷基、卤代的C3-C8环烷基、-CN、卤素。
在另一优选例中，当Z为具有1-2个取代基AA取代的C2-C4烯基，其中取代基AA选自下组：NR11R12、-(C1-C6亚烷基)-L1、C3-C8环烷基、-CN、卤素。
在另一优选例中，当Z具有下式结构：
Figure PCTCN2017087222-appb-000002
式中，RA、RB和RC独立地选自下组：H、上述取代基AA；
并且，RA、RB和RC不同时为H。
在另一优选例中，RA或RB为取代基AA。
在另一优选例中，RC为取代基AA。
在另一优选例中，所述的R1为下式的二环基团：
Figure PCTCN2017087222-appb-000003
式中，环A为取代或未取代的5元杂芳环；而环B为取代或未取代的6元杂芳环或取代或未取代的苯基。
在另一优选例中，所述的R1为下式的二环基团：
Figure PCTCN2017087222-appb-000004
在另一优选例中，所述的环A为含S的5元杂芳环。
在另一优选例中，所述的R1为下式的二环基团：
Figure PCTCN2017087222-appb-000005
在另一优选例中，所述的环A为含一个或两个N原子的5元杂芳环。
在另一优选例中，R1为取代的5-14元杂芳基或取代的6-14元芳基。
在另一优选例中，R1为取代的5-14元杂芳基或取代的6-14元芳基，且所述取代为具有选自下组的1个或多个取代基：C1-C6烷基、卤代的C1-C6烷基、C3-C8环烷基、卤代的C3-C8环烷基、羟基、C1-C6烷氧基、卤代的C1-C6烷氧基、-O-(C3-C8环烷基)、-O-(C3-C8的卤代环烷基)、-NR6R7、卤素、C(＝O)R8。
在另一优选例中，所述的R1为取代的5-14元杂芳基或取代的6-14元芳基，且所述取代为具有选自下组的1个或多个取代基：C1-C6烷基、羟基、C1-C6烷氧基、卤代的C1-C6烷氧基、-NH2、卤素。
在另一优选例中，对于R1，所述取代为具有至少一个选自下组的取代基：C1-C6烷氧基、卤代的C1-C6烷氧基。
在另一优选例中，对于R1，所述取代为具有至少一个选自下组的取代基：C1-C6烷氧基、卤代的C1-C6烷氧基、羟基和-NH2；以及具有至少一个选自下组的取代基：C1-C6烷基、C3-C8环烷基和卤素。
在另一优选例中，R1为取代的选自下组的基团：苯并噻吩基、苯并呋喃基、吲哚基、苯并咪唑基、苯并硒吩基、吲唑基和苯并噻唑基。
在另一优选例中，R1为取代的苯并噻吩基。
在另一优选例中，R1为取代的苯并噻吩基，并且在7位具有选自下组的取代基：C1-C6烷氧基、卤代的C1-C6烷氧基、羟基和-NH2。
在另一优选例中，所述的式I所示化合物选自表1所列化合物。
本发明的第二方面，提供了一种药物组合物，所述药物组合物包含治疗有效量的本发明第一方面所述的化合物或其立体异构体、几何异构体、互变异构体、其药用盐、其前药及其水合物或溶剂合物中的一种或多种以及任选的药学上可接受的载体。
在另一优选例中，所述药物组合物是用于预防和/或治疗癌症的药物组合物，或用于预防和/或治疗FGFR相关疾病的药物组合物。
在另一优选例中，所述的FGFR选自下组：FGFR1、FGFR2、FGFR3、FGFR4、或组合。
在另一优选例中，所述药物组合物用于预防和/或治疗FGF/FGFR信号通路异常表达相关疾病。
在另一优选例中，所述药物组合物的剂型选自下组：口服剂型、冻干制剂、注射剂。
本发明的第三方面，提供了一种本发明第一方面所述的化合物或其立体异构体、几何异构体、互变异构体、其药用盐、其前药及其水合物或溶剂合物或本发明第二方面所述药物组合物的用途，用于制备预防和/或治疗选自下组的疾病的药物：
a)肿瘤相关疾病；
b)蛋白酪氨酸激酶活性相关疾病。
在另一优选例中，所述肿瘤相关疾病选自下组：乳腺癌、肺癌、膀胱癌、胃癌、胰腺癌、 前列腺癌、结肠癌、多发性骨髓瘤AML、肝癌、黑色素瘤、头颈癌、甲状腺癌、肾细胞癌、胶质母细胞癌和睾丸癌。
在另一优选例中，所述肺癌为非小细胞肺癌。
在另一优选例中，所述蛋白酪氨酸激酶活性相关疾病选自下组：FGFR相关疾病。
本发明的第四方面，提供了一种FGFR抑制剂，所述FGFR抑制剂包含抑制有效量的本发明第一方面所述的化合物或其立体异构体、几何异构体、互变异构体、其药用盐、其前药及其水合物或溶剂合物中的一种或多种。
在另一优选例中，所述FGFR抑制剂为FGFR1抑制剂和FGFR4抑制剂。
本发明的第五方面，提供了一种本发明第一方面所述的化合物或其立体异构体、几何异构体、互变异构体、其药用盐、其前药及其水合物或溶剂合物的制备方法，所述方法包括如下步骤：
(i)在惰性溶剂中，将式4化合物或其盐与式15所示的酰卤化合物或者酸进行缩合反应，从而形成式I化合物；
Figure PCTCN2017087222-appb-000006
各式中，R1、R2、R3、R4、n、X1、X2、Y、E和Z的定义同上；
X选自下组：卤素、羟基。
在另一优选例中，步骤(i)所述反应在碱性条件下进行，优选为在三乙胺、吡啶、N，N-二异丙基乙胺、4-二甲氨基吡啶、N-甲基吗啡啉等存在的条件下进行。
在另一优选例中，所述酰卤化合物优选为酰氯化合物。
在另一优选例中，所述酸为羧酸。
本发明的第六方面，提供了一种本发明第二方面所述药物组合物的制备方法，包括步骤：将药学上可接受的载体与本发明第一方面所述的化合物或其立体异构体、几何异构体、互变异构体、其药用盐、其前药及其水合物或溶剂合物进行混合，从而形成药物组合物。
本发明的第七方面，提供了一种非诊断性、非治疗性抑制FGFR活性的方法，所述方法包括向所需患者施用抑制有效量的本发明第一方面所述的化合物或其立体异构体、几何异构体、互变异构体、其药用盐、其前药及其水合物或溶剂合物或本发明第二方面所述药物组合物。
应理解，在本发明范围内中，本发明的上述各技术特征和在下文(如实施例)中具体描述的各技术特征之间都可以互相组合，从而构成新的或优选的技术方案。限于篇幅，在此不再一一累述。
具体实施方式
本发明人经过长期而深入的研究，意外地制备得到一种结构新颖、FGFR抑制作用显著的式I化合物。与现有FGFR抑制剂相比，以本发明化合物制备的FGFR抑制剂可在nM水平上实现对FGFR1-4酶活性的明显抑制作用，并且所述抑制剂在细胞水平对FGFR1-4诱导的各种癌细胞增殖也具有显著抑制作用，这对于开发新型抗肿瘤药物具有重要意义。在此基础上，发明人完成了本发明。
术语
在本文中，除特别说明之处，术语“取代”指基团上的一个或多个氢原子被选自下组的取代基取代：C1-C6烷基、卤代的C1-C6烷基、C3-C8环烷基、卤代的C3-C8环烷基、羟基、C1-C6烷氧基、卤代的C1-C6烷氧基、-O-(C3-C8环烷基)、-O-(C3-C8的卤代环烷基)、卤素、含有1-3个选自S、O、N和Se杂原子的4-10元杂环基、氨基、苯基、氰基、C2-C6烯基、C2-C6炔基；所述的苯基包括未取代的苯基或具有1-3个取代基的取代苯基，所述取代基选自：卤素、C1-C6烷基、C3-C8环烷基、C1-C6烷氧基、OH、氰基、硝基、氨基。
除特别说明之处，本发明的所有化合物之中，各手性碳原子可以任选地为R构型或S构型，或R构型和S构型的混合物。
术语“5-14元杂芳基”指具有选自下组的1-3个杂原子的5-14元芳基失去一个氢原子形成的基团：N、S、O、Se，其中每个杂芳基的环状体系可以是单环或多环的；例如苯并噻吩基、苯并呋喃基、吲哚基、萘基、苯并咪唑基、苯并硒吩基、吡啶基、呋喃基、苯基、吲唑基、噻吩基、吡咯基、咪唑基、吡唑基、吡嗪基、噁唑基、噻唑基、苯并噻唑基、或类似基团。
术语“6-14元芳基”指6-14元芳基失去一个氢原子形成的基团；例如苯基、萘基，或类似基团。
术语“C1-C6烷基”指具有1-6个碳原子的直链或支链烷基，例如甲基、乙基、丙基、异丙基、丁基、异丁基、仲丁基、叔丁基、或类似基团。
术语“C1-C6烷氧基”指具有1-6个碳原子的直链或支链烷氧基，例如甲氧基、乙氧基、丙氧基、异丙氧基、丁氧基、异丁氧基、仲丁氧基、叔丁氧基、或类似基团。
术语“C2-C6烯基”指具有2-6个碳原子的烯烃失去一个或两个氢原子所形成的基团，所述的烯烃可以是单烯烃、二烯烃或三烯烃，例如-CH＝CH2、-C2H4＝CH2、-CH＝C2H4，或类似基团。
术语“卤素”指F、Cl、Br和I。
此外，在本发明中，术语“烷基”包括饱和或不饱和、直链、支链、环状的1-10个碳原子的全碳烷基或其中的1-3个碳原子被氧、氮、硫等杂原子取代的烷基，以及通过1个或1个以上碳原子连接的芳烷基。此外，所述的烷基是未取代的或取代的。
如本文所用，术语“芳基”包括稠合或非稠合的芳基，通常含有6-30个碳原子，代表性的芳基包括苯基、萘基，或含氧、氮、硫等杂原子的芳香基团。
除非特别说明，本发明所描述的结构式意在包括所有的同分异构形式(如对映异构，非对映异构和几何异构体(或构象异构体)：例如含有不对称中心的R、S构型，双键的(Z)、(E)异构体和(Z)、(E)的构象异构体。因此本发明的化合物的单个立体化学异构体或其对映异构体、非对映异构体或几何异构体(或构象异构体)的混合物都属于本发明的范围。
术语“互变异构体”表示具有不同能量的结构同分异构体可以超过低能垒，从而互相转化。比如，质子互变异构体(即质子移变)包括通过质子迁移进行互变，如1H-吲唑与2H-吲唑、1H-苯并[d]咪唑与3H-苯并[d]咪唑，化合价互变异构体包括通过一些成键电子重组而进行互变。
式I化合物
本发明人设计合成了一系列结构新颖的FGFR抑制剂化合物，通过结构优化，发现了在酶、细胞以及动物体内均有优异活性的小分子FGFR抑制剂，并且对FGFR1-4均有强烈抑制作用，该系列化合物有望在临床上用于治疗FGF/FGFR信号通路异常表达所引起的疾病，如癌症等。
具体地，本发明提供了一种如下式I所示化合物：
Figure PCTCN2017087222-appb-000007
在另一优选例中，R1、R2、R3、R4、R6、R7、R8、R9、R10、R11、R12、R13、R14、X1、X2、L1、L2、Y、E、Z中任一个分别为实施例中各具体化合物中所对应的基团。
在另一优选例中，所述化合物优选为实施例中所制备的化合物。
在另一优选例中，所述化合物NO.1-NO.56分别为实施例1-56制备所得化合物。
在另一优选例中，所述化合物选自表1所列化合物。
表1
Figure PCTCN2017087222-appb-000008
Figure PCTCN2017087222-appb-000009
Figure PCTCN2017087222-appb-000010
Figure PCTCN2017087222-appb-000011
Figure PCTCN2017087222-appb-000012
Figure PCTCN2017087222-appb-000013
Figure PCTCN2017087222-appb-000014
Figure PCTCN2017087222-appb-000015
Figure PCTCN2017087222-appb-000016
Figure PCTCN2017087222-appb-000017
Figure PCTCN2017087222-appb-000018
在另一优选例中，所述化合物具有显著的FGFR抑制活性。
在另一优选例中，所述化合物基本无EGFR抑制活性。
如本文所用，术语“药用盐”指本发明化合物与酸或碱所形成的适合用作药物的盐。所述药用盐包括无机盐和有机盐。一类优选的盐是本发明化合物与酸形成的盐。适合形成盐的酸包括但并不限于：盐酸、氢溴酸、氢氟酸、硫酸、硝酸、磷酸等无机酸；甲酸、乙酸、三氟乙酸、丙酸、草酸、丙二酸、琥珀酸、富马酸、马来酸、乳酸、苹果酸、酒石酸、柠檬酸、苦味酸、苯甲酸、甲磺酸、乙磺酸、甲烷磺酸、三氟甲烷磺酸、乙烷磺酸、对甲苯磺酸、苯磺酸、萘磺酸等有机酸；以及脯氨酸、苯丙氨酸、天冬氨酸、谷氨酸等氨基酸。
另一类优选的盐是本发明化合物与碱形成的盐，例如碱金属盐(例如钠盐或钾盐)、碱土金属盐(例如镁盐或钙盐)、铵盐(如低级的烷醇铵盐以及其它药学上可接受的胺盐)，例如甲胺盐、乙胺盐、丙胺盐、二甲基胺盐、三甲基胺盐、二乙基胺盐、三乙基胺盐、叔丁基胺盐、乙二胺盐、羟乙胺盐、二羟乙胺盐、三羟乙胺盐，以及分别由吗啉、哌嗪、赖氨酸形成的胺盐。
术语“溶剂合物”指本发明化合物与溶剂分子配位形成特定比例的配合物。“水合物”是指本发明化合物与水进行配位形成的配合物。
术语“前药”包括其本身可以是具有生物学活性的或非活性的，当用适当的方法服用后，其在人体内进行代谢或化学反应而转变成式I化合物的一类化合物，或式I的一个化合物所组成的盐或溶液。所述的前药包括(但不局限于)所述化合物的羧酸酯、碳酸酯、磷酸酯、硝酸酯、硫酸酯、砜酯、亚砜酯、氨基化合物、氨基甲酸盐、偶氮化合物、磷酰胺、葡萄糖苷、醚、乙缩醛等形式。
式I化合物制备方法
下面更具体地描述本发明式I结构化合物的制备方法，但这些具体方法不对本发明构成任何限制。本发明化合物还可以任选将在本说明书中描述的或本领域已知的各种合成方法组合起来而方便地制得，这样的组合可由本发明所属领域的技术人员容易地进行。
典型地，本发明化合物的制备工艺流程如下，其中所用原料和试剂如无特殊说明，均可通过商业途径购买。
Figure PCTCN2017087222-appb-000019
其中，R1、R2、R3、R4、n、X1、X2、Y、E和Z的定义同上，P是任何可做为氨基保护基的基团，L是发生亲核取代时的离去基团。
从化合物1出发通过光延反应或者亲核取代连接制成化合物2，然后偶联，去保护基后缩合得到目标产物。
具体的说，所述制备方法包括如下步骤：
1.化合物1通过光延反应或者亲核取代生成化合物2，或者化合物13通过光延反应或者亲核取代生成化合物14，化合物14与NHR2R3或其盐反应生成化合物2；
2.化合物2与相应的硼酸在钯催化剂作用下进行偶联反应得到化合物3；
3.化合物3在酸性条件下脱保护基得到化合物4；
4.化合物4与相应的酰氯化合物或者羧酸缩合得到目标产物。
其中，所述的中间体1可以通过本领域的常规方法制备，或通过市售途径获得。在本发明的一个优选例中，所述化合物(1a)由如下方法制备：
Figure PCTCN2017087222-appb-000020
其中R2、R3的定义同上。
从化合物5出发做成碘代物，再氨化得到中间体(1a)。
具体的说，所述制备方法包括如下步骤：
1.化合物5在N-碘代丁二酰亚胺条件下生成相应的碘代物6；
2.化合物6与适当的胺(氨)在适当的溶剂中加热或者微波反应得到中间体(1a)；
在本发明的另一个优选例中，所述化合物(1b)由如下方法制备：
Figure PCTCN2017087222-appb-000021
从化合物7出发环合后做成碘代物得到中间体(1b)。
具体的说，所述制备方法包括如下步骤：
1.化合物7在甲酰胺溶剂中180度环合生成相应的8；
2.化合物8与N-碘代丁二酰亚胺反应得到中间体(1b)；
在制备通式Ⅰ结构的方法中所需的相应硼酸可以通过本领域的常规方法制备，或通过市售途径获得。在本发明的一个优选例中，由如下方法制备：
Figure PCTCN2017087222-appb-000022
Ra是上述R1中的取代基，定义如上述。
从化合物9出发通过亲核取代连接制成化合物10，然后在氯苯在多聚磷酸条件下得到化合物11，11再在常规反应条件中(正丁基锂和三异丙基硼酸酯)制成相应硼酸。
具体的说，所述制备方法包括如下步骤：
1.化合物9通过亲核取代生成化合物10；
2.化合物10在氯苯等适当溶剂中与多聚磷酸回流得到化合物11；
3.化合物11在正丁基锂和三异丙基硼酸酯条件下得到硼酸化合物。
本发明化合物的制备方法具有反应条件温和、原料丰富易得、操作及后处理简单等优点。
药物组合物及其制备方法
本发明还提供了一种药物组合物，所述药物组合物包含治疗有效量的所述的化合物或其立体异构体、几何异构体、互变异构体、其药用盐、其前药及其水合物或溶剂合物中的一种或多种以及任选的药学上可接受的载体。
在另一优选例中，所述药物组合物是用于预防和/或治疗癌症的药物组合物，或用于预防和/或治疗FGFR相关疾病的药物组合物。
在另一优选例中，所述的FGFR选自下组：FGFR1、FGFR2、FGFR3、FGFR4、或组合。
在另一优选例中，所述药物组合物用于预防和/或治疗FGF/FGFR信号通路异常表达相关疾病。
在另一优选例中，所述药物组合物的剂型选自下组：口服剂型、冻干制剂、注射剂。
由于本发明化合物具有优异的对FGFR激酶(Kinase)例如FGFR1和FGFR4的抑制活性，因此本发明化合物及其各种晶型，药学上可接受的无机或有机盐，水合物或溶剂合物，以及含有本发明化合物为主要活性成分的药物组合物可用于治疗、预防以及缓解由与FGFR活性或表达量相关的疾病，例如预防和/或治疗与FGF/FGFR信号通路异常表达相关的疾病。根据现有技术，本发明化合物可用于治疗以下疾病：所述的癌症包括乳腺癌、肺癌、膀胱癌、胃癌、胰腺癌、前列腺癌、结肠癌、多发性骨髓瘤AML、肝癌、黑色素瘤、头颈癌、甲状腺癌、肾细胞癌、胶质母细胞癌和睾丸癌。更特别的是，这些癌症选 自：乳腺癌、非小细胞肺癌、膀胱癌、胃癌、胰腺癌、前列腺癌、结肠癌、多发性骨髓瘤、肝癌、黑色素瘤、头颈癌、甲状腺癌、肾细胞癌、胶质母细胞癌和睾丸癌。最特别的是，该癌症是非小细胞肺癌、胃癌或多发性骨髓瘤。
本发明的药物组合物包含安全有效量范围内的本发明化合物或其药理上可接受的盐及药理上可以接受的赋形剂或载体。其中“安全有效量”指的是：化合物的量足以明显改善病情，而不至于产生严重的副作用。通常，药物组合物含有1-2000mg本发明化合物/剂，更佳地，含有5-200mg本发明化合物/剂。较佳地，所述的“一剂”为一个胶囊或药片。
“药学上可接受的载体”指的是：一种或多种相容性固体或液体填料或凝胶物质，它们适合于人使用，而且必须有足够的纯度和足够低的毒性。“相容性”在此指的是组合物中各组份能和本发明的化合物以及它们之间相互掺和，而不明显降低化合物的药效。药学上可接受的载体部分例子有纤维素及其衍生物(如羧甲基纤维素钠、乙基纤维素钠、纤维素乙酸酯等)、明胶、滑石、固体润滑剂(如硬脂酸、硬脂酸镁)、硫酸钙、植物油(如豆油、芝麻油、花生油、橄榄油等)、多元醇(如丙二醇、甘油、甘露醇、山梨醇等)、乳化剂(如吐温
Figure PCTCN2017087222-appb-000023
)、润湿剂(如十二烷基硫酸钠)、着色剂、调味剂、稳定剂、抗氧化剂、防腐剂、无热原水等。
本发明化合物或药物组合物的施用方式没有特别限制，代表性的施用方式包括(但并不限于)：口服、瘤内、直肠、肠胃外(静脉内、肌肉内或皮下)和局部给药。
用于口服给药的固体剂型包括胶囊剂、片剂、丸剂、散剂和颗粒剂。在这些固体剂型中，活性化合物与至少一种常规惰性赋形剂(或载体)混合，如柠檬酸钠或磷酸二钙，或与下述成分混合：(a)填料或增容剂，例如，淀粉、乳糖、蔗糖、葡萄糖、甘露醇和硅酸；(b)粘合剂，例如，羟甲基纤维素、藻酸盐、明胶、聚乙烯基吡咯烷酮、蔗糖和阿拉伯胶；(c)保湿剂，例如，甘油；(d)崩解剂，例如，琼脂、碳酸钙、马铃薯淀粉或木薯淀粉、藻酸、某些复合硅酸盐和碳酸钠；(e)缓溶剂，例如石蜡；(f)吸收加速剂，例如，季胺化合物；(g)润湿剂，例如鲸蜡醇和单硬脂酸甘油酯；(h)吸附剂，例如，高岭土；和(i)润滑剂，例如，滑石、硬脂酸钙、硬脂酸镁、固体聚乙二醇、十二烷基硫酸钠，或其混合物。胶囊剂、片剂和丸剂中，剂型也可包含缓冲剂。
固体剂型如片剂、糖丸、胶囊剂、丸剂和颗粒剂可采用包衣和壳材制备，如肠衣和其它本领域公知的材料。它们可包含不透明剂，并且，这种组合物中活性化合物或化合物的释放可以延迟的方式在消化道内的某一部分中释放。可采用的包埋组分的实例是聚合物质和蜡类物质。必要时，活性化合物也可与上述赋形剂中的一种或多种形成微胶囊形式。
用于口服给药的液体剂型包括药学上可接受的乳液、溶液、悬浮液、糖浆或酊剂。除了活性化合物外，液体剂型可包含本领域中常规采用的惰性稀释剂，如水或其它溶剂，增溶剂和乳化剂，例知，乙醇、异丙醇、碳酸乙酯、乙酸乙酯、丙二醇、1,3-丁二醇、 二甲基甲酰胺以及油，特别是棉籽油、花生油、玉米胚油、橄榄油、蓖麻油和芝麻油或这些物质的混合物等。
除了这些惰性稀释剂外，组合物也可包含助剂，如润湿剂、乳化剂和悬浮剂、甜味剂、矫味剂和香料。
除了活性化合物外，悬浮液可包含悬浮剂，例如，乙氧基化异十八烷醇、聚氧乙烯山梨醇和脱水山梨醇酯、微晶纤维素、甲醇铝和琼脂或这些物质的混合物等。
用于肠胃外注射的组合物可包含生理上可接受的无菌含水或无水溶液、分散液、悬浮液或乳液，和用于重新溶解成无菌的可注射溶液或分散液的无菌粉末。适宜的含水和非水载体、稀释剂、溶剂或赋形剂包括水、乙醇、多元醇及其适宜的混合物。
用于局部给药的本发明化合物的剂型包括软膏剂、散剂、贴剂、喷射剂和吸入剂。活性成分在无菌条件下与生理上可接受的载体及任何防腐剂、缓冲剂，或必要时可能需要的推进剂一起混合。
本发明化合物可以单独给药，或者与其他治疗手段和/或其它治疗药物联用。
使用药物组合物时，是将安全有效量的本发明化合物适用于需要治疗的哺乳动物(如人)，其中施用时剂量为药学上认为的有效给药剂量，对于60kg体重的人而言，日给药剂量通常为1-2000mg，优选5-500mg。当然，具体剂量还应考虑给药途径、病人健康状况等因素，这些都是熟练医师技能范围之内的。
此外，本发明还提供了一种所述药物组合物的制备方法，包括步骤：将药学上可接受的载体与所述的化合物或其立体异构体、几何异构体、互变异构体、其药用盐、其前药及其水合物或溶剂合物进行混合，从而形成药物组合物。
用途
本发明还提供了一种所述的化合物或所述药物组合物的用途，用于制备预防和/或治疗选自下组的疾病的药物：
a)肿瘤相关疾病；
b)蛋白酪氨酸激酶活性相关疾病。
在另一优选例中，所述肿瘤相关疾病选自下组：乳腺癌、肺癌、膀胱癌、胃癌、胰腺癌、前列腺癌、结肠癌、多发性骨髓瘤AML、肝癌、黑色素瘤、头颈癌、甲状腺癌、肾细胞癌、胶质母细胞癌和睾丸癌。
在另一优选例中，所述肺癌为非小细胞肺癌。
在另一优选例中，所述蛋白酪氨酸激酶活性相关疾病选自下组：FGFR相关疾病。
此外，本发明提供了一种FGFR抑制剂，所述FGFR抑制剂包含抑制有效量的所述的化合物或其立体异构体、几何异构体、互变异构体、其药用盐、其前药及其水合物或溶剂合物中的一种或多种。
在另一优选例中，所述FGFR抑制剂为FGFR1抑制剂和FGFR4抑制剂。
治疗方法
本发明还提供了一种治疗FGFR相关疾病的方法，包括向所需患者施用抑制有效量的所述的化合物或其立体异构体、几何异构体、互变异构体、其药用盐、其前药及其水合物或溶剂合物或所述药物组合物。
本发明还提供了一种非诊断性、非治疗性抑制FGFR活性的方法，所述方法包括向所需患者施用抑制有效量的所述的化合物或其立体异构体、几何异构体、互变异构体、其药用盐、其前药及其水合物或溶剂合物或所述药物组合物。
与现有技术相比，本发明具有以下主要优点：
(1)以所述化合物制备所得抑制剂对FGFR酶活性有显著的抑制作用；
(2)所述化合物可在nM水平上实现对FGFR1-4酶活性的明显抑制作用；
(3)所述化合物在细胞水平上对FGFR1-4诱导的各种癌细胞增殖也具有显著抑制作用。
下面结合具体实施例，进一步阐述本发明。应理解，这些实施例仅用于说明本发明而不用于限制本发明的范围。下列实施例中未注明具体条件的实验方法，通常按照常规条件如Sambrook等人，分子克隆：实验室手册(New York:Cold Spring Harbor Laboratory Press,1989)中所述的条件，或按照制造厂商所建议的条件。除非另外说明，否则百分比和份数按重量计算。
除非另行定义，文中所使用的所有专业与科学用语与本领域熟练人员所熟悉的意义相同。此外，任何与所记载内容相似或均等的方法及材料皆可应用于本发明方法中。文中所述的较佳实施方法与材料仅作示范之用。
实施例1：
1-(3-(4-氨基-5-(7-甲氧基-5-甲基苯并噻吩-2-基)-7H-吡咯[2,3-d]嘧啶-7-基)吡咯烷-1-基)丙-2-烯-1-酮
Figure PCTCN2017087222-appb-000024
步骤1：制备2-甲氧基-4-甲基硝基苯
Figure PCTCN2017087222-appb-000025
将7.5克5-甲基-2-硝基苯酚和10.2克碳酸钾混悬于80毫升丙酮中，再加入8.3克碘甲烷，加热回流5小时后停止加热，硅藻土过滤掉不溶物，滤液旋干得7.7克橘色结晶，直接投下一步。
1H NMR(300MHz,DMSO-d6)δ7.76(d,J＝8.2Hz,1H),6.86(s,1H),6.79(d,J＝8.2Hz,1H),3.92(s,3H),2.39(s,3H).
步骤2：制备2-甲氧基-4-甲基苯胺
Figure PCTCN2017087222-appb-000026
将7.7克2-甲氧基-4-甲基硝基苯和32克二氯亚锡二水化合物溶于250毫升甲醇，加热回流3小时后停止反应，旋掉大部分甲醇反应液后用乙酸乙酯稀释，然后用饱和碳酸氢钠溶液中和，过滤掉不溶物后再用乙酸乙酯萃取，分液后有机相用饱和氯化钠水溶液洗涤后无水硫酸钠干燥，过滤浓缩得5.2克红褐色油状物，直接投下一步。
1H NMR(300MHz,DMSO-d6)δ6.45-6.63(m,3H),4.46(s,2H),3.72(s,3H),2.16(s,3H).
步骤3：制备2-甲氧基-4-甲基苯硫酚
Figure PCTCN2017087222-appb-000027
将5.2克2-甲氧基-4-甲基苯胺溶于32毫升水和11.4毫升浓盐酸，冰浴冷却至0-5℃，再将2.88克亚硝酸钠溶于9毫升水中滴加至上述冷却液中，冰浴下搅拌10分钟后再加入5.6克乙酸钠。将上述反应液滴加至热的(75℃左右)11克乙基黄原酸钾溶于51毫升水的溶液中，继续搅拌1小时后冷却至室温，用乙酸乙酯萃取，分液后有机相用饱和氯化钠水溶液洗涤后无水硫酸钠干燥，过滤浓缩后溶于1.3N氢氧化钾乙醇溶液中，加入3克葡萄糖一起加热回流3小时，浓缩反应液，冰浴冷却下用6N硫酸调PH至1左右，再加入5.7克锌粉50℃加热搅拌30分钟，过滤掉不溶物后乙酸乙酯萃取，分液后有机相用饱和氯化钠水溶液洗涤后无水硫酸钠干燥过滤浓缩，残余物柱层析(100％石油醚)分离得4.15克油状物，收率71％。
1H NMR(300MHz,DMSO-d6)δ7.17(s,1H),6.81(s,1H),6.66(s,1H),4.63(s,1H),3.80(s,3H),2.26(s,3H).
步骤4：制备(2,2-二乙氧基乙基)(2-甲氧基-4-甲基苯基)硫醚
Figure PCTCN2017087222-appb-000028
将4.15克2-甲氧基-4-甲基苯硫酚和5.91克2-溴-1,1-二乙氧基乙烷溶于40毫升N,N-二甲基甲酰胺，加入15.8克碳酸铯后室温搅拌过夜，加水后用乙酸乙酯萃取，有机相用饱和氯化钠水溶液洗涤后无水硫酸钠干燥过滤浓缩，残余物柱层析(100％石油醚)分离得6.6克油状物，收率90％。
1H NMR(300MHz,DMSO-d6)δ7.16(s,1H),6.82(s,1H),6.73(s,1H),4.55(t,J＝4.86Hz,1H),3.80(s,3H),3.52-3.64(m,2H)3.39-3.51(m,2H)2.96(m,2H),2.33(s,3H),1.09(t,j＝7.0Hz,6H).
步骤5：制备7-甲氧基-5-甲基苯并噻吩
Figure PCTCN2017087222-appb-000029
将17克多聚磷酸溶于150毫升氯苯中回流，将6.6克(2,2-二乙氧基乙基)(2-甲氧基-4-甲基苯基)硫醚滴加至反应液中回流过夜，次日倒出溶液，残留物用乙酸乙酯冲洗后合并有机相，浓缩柱层析(100％石油醚)分离得1.85克油状物，收率42％。
1H NMR(300MHz,DMSO-d6)δ7.68(s,1H),7.34(s,1H),7.28(s,1H),6.78(s,1H),3.93(s,3H),2.43(s,3H).
步骤6：制备(7-甲氧基-5-甲基苯并噻吩-2-基)硼酸
Figure PCTCN2017087222-appb-000030
将1.5克7-甲氧基-5-甲基苯并噻吩溶于20毫升干燥的四氢呋喃，冷却至-70℃后滴加1.5当量的正丁基锂溶液，保持-70℃搅拌1小时后，滴加3当量的三异丙基硼酸酯，保持-70℃搅拌1小时后缓慢升至室温，饱和氯化铵水溶液淬灭反应，室温搅拌半小时后用乙酸乙酯萃取，有机相用饱和氯化钠水溶液洗涤后无水硫酸钠干燥，过滤浓缩，残余物柱层析(二氯甲烷：甲醇＝98:2)分离得1.7克米色固体，收率94％。
步骤7：制备4-氯-5-碘-7H-吡咯[2,3-d]嘧啶
Figure PCTCN2017087222-appb-000031
将1.53克4-氯-7H-吡咯[2,3-d]嘧啶和2.7克N-碘代丁二酰亚胺溶于35毫升二氯甲烷中，室温搅拌1小时后用饱和硫代硫酸钠溶液洗，加入乙酸乙酯萃取，有机相用饱和氯化钠水溶液洗涤后无水硫酸钠干燥，过滤浓缩干得2.5克灰白色固体，直接投下一步。
1H NMR(300MHz,DMSO-d6)δ12.95(brs,1H),8.60(s,1H),7.91(d,J＝2.5Hz,1H).
步骤8：制备3-(4-氯-5-碘-7H-吡咯[2,3-d]嘧啶-7-基)吡咯烷-1-甲酸叔丁酯
Figure PCTCN2017087222-appb-000032
氩气氛围中将500毫克4-氯-5-碘-7H-吡咯[2,3-d]嘧啶，505毫克3-羟基吡咯烷-1-甲酸叔丁酯和945毫克三苯基膦溶于干燥的四氢呋喃溶液中，冰浴冷却下滴加0.7毫升偶氮二甲酸二异丙酯，滴加完后移至室温下搅拌2小时，反应液浓缩至干，残余物柱层析(乙酸乙酯：石油醚＝10:90)分离得1.05克淡黄色固体。
步骤9：制备3-(4-氨基-5-碘-7H-吡咯[2,3-d]嘧啶-7-基)吡咯烷-1-甲酸叔丁酯
Figure PCTCN2017087222-appb-000033
将120毫克3-(4-氯-5-碘-7H-吡咯[2,3-d]嘧啶-7-基)吡咯烷-1-甲酸叔丁酯溶于2毫升1,4-二氧六环和1毫升氨水的混合溶液中，封管100℃加热反应16小时，冷却至室温后加水，用乙酸乙酯萃取，有机相用饱和氯化钠水溶液洗涤后无水硫酸钠干燥，过滤浓缩，残余物柱层析(二氯甲烷：甲醇＝98:2)分离得113毫克米色固体，收率80％。
步骤10：制备3-(4-氨基-5-(7-甲氧基-5-甲基苯并噻吩-2-基)-7H-吡咯[2,3-d]嘧啶-7-基)吡咯烷-1-甲酸叔丁酯
Figure PCTCN2017087222-appb-000034
将178毫克的3-(4-氨基-5-碘-7H-吡咯[2,3-d]嘧啶-7-基)吡咯烷-1-甲酸叔丁酯，95毫克的(7-甲氧基-5-甲基苯并噻吩-2-基)硼酸，90毫克的碳酸钠和25毫克的四三苯基膦钯溶于4毫升的1,4-二氧六环和1毫升水的混合溶液中，置换氩气后80℃加热反应5小时，反应液浓缩至干，残余物柱层析(二氯甲烷：甲醇＝98:2)分离定量得200毫克淡黄色固体。
1H NMR(400MHz,CDCl3)δ8.34(s,1H),7.24(s,1H),7.23(s,1H),7.16(s, 1H),6.65(s,1H),5.68(s,2H),5.52–5.43(m,1H),4.01(s,3H),3.99–3.91(m,1H),3.81–3.53(m,3H),2.50(s,3H),2.48–2.38(m,1H),2.35–2.27(m,1H),1.49(s,9H).
步骤11：制备5-(7-甲氧基-5-甲基苯并噻吩-2-基)-7-(吡咯烷-3-基)-7H-吡咯[2,3-d]嘧啶-4-氨基盐酸盐
Figure PCTCN2017087222-appb-000035
将95毫克的3-(4-氨基-5-(7-甲氧基-5-甲基苯并噻吩-2-基)-7H-吡咯[2,3-d]嘧啶-7-基)吡咯烷-1-甲酸叔丁酯溶于4N的盐酸1,4-二氧六环溶液中，室温搅拌2小时，反应液浓缩至干，直接投下一反应。
步骤12：制备1-(3-(4-氨基-5-(7-甲氧基-5-甲基苯并噻吩-2-基)-7H-吡咯[2,3-d]嘧啶-7-基)吡咯烷-1-基)丙-2-烯-1-酮
将步骤11制得的5-(7-甲氧基-5-甲基苯并噻吩-2-基)-7-(吡咯烷-3-基)-7H-吡咯[2,3-d]嘧啶-4-氨基盐酸盐混悬于2毫升干燥的二氯甲烷中，加入0.15毫升的三乙胺，冰浴冷却下滴加17微升的丙烯酰氯，滴加完后移至室温搅拌过夜，次日反应液浓缩至干，残余物柱层析(二氯甲烷：甲醇＝97:3)分离得59毫克白色固体，收率70％。
1H NMR(400MHz,CDCl3)δ8.33(s,1H),7.23–7.19(m,2H),7.11(d,J＝14.3Hz,1H),6.64(s,1H),6.51–6.39(m,2H),5.79–5.69(m,1H),5.50(m,3H),4.22–4.09(m,1H),4.04–3.90(m,1H),3.99(s,3H),3.86–3.75(m,2H),2.68–2.53(m,1H),2.48(s,3H),2.43(m,1H).
实施例2：
1-(3-(4-氨基-3-(7-甲氧基-5-甲基苯并噻吩-2-基)-1H-吡唑[3,4-d]嘧啶-1-基)吡咯烷-1-基)丙-2-烯-1-酮
Figure PCTCN2017087222-appb-000036
步骤1：制备1H-吡唑[3,4-d]嘧啶-4-胺
Figure PCTCN2017087222-appb-000037
将1克3-氨基-4-氰基吡唑溶于甲酰胺中，氩气保护下加热至170℃反应5小时，冷却至室温后倒入30毫升水中打浆，过滤后滤饼干燥得1.15克棕色固体。
1H NMR(300MHz,DMSO-d6)δ13.34(s,1H),8.13(s,1H),8.07(s,1H),7.61(s,2H).
步骤2：制备3-碘-1H-吡唑[3,4-d]嘧啶-4-胺
Figure PCTCN2017087222-appb-000038
将500毫克1H-吡唑[3,4-d]嘧啶-4-胺和1.25克N-碘代丁二酰亚胺溶于5毫升N,N-二甲基甲酰胺，氩气保护下加热至80℃反应18小时，冷却至室温后倒入20毫升饱和硫代硫酸钠水溶液中打浆，过滤，滤饼干燥得1.2克棕色固体。
1H NMR(300MHz,DMSO-d6)δ13.81(s,1H),8.13(s,1H),8.17(s,1H).
步骤3：制备3-((甲磺酰基)氧)吡咯烷-1-甲酸叔丁酯
Figure PCTCN2017087222-appb-000039
将136毫克3-羟基吡咯烷-1-甲酸叔丁酯溶于1毫升二氯甲烷中，加入0.15毫升的三乙胺，冰浴冷却下滴加67微升的甲磺酰氯，加料完毕后移至室温下搅拌2小时停止反应。加水后用乙酸乙酯萃取，有机相用饱和氯化钠水溶液洗涤后无水硫酸钠干燥，过滤浓缩后定量得190毫克油状物。
1H NMR(300MHz,CDCl3)δ5.25-5.28(m,1H),3.49-3.68(m,4H),3.05(s,3H),2.12-2.28(m,2H),1.47(s,9H).
步骤4：制备3-(4-氨基-3-碘-1H-吡唑[3,4-d]嘧啶-1-基)吡咯烷-1-甲酸叔丁酯
Figure PCTCN2017087222-appb-000040
将110毫克3-碘-1H-吡唑[3,4-d]嘧啶-4-胺和110毫克3-((甲磺酰基)氧)吡咯烷-1-甲酸叔丁酯溶于2毫升N,N-二甲基甲酰胺，再加入270毫克碳酸铯，氩气保护下加热至90℃过夜，次日加水后用乙酸乙酯萃取，有机相用饱和氯化钠水溶液洗涤后无水硫酸钠干燥， 过滤浓缩，残余物柱层析(二氯甲烷：甲醇＝97:3)分离得100毫克黄色固体，收率57％。
1H NMR(300MHz,CDCl3)δ8.32(s,1H),6.05(s,2H),5.40-5.39(m,1H),3.90-3.68(m,3H),3.58-3.47(m,1H),2.58-2.26(m,2H),1.48(s,9H).
步骤5：制备3-(4-氨基-3-(7-甲氧基-5-甲基苯并噻吩-2-基)-1H-吡唑[3,4-d]嘧啶-1-基)吡咯烷-1-甲酸叔丁酯
Figure PCTCN2017087222-appb-000041
将3-(4-氨基-5-碘-7H-吡咯[2,3-d]嘧啶-7-基)吡咯烷-1-甲酸叔丁酯替换成3-(4-氨基-3-碘-1H-吡唑[3,4-d]嘧啶-1-基)吡咯烷-1-甲酸叔丁酯，其余所需原料、试剂及制备方法同实施例1中的步骤10，得3-(4-氨基-3-(7-甲氧基-5-甲基苯并噻吩-2-基)-1H-吡唑[3,4-d]嘧啶-1-基)吡咯烷-1-甲酸叔丁酯。
步骤6：制备3-(7-甲氧基-5-甲基苯并噻吩-2-基)-1-(吡咯烷-3-基)-1H-吡唑[3,4-d]嘧啶-4-胺盐酸盐
Figure PCTCN2017087222-appb-000042
将3-(4-氨基-5-(7-甲氧基-5-甲基苯并噻吩-2-基)-7H-吡咯[2,3-d]嘧啶-7-基)吡咯烷-1-甲酸叔丁酯替换成3-(4-氨基-3-(7-甲氧基-5-甲基苯并噻吩-2-基)-1H-吡唑[3,4-d]嘧啶-1-基)吡咯烷-1-甲酸叔丁酯，其余所需原料、试剂及制备方法同实施例1中的步骤11，得3-(7-甲氧基-5-甲基苯并噻吩-2-基)-1-(吡咯烷-3-基)-1H-吡唑[3,4-d]嘧啶-4-胺盐酸盐。
步骤7：制备1-(3-(4-氨基-3-(7-甲氧基-5-甲基苯并噻吩-2-基)-1H-吡唑[3,4-d]嘧啶-1-基)吡咯烷-1-基)丙基-2-烯-1-酮
将5-(7-甲氧基-5-甲基苯并噻吩-2-基)-7-(吡咯烷-3-基)-7H-吡咯[2,3-d]嘧啶-4-氨基盐酸盐替换成3-(7-甲氧基-5-甲基苯并噻吩-2-基)-1-(吡咯烷-3-基)-1H-吡唑[3,4-d]嘧啶-4-胺盐酸盐，其余所需原料、试剂及制备方法同实施例1中的步骤12，得 1-(3-(4-氨基-3-(7-甲氧基-5-甲基苯并噻吩-2-基)-1H-吡唑[3,4-d]嘧啶-1-基)吡咯烷-1-基)丙-2-烯-1-酮，收率53％。
1H NMR(300MHz,DMSO-d6)δ8.29(s,1H),7.67(s,1H),7.34(s,1H),6.85(s,1H),6.74–6.51(m,1H),6.17(d,J＝16.4Hz,1H),5.68(dd,J＝16.9,11.0Hz,1H),5.51(d,J＝21.5Hz,1H),4.18–4.05(m,1H),3.95(s,3H),3.84(m,3H),2.44(s,3H),2.39(m,2H).
实施例3：
1-(3-(4-氨基-5-(7-甲氧基-5-甲基苯并噻吩-2-基)-7H-吡咯[2,3-d]嘧啶-7-基)吡咯烷-1-基)丙-2-炔-1-酮
Figure PCTCN2017087222-appb-000043
将100毫克5-(7-甲氧基-5-甲基苯并噻吩-2-基)-7-(吡咯烷-3-基)-7H-吡咯[2,3-d]嘧啶-4-氨基盐酸，20微升丙炔酸和145毫克2-(7-偶氮苯并三氮唑)-N,N,N',N'-四甲基脲六氟磷酸酯溶于2毫升N,N-二甲基甲酰胺，滴加0.2毫升的N,N-二异丙基乙胺，室温下搅拌8小时停止反应，加水后用乙酸乙酯萃取，有机相用饱和氯化钠水溶液洗涤后无水硫酸钠干燥，过滤浓缩，残余物柱层析(二氯甲烷：甲醇＝97:3)分离得48毫克黄色固体，收率44％。
1H NMR(300MHz,CDCl3)δ8.33(s,1H),7.22(s,2H),7.11(d,J＝9.3Hz,1H),6.65(s,1H),5.69(s,2H),5.59–5.40(m,1H),4.37–3.57(m,4H),4.00(s,3H),3.09(d,J＝15.2Hz,1H),2.64–2.24(m,2H),2.49(s,3H).
实施例4：
(E)1-(3-(4-氨基-5-(7-甲氧基-5-甲基苯并噻吩-2-基)-7H-吡咯[2,3-d]嘧啶-7-基)吡咯烷-1-基)-4-(二甲氨基)丁-2-烯-1-酮
Figure PCTCN2017087222-appb-000044
将丙炔酸替换成反式-4-二甲基胺基巴豆酸盐酸盐，其余所需原料、试剂及制备方法同实施例3，得(E)1-(3-(4-氨基-5-(7-甲氧基-5-甲基苯并噻吩-2-基)-7H-吡咯[2,3-d]嘧啶-7-基)吡咯烷-1-基)-4-(二甲氨基)丁-2-烯-1-酮，收率43％。
1H NMR(300MHz,CDCl3)δ8.32(s,1H),7.22(s,2H),7.11(d,J＝11.4Hz,1H),7.03–6.86(m,1H),6.78(d,J＝15.9Hz,0.5H),6.61(d,J＝17.7Hz,1.5H),5.57(s,2H),5.53–5.39(m,1H),4.33–3.81(m,4H),3.99(s,3H),3.52(m,2H),2.63(s,3H),2.57(s,3H),2.53–2.44(m,5H).
实施例5：
(E)1-(3-(4-氨基-5-(7-甲氧基-5-甲基苯并噻吩-2-基)-7H-吡咯[2,3-d]嘧啶-7-基)吡咯烷-1-基)-4-(乙基(甲基)氨基)丁-2-烯-1-酮
Figure PCTCN2017087222-appb-000045
将丙炔酸替换成反式-4-(乙基(甲基)胺基)巴豆酸盐酸盐，其余所需原料、试剂及制备方法同实施例3，得(E)1-(3-(4-氨基-5-(7-甲氧基-5-甲基苯并噻吩-2-基)-7H-吡咯[2,3-d]嘧啶-7-基)吡咯烷-1-基)-4-(乙基(甲基)氨基)丁-2-烯-1-酮，收率45％。
1H NMR(300MHz,CDCl3)δ8.32(s,1H),7.22(s,2H),7.11(d,J＝12.3Hz,1H),6.99–6.71(m,2H),6.64(s,1H),5.61(s,2H),5.56–5.38(m,1H),4.40–3.88(m,4H),3.99(s,3H),3.81–3.62(m,2H),2.75–2.59(m,3H),2.48(s,3H),2.62–2.34(m,2H),1.52–1.33(m,3H).
实施例6：
(E)1-(3-(4-氨基-5-(7-甲氧基-5-甲基苯并噻吩-2-基)-7H-吡咯[2,3-d]嘧啶-7-基)吡咯烷-1-基)-4-(异丙基基(甲基)氨基)丁-2-烯-1-酮
Figure PCTCN2017087222-appb-000046
将丙炔酸替换成反式-4-(异丙基(甲基)胺基)巴豆酸盐酸盐，其余所需原料、试剂及制备方法同实施例3，得(E)1-(3-(4-氨基-5-(7-甲氧基-5-甲基苯并噻吩-2-基)-7H-吡咯[2,3-d]嘧啶-7-基)吡咯烷-1-基)-4-(异丙基(甲基)氨基)丁-2-烯-1-酮，收率42％。
1H NMR(300MHz,CDCl3)δ8.32(s,1H),7.21(s,2H),7.11(d,J＝10.8Hz,1H),6.98–6.85(m,1.5H),6.78–6.67(m,0.5H),6.64(s,1H),5.54(s,2H),5.50–5.38(m,1H),4.33–3.84(m,4H),3.99(s,3H),3.71–3.52(m,2H),3.45–3.22(m,1H),2.54(d,J＝16.3Hz,3H),2.48(s,3H),1.36–1.21(m,6H).
实施例7：
(E)1-(3-(4-氨基-5-(7-甲氧基-5-甲基苯并噻吩-2-基)-7H-吡咯[2,3-d]嘧啶-7-基)吡咯烷-1-基)-4-(吡咯烷-1-基)丁-2-烯-1-酮
Figure PCTCN2017087222-appb-000047
将丙炔酸替换成反式-4-(吡咯烷-1-基)巴豆酸盐酸盐，其余所需原料、试剂及制备方法同实施例3，得(E)1-(3-(4-氨基-5-(7-甲氧基-5-甲基苯并噻吩-2-基)-7H-吡咯[2,3-d]嘧啶-7-基)吡咯烷-1-基)-4-(吡咯烷-1-基)丁-2-烯-1-酮，收率43％。
1H NMR(300MHz,CDCl3)δ8.32(s,1H),7.22(s,2H),7.11(d,J＝11.1Hz,1H),6.92(m,1H),6.79(d,J＝17.6Hz,0.5H),6.61(d,J＝12.3Hz,1.5H),5.54–5.40(m,3H),4.32–3.81(m,4H),3.99(s,3H),3.61(dd,J＝19.1,6.0Hz,2H),3.11–2.85(m,4H),2.53–2.40(m,5H),2.11–1.77(m,4H).
实施例8：
(E)1-(3-(4-氨基-5-(7-甲氧基-5-甲基苯并噻吩-2-基)-7H-吡咯[2,3-d]嘧啶-7-基)吡咯烷-1-基)-4-(哌啶-1-基)丁-2-烯-1-酮
Figure PCTCN2017087222-appb-000048
将丙炔酸替换成反式-4-(哌啶-1-基)巴豆酸盐酸盐，其余所需原料、试剂及制备方法同实施例3，得(E)1-(3-(4-氨基-5-(7-甲氧基-5-甲基苯并噻吩-2-基)-7H-吡咯[2,3-d]嘧啶-7-基)吡咯烷-1-基)-4-(哌啶-1-基)丁-2-烯-1-酮，收率44％。
1H NMR(300MHz,DMSO-d6)δ8.19(s,1H),7.48(s,1H),7.31(d,J＝6.6Hz,1H),7.27(s,1H),6.80(s,1H),6.48(s,2H),5.46-5.35(m,2H),5.29-5.26(m,1H),4.05–3.62(m,4H),3.94(s,3H),3.40–3.20(m,6H),2.43(s,3H),2.42(s,2H),1.42-1.18(m,6H).
实施例9：
5-(7-甲氧基-5-甲基苯并噻吩-2-基)-7-(吡咯烷-3-基)-7H-吡咯[2,3-d]嘧啶-4-胺
Figure PCTCN2017087222-appb-000049
将95毫克的3-(4-氨基-5-(7-甲氧基-5-甲基苯并噻吩-2-基)-7H-吡咯[2,3-d]嘧啶-7-基)吡咯烷-1-甲酸叔丁酯溶于4N的盐酸1,4-二氧六环溶液中，室温搅拌2小时，反应液浓缩至干，用饱和碳酸氢钠水溶液洗，加入乙酸乙酯萃取，有机相用饱和氯化钠水溶液洗涤后无水硫酸钠干燥，过滤浓缩，残余物柱层析(二氯甲烷：甲醇(含1％氨水)＝95:5)分离得48毫克黄色固体，收率44％。
1H NMR(300MHz,DMSO-d6)δ8.20(s,1H),7.59(d,J＝17.3Hz,1H),7.32(s,1H),7.27(s,1H),6.79(s,1H),6.49(s,2H),5.46–5.26(m,1H),4.19–4.06(m,0.5H),3.94(s,3H),3.85-4.0(m,1H),3.80–3.68(m,1.5H),3.58-3.45(m,1H),2.43(s,3H),2.43–2.34(m,2H).
实施例10：
3-(4-氨基-5-(7-甲氧基-5-甲基苯并噻吩-2-基)-7H-吡咯[2,3-d]嘧啶-7-基)吡咯烷-1-甲酸叔丁酯
Figure PCTCN2017087222-appb-000050
制备同实施例1步骤10。
1H NMR(300MHz,DMSO-d6)δ8.20(s,1H),7.59(d,J＝17.3Hz,1H),7.32(s,1H),7.27(s,1H),6.79(s,1H),6.49(s,2H),5.46–5.26(m,1H),4.19–4.06(m,0.5H),3.94(s,3H),3.85-4.0(m,1H),3.80–3.68(m,1.5H),3.58-3.45(m,1H),2.43(s,3H),2.43–2.34(m,2H),1.48(s,9H).
实施例11：
(R)-1-(3-(4-氨基-5-(7-甲氧基-5-甲基苯并噻吩-2-基)-7H-吡咯[2,3-d]嘧啶-7-基)吡咯烷-1-基)丙-2-烯-1-酮
Figure PCTCN2017087222-appb-000051
步骤1：制备(R)-3-(4-氯-5-碘-7H-吡咯[2,3-d]嘧啶-7-基)吡咯烷-1-甲酸叔丁酯
Figure PCTCN2017087222-appb-000052
将3-羟基吡咯烷-1-甲酸叔丁酯替换成(S)-3-羟基吡咯烷-1-甲酸叔丁酯，其余所需原料、试剂及制备方法同实施例1中的步骤8，得(R)-3-(4-氯-5-碘-7H-吡咯[2,3-d]嘧啶-7-基)吡咯烷-1-甲酸叔丁酯
步骤2：制备(R)-3-(4-氨基-5-碘-7H-吡咯[2,3-d]嘧啶-7-基)吡咯烷-1-甲酸叔丁酯
Figure PCTCN2017087222-appb-000053
将3-(4-氯-5-碘-7H-吡咯[2,3-d]嘧啶-7-基)吡咯烷-1-甲酸叔丁酯替换成(R)-3-(4-氯-5-碘-7H-吡咯[2,3-d]嘧啶-7-基)吡咯烷-1-甲酸叔丁酯，其余所需原料、试剂及制备方法同实施例1中的步骤9，得(R)-3-(4-氨基-5-碘-7H-吡咯[2,3-d]嘧啶-7-基)吡咯烷-1-甲酸叔丁酯。
步骤3：制备(R)3-(4-氨基-5-(7-甲氧基-5-甲基苯并噻吩-2-基)-7H-吡咯[2,3-d]嘧啶-7-基)吡咯烷-1-甲酸叔丁酯
Figure PCTCN2017087222-appb-000054
将3-(4-氨基-5-碘-7H-吡咯[2,3-d]嘧啶-7-基)吡咯烷-1-甲酸叔丁酯替换成(R)-3-(4-氨基-5-碘-7H-吡咯[2,3-d]嘧啶-7-基)吡咯烷-1-甲酸叔丁酯，其余所需原料、试剂及制备方法同实施例1中的步骤10，得(R)3-(4-氨基-5-(7-甲氧基-5-甲基苯并噻吩-2-基)-7H-吡咯[2,3-d]嘧啶-7-基)吡咯烷-1-甲酸叔丁酯。
步骤4：制备(R)-5-(7-甲氧基-5-甲基苯并噻吩-2-基)-7-(吡咯烷-3-基)-7H-吡咯[2,3-d]嘧啶-4-氨基盐酸盐
Figure PCTCN2017087222-appb-000055
将3-(4-氨基-5-(7-甲氧基-5-甲基苯并噻吩-2-基)-7H-吡咯[2,3-d]嘧啶-7-基)吡咯烷-1-甲酸叔丁酯替换成(S)-3-(4-氨基-5-(7-甲氧基-5-甲基苯并噻吩-2-基)-7H-吡咯[2,3-d]嘧啶-7-基)吡咯烷-1-甲酸叔丁酯，其余所需原料、试剂及制备方法同实施例1中的步骤11，得(R)-5-(7-甲氧基-5-甲基苯并噻吩-2-基)-7-(吡咯烷-3-基)-7H-吡咯[2,3-d]嘧啶-4-氨基盐酸盐。
步骤5：制备(R)-1-(3-(4-氨基-5-(7-甲氧基-5-甲基苯并噻吩-2-基)-7H-吡咯[2,3-d]嘧啶-7-基)吡咯烷-1-基)丙-2-烯-1-酮
将5-(7-甲氧基-5-甲基苯并噻吩-2-基)-7-(吡咯烷-3-基)-7H-吡咯[2,3-d]嘧啶-4-氨基盐酸盐替换成(R)-5-(7-甲氧基-5-甲基苯并噻吩-2-基)-7-(吡咯烷-3-基)-7H-吡咯[2,3-d]嘧啶-4-氨基盐酸盐，其余所需原料、试剂及制备方法同实施例1中的步骤12，得(R)-1-(3-(4-氨基-5-(7-甲氧基-5-甲基苯并噻吩-2-基)-7H-吡咯[2,3-d]嘧啶-7-基)吡咯烷-1-基)丙-2-烯-1-酮，收率51％。
1H NMR(400MHz,CDCl3)δ8.33(s,1H),7.23–7.19(m,2H),7.11(d,J＝14.3Hz,1H),6.64(s,1H),6.51–6.39(m,2H),5.79–5.69(m,1H),5.50(m,3H),4.22–4.09(m,1H),4.04–3.90(m,1H),3.99(s,3H),3.86–3.75(m,2H),2.68–2.53(m,1H),2.48(s,3H),2.43(m,1H).
实施例12：
(S)-1-(3-(4-氨基-5-(7-甲氧基-5-甲基苯并噻吩-2-基)-7H-吡咯[2,3-d]嘧啶-7-基)吡咯烷-1-基)丙-2-烯-1-酮
Figure PCTCN2017087222-appb-000056
步骤1：制备(S)-3-(4-氯-5-碘-7H-吡咯[2,3-d]嘧啶-7-基)吡咯烷-1-甲酸叔丁酯
Figure PCTCN2017087222-appb-000057
将3-羟基吡咯烷-1-甲酸叔丁酯替换成(R)-3-羟基吡咯烷-1-甲酸叔丁酯，其余所需原料、试剂及制备方法同实施例1中的步骤8，得(S)-3-(4-氯-5-碘-7H-吡咯[2,3-d]嘧啶-7-基)吡咯烷-1-甲酸叔丁酯
步骤2：制备(S)-3-(4-氨基-5-碘-7H-吡咯[2,3-d]嘧啶-7-基)吡咯烷-1-甲酸叔丁酯
Figure PCTCN2017087222-appb-000058
将3-(4-氯-5-碘-7H-吡咯[2,3-d]嘧啶-7-基)吡咯烷-1-甲酸叔丁酯替换成(S)-3-(4-氯-5-碘-7H-吡咯[2,3-d]嘧啶-7-基)吡咯烷-1-甲酸叔丁酯，其余所需原料、试剂及制备方法同实施例1中的步骤9，得(S)-3-(4-氨基-5-碘-7H-吡咯[2,3-d]嘧啶-7-基)吡咯烷-1-甲酸叔丁酯。
步骤3：制备(S)3-(4-氨基-5-(7-甲氧基-5-甲基苯并噻吩-2-基)-7H-吡咯[2,3-d]嘧啶-7-基)吡咯烷-1-甲酸叔丁酯
Figure PCTCN2017087222-appb-000059
将3-(4-氨基-5-碘-7H-吡咯[2,3-d]嘧啶-7-基)吡咯烷-1-甲酸叔丁酯替换成(S)-3-(4-氨基-5-碘-7H-吡咯[2,3-d]嘧啶-7-基)吡咯烷-1-甲酸叔丁酯，其余所需原料、试剂及制备方法同实施例1中的步骤10，得(S)3-(4-氨基-5-(7-甲氧基-5-甲基苯并噻吩-2-基)-7H-吡咯[2,3-d]嘧啶-7-基)吡咯烷-1-甲酸叔丁酯。
步骤4：制备(S)-5-(7-甲氧基-5-甲基苯并噻吩-2-基)-7-(吡咯烷-3-基)-7H-吡咯[2,3-d]嘧啶-4-氨基盐酸盐
Figure PCTCN2017087222-appb-000060
将3-(4-氨基-5-(7-甲氧基-5-甲基苯并噻吩-2-基)-7H-吡咯[2,3-d]嘧啶-7-基)吡咯烷-1-甲酸叔丁酯替换成(S)-3-(4-氨基-5-(7-甲氧基-5-甲基苯并噻吩-2-基)-7H-吡咯[2,3-d]嘧啶-7-基)吡咯烷-1-甲酸叔丁酯，其余所需原料、试剂及制备方法同实施例1中的步骤11，得(S)-5-(7-甲氧基-5-甲基苯并噻吩-2-基)-7-(吡咯烷-3-基)-7H-吡咯[2,3-d]嘧啶-4-氨基盐酸盐。
步骤5：制备(S)-1-(3-(4-氨基-5-(7-甲氧基-5-甲基苯并噻吩-2-基)-7H-吡咯[2,3-d]嘧啶-7-基)吡咯烷-1-基)丙-2-烯-1-酮
将5-(7-甲氧基-5-甲基苯并噻吩-2-基)-7-(吡咯烷-3-基)-7H-吡咯[2,3-d]嘧啶-4-氨基盐酸盐替换成(S)-5-(7-甲氧基-5-甲基苯并噻吩-2-基)-7-(吡咯烷-3-基)-7H-吡咯[2,3-d]嘧啶-4-氨基盐酸盐，其余所需原料、试剂及制备方法同实施例1中的步骤12，得(S)-1-(3-(4-氨基-5-(7-甲氧基-5-甲基苯并噻吩-2-基)-7H-吡咯[2,3-d]嘧啶-7-基)吡咯烷-1-基)丙-2-烯-1-酮,收率53％。
1H NMR(400MHz,CDCl3)δ8.33(s,1H),7.23–7.19(m,2H),7.11(d,J＝14.3Hz,1H),6.64(s,1H),6.51–6.39(m,2H),5.79–5.69(m,1H),5.50(m,3H),4.22–4.09(m,1H),4.04–3.90(m,1H),3.99(s,3H),3.86–3.75(m,2H),2.68–2.53(m,1H),2.48(s,3H),2.43(m,1H).
实施例13：
1-(3-(4-氨基-5-(7-甲氧基-5-甲基苯并噻吩-2-基)-7H-吡咯[2,3-d]嘧啶-7-基)氮杂环丁烷-1-基)丙-2-烯-1-酮
Figure PCTCN2017087222-appb-000061
步骤1：制备3-(4-氯-5-碘-7H-吡咯[2,3-d]嘧啶-7-基)氮杂环丁烷-1-甲酸叔丁酯
Figure PCTCN2017087222-appb-000062
将3-羟基吡咯烷-1-甲酸叔丁酯替换成3-羟基氮杂环丁烷-1-甲酸叔丁酯，其余所需原料、试剂及制备方法同实施例1中的步骤8，得3-(4-氯-5-碘-7H-吡咯[2,3-d]嘧啶-7-基)氮杂环丁烷-1-甲酸叔丁酯。
步骤2：制备3-(4-氨基-5-碘-7H-吡咯[2,3-d]嘧啶-7-基)氮杂环丁烷-1-甲酸叔丁酯
Figure PCTCN2017087222-appb-000063
将3-(4-氯-5-碘-7H-吡咯[2,3-d]嘧啶-7-基)吡咯烷-1-甲酸叔丁酯替换成3-(4-氯-5-碘-7H-吡咯[2,3-d]嘧啶-7-基)氮杂环丁烷-1-甲酸叔丁酯，其余所需原料、试剂及制备方法同实施例1中的步骤9，得3-(4-氨基-5-碘-7H-吡咯[2,3-d]嘧啶-7-基)氮杂环丁烷-1-甲酸叔丁酯。
1H NMR(300MHz,CDCl3)δ8.22(d,J＝1.7Hz,1H),7.35(d,J＝1.3Hz,1H),6.02–5.86(m,2H),5.50(d,J＝4.0Hz,1H),4.45(m,2H),4.15(m,2H),1.47(s,9H).
步骤3：制备3-(4-氨基-5-(7-甲氧基-5-甲基苯并噻吩-2-基)-7H-吡咯[2,3-d]嘧啶-7-基)氮杂环丁烷-1-甲酸叔丁酯
Figure PCTCN2017087222-appb-000064
将3-(4-氨基-5-碘-7H-吡咯[2,3-d]嘧啶-7-基)吡咯烷-1-甲酸叔丁酯替换成3-(4-氨基-5-碘-7H-吡咯[2,3-d]嘧啶-7-基)氮杂环丁烷-1-甲酸叔丁酯，其余所需原料、试剂及制备方法同实施例1中的步骤10，得3-(4-氨基-5-(7-甲氧基-5-甲基苯并噻吩-2-基)-7H-吡咯[2,3-d]嘧啶-7-基)氮杂环丁烷-1-甲酸叔丁酯。
步骤4：制备5-(7-甲氧基-5-甲基苯并噻吩-2-基)-7-(氮杂环丁烷-3-基)-7H-吡咯[2,3-d]嘧啶-4-胺盐酸盐
Figure PCTCN2017087222-appb-000065
将3-(4-氨基-5-(7-甲氧基-5-甲基苯并噻吩-2-基)-7H-吡咯[2,3-d]嘧啶-7-基)吡 咯烷-1-甲酸叔丁酯替换成3-(4-氨基-5-(7-甲氧基-5-甲基苯并噻吩-2-基)-7H-吡咯[2,3-d]嘧啶-7-基)氮杂环丁烷-1-甲酸叔丁酯，其余所需原料、试剂及制备方法同实施例1中的步骤11，得5-(7-甲氧基-5-甲基苯并噻吩-2-基)-7-(氮杂环丁烷-3-基)-7H-吡咯[2,3-d]嘧啶-4-胺盐酸盐。
步骤5：制备1-(3-(4-氨基-5-(7-甲氧基-5-甲基苯并噻吩-2-基)-7H-吡咯[2,3-d]嘧啶-7-基)氮杂环丁烷-1-基)丙-2-烯-1-酮
将5-(7-甲氧基-5-甲基苯并噻吩-2-基)-7-(吡咯烷-3-基)-7H-吡咯[2,3-d]嘧啶-4-氨基盐酸盐替换成5-(7-甲氧基-5-甲基苯并噻吩-2-基)-7-(氮杂环丁烷-3-基)-7H-吡咯[2,3-d]嘧啶-4-氨基盐酸盐，其余所需原料、试剂及制备方法同实施例1中的步骤12，得1-(3-(4-氨基-5-(7-甲氧基-5-甲基苯并噻吩-2-基)-7H-吡咯[2,3-d]嘧啶-7-基)氮杂环丁烷-1-基)丙-2-烯-1-酮，收率61％。
1H NMR(300MHz,CDCl3)δ8.30(s,1H),7.35(s,1H),7.23(d,J＝2.8Hz,2H),6.64(s,1H),6.47–6.37(m,1H),6.24(dd,J＝17.0,10.1Hz,1H),5.71(dd,J＝25.2,12.1Hz,4H),4.78(s,1H),4.66(s,1H),4.54(d,J＝14.2Hz,2H),3.99(s,3H),2.48(s,3H)..
实施例14：
1-(4-(4-氨基-5-(7-甲氧基-5-甲基苯并噻吩-2-基)-7H-吡咯[2,3-d]嘧啶-7-基)哌啶-1-基)丙-2-烯-1-酮
Figure PCTCN2017087222-appb-000066
步骤1：制备4-(4-氯-5-碘-7H-吡咯[2,3-d]嘧啶-7-基)哌啶-1-甲酸叔丁酯
Figure PCTCN2017087222-appb-000067
将3-羟基吡咯烷-1-甲酸叔丁酯替换成4-羟基哌啶-1-甲酸叔丁酯，其余所需原料、试剂及制备方法同实施例1中的步骤8，得4-(4-氯-5-碘-7H-吡咯[2,3-d]嘧啶-7-基)哌啶-1-甲酸叔丁酯。
步骤2：制备4-(4-氨基-5-碘-7H-吡咯[2,3-d]嘧啶-7-基)哌啶-1-甲酸叔丁酯
Figure PCTCN2017087222-appb-000068
将3-(4-氯-5-碘-7H-吡咯[2,3-d]嘧啶-7-基)吡咯烷-1-甲酸叔丁酯替换成4-(4-氯-5-碘-7H-吡咯[2,3-d]嘧啶-7-基)哌啶-1-甲酸叔丁酯，其余所需原料、试剂及制备方法同实施例1中的步骤9，得4-(4-氨基-5-碘-7H-吡咯[2,3-d]嘧啶-7-基)哌啶-1-甲酸叔丁酯。
1H NMR(300MHz,CDCl3)δ8.26(s,1H),7.09(s,1H),5.64(s,2H),4.75-4.85(m,1H),4.25-4.35(m,2H),2.85-2.95(m,2H),1.98-2.04(m,2H),1.78-1.90(m,2H),1.48(s,9H).
步骤3：制备4-(4-氨基-5-(7-甲氧基-5-甲基苯并噻吩-2-基)-7H-吡咯[2,3-d]嘧啶-7-基)哌啶-1-甲酸叔丁酯
Figure PCTCN2017087222-appb-000069
将3-(4-氨基-5-碘-7H-吡咯[2,3-d]嘧啶-7-基)吡咯烷-1-甲酸叔丁酯替换成4-(4-氨基-5-碘-7H-吡咯[2,3-d]嘧啶-7-基)哌啶-1-甲酸叔丁酯，其余所需原料、试剂及制备方法同实施例1中的步骤10，得4-(4-氨基-5-(7-甲氧基-5-甲基苯并噻吩-2-基)-7H-吡咯[2,3-d]嘧啶-7-基)哌啶-1-甲酸叔丁酯。
1H NMR(300MHz,CDCl3)δ8.32(s,1H),7.20(s,2H),7.16(s,1H),6.63(s,1H),5.63(s,2H),4.82-4.92(m,1H),4.25-4.40(m,2H),3.99(s,3H),2.88-3.02(m,2H),2.48(s,3H),2.15-1.80(m,4H),1.48(s,9H).
步骤4：制备5-(7-甲氧基-5-甲基苯并噻吩-2-基)-7-(哌啶-4-基)-7H-吡咯[2,3-d]嘧啶-4-氨基盐酸盐
Figure PCTCN2017087222-appb-000070
将3-(4-氨基-5-(7-甲氧基-5-甲基苯并噻吩-2-基)-7H-吡咯[2,3-d]嘧啶-7-基)吡咯烷-1-甲酸叔丁酯替换成4-(4-氨基-5-(7-甲氧基-5-甲基苯并噻吩-2-基)-7H-吡咯[2,3-d]嘧啶-7-基)哌啶-1-甲酸叔丁酯，其余所需原料、试剂及制备方法同实施例1中的步骤11，得5-(7-甲氧基-5-甲基苯并噻吩-2-基)-7-(哌啶-4-基)-7H-吡咯[2,3-d]嘧啶-4-氨基盐酸盐。
步骤5：制备1-(4-(4-氨基-5-(7-甲氧基-5-甲基苯并噻吩-2-基)-7H-吡咯[2,3-d]嘧啶-7-基)哌啶-1-基)丙-2-烯-1-酮
将5-(7-甲氧基-5-甲基苯并噻吩-2-基)-7-(吡咯烷-3-基)-7H-吡咯[2,3-d]嘧啶-4-氨基盐酸盐替换成5-(7-甲氧基-5-甲基苯并噻吩-2-基)-7-(哌啶-4-基)-7H-吡咯[2,3-d]嘧啶-4-氨基盐酸盐，其余所需原料、试剂及制备方法同实施例1中的步骤12，得1-(4-(4-氨基-5-(7-甲氧基-5-甲基苯并噻吩-2-基)-7H-吡咯[2,3-d]嘧啶-7-基)哌啶-1-基)丙-2-烯-1-酮，收率56％。
1H NMR(300MHz,CDCl3)δ8.32(s,1H),7.20(d,J＝4.1Hz,2H),7.14(s,1H),6.62(m,2H),6.32(m,1H),5.73(m,1H),5.62(s,2H),4.97(m,2H),4.18(d,J＝11.4Hz,1H),3.99(s,3H),3.32(s,1H),2.85(s,1H),2.48(s,3H),1.99(m,4H).
实施例15：
1-(3-(4-氨基-5-(7-甲氧基-5-甲基苯并噻吩-2-基)-7H-吡咯[2,3-d]嘧啶-7-基)哌啶-1-基)丙-2-烯-1-酮
Figure PCTCN2017087222-appb-000071
步骤1：制备3-(4-氯-5-碘-7H-吡咯[2,3-d]嘧啶-7-基)哌啶-1-甲酸叔丁酯
Figure PCTCN2017087222-appb-000072
将3-羟基吡咯烷-1-甲酸叔丁酯替换成3-羟基哌啶-1-甲酸叔丁酯，其余所需原料、试剂及制备方法同实施例1中的步骤8，得3-(4-氯-5-碘-7H-吡咯[2,3-d]嘧啶-7-基)哌啶-1-甲酸叔丁酯。
步骤2：制备3-(4-氨基-5-碘-7H-吡咯[2,3-d]嘧啶-7-基)哌啶-1-甲酸叔丁酯
Figure PCTCN2017087222-appb-000073
将3-(4-氯-5-碘-7H-吡咯[2,3-d]嘧啶-7-基)吡咯烷-1-甲酸叔丁酯替换成3-(4-氯-5-碘-7H-吡咯[2,3-d]嘧啶-7-基)哌啶-1-甲酸叔丁酯，其余所需原料、试剂及制备方法同实施例1中的步骤9，得3-(4-氨基-5-碘-7H-吡咯[2,3-d]嘧啶-7-基)哌啶-1-甲酸叔丁酯。
步骤3：制备3-(4-氨基-5-(7-甲氧基-5-甲基苯并噻吩-2-基)-7H-吡咯[2,3-d]嘧啶-7-基)哌啶-1-甲酸叔丁酯
Figure PCTCN2017087222-appb-000074
将3-(4-氨基-5-碘-7H-吡咯[2,3-d]嘧啶-7-基)吡咯烷-1-甲酸叔丁酯替换成3-(4-氨基-5-碘-7H-吡咯[2,3-d]嘧啶-7-基)哌啶-1-甲酸叔丁酯，其余所需原料、试剂及制备方法同实施例1中的步骤10，得3-(4-氨基-5-(7-甲氧基-5-甲基苯并噻吩-2-基)-7H-吡咯[2,3-d]嘧啶-7-基)哌啶-1-甲酸叔丁酯。
步骤4：制备5-(7-甲氧基-5-甲基苯并噻吩-2-基)-7-(哌啶-3-基)-7H-吡咯[2,3-d]嘧啶-4-氨基盐酸盐
Figure PCTCN2017087222-appb-000075
将3-(4-氨基-5-(7-甲氧基-5-甲基苯并噻吩-2-基)-7H-吡咯[2,3-d]嘧啶-7-基)吡咯烷-1-甲酸叔丁酯替换成3-(4-氨基-5-(7-甲氧基-5-甲基苯并噻吩-2-基)-7H-吡咯[2,3-d]嘧啶-7-基)哌啶-1-甲酸叔丁酯，其余所需原料、试剂及制备方法同实施例1中的步骤11，得5-(7-甲氧基-5-甲基苯并噻吩-2-基)-7-(哌啶-3-基)-7H-吡咯[2,3-d]嘧啶-4-氨基盐酸盐。
步骤5：制备1-(3-(4-氨基-5-(7-甲氧基-5-甲基苯并噻吩-2-基)-7H-吡咯[2,3-d]嘧啶-7-基)哌啶-1-基)丙-2-烯-1-酮
将5-(7-甲氧基-5-甲基苯并噻吩-2-基)-7-(吡咯烷-3-基)-7H-吡咯[2,3-d]嘧啶-4-氨基盐酸盐替换成5-(7-甲氧基-5-甲基苯并噻吩-2-基)-7-(哌啶-3-基)-7H-吡咯[2,3-d]嘧啶-4-氨基盐酸盐，其余所需原料、试剂及制备方法同实施例1中的步骤12，得1-(3-(4-氨基-5-(7-甲氧基-5-甲基苯并噻吩-2-基)-7H-吡咯[2,3-d]嘧啶-7-基)哌啶-1-基)丙-2-烯-1-酮，收率50％。
1H NMR(300MHz,CDCl3)δ8.33(s,1H),7.22(d,J＝3.2Hz,2H),7.17(s,1H),6.63(q,J＝10.3Hz,2H),6.32(d,J＝16.9Hz,1H),5.72(d,J＝10.1Hz,1H),5.53(s,2H),4.74(s,2H),4.32(s,1H),3.25(s,1H),2.84(s,1H),2.28(s,1H),2.17(s,1H),1.95(s,1H),1.77(s,4H).
实施例16：
1-(3-(4-氨基-5-(7-甲氧基-5-甲基苯并噻吩-2-基)-7H-吡咯[2,3-d]嘧啶-7-基)吡咯烷-1-基)丙-1-酮
Figure PCTCN2017087222-appb-000076
将丙烯酰氯替换成丙酰氯，其余所需原料、试剂及制备方法同实施例1中的步骤12， 得1-(3-(4-氨基-5-(7-甲氧基-5-甲基苯并噻吩-2-基)-7H-吡咯[2,3-d]嘧啶-7-基)吡咯烷-1-基)丙-1-酮，收率46％。
1H NMR(300MHz,CDCl3)δ8.33(s,1H),7.22(s,2H),7.10(d,J＝14.8Hz,1H),6.64(s,1H),5.55(s,2H),5.52–5.39(m,1H),4.10–4.02(m,1H),4.00(s,3H),3.86(dd,J＝16.1,7.0Hz,1H),3.74–3.62(m,2H),2.49(s,3H),2.44–2.22(m,4H),1.18(m,3H).
实施例17：
(E)-1-(3-(4-氨基-5-(7-甲氧基-5-甲基苯并噻吩-2-基)-7H-吡咯[2,3-d]嘧啶-7-基)氮杂环丁烷-1-基)-4-(吡咯烷-1-基)丁-2-烯-1-酮
Figure PCTCN2017087222-appb-000077
将80毫克5-(7-甲氧基-5-甲基苯并噻吩-2-基)-7-(氮杂环丁烷-3-基)-7H-吡咯[2,3-d]嘧啶-4-氨基盐酸盐，116毫克反式-4-(吡咯烷-1-基)巴豆酸盐酸盐和228毫克2-(7-偶氮苯并三氮唑)-N,N,N',N'-四甲基脲六氟磷酸酯溶于2毫升N,N-二甲基甲酰胺，滴加0.26毫升的N,N-二异丙基乙胺，室温下搅拌过夜，次日加水后用乙酸乙酯萃取，有机相用饱和氯化钠水溶液洗涤后无水硫酸钠干燥，过滤浓缩，残余物柱层析(二氯甲烷：甲醇(含1％氨水)＝95:5)分离得30毫克黄色固体，收率23％。
1H NMR(300MHz,CD3OD)δ8.13(s,1H),7.56(s,1H),7.22(s,1H),7.16(s,1H),6.76(dd,J＝14.8,7.5Hz,1H),6.69(s,1H),6.46(d,J＝15.1Hz,1H),5.60–5.50(m,1H),4.80(d,J＝8.2Hz,2H),4.55(dd,J＝17.1,8.5Hz,2H),3.94(s,3H),3.83(d,J＝6.8Hz,2H),3.18(s,4H),2.42(s,3H),2.01(d,J＝3.7Hz,4H).
实施例18：
(E)-1-(3-(4-氨基-5-(7-甲氧基-5-甲基苯并噻吩-2-基)-7H-吡咯[2,3-d]嘧啶-7-基)吡咯烷-1-基)丁-2-烯-1-酮
Figure PCTCN2017087222-appb-000078
将丙烯酰氯替换成巴豆酰氯，其余所需原料、试剂及制备方法同实施例1中的步骤12，得(E)-1-(3-(4-氨基-5-(7-甲氧基-5-甲基苯并噻吩-2-基)-7H-吡咯[2,3-d]嘧啶-7-基)吡咯烷-1-基)丁-2-烯-1-酮，收率36％。
1H NMR(300MHz,CDCl3)δ8.31(s,1H),7.20(s,2H),7.09(dd,J＝13.6,4.4Hz,1H),7.04–6.88(m,1H),6.62(s,1H),6.12(dd,J＝22.6,15.1Hz,1H),5.73(s,2H),5.55–5.38(m,1H),4.20–4.04(m,1H),3.98(s,4H),3.81(dd,J＝13.8,7.8Hz,1H),3.74(dd,J＝13.8,6.7Hz,2H),2.49(d,J＝10.5Hz,4H),2.45–2.33(m,2H),1.88(dd,J＝10.8,7.5Hz,3H).
实施例19：
(E)-1-(3-(4-氨基-5-(7-甲氧基-5-甲基苯并噻吩-2-基)-7H-吡咯[2,3-d]嘧啶-7-基)氮杂环丁烷-1-基)-4-(二甲氨基)丁-2-烯-1-酮
Figure PCTCN2017087222-appb-000079
将反式-4-(吡咯烷-1-基)巴豆酸盐酸盐替换成反式-4-(二甲氨基)巴豆酸盐酸盐，其余所需原料、试剂及制备方法同实施例17，得(E)-1-(3-(4-氨基-5-(7-甲氧基-5-甲基苯并噻吩-2-基)-7H-吡咯[2,3-d]嘧啶-7-基)氮杂环丁烷-1-基)-4-(二甲氨基)丁-2-烯-1-酮，收率40％。
1H NMR(300MHz,CDCl3)δ8.29(s,1H),7.35(s,1H),7.23(s,2H),7.00–6.86(m,1H),6.64(s,1H),6.32(d,J＝15.8Hz,1H),5.65(s,2H),4.82(s,1H),4.65(s,2H),4.49(s,1H),3.99(s,3H),3.36(d,J＝6.0Hz,2H),2.47(s,9H),1.25(s,4H).
实施例20：
1-(3-((4-氨基-5-(7-甲氧基-5-甲基苯并噻吩-2-基)-7H-吡咯[2,3-d]嘧啶-7-基)甲基)吡咯烷-1-基)丙-2-烯-1-酮
Figure PCTCN2017087222-appb-000080
步骤1：制备3-((4-氯-5-碘-7H-吡咯[2,3-d]嘧啶-7-基)甲基)吡咯烷-1-甲酸叔丁酯
Figure PCTCN2017087222-appb-000081
将3-羟基吡咯烷-1-甲酸叔丁酯替换成3-羟甲基吡咯烷-1-甲酸叔丁酯，其余所需原料、试剂及制备方法同实施例1中的步骤8，得3-((4-氯-5-碘-7H-吡咯[2,3-d]嘧啶-7-基)甲基)吡咯烷-1-甲酸叔丁酯。
步骤2：制备3-((4-氨基-5-碘-7H-吡咯[2,3-d]嘧啶-7-基)甲基)吡咯烷-1-甲酸叔丁酯
Figure PCTCN2017087222-appb-000082
3-(4-氯-5-碘-7H-吡咯[2,3-d]嘧啶-7-基)吡咯烷-1-甲酸叔丁酯替换成3-((4-氯-5-碘-7H-吡咯[2,3-d]嘧啶-7-基)甲基)吡咯烷-1-甲酸叔丁酯，其余所需原料、试剂及制备方法同实施例1中的步骤9，得3-((4-氨基-5-碘-7H-吡咯[2,3-d]嘧啶-7-基)甲基)吡咯烷-1-甲酸叔丁酯。
步骤3：制备3-((4-氨基-5-(7-甲氧基-5-甲基苯并噻吩-2-基)-7H-吡咯[2,3-d]嘧啶-7-基)甲基)吡咯烷-1-甲酸叔丁酯
Figure PCTCN2017087222-appb-000083
将3-(4-氨基-5-碘-7H-吡咯[2,3-d]嘧啶-7-基)吡咯烷-1-甲酸叔丁酯替换成3-((4-氨基-5-碘-7H-吡咯[2,3-d]嘧啶-7-基)甲基)吡咯烷-1-甲酸叔丁酯，其余所需原料、试剂及制备方法同实施例1中的步骤10，得3-((4-氨基-5-(7-甲氧基-5-甲基苯并噻吩-2-基)-7H-吡咯[2,3-d]嘧啶-7-基)甲基)吡咯烷-1-甲酸叔丁酯。
步骤4：制备5-(7-甲氧基-5-甲基苯并噻吩-2-基)-7-(吡咯烷-3-亚甲基)-7H-吡咯[2,3-d]嘧啶-4-胺盐酸盐
Figure PCTCN2017087222-appb-000084
将3-(4-氨基-5-(7-甲氧基-5-甲基苯并噻吩-2-基)-7H-吡咯[2,3-d]嘧啶-7-基)吡咯烷-1-甲酸叔丁酯替换成3-((4-氨基-5-(7-甲氧基-5-甲基苯并噻吩-2-基)-7H-吡咯[2,3-d]嘧啶-7-基)甲基)吡咯烷-1-甲酸叔丁酯，其余所需原料、试剂及制备方法同实施例1中的步骤11，得5-(7-甲氧基-5-甲基苯并噻吩-2-基)-7-(吡咯烷-3-亚甲基)-7H-吡咯[2,3-d]嘧啶-4-胺盐酸盐。
步骤5：制备1-(3-((4-氨基-5-(7-甲氧基-5-甲基苯并噻吩-2-基)-7H-吡咯[2,3-d]嘧啶-7-基)甲基)吡咯烷-1-基)丙-2-烯-1-酮
将5-(7-甲氧基-5-甲基苯并噻吩-2-基)-7-(吡咯烷-3-基)-7H-吡咯[2,3-d]嘧啶-4-氨基盐酸盐替换成5-(7-甲氧基-5-甲基苯并噻吩-2-基)-7-(吡咯烷-3-亚甲基)-7H-吡咯[2,3-d]嘧啶-4-胺盐酸盐，其余所需原料、试剂及制备方法同实施例1中的步骤12，得1-(3-((4-氨基-5-(7-甲氧基-5-甲基苯并噻吩-2-基)-7H-吡咯[2,3-d]嘧啶-7-基)甲基)吡咯烷-1-基)丙-2-烯-1-酮，收率37％。
1H NMR(300MHz,CDCl3)δ8.32(s,1H),7.22(s,2H),7.11(d,J＝7.2Hz,1H),6.64(s,1H),6.39(dd,J＝14.4,7.6Hz,2H),5.67(dd,J＝9.9,5.4Hz,1H),5.55(s,2H),4.38–4.15(m,2H),4.00(s,3H),3.75(s,2H),3.65–3.47(m,1H),3.38(m,1H),2.89(m,1H),2.49(s,3H),2.17–1.96(m,1H).
实施例21：
1-(3-(4-氨基-5-(6-甲氧基苯并噻吩-2-基)-7H-吡咯[2,3-d]嘧啶-7-基)氮杂环丁烷-1-基)丙-2-烯-1-酮
Figure PCTCN2017087222-appb-000085
步骤1：制备(2,2-二乙氧基乙基)(3-甲氧基苯基)硫醚
Figure PCTCN2017087222-appb-000086
将2-甲氧基-4-甲基苯硫酚替换成3-甲氧基苯硫酚，其余所需原料、试剂及制备方法同实施例1中的步骤4，得(2,2-二乙氧基乙基)(3-甲氧基苯基)硫醚。
1H NMR(300MHz,CDCl3)7.19(t,J＝7.9Hz,1H),6.94(m,2H),6.75–6.68(m,1H),4.65(t,J＝5.5Hz,1H),3.79(s,3H),3.68(m,2H),3.55(m,2H),3.14(d,J＝5.6Hz,2H),1.20(t,J＝7.0Hz,6H).
步骤2：制备6-甲氧基苯并噻吩
Figure PCTCN2017087222-appb-000087
将(2,2-二乙氧基乙基)(2-甲氧基-4-甲基苯基)硫醚替换成(2,2-二乙氧基乙基)(3-甲氧基苯基)硫醚，其余所需原料、试剂及制备方法同实施例1中的步骤5，得6-甲氧基苯并噻吩。
1H NMR(300MHz,CDCl3)δ7.70(d,J＝8.7Hz,1H),7.36(d,J＝2.0Hz,1H),7.00(dd,J＝8.7,2.0Hz,1H),3.88(s,3H).
步骤3：制备(6-甲氧基苯并噻吩-2-基)硼酸
Figure PCTCN2017087222-appb-000088
将7-甲氧基-5-甲基苯并噻吩替换成6-甲氧基苯并噻吩，其余所需原料、试剂及制备方法同实施例1中的步骤6，得(6-甲氧基苯并噻吩-2-基)硼酸。
步骤4：制备3-(4-氨基-5-(6-甲氧基苯并噻吩-2-基)-7H-吡咯[2,3-d]嘧啶-7-基)氮杂环丁烷-1-甲酸叔丁酯
Figure PCTCN2017087222-appb-000089
将3-(4-氨基-5-碘-7H-吡咯[2,3-d]嘧啶-7-基)吡咯烷-1-甲酸叔丁酯替换成3-(4-氨基-5-碘-7H-吡咯[2,3-d]嘧啶-7-基)氮杂环丁烷-1-甲酸叔丁酯，(7-甲氧基-5-甲基苯并噻吩-2-基)硼酸替换成(6-甲氧基苯并噻吩-2-基)硼酸，其余所需原料、试剂及制备方法同实施例1中的步骤10，得3-(4-氨基-5-(6-甲氧基苯并噻吩-2-基)-7H-吡咯[2,3-d]嘧啶-7-基)氮杂环丁烷-1-甲酸叔丁酯。
步骤5：制备5-(6-甲氧基苯并噻吩-2-基)-7-(氮杂环丁烷-3-基)-7H-吡咯[2,3-d]嘧啶-4-胺盐酸盐
Figure PCTCN2017087222-appb-000090
将3-(4-氨基-5-(7-甲氧基-5-甲基苯并噻吩-2-基)-7H-吡咯[2,3-d]嘧啶-7-基)吡咯烷-1-甲酸叔丁酯替换成3-(4-氨基-5-(6-甲氧基苯并噻吩-2-基)-7H-吡咯[2,3-d]嘧啶-7-基)氮杂环丁烷-1-甲酸叔丁酯，其余所需原料、试剂及制备方法同实施例1中的步骤11，得5-(6-甲氧基苯并噻吩-2-基)-7-(氮杂环丁烷-3-基)-7H-吡咯[2,3-d]嘧啶-4-胺盐酸盐。
步骤6：制备1-(3-(4-氨基-5-(6-甲氧基苯并噻吩-2-基)-7H-吡咯[2,3-d]嘧啶-7-基)氮杂环丁烷-1-基)丙-2-烯-1-酮
将5-(7-甲氧基-5-甲基苯并噻吩-2-基)-7-(吡咯烷-3-基)-7H-吡咯[2,3-d]嘧啶-4-氨基盐酸盐替换成5-(6-甲氧基苯并噻吩-2-基)-7-(氮杂环丁烷-3-基)-7H-吡咯[2,3-d]嘧啶-4-胺盐酸盐，其余所需原料、试剂及制备方法同实施例1中的步骤12，得1-(3-(4-氨基-5-(6-甲氧基苯并噻吩-2-基)-7H-吡咯[2,3-d]嘧啶-7-基)氮杂环丁烷-1-基)丙-2-烯-1-酮，收率65％。
1H NMR(300MHz,CDCl3)δ8.32(s,1H),7.68(d,J＝8.7Hz,1H),7.34(s, 2H),7.24(s,1H),7.07–7.00(m,1H),6.42(d,J＝17.0Hz,1H),6.25(dd,J＝16.9,10.3Hz,1H),5.73(dd,J＝16.6,8.0Hz,2H),5.52(s,2H),4.77(d,J＝7.6Hz,1H),4.72–4.62(m,1H),4.53(s,2H),3.90(s,3H).
实施例22：
1-(3-(4-氨基-5-(7-甲氧基苯并噻吩-2-基)-7H-吡咯[2,3-d]嘧啶-7-基)氮杂环丁烷-1-基)丙-2-烯-1-酮
Figure PCTCN2017087222-appb-000091
步骤1：制备(2,2-二乙氧基乙基)(2-甲氧基苯基)硫醚
Figure PCTCN2017087222-appb-000092
将2-甲氧基-4-甲基苯硫酚替换成2-甲氧基苯硫酚，其余所需原料、试剂及制备方法同实施例1中的步骤4，得(2,2-二乙氧基乙基)(2-甲氧基苯基)硫醚。
步骤2：制备7-甲氧基苯并噻吩
Figure PCTCN2017087222-appb-000093
将(2,2-二乙氧基乙基)(2-甲氧基-4-甲基苯基)硫醚替换成(2,2-二乙氧基乙基)(2-甲氧基苯基)硫醚，其余所需原料、试剂及制备方法同实施例1中的步骤5，得7-甲氧基苯并噻吩。
1H NMR(300MHz,CDCl3)δ7.49–7.41(m,2H),7.33(m,2H),6.78(d,J＝7.9Hz,1H),4.01(s,3H).
步骤3：制备(7-甲氧基苯并噻吩-2-基)硼酸
Figure PCTCN2017087222-appb-000094
将7-甲氧基-5-甲基苯并噻吩替换成7-甲氧基苯并噻吩，其余所需原料、试剂及制备方法同实施例1中的步骤6，得(6-甲氧基苯并噻吩-2-基)硼酸。
步骤4：制备3-(4-氨基-5-(6-甲氧基苯并噻吩-2-基)-7H-吡咯[2,3-d]嘧啶-7-基)氮 杂环丁烷-1-甲酸叔丁酯
Figure PCTCN2017087222-appb-000095
将3-(4-氨基-5-碘-7H-吡咯[2,3-d]嘧啶-7-基)吡咯烷-1-甲酸叔丁酯替换成3-(4-氨基-5-碘-7H-吡咯[2,3-d]嘧啶-7-基)氮杂环丁烷-1-甲酸叔丁酯，(7-甲氧基-5-甲基苯并噻吩-2-基)硼酸替换成(7-甲氧基苯并噻吩-2-基)硼酸，其余所需原料、试剂及制备方法同实施例1中的步骤10，得3-(4-氨基-5-(7-甲氧基苯并噻吩-2-基)-7H-吡咯[2,3-d]嘧啶-7-基)氮杂环丁烷-1-甲酸叔丁酯。
步骤5：制备5-(7-甲氧基苯并噻吩-2-基)-7-(氮杂环丁烷-3-基)-7H-吡咯[2,3-d]嘧啶-4-胺盐酸盐
Figure PCTCN2017087222-appb-000096
将3-(4-氨基-5-(7-甲氧基-5-甲基苯并噻吩-2-基)-7H-吡咯[2,3-d]嘧啶-7-基)吡咯烷-1-甲酸叔丁酯替换成3-(4-氨基-5-(7-甲氧基苯并噻吩-2-基)-7H-吡咯[2,3-d]嘧啶-7-基)氮杂环丁烷-1-甲酸叔丁酯，其余所需原料、试剂及制备方法同实施例1中的步骤11，得5-(7-甲氧基苯并噻吩-2-基)-7-(氮杂环丁烷-3-基)-7H-吡咯[2,3-d]嘧啶-4-胺盐酸盐。
步骤6：制备1-(3-(4-氨基-5-(7-甲氧基苯并噻吩-2-基)-7H-吡咯[2,3-d]嘧啶-7-基)氮杂环丁烷-1-基)丙-2-烯-1-酮
将5-(7-甲氧基-5-甲基苯并噻吩-2-基)-7-(吡咯烷-3-基)-7H-吡咯[2,3-d]嘧啶-4-氨基盐酸盐替换成5-(7-甲氧基苯并噻吩-2-基)-7-(氮杂环丁烷-3-基)-7H-吡咯[2,3-d]嘧啶-4-胺盐酸盐，其余所需原料、试剂及制备方法同实施例1中的步骤12，得1-(3-(4-氨基-5-(7-甲氧基苯并噻吩-2-基)-7H-吡咯[2,3-d]嘧啶-7-基)氮杂环丁烷-1-基)丙-2-烯-1-酮，收率55％。
1H NMR(300MHz,CDCl3)δ8.31(s,1H),7.42(d,J＝7.6Hz,1H),7.37(t,J＝3.8Hz,2H),7.33(d,J＝3.3Hz,1H),6.81(d,J＝7.5Hz,1H),6.42(dd, J＝16.9,1.6Hz,1H),6.24(dd,J＝17.0,10.2Hz,1H),5.80–5.57(m,4H),4.78–4.62(m,2H),4.55(m,2H),4.02(s,3H).
实施例23：
1-(3-(4-氨基-5-(7-甲氧基-5-甲基苯并呋喃-2-基)-7H-吡咯[2,3-d]嘧啶-7-基)氮杂环丁烷-1-基)丙-2-烯-1-酮
Figure PCTCN2017087222-appb-000097
步骤1：制备1-(2,2-二乙氧基乙氧基)-2-甲氧基-4-甲基苯
Figure PCTCN2017087222-appb-000098
将2-甲氧基-4-甲基苯硫酚替换成2-甲氧基-4-甲基苯酚，其余所需原料、试剂及制备方法同实施例1中的步骤4，得1-(2,2-二乙氧基乙氧基)-2-甲氧基-4-甲基苯。
1H NMR(300MHz,CDCl3)δ6.83(d,J＝7.9Hz,1H),6.68(d,J＝10.8Hz,2H),4.86(t,J＝5.1Hz,1H),4.03(d,J＝5.2Hz,2H),3.83(s,3H),3.74–3.57(m,4H),2.29(s,3H),1.23(t,J＝7.0Hz,6H).
步骤2：制备7-甲氧基-5-甲基苯并呋喃
Figure PCTCN2017087222-appb-000099
将(2,2-二乙氧基乙基)(2-甲氧基-4-甲基苯基)硫醚替换成1-(2,2-二乙氧基乙氧基)-2-甲氧基-4-甲基苯，其余所需原料、试剂及制备方法同实施例1中的步骤5，得7-甲氧基-5-甲基苯并呋喃。
步骤3：制备(7-甲氧基-5-甲基苯并呋喃-2-基)硼酸
Figure PCTCN2017087222-appb-000100
将7-甲氧基-5-甲基苯并噻吩替换成7-甲氧基苯并呋喃，其余所需原料、试剂及制备方法同实施例1中的步骤6，得(7-甲氧基-5-甲基苯并呋喃-2-基)硼酸。
步骤4：制备3-(4-氨基-5-(7-甲氧基-5-甲基苯并呋喃-2-基)-7H-吡咯[2,3-d]嘧啶-7-基)氮杂环丁烷-1-甲酸叔丁酯
Figure PCTCN2017087222-appb-000101
将3-(4-氨基-5-碘-7H-吡咯[2,3-d]嘧啶-7-基)吡咯烷-1-甲酸叔丁酯替换成3-(4-氨基-5-碘-7H-吡咯[2,3-d]嘧啶-7-基)氮杂环丁烷-1-甲酸叔丁酯，(7-甲氧基-5-甲基苯并噻吩-2-基)硼酸替换成(7-甲氧基-5-甲基苯并呋喃-2-基)硼酸，其余所需原料、试剂及制备方法同实施例1中的步骤10，得3-(4-氨基-5-(7-甲氧基-5-甲基苯并呋喃-2-基)-7H-吡咯[2,3-d]嘧啶-7-基)氮杂环丁烷-1-甲酸叔丁酯。
步骤5：制备5-(7-甲氧基-5-甲基苯并呋喃-2-基)-7-(氮杂环丁烷-3-基)-7H-吡咯[2,3-d]嘧啶-4-胺盐酸盐
Figure PCTCN2017087222-appb-000102
将3-(4-氨基-5-(7-甲氧基-5-甲基苯并噻吩-2-基)-7H-吡咯[2,3-d]嘧啶-7-基)吡咯烷-1-甲酸叔丁酯替换成3-(4-氨基-5-(7-甲氧基-5-甲基苯并呋喃-2-基)-7H-吡咯[2,3-d]嘧啶-7-基)氮杂环丁烷-1-甲酸叔丁酯，其余所需原料、试剂及制备方法同实施例1中的步骤11，得5-(7-甲氧基-5-甲基苯并呋喃-2-基)-7-(氮杂环丁烷-3-基)-7H-吡咯[2,3-d]嘧啶-4-胺盐酸盐。
步骤6：制备1-(3-(4-氨基-5-(7-甲氧基-5-甲基苯并呋喃-2-基)-7H-吡咯[2,3-d]嘧啶-7-基)氮杂环丁烷-1-基)丙-2-烯-1-酮
将5-(7-甲氧基-5-甲基苯并噻吩-2-基)-7-(吡咯烷-3-基)-7H-吡咯[2,3-d]嘧啶-4-氨基盐酸盐替换成5-(7-甲氧基-5-甲基苯并呋喃-2-基)-7-(氮杂环丁烷-3-基)-7H-吡咯[2,3-d]嘧啶-4-胺盐酸盐，其余所需原料、试剂及制备方法同实施例1中的步骤12，得1-(3-(4-氨基-5-(7-甲氧基-5-甲基苯并呋喃-2-基)-7H-吡咯[2,3-d]嘧啶-7-基)氮杂环丁烷-1-基)丙-2-烯-1-酮，收率56％。
1H NMR(300MHz,CD3OD)δ8.14(s,1H),8.00(s,1H),6.94(s,2H),6.71(s,1H),6.38(m,2H),5.79(d,J＝10.0Hz,1H),5.63(s,1H),4.85-4.54(m,4H),3.98(s,3H),2.42(s,3H).
实施例24：
1-(2-((4-氨基-5-(7-甲氧基-5-甲基苯并噻吩-2-基)-7H-吡咯[2,3-d]嘧啶-7-基)甲基)吡咯烷-1-基)丙-2-烯-1-酮
Figure PCTCN2017087222-appb-000103
步骤1：制备2-((4-氯-5-碘-7H-吡咯[2,3-d]嘧啶-7-基)甲基)吡咯烷-1-甲酸叔丁酯
Figure PCTCN2017087222-appb-000104
将3-羟基吡咯烷-1-甲酸叔丁酯替换成2-羟甲基吡咯烷-1-甲酸叔丁酯，其余所需原料、试剂及制备方法同实施例1中的步骤8，得2-((4-氯-5-碘-7H-吡咯[2,3-d]嘧啶-7-基)甲基)吡咯烷-1-甲酸叔丁酯。
步骤2：制备2-((4-氨基-5-碘-7H-吡咯[2,3-d]嘧啶-7-基)甲基)吡咯烷-1-甲酸叔丁酯
Figure PCTCN2017087222-appb-000105
3-(4-氯-5-碘-7H-吡咯[2,3-d]嘧啶-7-基)吡咯烷-1-甲酸叔丁酯替换成2-((4-氯-5-碘-7H-吡咯[2,3-d]嘧啶-7-基)甲基)吡咯烷-1-甲酸叔丁酯，其余所需原料、试剂及制备方法同实施例1中的步骤9，得2-((4-氨基-5-碘-7H-吡咯[2,3-d]嘧啶-7-基)甲基)吡咯烷-1-甲酸叔丁酯。
步骤3：制备2-((4-氨基-5-(7-甲氧基-5-甲基苯并噻吩-2-基)-7H-吡咯[2,3-d]嘧啶-7-基)甲基)吡咯烷-1-甲酸叔丁酯
Figure PCTCN2017087222-appb-000106
将3-(4-氨基-5-碘-7H-吡咯[2,3-d]嘧啶-7-基)吡咯烷-1-甲酸叔丁酯替换成2-((4-氨基-5-碘-7H-吡咯[2,3-d]嘧啶-7-基)甲基)吡咯烷-1-甲酸叔丁酯，其余所需原料、试剂及制备方法同实施例1中的步骤10，得2-((4-氨基-5-(7-甲氧基-5-甲基苯并噻吩-2-基)-7H-吡咯[2,3-d]嘧啶-7-基)甲基)吡咯烷-1-甲酸叔丁酯。
步骤4：制备5-(7-甲氧基-5-甲基苯并噻吩-2-基)-7-(吡咯烷-2-亚甲基)-7H-吡咯[2,3-d]嘧啶-4-胺盐酸盐
Figure PCTCN2017087222-appb-000107
将3-(4-氨基-5-(7-甲氧基-5-甲基苯并噻吩-2-基)-7H-吡咯[2,3-d]嘧啶-7-基)吡咯烷-1-甲酸叔丁酯替换成2-((4-氨基-5-(7-甲氧基-5-甲基苯并噻吩-2-基)-7H-吡咯[2,3-d]嘧啶-7-基)甲基)吡咯烷-1-甲酸叔丁酯，其余所需原料、试剂及制备方法同实施例1中的步骤11，得5-(7-甲氧基-5-甲基苯并噻吩-2-基)-7-(吡咯烷-2-亚甲基)-7H-吡咯[2,3-d]嘧啶-4-胺盐酸盐。
步骤5：制备1-(2-((4-氨基-5-(7-甲氧基-5-甲基苯并噻吩-2-基)-7H-吡咯[2,3-d]嘧啶-7-基)甲基)吡咯烷-1-基)丙-2-烯-1-酮
将5-(7-甲氧基-5-甲基苯并噻吩-2-基)-7-(吡咯烷-3-基)-7H-吡咯[2,3-d]嘧啶-4-氨基盐酸盐替换成5-(7-甲氧基-5-甲基苯并噻吩-2-基)-7-(吡咯烷-2-亚甲基)-7H-吡咯[2,3-d]嘧啶-4-胺盐酸盐，其余所需原料、试剂及制备方法同实施例1中的步骤12，得1-(2-((4-氨基-5-(7-甲氧基-5-甲基苯并噻吩-2-基)-7H-吡咯[2,3-d]嘧啶-7-基)甲基)吡咯烷-1-基)丙-2-烯-1-酮，收率61％。
1H NMR(300MHz,CDCl3)δ8.34(d,J＝9.2Hz,1H),7.23–7.16(m,3H),6.63(s,1H),6.47(d,J＝8.7Hz,2H),5.75(dd,J＝8.2,3.6Hz,1H),5.55(m,3H),4.53(s,3H),3.99(s,3H),3.55–3.43(m,2H),2.48(s,3H),1.93(m,4H).
实施例25：
2-(3-(4-氨基-5-(7-甲氧基-5-甲基苯并噻吩-2-基)-7H-吡咯[2,3-d]嘧啶-7-基)吡咯烷-1-羰基)-3-环丙基丙烯腈
Figure PCTCN2017087222-appb-000108
步骤1：制备3-(3-(4-氨基-5-(7-甲氧基-5-甲基苯并噻吩-2-基)-7H-吡咯[2,3-d]嘧啶-7-基)吡咯烷-1-基)-3-氧代丙腈
Figure PCTCN2017087222-appb-000109
将反式-4-(吡咯烷-1-基)巴豆酸盐酸盐替换成氰乙酸，其余所需原料、试剂及制备方法同实施例17，得3-(3-(4-氨基-5-(7-甲氧基-5-甲基苯并噻吩-2-基)-7H-吡咯[2,3-d]嘧啶-7-基)吡咯烷-1-基)-3-氧代丙腈。
1H NMR(300MHz,DMSO-d6)δ8.29(s,1H),7.69m,1H),7.35(s,1H),7.28(s,1H),6.96(s,2H),6.81(s,1H),5.39(m,1H),4.05-3.30(m,11H),2.44(s,3H).
步骤2：制备2-(3-(4-氨基-5-(7-甲氧基-5-甲基苯并噻吩-2-基)-7H-吡咯[2,3-d]嘧啶-7-基)氮杂环丁烷-1-羰基)-3-环丙基丙烯腈
将50毫克3-(3-(4-氨基-5-(7-甲氧基-5-甲基苯并噻吩-2-基)-7H-吡咯[2,3-d]嘧啶-7-基)吡咯烷-1-基)-3-氧代丙腈，17微升环丙基甲醛和10微升哌啶溶于1毫升甲醇，室温下搅拌过夜，次日有固体析出，过滤，滤饼干燥得13毫克米色固体，收率80％。
1H NMR(300MHz,CDCl3)δ8.32(s,1H),7.22(s,2H),7.11(s,1H),6.91(m,1H),6.64(s,1H),5.62(s,2H),5.48(s,1H),4.38–4.34(m,0.5H),4.00(s,3H),3.85-4.1(m,2.5H),3.80–3.68(m,1H),,2.48(s,3H),2.6–2.43(m,2H),1.3-1.25(m,1H),1.0-0.8(m,4H).
实施例26：
1-(3-(4-氨基-3-(7-甲氧基-5-甲基苯并噻吩-2-基)-1H-吡唑[3,4-d]嘧啶-1-基)氮杂环丁烷-1-基)丙-2-烯-1-酮
Figure PCTCN2017087222-appb-000110
步骤1：制备3-((甲磺酰基)氧)氮杂环丁烷-1-甲酸叔丁酯
Figure PCTCN2017087222-appb-000111
将3-羟基吡咯烷-1-甲酸叔丁酯替换成3-羟基氮杂环丁烷-1-甲酸叔丁酯，其余所需原料、试剂及制备方法同实施例2中的步骤3，得3-((甲磺酰基)氧)氮杂环丁烷-1-甲酸叔丁酯。
1H NMR(300MHz,CDCl3)δ5.23–5.04(m,1H),4.23(m,2H),4.05(m,2H),3.03(s,3H),1.40(s,9H).
步骤2：制备3-(4-氨基-3-碘-1H-吡唑[3,4-d]嘧啶-1-基)氮杂环丁烷-1-甲酸叔丁酯
Figure PCTCN2017087222-appb-000112
将3-((甲磺酰基)氧)吡咯烷-1-甲酸叔丁酯替换成3-((甲磺酰基)氧)氮杂环丁烷-1-甲酸叔丁酯，其余所需原料、试剂及制备方法同实施例2中的步骤4，得3-(4-氨基-3-碘-1H-吡唑[3,4-d]嘧啶-1-基)氮杂环丁烷-1-甲酸叔丁酯。
步骤3：制备3-(4-氨基-3-(7-甲氧基-5-甲基苯并噻吩-2-基)-1H-吡唑[3,4-d]嘧啶-1-基)氮杂环丁烷-1-甲酸叔丁酯
Figure PCTCN2017087222-appb-000113
将3-(4-氨基-5-碘-7H-吡咯[2,3-d]嘧啶-7-基)吡咯烷-1-甲酸叔丁酯替换成3-(4-氨基-3-碘-1H-吡唑[3,4-d]嘧啶-1-基)氮杂环丁烷-1-甲酸叔丁酯，其余所需原料、试剂及制备方法同实施例1中的步骤10，得3-(4-氨基-3-(7-甲氧基-5-甲基苯并噻吩-2-基)-1H-吡唑[3,4-d]嘧啶-1-基)氮杂环丁烷-1-甲酸叔丁酯。
步骤4：制备3-(7-甲氧基-5-甲基苯并噻吩-2-基)-1-(氮杂环丁烷-3-基)-1H-吡唑[3,4-d]嘧啶-4-胺盐酸盐
Figure PCTCN2017087222-appb-000114
将3-(4-氨基-5-(7-甲氧基-5-甲基苯并噻吩-2-基)-7H-吡咯[2,3-d]嘧啶-7-基)吡咯烷-1-甲酸叔丁酯替换成3-(4-氨基-3-(7-甲氧基-5-甲基苯并噻吩-2-基)-1H-吡唑[3,4-d]嘧啶-1-基)氮杂环丁烷-1-甲酸叔丁酯，其余所需原料、试剂及制备方法同实施例1中的步骤11，得3-(7-甲氧基-5-甲基苯并噻吩-2-基)-1-(氮杂环丁烷-3-基)-1H-吡唑[3,4-d]嘧啶-4-胺盐酸盐。
步骤5：制备1-(3-(4-氨基-3-(7-甲氧基-5-甲基苯并噻吩-2-基)-1H-吡唑[3,4-d]嘧啶-1-基)氮杂环丁烷-1-基)丙基-2-烯-1-酮
将5-(7-甲氧基-5-甲基苯并噻吩-2-基)-7-(吡咯烷-3-基)-7H-吡咯[2,3-d]嘧啶-4-氨基盐酸盐替换成3-(7-甲氧基-5-甲基苯并噻吩-2-基)-1-(氮杂环丁烷-3-基)-1H-吡唑[3,4-d]嘧啶-4-胺盐酸盐，其余所需原料、试剂及制备方法同实施例1中的步骤12，得1-(3-(4-氨基-3-(7-甲氧基-5-甲基苯并噻吩-2-基)-1H-吡唑[3,4-d]嘧啶-1-基)氮杂环丁烷-1-基)丙基-2-烯-1-酮，收率54％。
1H NMR(300MHz,DMSO-d6)δ8.29(s,1H),7.71(s,1H),7.36(s,1H),6.86(s,1H),6.40(dd,J＝17.0,10.5Hz,1H),6.17(d,J＝17.2Hz,1H),5.73(d,J＝11.7Hz,2H),4.76-4.67(m,2H),4.53–4.35(m,2H),3.96(s,3H),2.45(s, 3H).
实施例27：
1-(3-(4-氨基-3-(7-甲氧基-5-甲基苯并噻吩-2-基)-1H-吡唑[3,4-d]嘧啶-1-基)氮杂环丁烷-1-基)丙-2-炔-1-酮
Figure PCTCN2017087222-appb-000115
将100毫克3-(7-甲氧基-5-甲基苯并噻吩-2-基)-1-(氮杂环丁烷-3-基)-1H-吡唑[3,4-d]嘧啶-4-胺盐酸盐，20微升丙炔酸和145毫克苯并三唑-1-三(三甲氨基)-三氟磷酸酯溶于2毫升N,N-二甲基甲酰胺，滴加0.2毫升的N,N-二异丙基乙胺，室温下搅拌8小时停止反应，加水后用乙酸乙酯萃取，有机相用饱和氯化钠水溶液洗涤后无水硫酸钠干燥，过滤浓缩，残余物柱层析(二氯甲烷：甲醇＝97:3)分离得48毫克淡黄色固体，收率44％。
1H NMR(300MHz,DMSO-d6)δ8.30(s,1H),7.72(s,1H),7.36(s,1H),6.87(s,1H),5.76(s,1H),4.73-4.58(m,2H),4.55(s,1H),4.53–4.37(m,1H),3.97(s,3H),2.45(s,3H).
实施例28：
5-(7-甲氧基-5-甲基苯并噻吩-2-基)-7-(1-(乙烯磺酰基)氮杂环丁烷-3-基)-7H-吡咯[2,3-d]嘧啶-4-胺
Figure PCTCN2017087222-appb-000116
将100毫克5-(7-甲氧基-5-甲基苯并噻吩-2-基)-7-(氮杂环丁烷-3-基)-7H-吡咯[2,3-d]嘧啶-4-胺盐酸盐混悬于2毫升干燥的二氯甲烷中，加入0.18毫升的三乙胺，冰浴冷却下滴加31微升的2-氯乙基磺酰氯，滴加完后移至室温搅拌过夜，次日反应液 浓缩至干，残余物柱层析(二氯甲烷：甲醇＝97:3)分离得46毫克白色固体，收率52％。
1H NMR(300MHz,CDCl3)δ8.30(s,1H),7.39(s,1H),7.24(s,2H),6.73(dd,J＝16.6,9.9Hz,1H),6.65(s,1H),6.43(d,J＝16.6Hz,1H),6.24(d,J＝9.9Hz,1H),5.62-5.54(m,3H),4.45-4.35(m,4H),4.01(s,3H),2.49(s,3H).
实施例29：
1-(3-(4-氨基-5-(7-甲氧基-5-甲基苯并噻吩-2-基)-7H-吡咯[2,3-d]嘧啶-7-基)氮杂环丁烷-1-基)-2-氟-丙-2-炔-1-酮
Figure PCTCN2017087222-appb-000117
将60毫克5-(7-甲氧基-5-甲基苯并噻吩-2-基)-7-(氮杂环丁烷-3-基)-7H-吡咯[2,3-d]嘧啶-4-胺盐酸盐，17毫克2-氟-丙炔酸和80毫克苯并三唑-1-三(三甲氨基)-三氟磷酸酯溶于2毫升N,N-二甲基甲酰胺，滴加0.12毫升的N,N-二异丙基乙胺，室温下搅拌8小时停止反应，加水后用乙酸乙酯萃取，有机相用饱和氯化钠水溶液洗涤后无水硫酸钠干燥，过滤浓缩，残余物柱层析(二氯甲烷：甲醇＝97:3)分离得46毫克淡黄色固体，收率61％。
1H NMR(300MHz,DMSO-d6)δ8.19(s,1H),7.97(s,1H),7.35(s,1H),7.28(s,1H),6.80(s,1H),6.53(s,2H),5.70-60(m,2H),5.39–5.26(m,1H),4.85(s,2H),4.56–4.33(m,2H),3.95(s,3H),2.43(s,3H).
实施例30：
1-(3-(4-氨基-5-(1H-吲哚-2-基)-7H-吡咯[2,3-d]嘧啶-7-基)氮杂环丁烷-1-基)丙-2-烯-1-酮
Figure PCTCN2017087222-appb-000118
步骤1：制备1H-吲哚-1-甲酸叔丁酯
Figure PCTCN2017087222-appb-000119
将200毫克吲哚，0.7毫升三乙胺和48毫克的4-二甲氨基吡啶溶于2毫升二氯甲烷，再加入375毫克的二碳酸二叔丁酯，放至室温搅拌1小时候后浓缩反应液，柱层析(石油醚100％)分得370毫克油状物。
1H NMR(300MHz,CDCl3)δ8.16(d,J＝8.0Hz,1H),7.59(dd,J＝10.5,5.6Hz,2H),7.32(td,J＝8.0,1.3Hz,1H),7.27–7.19(m,1H),6.58(d,J＝3.7Hz,1H),1.68(s,9H).
步骤2：制备(1-甲酸叔丁酯-1H-吲哚-2-基)硼酸
Figure PCTCN2017087222-appb-000120
氩气保护下将370毫克的1H-吲哚-1-甲酸叔丁酯和0.6毫升三异丙基硼酸酯溶于3毫升干燥的四氢呋喃，冰浴冷却下滴加1.2当量的二异丙基氨基锂，冰浴下搅拌2小时，用3毫升的1N盐酸水溶液淬灭，加水后用乙酸乙酯萃取，有机相用饱和氯化钠水溶液洗涤后无水硫酸钠干燥，过滤浓缩，直接投下一步。
步骤3：制备2-(4-氨基-7-(1-(甲酸叔丁酯)氮杂环丁烷-3-基)-7H-吡咯[2,3-d]嘧啶-5-基)-1H-吲哚-1-甲酸叔丁酯
Figure PCTCN2017087222-appb-000121
将3-(4-氨基-5-碘-7H-吡咯[2,3-d]嘧啶-7-基)吡咯烷-1-甲酸叔丁酯替换成3-(4-氨基-5-碘-7H-吡咯[2,3-d]嘧啶-7-基)氮杂环丁烷-1-甲酸叔丁酯，(7-甲氧基-5-甲基苯并噻吩-2-基)硼酸替换成(1-甲酸叔丁酯-1H-吲哚-2-基)硼酸，其余所需原料、试剂及制备方法同实施例1中的步骤10，得2-(4-氨基-7-(1-(甲酸叔丁酯)氮杂环丁烷-3-基)-7H-吡咯[2,3-d]嘧啶-5-基)-1H-吲哚-1-甲酸叔丁酯。
1H NMR(300MHz,CDCl3)δ8.32(s,1H),8.29(d,J＝8.2Hz,1H),7.59(d,J＝7.3Hz,1H),7.42–7.29(m,3H),6.68(s,1H),5.65-5.56(m,1H),4.97(s,2H),4.55-4.49(m,2H),4.27-4.22(m,2H),1.48(s,9H),1.19(s,9H).
步骤4：制备7-(氮杂环丁烷-3-基)-5-(1H-吲哚-2-基)-7H-吡咯[2,3-d]嘧啶-4-胺 盐酸盐
Figure PCTCN2017087222-appb-000122
将3-(4-氨基-5-(7-甲氧基-5-甲基苯并噻吩-2-基)-7H-吡咯[2,3-d]嘧啶-7-基)吡咯烷-1-甲酸叔丁酯替换成2-(4-氨基-7-(1-(甲酸叔丁酯)氮杂环丁烷-3-基)-7H-吡咯[2,3-d]嘧啶-5-基)-1H-吲哚-1-甲酸叔丁酯，其余所需原料、试剂及制备方法同实施例1中的步骤11，得7-(氮杂环丁烷-3-基)-5-(1H-吲哚-2-基)-7H-吡咯[2,3-d]嘧啶-4-胺盐酸盐。
步骤5：制备1-(3-(4-氨基-5-(1H-吲哚-2-基)-7H-吡咯[2,3-d]嘧啶-7-基)氮杂环丁烷-1-基)丙-2-烯-1-酮
将5-(7-甲氧基-5-甲基苯并噻吩-2-基)-7-(吡咯烷-3-基)-7H-吡咯[2,3-d]嘧啶-4-氨基盐酸盐替换成7-(氮杂环丁烷-3-基)-5-(1H-吲哚-2-基)-7H-吡咯[2,3-d]嘧啶-4-胺盐酸盐，其余所需原料、试剂及制备方法同实施例1中的步骤12，得1-(3-(4-氨基-5-(1H-吲哚-2-基)-7H-吡咯[2,3-d]嘧啶-7-基)氮杂环丁烷-1-基)丙-2-烯-1-酮，33％。
1H NMR(300MHz,DMSO-d6)δ11.38(s,1H),8.20(s,1H),7.88(s,1H),7.56(d,J＝7.9Hz,1H),7.39(d,J＝7.8Hz,1H),7.10(t,J＝7.5Hz,1H),7.02(t,J＝7.3Hz,1H),6.61(s,1H),6.55(s,1H),6.40(dd,J＝17.0,10.3Hz,1H),6.17(d,J＝16.8Hz,1H),5.78–5.59(m,3H),4.79(t,J＝8.7Hz,1H),4.65(d,J＝6.1Hz,1H),4.55–4.44(m,1H),4.37(d,J＝5.9Hz,1H).
实施例31：
1-(3-(4-氨基-5-(7-甲氧基-5-甲基苯并噻吩-2-基)-7H-吡咯[2,3-d]嘧啶-7-基)氮杂环丁烷-1-基)-2-(吗啡啉亚甲基)丙-2-烯-1-酮
Figure PCTCN2017087222-appb-000123
步骤1：制备2-(吗啡啉亚甲基)丙烯酸
Figure PCTCN2017087222-appb-000124
将0.66克多聚甲醛和1克丙二酸溶于1,4二氧六环溶液中，再加入吗啡啉，加热至70℃反应2小时，反应液浓缩柱层析(二氯甲烷：甲醇＝98:2)，分离得850毫克白色固体，产率52％
1H NMR(300MHz,CDCl3)δ11.15(s,1H),6.38(s,1H),5.62(s,1H),3.78(s,4H),3.37(s,2H),2.66(s,4H).
步骤2：制备1-(3-(4-氨基-5-(7-甲氧基-5-甲基苯并噻吩-2-基)-7H-吡咯[2,3-d]嘧啶-7-基)氮杂环丁烷-1-基)-2-(吗啡啉亚甲基)丙-2-烯-1-酮
将2-氟-丙炔酸替换成2-(吗啡啉亚甲基)丙烯酸，其余所需原料、试剂及制备方法同实施例28，得1-(3-(4-氨基-5-(7-甲氧基-5-甲基苯并噻吩-2-基)-7H-吡咯[2,3-d]嘧啶-7-基)氮杂环丁烷-1-基)-2-(吗啡啉亚甲基)丙-2-烯-1-酮，45％。
1H NMR(300MHz,DMSO-d6)δ8.19(s,1H),7.83(s,1H),7.35(s,1H),7.27(s,1H),6.80(s,1H),6.54(s,2H),5.59(d,J＝20.1Hz,3H),4.66(s,2H),4.50–4.32(m,2H),3.95(s,3H),3.55(s,4H),3.11(s,2H),2.43(s,3H),2.37(s,4H).
实施例32：
(E)-1-(3-(4-氨基-5-(7-甲氧基-5-甲基苯并噻吩-2-基)-7H-吡咯[2,3-d]嘧啶-7-基)吡咯烷-1-基)-4-(4-乙基哌嗪-1-基)丁-2-烯-1-酮
Figure PCTCN2017087222-appb-000125
将丙炔酸替换成反式-4-(4-乙基哌嗪-1-基)巴豆酸盐酸盐，其余所需原料、试剂及制备方法同实施例3，得(E)-1-(3-(4-氨基-5-(7-甲氧基-5-甲基苯并噻吩-2-基)-7H-吡咯[2,3-d]嘧啶-7-基)吡咯烷-1-基)-4-(4-乙基哌嗪-1-基)丁-2-烯-1-酮，收率44％。
1H NMR(300MHz,CDCl3)δ8.33(s,1H),7.21(d,J＝2.5Hz,2H),7.10(d,J＝13.3Hz,1H),6.94(dd,J＝15.0,6.2Hz,1H),6.63(s,1H),6.30(dd,J＝23.5,15.2Hz,1H),5.50(d,J＝5.5Hz,3H),3.99(s,3H),4.32–3.58(m,4H),3.19(dd,J＝11.9,6.1Hz,2H),2.72(s,3H),2.83–2.59(m,10H),2.58–2.29(m, 2H),2.48(s,3H),1.30–1.07(m,3H).
实施例33：
(E)-1-(3-(4-氨基-5-(7-甲氧基-5-甲基苯并噻吩-2-基)-7H-吡咯[2,3-d]嘧啶-7-基)吡咯烷-1-基)-4-(氮杂环丁烷-1-基)丁-2-烯-1-酮
Figure PCTCN2017087222-appb-000126
将丙炔酸替换成反式-4-(氮杂环丁烷-1-基)巴豆酸盐酸盐，其余所需原料、试剂及制备方法同实施例3，得(E)-1-(3-(4-氨基-5-(7-甲氧基-5-甲基苯并噻吩-2-基)-7H-吡咯[2,3-d]嘧啶-7-基)吡咯烷-1-基)-4-(氮杂环丁烷-1-基)丁-2-烯-1-酮，收率47％。
1H NMR(300MHz,CDCl3)δ8.33(s,1H),7.24–7.18(m,2H),7.10(d,J＝11.0Hz,1H),6.94–6.79(m,1H),6.64(s,1H),6.37(dd,J＝38.3,15.0Hz,1H),5.56–5.39(m,3H),4.29–3.65(m,4H),3.99(s,3H),3.48–3.20(m,6H),2.48(s,3H),2.65–2.09(m,4H).
实施例34：
1-(3-(4-氨基-5-(7-甲氧基-5-甲基苯并噻吩-2-基)-7H-吡咯[2,3-d]嘧啶-7-基)吡咯烷-1-基)丁-2-炔-1-酮
Figure PCTCN2017087222-appb-000127
将丙炔酸替换成2-丁炔酸，其余所需原料、试剂及制备方法同实施例3，得1-(3-(4-氨基-5-(7-甲氧基-5-甲基苯并噻吩-2-基)-7H-吡咯[2,3-d]嘧啶-7-基)吡咯烷-1-基)丁-2-炔-1-酮，收率33％。
1H NMR(300MHz,CDCl3)δ8.32(d,J＝3.8Hz,1H),7.24–7.19(m,2H),7.11(d,J＝11.6Hz,1H),6.64(s,1H),5.62(s,2H),5.55–5.40(m,1H),4.32–3.61 (m,4H),4.00(s,3H),2.49(s,3H),2.68–2.22(m,2H),2.00(d,J＝15.2Hz,3H).
实施例35：
(S)-1-(3-(4-氨基-5-(7-甲氧基-5-甲基苯并噻吩-2-基)-7H-吡咯[2,3-d]嘧啶-7-基)吡咯烷-1-基)丙-2-炔-1-酮
Figure PCTCN2017087222-appb-000128
将100毫克(S)-5-(7-甲氧基-5-甲基苯并噻吩-2-基)-7-(吡咯烷-3-基)-7H-吡咯[2,3-d]嘧啶-4-氨基盐酸，20微升丙炔酸和145毫克2-(7-偶氮苯并三氮唑)-N,N,N',N'-四甲基脲六氟磷酸酯溶于2毫升N,N-二甲基甲酰胺，滴加0.2毫升的N,N-二异丙基乙胺，室温下搅拌8小时停止反应，加水后用乙酸乙酯萃取，有机相用饱和氯化钠水溶液洗涤后无水硫酸钠干燥，过滤浓缩，残余物柱层析(二氯甲烷：甲醇＝97:3)分离得48毫克黄色固体，收率44％。
1H NMR(300MHz,CDCl3)δ8.33(s,1H),7.22(s,2H),7.11(d,J＝9.3Hz,1H),6.65(s,1H),5.69(s,2H),5.59–5.40(m,1H),4.37–3.57(m,4H),4.00(s,3H),3.09(d,J＝15.2Hz,1H),2.64–2.24(m,2H),2.49(s,3H).
实施例36：
1-(3-(4-氨基-5-(2-甲基-1H-苯并咪唑-5-基)-7H-吡咯[2,3-d]嘧啶-7-yl)吡咯烷-1-基)丙-2-烯-1-酮
Figure PCTCN2017087222-appb-000129
将(7-甲氧基-5-甲基苯并噻吩-2-基)硼酸替换成2-甲基-5-(4,4,5,5-四甲基-1,3,2-二氧硼烷-2-基)-1H-苯并咪唑-1-甲酸叔丁酯(制备方法见专利WO2011055215)，其余所需原料、试剂及制备方法同实施例1，收率37％。
1H NMR(400MHz,DMSO)δ8.17(s,1H),7.55(d,J＝8.1Hz,1H),7.50(s,1H),7.39(s,0.5H),7.34(s,0.5H),7.23(d,J＝8.1Hz,1H),6.62(m,1H),6.22-6.12(m,1H),5.73-5.63(m,1H),5.45–5.30(m,1H),3.85–3.75(m,1H),3.65–3.48(m,3H),2.50(s,3H),2.48–2.25(m,2H).
实施例37
(S)-1-(3-(5-(7-甲氧基-5-甲基苯并噻吩-2-基)-4-甲氨基-7H-吡咯[2,3-d]嘧啶-7-基)吡咯烷-1-基)丙-2-烯-1-酮
Figure PCTCN2017087222-appb-000130
制备方法同实施例12，收率55％。
1H NMR(300MHz,DMSO-d)δ8.28(s,1H),7.55(d,J＝17.7Hz,1H),7.28(d,J＝6.9Hz,2H),6.79(s,1H),6.71–6.53(m,1H),6.16(d,J＝16.8Hz,2H),5.71(dd,J＝22.5,9.6Hz,1H),5.37(d,J＝25.8Hz,1H),4.21–3.99(m,1H),3.94(s,3H),3.73(d,J＝6.9Hz,2H),3.54(dd,J＝15.4,8.9Hz,1H),2.95(d,J＝3.7Hz,3H),2.46–2.31(m,5H).
实施例38
(S,E)-1-(3-(4-氨基-5-(7-甲氧基-5-甲基苯并噻吩-2-基)-7H-吡咯[2,3-d]嘧啶-7-基)吡咯烷-1-基)-4-二甲氨基-丁-2-烯-1-酮
Figure PCTCN2017087222-appb-000131
制备方法同实施例26，收率47％。
1H NMR(300MHz,DMSO-d)δ8.29(s,1H),7.67(s,1H),7.34(s,1H),6.85(s,1H),6.69–6.42(m,2H),5.59–5.41(m,1H),4.10(s,1H),3.95(s,3H),3.86 (d,J＝7.2Hz,3H),3.23(d,J＝12.7Hz,2H),2.47–2.34(m,5H),2.30(s,3H),2.27(s,3H).
实施例39
(S)-1-(3-(4-氨基-3-(7-甲氧基-5-甲基苯并噻吩-2-基)1H-吡唑并[3,4-d]嘧啶-1-基)吡咯烷-1-基)丙-2-炔-1-酮
Figure PCTCN2017087222-appb-000132
制备方法同实施例26，收率39％。
1H NMR(300MHz,DMSO-d)δ8.29(s,1H),7.68(s,1H),7.35(s,1H),6.85(s,1H),5.52(s,1H),4.51(d,J＝23.6Hz,1H),4.10(d,J＝16.8Hz,1H),3.96(s,3H),3.91–3.51(m,3H),2.44(s,5H).
实施例40
(S)-3-(7-甲氧基-5-甲基苯并噻吩-2-基)-1-(1-乙烯磺酰基)吡咯烷-3-基)-1H-吡唑[3,4-d]嘧啶-4-胺)
Figure PCTCN2017087222-appb-000133
制备方法同实施例27，收率46％。
1H NMR(300MHz,DMSO-d)δ8.29(s,1H),7.69(s,1H),7.36(s,1H),6.88(dd,J＝17.9,8.4Hz,2H),6.10(dd,J＝16.8,13.4Hz,2H),5.48(dt,J＝10.7,5.4Hz,1H),3.97(s,3H),3.79(dd,J＝10.6,7.1Hz,1H),3.59(dd,J＝15.8,6.2Hz,2H),3.46(dd,J＝9.9,6.8Hz,1H),2.47–2.35(m,5H).
实施例41
1-(3-(4-氨基-5-(4,7-二甲氧基苯并噻吩-2-基)-7H-吡咯[2,3-d]嘧啶-7-基)氮杂环丁烷-1-基)丙-2-烯-1-酮
Figure PCTCN2017087222-appb-000134
制备方法同实施例13，收率60％。
1H NMR(300MHz,DMSO-d)δ8.19(s,1H),7.95(s,1H),7.40(s,1H),6.86(s,2H),6.54(s,2H),6.45–6.30(m,1H),6.15(d,J＝17.1Hz,1H),5.71(d,J＝10.2Hz,1H),5.62(s,1H),4.72(d,J＝8.2Hz,2H),4.42(d,J＝9.2Hz,2H),3.91(s,3H),3.88(s,3H).
实施例42
1-(3-(4-氨基-5-(6-甲氧基苯并噻吩-2-基)-7H-吡咯[2,3-d嘧啶-7-基)氮杂环丁烷-1-基)丙-2-烯-1-酮
Figure PCTCN2017087222-appb-000135
制备方法同实施例1，收率45％。
1H NMR(300MHz,DMSO-d)δ8.18(s,1H),7.93(d,J＝8.8Hz,2H),7.87(d,J＝9.1Hz,1H),7.80(s,1H),7.63(d,J＝8.3Hz,1H),7.38(s,1H),7.21(d,J＝8.9Hz,1H),6.38(dd,J＝17.1,10.3Hz,1H),6.16(d,J＝17.1Hz,1H),5.71(d,J＝12.4Hz,1H),5.62(d,J＝8.7Hz,1H),4.72(m,J＝2H),4.42(m,2H),3.89(s,3H).
实施例43
1-(3-(4-氨基-5-(苯并噻吩-2-基)-7H-吡咯[2,3-d]嘧啶-7-基)氮杂环丁基-1-基)丙-2-烯-1-酮
Figure PCTCN2017087222-appb-000136
制备方法同实施例1，收率43％。
1H NMR(300MHz,DMSO-d)δ8.18(s,1H),7.97(d,J＝7.3Hz,1H),7.90(s,1H),7.85(d,J＝7.0Hz,1H),7.44(s,1H),7.36(td,J＝13.1,6.5Hz,2H),6.54(s,2H),6.36(dd,J＝17.0,10.2Hz,1H),6.19–6.08(m,1H),5.74(s,1H),5.72–5.67(m,1H),5.61(dt,J＝13.8,6.8Hz,1H),4.77–4.64(m,2H),4.48–4.32(m,2H).
实施例44
1-(3-(4-氨基-5-(7-甲氧基-5-甲基苯并噻吩-2-基)-7H-吡咯[2,3-d]嘧啶-7-基)氮杂环丁烷-1-基)-2-(吡咯烷-1-亚甲基)丙-2-烯-1-酮
Figure PCTCN2017087222-appb-000137
制备方法同实施例31，收率45％。
1H NMR(300MHz,DMSO-d6)δ8.19(s,1H),7.93(s,1H),7.35(s,1H),7.28(s,1H),6.81(s,1H),6.52(s,2H),5.59(d,J＝20.1Hz,3H),4.77(s,2H),4.50–4.35(m,2H),3.95(s,3H),3.4-3.2(6H),2.44(s,3H),1.89(s,4H).
实施例45
(S)1-(3-(4-氨基-5-(7-甲氧基-5-甲基苯并噻吩-2-基)-7H-吡咯[2,3-d]嘧啶-7-基)吡咯烷-1-基)-2-(吗啡啉亚甲基)丙-2-烯-1-酮
Figure PCTCN2017087222-appb-000138
制备方法同实施例31，收率47％。
1H NMR(300MHz,DMSO-d6)δ8.19(s,1H),7.50(d,J＝9.4Hz,1H),7.31(d,J＝6.6Hz,1H),7.27(s,1H),6.80(s,1H),6.48(s,2H),5.46-5.35(m,2H),5.29-5.26(m,1H),4.05–3.62(m,4H),3.94(s,3H),3.40–3.20(m,6H),2.43(s,3H),2.37(s,4H),2.24(s,2H).
实施例46
(S)1-(3-(4-氨基-5-(7-甲氧基-5-甲基苯并噻吩-2-基)-7H-吡咯[2,3-d]嘧啶-7-基)吡咯烷-1-基)-2-((二乙胺基)亚甲基)丙-2-烯-1-酮
Figure PCTCN2017087222-appb-000139
制备方法同实施例31，收率49％。
1H NMR(300MHz,DMSO-d6)δ8.19(s,1H),7.50(d,J＝9.4Hz,1H),7.31(d,J＝6.6Hz,1H),7.27(s,1H),6.80(s,1H),6.48(s,2H),5.46-5.35(m,2H),5.29-5.26(m,1H),4.05–3.62(m,4H),3.94(s,3H),3.40–3.20(m,6H),2.43(s,3H),2.40(s,2H),1.02-0.68(m,6H).
实施例47
(S)1-(3-(4-氨基-5-(7-甲氧基-5-甲基苯并噻吩-2-基)-7H-吡咯[2,3-d]嘧啶-7-基)吡咯烷-1-基)-2-((二甲胺基)亚甲基)丙-2-烯-1-酮
Figure PCTCN2017087222-appb-000140
制备方法同实施例31，收率43％
1H NMR(300MHz,DMSO-d6)δ8.19(s,1H),7.53(d,J＝9.4Hz,1H),7.31(d,J＝6.6Hz,1H),7.27(s,1H),6.80(s,1H),6.48(s,2H),5.52-5.41(m,2H),5.38-5.29(m,1H),4.05–3.62(m,4H),3.94(s,3H),3.40–3.20(m,6H),2.43(s,3H),2.40(s,2H),2.22(s,3H),2.11(s,3H).
实施例48
(S)1-(3-(4-氨基-5-(7-甲氧基-5-甲基苯并噻吩-2-基)-7H-吡咯[2,3-d]嘧啶-7-基)吡咯烷-1-基)-2-(哌啶-1-亚甲基)丙-2-烯-1-酮
Figure PCTCN2017087222-appb-000141
制备方法同实施例31，收率44％。
1H NMR(300MHz,DMSO-d6)δ8.19(s,1H),7.48(s,1H),7.31(d,J＝6.6Hz,1H),7.27(s,1H),6.80(s,1H),6.48(s,2H),5.46-5.35(m,2H),5.29-5.26(m,1H),4.05–3.62(m,4H),3.94(s,3H),3.40–3.20(m,6H),2.43(s,3H),2.42(s,2H),1.42-1.18(m,6H).
实施例49
(S)1-(3-(4-氨基-5-(7-甲氧基-5-甲基苯并噻吩-2-基)-7H-吡咯[2,3-d]嘧啶-7-基)吡咯烷-1-基)-2-((4-甲基哌嗪-1-基)-亚甲基)丙-2-烯-1-酮
Figure PCTCN2017087222-appb-000142
制备方法同实施例31，收率49％
1H NMR(300MHz,DMSO-d6)δ8.19(s,1H),7.48(d,J＝9.4Hz,1H),7.31(d,J＝6.6Hz,1H),7.27(s,1H),6.80(s,1H),6.48(s,2H),5.46-5.35(m,2H),5.29-5.26(m,1H),4.05–3.62(m,4H),3.94(s,3H),3.40–3.20(m,2H),2.43(s,3H),2.50(m,8H),2.24(s,3H).
实施例50
N-(2-(4-氨基-3-(7-甲氧基-5-甲基苯并噻吩-2-基)-1H-吡唑[3,4-d]嘧啶-1-基)乙基)丙烯酰胺
Figure PCTCN2017087222-appb-000143
制备方法同实施例31，收率46％。
1H NMR(300MHz,DMSO-d6)δ8.37(s,1H),7.53(s,1H),7.27(s,1H),6.91(s,1H),6.69(s,1H),6.26(d,J＝17.2Hz,1H),6.15–6.01(m,3H),5.63(d,J＝10.2Hz,1H),4.69–4.59(m,2H),4.01(s,3H),3.88(dd,J＝10.2,5.3Hz,2H),2.50(s,3H).
实施例51
1-(3-(4-氨基-5-(7-甲氧基-5-甲基苯并噻吩-2-基)-7H-吡咯[2,3-d]嘧啶-7-基)氮杂环丁烷-1-基)-2-((二乙胺基)亚甲基)丙-2-烯-1-酮
Figure PCTCN2017087222-appb-000144
制备方法同实施例31
1H NMR(300MHz,DMSO-d6)δ8.18(s,1H),7.83(s,1H),7.34(s,1H),7.28(s,1H),6.80(s,1H),6.51(s,2H),5.65-5.45(m,3H),4.67(s,2H),4.45–4.37(m,2H),3.94(s,3H),3.40–3.20(m,6H),2.43(s,3H),1.02-0.68(m,6H).
实施例52
1-(3-(4-氨基-5-(7-甲氧基-5-甲基苯并噻吩-2-基)-7H-吡咯[2,3-d]嘧啶-7-基)氮杂环丁烷-1-基)-2-((4-甲基哌嗪-1-基)亚甲基)丙-2-烯-1-酮
Figure PCTCN2017087222-appb-000145
制备方法同实施例31
1H NMR(300MHz,DMSO-d6)δ8.19(s,1H),7.78(s,1H),7.34(s,1H),7.28(s,1H),6.80(s,1H),6.51(s,2H),5.60-5.53(m,3H),4.70-4.62(m,2H),4.48–4.31(m,2H),3.94(s,3H),3.40–3.20(m,4H),3.12(s,2H),2.43(s,7H),2.18(s,3H).
实施例53
1-(3-(4-氨基-5-(7-甲氧基-5-甲基苯并噻吩-2-基)-7H-吡咯[2,3-d]嘧啶-7-基)氮杂环丁烷-1-基)-2-((二甲胺基)亚甲基)丙-2-烯-1-酮
Figure PCTCN2017087222-appb-000146
制备方法同实施例31，收率47％
1H NMR(300MHz,DMSO-d6)δ8.18(s,1H),7.85(s,1H),7.35(s,1H),7.28(s,1H),6.81(s,1H),6.51(s,2H),5.75-5.68(m,2H),5.60-5.52(m,1H),4.72-4.71(m,2H),4.50–4.42(m,1H),4.38–4.32(m,1H),3.95(s,3H),3.40–3.20(m,2H),2.44(s,3H),2.31(s,6H).
实施例54
(S)1-(3-(4-氨基-3-(7-甲氧基-5-甲基苯并噻吩-2-基)-1H-吡唑[3,4-d]嘧啶-1-基)吡咯烷-1-基)-2-(吗啡啉亚甲基)丙-2-烯-1-酮
Figure PCTCN2017087222-appb-000147
制备方法同实施例31，收率46％。
1H NMR(300MHz,DMSO-d6)δ8.29(s,1H),7.67(s,1H),7.34(s,1H),7.25(s,2H),6.85(s,1H),5.57–5.30(m,3H),4.11(m,1H),3.96(s,3H),3.81(m,3H),3.40–3.20(m,6H),2.44(s,3H),2.43–2.34(m,2H),2.37(m,4H).
实施例55
1-(3-(4-氨基-3-(7-甲氧基-5-甲基苯并噻吩-2-基)-1H-吡唑[3,4-d]嘧啶-1-基)氮杂环丁烷-1-基)-2-(吗啡啉亚甲基)丙-2-烯-1-酮
Figure PCTCN2017087222-appb-000148
制备方法同实施例31，收率45％
1H NMR(300MHz,DMSO-d6)δ8.29(s,1H),7.71(s,1H),7.36(s,1H),7.25(s,2H),6.86(s,1H),5.75–5.65(m,1H),5.75(s,2H),4.75-4.67(m,2H),4.48-4.38(m,2H),3.96(s,3H),3.59(s,3H),3.40–3.20(m,2H),2.45(s,3H),2.37(m,4H).
实施例56
(S)1-(3-(4-氨基-3-(7-甲氧基-5-甲基苯并噻吩-2-基)-1H-吡唑[3,4-d]嘧啶-1-基)吡咯烷-1-基)丙-2-烯-1-酮
Figure PCTCN2017087222-appb-000149
制备方法同实施例2，收率55％。
1H NMR(300MHz,DMSO-d6)δ8.29(s,1H),7.67(s,1H),7.34(s,1H),7.25(s,2H),6.85(s,1H),6.75–6.48(m,2H),6.16(d,J＝16.9Hz,1H),5.75–5.61(m,2H),5.51(d,J＝20.1Hz,1H),4.11(m,1H),3.94(d,J＝9.9Hz,3H),3.81(m,3H),2.44(s,3H),2.43–2.34(m,2H).
实施例57
1-(3-(4-氨基-5-(5,7-二甲氧基苯并噻吩-2-基)-7H-吡咯[2,3-d]嘧啶-7-基)吡咯烷-1-基)丙-2-烯-1-酮
Figure PCTCN2017087222-appb-000150
步骤1：将实施例1步骤4中的原料2-甲氧基-4-甲基苯硫酚替换成2,4-二甲氧基苯硫酚，其余步骤同实施例1,可制备得到标题化合物1-(3-(4-氨基-5-(5,7-二甲氧基苯并噻吩-2-基)-7H-吡咯[2,3-d]嘧啶-7-基)吡咯烷-1-基)丙-2-烯-1-酮。
1H NMR(300MHz,CDCl3)δ8.33(s,1H),7.21(s,1H),7.11(d,J＝10.2Hz,1H),6.87(s,1H),6.48(s,2H),6.42(d,J＝5.6Hz,1H),5.74(s,1H),5.60(s,2H),5.49(s,1H),4.05(s,2H),3.97(s,3H),3.88(s,3H),3.81(s,2H),2.45(s,3H).
实施例58
1-(3-(4-氨基-5-(7-甲氧基-5-(三氟甲基)苯并噻吩-2-基)-7H-吡咯[2,3-d]嘧啶-7-基)吡咯烷-1-基)丙-2-烯-1-酮
Figure PCTCN2017087222-appb-000151
步骤1：制备2-碘-5-(三氟甲基)苯酚
Figure PCTCN2017087222-appb-000152
将495mg钠氢(60％)混选于40ml干燥得甲苯中，冰浴冷却。缓慢滴加3-三氟甲基苯酚，滴加完后搅拌10分钟后再加入2.08g单质碘。放置室温搅拌16小时。3N盐酸水溶液稀释，乙酸乙酯萃取，硫代硫酸钠水溶液洗，合并有机相浓缩柱层析(石油醚：乙酸乙酯＝98:2)可获得2.35g油状物。
1H NMR(300MHz,CDCl3)δ5.68(s,1H),6.93(dd,J＝8.4,2.1Hz,1H),7.22(d,J＝ 2.1Hz,1H),7.79(d,J＝8.4Hz,1H).
步骤2：制备1-碘-2-甲氧基-4-(三氟甲基)苯
Figure PCTCN2017087222-appb-000153
将步骤1获得的2-碘-5-(三氟甲基)苯酚2.35g溶于25ml乙醇，加入3.9g碳酸钾和1.4g碘甲烷，氩气保护下回流13小时。硅藻土过滤，滤液浓缩干柱层析(石油醚100％)可定量获得2g油状物。
1H NMR(300MHz,CDCl3)δ3.92(s,3H),6.94(d,J＝8.1Hz,1H),6.99(s,1H),7.86(d,J＝8.1Hz,1H).
步骤3：制备(2,2-二乙氧基)(2-甲氧基-4-(三氟甲基)苯)硫烷
Figure PCTCN2017087222-appb-000154
以步骤2获得的1-碘-2-甲氧基-4-(三氟甲基)苯为原料，通过Adv.Synth.Catal.2015,357,2205–2212报道的方法可获得(2,2-二乙氧基)(2-甲氧基-4-(三氟甲基)苯)硫烷。
1H NMR(300MHz,CDCl3)δ7.37(d,J＝8.1Hz,1H),7.17(d,J＝8.1Hz,1H),7.00(s,1H),4.68(t,J＝5.5Hz,1H),3.93(s,3H),3.69(tt,J＝14.1,7.1Hz,2H),3.62–3.46(m,2H),3.14(d,J＝5.5Hz,2H),1.20(t,J＝7.1Hz,6H).
步骤4：将实施例1步骤5的原料(2,2-二乙氧基乙基)(2-甲氧基-4-甲基苯基)硫醚替换成(2,2-二乙氧基)(2-甲氧基-4-(三氟甲基)苯)硫烷，按照实施例1的步骤可获得标题化合物。
1H NMR(300MHz,CDCl3)δ8.35(s,1H),7.70(s,1H),7.35(s,1H),7.15(d,J＝10.1Hz,1H),6.96(s,1H),6.49(d,J＝8.6Hz,1H),6.43(d,J＝7.1Hz,1H),5.80–5.68(m,1H),5.53(m,3H),4.24–4.09(m,1H),4.07(s,3H),3.85(m,3H),2.59(m,1H),2.44(m,1H).
实施例59
(S)-1-(3-(4-氨基-5-(5-氯-7-甲氧基苯并噻吩-2-基)-7H-吡咯[2,3-d]嘧啶-7-基)吡咯烷-1-基)丙-2-烯-1-酮
Figure PCTCN2017087222-appb-000155
步骤1：制备4-氯-2-甲氧基苯胺
Figure PCTCN2017087222-appb-000156
将3g的铁粉和375mg的氯化铵固体在50ml水中加热回流15分钟，再加入2g的4-氯-2-甲氧基硝基苯。回流反应1.5个小时后冷至室温，饱和碳酸氢钠水溶液调节PH至中性。硅藻土过滤后，滤液用乙酸乙酯萃取，无水硫酸钠干燥得1.45g产物，收率86％。
步骤2：制备4-氯-1-碘-2-甲氧基苯
Figure PCTCN2017087222-appb-000157
将1.45g的4-氯-2-甲氧基苯胺溶于60ml水和10ml浓硫酸的溶液中，冰浴冷却至5摄氏度以下。再将634mg亚硝酸钠溶于1ml水中缓慢滴加至反应液，冰浴冷却下搅拌45分钟后，再滴加1.98g的碘化钾水溶液5ml。移至室温搅拌1小时后，乙酸乙酯萃取，饱和硫代硫酸钠水溶液洗，无水硫酸钠干燥后柱层析(PE100％)得2.17g液体，收率88％。
步骤3：制备(4-氯-2-甲氧基苯)(2,2-二乙氧基乙基)硫醚
Figure PCTCN2017087222-appb-000158
以步骤2获得的4-氯-1-碘-2-甲氧基苯为原料，通过Adv.Synth.Catal.2015,357,2205–2212报道的方法可获得(4-氯-2-甲氧基苯)(2,2-二乙氧基乙基)硫醚。
1H NMR(300MHz,CDCl3)δ7.27(d,J＝8.3Hz,1H),6.89(dd,J＝8.3,2.1Hz,1H),6.83(d,J＝2.0Hz,1H),4.62(t,J＝5.6Hz,1H),3.88(s,3H),3.72–3.47(m,4H),3.06(d,J＝5.6Hz,2H),1.20(m,6H).
步骤4：将实施例1步骤5的原料(2,2-二乙氧基乙基)(2-甲氧基-4-甲基苯基)硫醚替换成(4-氯-2-甲氧基苯)(2,2-二乙氧基乙基)硫醚，按照实施例1的步骤可获得标题化合物。
1H NMR(300MHz,CDCl3)δ8.34(s,1H),7.40(s,1H),7.22(s,1H),7.12(d,J＝10.4Hz,1H),6.78(s,1H),6.48(d,J＝8.5Hz,1H),6.42(d,J＝5.0Hz,1H),5.70-5.80(m,1H),5.57(s,2H),5.45-5.55(m,1H),4.05-4.14(m,2H),4.00(s,3H),3.75-3.90(m,3H),2.40-2.60(m,2H).
以下化合物的制备方法与化合物59类似。
Figure PCTCN2017087222-appb-000159
Figure PCTCN2017087222-appb-000160
实施例61
1-(3-(4-氨基-2-氯-5-(7-甲氧基-5-甲基苯并噻吩-2-基)-7H-吡咯[2,3-d]嘧啶-7-基)氮杂环丁烷-1-基)丙-2-烯-1-酮
Figure PCTCN2017087222-appb-000161
将实施例13中步骤1的原料4-氯-7H-吡咯[2,3-d]嘧啶替换成2,4-二氯-7H-吡咯[2,3-d]嘧啶，操作步骤同实施例13。可制备得1-(3-(4-氨基-2-氯-5-(7-甲氧基-5-甲基苯并噻吩-2-基)-7H-吡咯[2,3-d]嘧啶-7-基)氮杂环丁烷-1-基)丙-2-烯-1-酮，收率45％。
1H NMR(300MHz,CDCl3)δ7.36(s,1H),7.23(s,2H),6.66(s,1H),6.42(d,J＝16.9Hz,1H),6.27(d,J＝10.3Hz,1H),5.80–5.65(m,4H),4.73(m,2H),4.45(s,2H),4.00(s,3H),2.49(s,3H).
以下化合物的制备方法与化合物61类似。
Figure PCTCN2017087222-appb-000162
实施例63
1-(3-(4-(二甲氨基)-5-(7-甲氧基-5-甲基苯并噻吩-2-基)-7H-吡咯[2,3-d]嘧啶-7-基)吡咯烷-1-基)丙-2-烯-1-酮
Figure PCTCN2017087222-appb-000163
将实施例12中步骤2的原料氨水替换成二甲胺的水溶液，其余步骤同实施例12。制备得1-(3-(4-(二甲氨基)-5-(7-甲氧基-5-甲基苯并噻吩-2-基)-7H-吡咯[2,3-d]嘧啶-7-基)吡咯烷-1-基)丙-2-烯-1-酮。收率59％。
1H NMR(300MHz,CDCl3)δ8.42(s,1H),7.22–7.08(m,3H),6.62(s,1H),6.45(m,2H),5.73(m,1H),5.53(m,1H),4.25–4.03(m,2H),4.00(s,3H),3.79(s,2H),2.94(s,6H),2.54(s,1H),2.48(s,3H),2.44–2.36(m,1H).
实施例66
1-(3-(4-氨基-5-(7-甲氧基-5-甲基苯并噻吩-2-基)-2-(2-吗啡啉乙氧基)-7H-吡咯[2,3-d]嘧啶-7-基)吡咯烷-1-基)丙-2-烯-1-酮
Figure PCTCN2017087222-appb-000164
步骤1：制备6-氨基-5-(2,2-二乙氧基)-2-巯基嘧啶-4-醇
Figure PCTCN2017087222-appb-000165
将2.15g的2-氰基-4,4-二乙氧基丁酸乙酯溶于5.3ml的20％乙醇钠乙醇溶液中，再加入1.2g硫脲，加热回流过夜。次日浓缩干反应液，再用38ml的水稀释，乙醚洗涤。水相用醋酸调节PH至中性，大量固体析出，过滤，收集固体干燥，得1.5g浅黄色固体，收率62％。
1H NMR(300MHz,DMSO-d6)δ11.75(s,1H),11.44(s,1H),6.07(s,2H),4.50(t,J＝5.6,1H),3.59(dq,J＝7.0,9.5,2H),3.40(dq,J＝7.0,9.6,2H),2.44(d,J＝5.6,2H),1.07(t,J＝7.0,6H)；
步骤2：制备2-硫代-1,2,3,7-四氢-4H-吡咯[2,3-d]嘧啶-4-酮。
Figure PCTCN2017087222-appb-000166
将步骤2得到的6-氨基-5-(2,2-二乙氧基)-2-巯基嘧啶-4-醇1g于12ml的0.2N HCl水溶液中室温搅拌24小时，大量固体析出，过滤干燥定量得到2-硫代-1,2,3,7-四氢-4H-吡咯[2,3-d]嘧啶-4-酮。
1H NMR(300MHz,DMSO-d6)δ13.20(s,1H),11.86(s,1H),11.26(s,1H),6.72(s, 1H),6.33(d,J＝2.8Hz,1H).
步骤3：制备2-(甲硫基)-3,7-二氢-4H-吡咯[2,3-d]嘧啶-4-酮。
Figure PCTCN2017087222-appb-000167
320mg氢氧化钠溶于15ml乙醇中，将步骤3得到的2-硫代-1,2,3,7-四氢-4H-吡咯[2,3-d]嘧啶-4-酮660mg溶于该溶液中，冰浴冷却下滴加0.25ml碘甲烷。完毕后恢复室温，搅拌3小时。浓缩反应液，少量水溶解，用5N的HCl调节PH至弱酸性，大量固体析出。过滤收集固体，干燥得700mg白色固体。收率97％。
1H NMR(300MHz,DMSO-d6)δ12.02(s,1H),11.74(s,1H),6.89-6.91(m,1H),6.34-6.36(m,1H),2.09(s,3H).
步骤4：制备4-氯-2-(甲硫基)-7H-吡咯[2,3-d]嘧啶。
Figure PCTCN2017087222-appb-000168
将700mg步骤3得到的2-(甲硫基)-3,7-二氢-4H-吡咯[2,3-d]嘧啶-4-酮于10ml三氯氧磷中加热回流过夜，次日浓缩反应液，加水稀释后乙酸乙酯萃取，饱和氯化钠水洗，无水硫酸钠干燥有机相，浓缩干定量得4-氯-2-(甲硫基)-7H-吡咯[2,3-d]嘧啶。
1H NMR(300MHz,DMSO-d6)δ12.40(s,1H),7.52(d,J＝3.4Hz,1H),6.51(s,1H),2.55(s,3H).
步骤5：制备4-氯-5-碘-2-(甲硫基)-7H-吡咯[2,3-d]嘧啶。
Figure PCTCN2017087222-appb-000169
将上述制备的4-氯-2-(甲硫基)-7H-吡咯[2,3-d]嘧啶100mg和146mgN-碘代丁二酰亚胺溶于2mlN,N-二甲基甲酰胺中，室温搅拌18小时。用水稀释，大量固体析出，过滤干燥的140mg紫色固体，收率91％。
1H NMR(300MHz,DMSO-d6)δ12.72(s,1H),7.75(s,1H),2.55(s,3H).
步骤6：制备3-(4-氯-5-碘-2-(甲硫基)-7H-吡咯[2,3-d]嘧啶-7-基)吡咯烷-1-甲酸叔丁酯。
Figure PCTCN2017087222-appb-000170
将实施例1中步骤8的4-氯-5-碘-7H-吡咯[2,3-d]嘧啶替换成4-氯-5-碘-2-(甲硫基)-7H-吡咯[2,3-d]嘧啶，其余所需原料、操作步骤同实施例1步骤8，定量得到3-(4-氯-5-碘-2-(甲硫基)-7H-吡咯[2,3-d]嘧啶-7-基)吡咯烷-1-甲酸叔丁酯。
1H NMR(300MHz,CDCl3)δ7.20(s,1H),5.37(s,1H),3.88(dd,J＝11.7,6.9Hz,1H),3.55(s,3H),2.60(s,3H),2.42(td,J＝13.9,7.2Hz,1H),2.22(td,J＝13.2,6.7Hz,1H),1.48(s,9H).
步骤7：制备3-(4-氨基-5-碘-2-(甲硫基)-7H-吡咯[2,3-d]嘧啶-7-基)吡咯烷-1-甲酸叔丁酯。
Figure PCTCN2017087222-appb-000171
制备方法同实施例1中的步骤9。
1H NMR(300MHz,CDCl3)δ6.91(s,1H),5.55(s,2H),5.31(s,1H),3.91–3.77(m,1H),3.53(s,3H),2.55(s,3H),2.37(dd,J＝13.5,6.5Hz,1H),2.28–2.13(m,1H),1.48(s,9H).
步骤8：制备3-(4-氨基-5-碘-2-(甲磺酰基)-7H-吡咯[2,3-d]嘧啶-7-基)吡咯烷-1-甲酸叔丁酯。
Figure PCTCN2017087222-appb-000172
将上述制备的3-(4-氨基-5-碘-2-(甲硫基)-7H-吡咯[2,3-d]嘧啶-7-基)吡咯烷-1-甲酸叔丁酯1.2g溶于乙醇中，分批加入2.5当量的间氯过氧苯甲酸室温搅拌过夜。次日浓缩干柱层析(二氯甲烷：甲醇＝98:2)，得1.28g的3-(4-氨基-5-碘-2-(甲磺酰基)-7H-吡咯[2,3-d]嘧啶-7-基)吡咯烷-1-甲酸叔丁酯。
1H NMR(300MHz,CDCl3)δ7.21(s,1H),6.20(s,2H),5.46(s,1H),3.87(s,1H), 3.70-3.50(m,3H),3.30(s,3H),2.42(s,1H),2.19(s,1H),1.49(s,9H).
步骤9：制备3-(4-氨基-5-碘-2-(2-吗啡啉乙氧基)-7H-吡咯[2,3-d]嘧啶-7-基)吡咯烷-1-甲酸叔丁酯。
Figure PCTCN2017087222-appb-000173
将413mg的2-吗啡啉乙醇和384mg叔丁醇钾溶于四氢呋喃，室温搅拌10分钟后加入3-(4-氨基-5-碘-2-(甲磺酰基)-7H-吡咯[2,3-d]嘧啶-7-基)吡咯烷-1-甲酸叔丁酯800mg，80℃封管过夜。次日浓缩干反应液，柱层析(二氯甲烷：甲醇＝98:2+0.1％氨水)，得到780mg固体，收率89％。
1H NMR(300MHz,CDCl3)δ6.87(s,1H),5.56(s,2H),5.24(s,1H),4.45(t,J＝6.0Hz,2H),3.80(s,1H),3.75–3.69(m,4H),3.65-3.45(m,3H),2.80(t,J＝6.0Hz,2H),2.64–2.50(m,4H),2.41–2.25(m,1H),2.18(dd,J＝13.5,6.6Hz,1H),1.48(s,9H).
步骤10：将实施例1步骤10中的3-(4-氨基-5-碘-7H-吡咯[2,3-d]嘧啶-7-基)吡咯烷-1-甲酸叔丁酯替换成3-(4-氨基-5-碘-2-(2-吗啡啉乙氧基)-7H-吡咯[2,3-d]嘧啶-7-基)吡咯烷-1-甲酸叔丁酯，其余步骤与实施例1类似，可以制备得到标题化合物。
1H NMR(300MHz,CDCl3)δ7.19(d,J＝4.4Hz,2H),6.93(d,J＝9.2Hz,1H),6.62(s,1H),6.51–6.37(m,2H),5.78–5.65(m,1H),5.44(s,2H),5.41–5.26(m,1H),4.48(t,J＝5.8Hz,2H),4.17–4.03(m,1H),3.98(s,3H),3.79(d,J＝8.6Hz,3H),3.72(s,4H),2.82(t,J＝5.8Hz,2H),2.59(s,4H),2.47(s,3H),2.44–2.31(m,2H).
实施例68
1-(3-(4-氨基-5-(7-甲基苯并噻吩-2-基)-7H-吡咯[2,3-d]嘧啶-7-基)吡咯烷-1-基)丙-2-烯-1-酮
Figure PCTCN2017087222-appb-000174
将实施例22中步骤1的原料2-甲氧基苯硫酚替换成2-甲基苯硫酚，其余操作步骤类似实施例22。制备得1-(3-(4-氨基-5-(7-甲基苯并噻吩-2-基)-7H-吡咯[2,3-d]嘧啶-7-基)吡咯烷-1-基)丙-2-烯-1-酮，收率56％。
1H NMR(300MHz,CDCl3)δ8.34(s,1H),7.65(d,J＝8.0Hz,1H),7.33(m,2H),7.20–7.09(m,2H),6.53–6.45(m,1H),6.45–6.39(m,1H),5.78–5.68(m,1H),5.58(s,2H),5.51(dd,J＝12.1,5.9Hz,1H),4.26–4.04(m,2H),3.88–3.76(m,2H),2.58(s,3H),2.55–2.35(m,2H).
实施例69
(E)-1-(3-(4-氨基-5-(7-甲氧基-5-甲基苯并噻吩-2-基)-7H-吡咯[2,3-d]嘧啶-7-基)吡咯烷-1-基)-4-(3-氟吡咯烷-1-基)丁-2-烯-1-酮
Figure PCTCN2017087222-appb-000175
将实施例7中的反式-4-(吡咯烷-1-基)巴豆酸盐酸盐替换成反式-4-(3-氟吡咯烷-1-基)丁-2-烯酸盐酸盐，其余步骤与实施例7类似。制备得(E)-1-(3-(4-氨基-5-(7-甲氧基-5-甲基苯并噻吩-2-基)-7H-吡咯[2,3-d]嘧啶-7-基)吡咯烷-1-基)-4-(3-氟吡咯烷-1-基)丁-2-烯-1-酮
以下化合物的制备方法与化合物69类似。
Figure PCTCN2017087222-appb-000176
Figure PCTCN2017087222-appb-000177
实施例70
1-(4-(4-氨基-5-(7-甲氧基-5-甲基苯并噻吩-2-基)-7H-吡咯[2,3-d]嘧啶-7-基)-2-(羟甲基)吡咯烷-1-基)丙-2-烯-1-酮
Figure PCTCN2017087222-appb-000178
步骤1：制备2-(((叔丁基二苯基硅)氧)甲基)-4-羟基吡咯烷-1-甲酸叔丁酯
Figure PCTCN2017087222-appb-000179
将4-羟基-2-(羟甲基)吡咯烷-1-甲酸叔丁酯500mg和164mg咪唑溶于N,N-二甲基甲酰胺中。冰浴冷却下滴加叔丁基二苯基氯硅烷0.62ml。室温下搅拌1小时后加水淬灭反应，乙酸乙酯萃取后柱层析(石油醚：乙酸乙酯＝85:15)，得900mg油状物，收率86％。[M+H]+:456
步骤2：制备4-(4-氨基-5-(7-甲氧基-5-甲基苯并噻吩-2-基)-7H-吡咯[2,3-d]嘧啶-7-基)-2-(((叔丁基二苯基硅)氧)亚甲基)吡咯烷-1-甲酸叔丁酯。
Figure PCTCN2017087222-appb-000180
将实施例1中步骤8的3-羟基吡咯烷-1-甲酸叔丁酯替换成2-(((叔丁基二苯基硅)氧) 甲基)-4-羟基吡咯烷-1-甲酸叔丁酯，其余步骤与实施例1中步骤8至步骤10类似。[M+H]+:748
步骤3：制备4-(4-氨基-5-(7-甲氧基-5-甲基苯并噻吩-2-基)-7H-吡咯[2,3-d]嘧啶-7-基)-2-(羟甲基)吡咯烷-1-甲酸叔丁酯。
Figure PCTCN2017087222-appb-000181
将4-(4-氨基-5-(7-甲氧基-5-甲基苯并噻吩-2-基)-7H-吡咯[2,3-d]嘧啶-7-基)-2-(((叔丁基二苯基硅)氧)亚甲基)吡咯烷-1-甲酸叔丁酯300mg溶于3ml四氢呋喃，再加入2.5当量的四丁基氟化铵，室温搅拌过夜。次日浓缩干柱层析(二氯甲烷：甲醇＝98:2)得170mg固体，收率83％。[M+H]+:510
步骤4：以步骤3获得的4-(4-氨基-5-(7-甲氧基-5-甲基苯并噻吩-2-基)-7H-吡咯[2,3-d]嘧啶-7-基)-2-(羟甲基)吡咯烷-1-甲酸叔丁酯为原料，制备方法同实施例1步骤11至步骤12可获得标题化合物。
1H NMR(300MHz,CDCl3)δ8.30(s,1H),7.20(m,3H),6.64(s,1H),6.43(m,2H),5.80–5.73(m,1H),5.62(s,2H),5.26(m,1H),4.50–4.29(m,2H),3.99(s,3H),3.94(m,2H),3.88–3.76(m,2H),2.72-2.65(m,1H),2.48(s,3H),2.40-2.25(m,1H).
实施例71
1-(3-(4-氨基-6-氯-5-(7-甲氧基-5-甲基苯并噻吩-2-基)-7H-吡咯[2,3-d]嘧啶-7-基)嘧啶-1-基)丙-2-烯-1-酮
Figure PCTCN2017087222-appb-000182
步骤1：制备3-(4-氯-5-(7-甲氧基-5-甲基苯并噻吩-2-基)-7H-吡咯[2,3-d]嘧啶-7-基)吡咯烷-1-甲酸叔丁酯
Figure PCTCN2017087222-appb-000183
将实施例1中步骤10中的3-(4-氨基-5-碘-7H-吡咯[2,3-d]嘧啶-7-基)吡咯烷-1-甲酸叔丁酯替换成3-(4-氯-5-碘-7H-吡咯[2,3-d]嘧啶-7-基)吡咯烷-1-甲酸叔丁酯，其他原料不变可制备得3-(4-氯-5-(7-甲氧基-5-甲基苯并噻吩-2-基)-7H-吡咯[2,3-d]嘧啶-7-基)吡咯烷-1-甲酸叔丁酯。[M+H]＝499
步骤2：制备3-(4,6-二氯-5-(7-甲氧基-5-甲基苯并噻吩-2-基)-7H-吡咯[2,3-d]嘧啶-7-基)吡咯烷-1-甲酸叔丁酯
Figure PCTCN2017087222-appb-000184
将步骤1得到的3-(4-氯-5-(7-甲氧基-5-甲基苯并噻吩-2-基)-7H-吡咯[2,3-d]嘧啶-7-基)吡咯烷-1-甲酸叔丁酯50mg和3mg的三苯硫磷溶于1ml二氯甲烷中，室温搅拌5分钟后加入16mg的N-氯代丁二酰亚胺，室温搅拌1小时。浓缩干后柱层析(PE：EA)＝55:45，可以得到3-(4,6-二氯-5-(7-甲氧基-5-甲基苯并噻吩-2-基)-7H-吡咯[2,3-d]嘧啶-7-基)吡咯烷-1-甲酸叔丁酯。[M+H]＝533
步骤3：制备3-(4-氨基-6-氯-5-(7-甲氧基-5-甲基苯并噻吩-2-基)-7H-吡咯[2,3-d]嘧啶-7-基)吡咯烷-1-甲酸叔丁酯
Figure PCTCN2017087222-appb-000185
将步骤2得到的3-(4,6-二氯-5-(7-甲氧基-5-甲基苯并噻吩-2-基)-7H-吡咯[2,3-d]嘧啶-7-基)吡咯烷-1-甲酸叔丁酯40mg于2ml氨水和2ml二氧六环混合溶液中110℃加热反应48小时。浓缩干后柱层析(二氯甲烷：甲醇＝98:2)，以78％收率得到3-(4-氨基-6-氯-5-(7-甲氧基-5-甲基苯并噻吩-2-基)-7H-吡咯[2,3-d]嘧啶-7-基)吡咯烷-1-甲酸叔丁酯。[M+H]＝514
步骤4：根据实施例1，将实施例1步骤11的原料3-(4-氨基-5-(7-甲氧基-5-甲基苯并噻吩-2-基)-7H-吡咯[2,3-d]嘧啶-7-基)吡咯烷-1-甲酸叔丁酯替换成3-(4-氨基-6-氯-5-(7-甲氧基-5-甲基苯并噻吩-2-基)-7H-吡咯[2,3-d]嘧啶-7-基)吡咯烷-1-甲酸叔丁酯可制备得到标题化合物。
1H NMR(300MHz,CDCl3)δ8.34(s,1H),7.41(d,J＝2.0Hz,1H),7.14(d,J＝11.4Hz,1H),6.68(s,1H),6.51–6.39(m,2H),5.80–5.69(m,1H),5.57(s,2H),5.54–5.43(m,1H),4.24–4.03(m,2H),3.98(s,3H),3.91–3.75(m,2H),2.60–2.52(m,1H),2.51(s,3H),2.45(s,1H).
实施例72
N-(2-(4-氨基-5-(7-甲氧基-5-甲基苯并噻吩-2-基)-7H-吡咯[2,3-d]嘧啶-7-基)乙基)丙烯酰胺
Figure PCTCN2017087222-appb-000186
步骤1：制备(2-(4-氯-5-碘-7H-吡咯[2,3-d]嘧啶-7-基)乙基)氨基甲酸叔丁酯
Figure PCTCN2017087222-appb-000187
将实施例1步骤7得到的4-氯-5-碘-7H-吡咯[2,3-d]嘧啶100mg，108mg的N-Boc-溴乙胺和200mg碳酸铯于2ml的N,N-二甲基甲酰胺中加热80℃反应6小时。加水稀释后用乙酸乙酯萃取，有机相用无水硫酸钠干燥。浓缩干后柱层析(PE：EA＝85:15)，得145mg(2-(4-氯-5-碘-7H-吡咯[2,3-d]嘧啶-7-基)乙基)氨基甲酸叔丁酯，收率85％。
1H NMR(300MHz,CDCl3)δ8.60(s,1H),7.38(s,1H),4.75(s,1H),4.40(s,2H),3.52(dd,J＝12.9,7.0Hz,2H),1.40(s,9H).
步骤2：制备标题化合物
根据实施例1，将步骤1获得的中间体为原料替换实施例1步骤9中的3-(4-氯-5-碘-7H-吡咯[2,3-d]嘧啶-7-基)吡咯烷-1-甲酸叔丁酯，可制备得到标题化合物。
1H NMR(300MHz,CDCl3)δ8.31(s,1H),7.20(d,J＝3.8Hz,2H),7.11(s,2H),6.64(s,1H),6.25(d,J＝17.3Hz,1H),6.14–6.01(m,1H),5.63(d,J＝16.7Hz,3H),4.47–4.36(m,2H),4.00(s,3H),3.77(dd,J＝10.4,4.9Hz,2H),2.48(s,3H).
实施例73
N-(4-(4-氨基-5-(7-甲氧基-5-甲基苯并噻吩-2-基)-7H-吡咯[2,3-d]嘧啶-7-基)环己基)丙烯酰胺
Figure PCTCN2017087222-appb-000188
将实施例1中步骤8的3-羟基吡咯烷-1-甲酸叔丁酯替换成(4-羟基环己基)氨基甲酸叔丁酯，其他步骤同实施例1可获得标题化合物。
1H NMR(300MHz,CDCl3)δ8.31(s,1H),7.20(m,3H),6.64(s,1H),6.35(d,J＝16.6Hz,1H),6.24(d,J＝10.1Hz,1H),6.18(d,J＝9.9Hz,1H),5.69(d,J＝10.1Hz,1H),5.55(s,2H),4.62(s,1H),4.34(s,1H),3.99(s,3H),2.48(s,3H),2.06(s,4H),1.85(s,4H).
实施例75
(4S)-1-丙烯酰胺-4-(4-氨基-3-(7-甲氧基-5-甲基苯并噻吩-2-基)-1H-吡唑[3,4-d]嘧啶-1-基)-N-(2-(二甲胺)乙基-N-甲基吡咯烷-2-甲酰胺
Figure PCTCN2017087222-appb-000189
步骤1：制备1-(叔丁基2-甲基(4S)-4-(4-氨基-3-(7-甲氧基-5-甲基苯并噻吩-2-基)-1H-吡唑[3,4-d]嘧啶-1-基)吡咯烷-1,2-二甲酸酯
Figure PCTCN2017087222-appb-000190
将实施例2步骤3中的3-羟基吡咯烷-1-甲酸叔丁酯替换成N-Boc-反式-4-羟基-L-脯氨酸甲酯，其余步骤同实施例2得1-(叔丁基2-甲基(4S)-4-(4-氨基-3-(7-甲氧基-5-甲基苯并噻吩-2-基)-1H-吡唑[3,4-d]嘧啶-1-基)吡咯烷-1,2-二甲酸酯。[M+H]+:539
步骤2：制备(4S)-4-(4-氨基-3-(7-甲氧基-5-甲基苯并噻吩-2-基)-1H-吡唑[3,4-d]嘧啶-1-基)-1-(甲酸叔丁酯)吡咯烷-2-甲酸。
Figure PCTCN2017087222-appb-000191
将步骤1得到的1-(叔丁基2-甲基(4S)-4-(4-氨基-3-(7-甲氧基-5-甲基苯并噻吩-2-基)-1H-吡唑[3,4-d]嘧啶-1-基)吡咯烷-1,2-二甲酸酯670mg溶于6ml甲醇，冰浴冷却。滴加4N氢氧化钠水溶液3.8ml，室温搅拌5小时。稀盐酸调节PH至弱酸性，乙酸乙酯萃取，无水硫酸钠干燥。浓缩干得530mg粗品。
步骤3：制备(4S)-4-(4-氨基-3-(7-甲氧基-5-甲基苯并噻吩-2-基)-1H-吡唑[3,4-d]嘧啶-1-基)-2-((2-(二甲氨基)乙基)(甲基)甲酰胺)吡咯烷-1-甲酸叔丁酯
Figure PCTCN2017087222-appb-000192
将步骤2获得的(4S)-4-(4-氨基-3-(7-甲氧基-5-甲基苯并噻吩-2-基)-1H-吡唑[3,4-d]嘧啶-1-基)-1-(甲酸叔丁酯)吡咯烷-2-甲酸200mg、N,N,N’-三甲基乙烷-1,2-二胺50微升、BOP252mg和0.12ml二异丙基乙胺溶于5ml乙腈中，室温搅拌过夜。次日浓缩干，柱层析(二氯甲烷：甲醇＝95:5+0.1％氨水)得100mg黄色固体。[M+H]+:609
步骤4：将步骤3获得的(4S)-4-(4-氨基-3-(7-甲氧基-5-甲基苯并噻吩-2-基)-1H-吡唑[3,4-d]嘧啶-1-基)-2-((2-(二甲氨基)乙基)(甲基)甲酰胺)吡咯烷-1-甲酸叔丁酯替换掉实施例2步骤6的3-(4-氨基-3-(7-甲氧基-5-甲基苯并噻吩-2-基)-1H-吡唑[3,4-d]嘧啶-1-基)吡咯烷-1-甲酸叔丁酯，获得标题化合物。[M+H]+:563
实施例76
5-(7-甲氧基-5-甲基苯并噻吩-2-基)-7-(2-吗啡啉乙基)-7H-吡咯[2,3-d]嘧啶-4-氨
Figure PCTCN2017087222-appb-000193
步骤1：制备2-吗啉基乙基甲磺酸酯
Figure PCTCN2017087222-appb-000194
将实施例2步骤3中的3-羟基吡咯烷-1-甲酸叔丁酯替换成2-吗啉乙醇获得2-吗啉基乙基甲磺酸酯，直接投下一步。
步骤2：制备4-(2-(4-氯-5-碘-7H-吡咯[2,3-d]嘧啶-7-基)乙基)吗啉
Figure PCTCN2017087222-appb-000195
将200毫克4-氯-5-碘-7H-吡咯[2,3-d]嘧啶和步骤1获得的480毫克2-吗啉基乙基甲磺酸酯溶于2毫升N,N-二甲基甲酰胺，再加入456毫克碳酸铯，氩气保护下加热至90℃过夜，次日加水后用乙酸乙酯萃取，有机相用饱和氯化钠水溶液洗涤后无水硫酸钠干燥，过滤浓缩，残余物柱层析(二氯甲烷：甲醇＝97:3)分离得38毫克黄色固体。
1H NMR(300MHz,CDCl3)δ8.60(s,1H),7.50(s,1H),4.37(t,J＝6.1Hz,2H),3.67(s,4H),2.74(t,J＝6.3Hz,2H),2.50(s,4H).
步骤3将实施例1步骤9中的3-(4-氯-5-碘-7H-吡咯[2,3-d]嘧啶-7-基)吡咯烷-1-甲酸叔丁酯替换成步骤2获得的4-(2-(4-氯-5-碘-7H-吡咯[2,3-d]嘧啶-7-基)乙基)吗啉，按照实施例1步骤9至步骤10可获得标题化合物。
1H NMR(300MHz,CDCl3)δ8.33(s,1H),7.25(s,1H),7.22(d,J＝2.3Hz,2H),6.64(s,1H),5.56(s,2H),4.34(t,J＝6.3Hz,2H),4.00(s,3H),3.74–3.65(m,4H),2.79(t,J＝6.3Hz,2H),2.53(s,4H),2.49(s,3H).
实施例78
N-(3-((4-氨基-3-(7-甲氧基-5-甲基苯并噻吩-2-基)-1H-吡唑[3,4-d]嘧啶-1-基)甲基)苯基)丙烯酰胺
Figure PCTCN2017087222-appb-000196
步骤1：制备1-(3-氨基苄基)-3-碘-1H-吡唑[3,4-d]嘧啶-4-胺
Figure PCTCN2017087222-appb-000197
将实施例1步骤8中的原料4-氯-5-碘-7H-吡咯[2,3-d]嘧啶替换成3-碘-1H-吡唑[3,4-d] 嘧啶-4-胺，3-羟基吡咯烷-1-甲酸叔丁酯替换成(3-氨基苯基)甲醇，可获得1-(3-氨基苄基)-3-碘-1H-吡唑[3,4-d]嘧啶-4-胺。[M+H]+:367
步骤2：将实施例1步骤10的原料3-(4-氨基-5-碘-7H-吡咯[2,3-d]嘧啶-7-基)吡咯烷-1-甲酸叔丁酯替换成步骤1得到的1-(3-氨基苄基)-3-碘-1H-吡唑[3,4-d]嘧啶-4-胺，其余步骤同实施例1，可获得标题化合物N-(3-((4-氨基-3-(7-甲氧基-5-甲基苯并噻吩-2-基)-1H-吡唑[3,4-d]嘧啶-1-基)甲基)苯基)丙烯酰胺。
1H NMR(300MHz,DMSO-d6)δ10.14(s,1H),8.31(s,1H),7.67(d,J＝11.4Hz,2H),7.50(s,1H),7.35(s,1H),7.33–7.25(m,2H),7.03(d,J＝7.8Hz,1H),6.85(s,1H),6.39(dd,J＝16.9,9.9Hz,2H),6.22(d,J＝16.7Hz,1H),5.72(d,J＝9.9Hz,1H),5.54(s,2H),3.95(s,3H),2.44(s,3H).
实施例79
1-(3-(4-氨基-5-(3,5-二甲氧基苯)-7H-吡咯[2,3-d]嘧啶-7-基)吡咯烷-1-基)丙-2-烯-1-酮
Figure PCTCN2017087222-appb-000198
步骤1：将实施例1步骤10的原料(7-甲氧基-5-甲基苯并噻吩-2-基)硼酸替换成3,5-二甲氧基苯基硼酸频那醇酯，其余步骤同实施例1，可获得标题化合物1-(3-(4-氨基-5-(3,5-二甲氧基苯)-7H-吡咯[2,3-d]嘧啶-7-基)吡咯烷-1-基)丙-2-烯-1-酮。
以下化合物的制备方法与化合物79类似。
Figure PCTCN2017087222-appb-000199
Figure PCTCN2017087222-appb-000200
实施例80
1-(3-(4-氨基-5-(3-氨基-1H-吲唑-6-基)-7H-吡咯[2,3-d]嘧啶-7-基)吡咯烷-1-基)丙-2-烯-1-酮
Figure PCTCN2017087222-appb-000201
步骤1：制备3-(4-氨基-5-(4-氰基-3-氟苯)-7H-吡咯[2,3-d]嘧啶-7-基)吡咯烷-1-甲酸叔丁酯
Figure PCTCN2017087222-appb-000202
将200毫克的3-(4-氨基-5-碘-7H-吡咯[2,3-d]嘧啶-7-基)吡咯烷-1-甲酸叔丁酯，91毫克的4-氰基-3-氟苯硼酸，98毫克的碳酸钠和27毫克的四三苯基膦钯溶于4毫升的1,4-二氧六环和1毫升水的混合溶液中，置换氩气后80℃加热反应过夜，反应液浓缩至干，残余物柱层析(二氯甲烷：甲醇＝98:2)分离定量得150毫克淡黄色固体。[M+H]+:423
步骤2：制备3-(4-氨基-5-(3-氨基-1H-吲唑-6-基)-7H-吡咯[2,3-d]嘧啶-7-基)吡咯烷-1-甲酸叔丁酯
Figure PCTCN2017087222-appb-000203
将上一步得到的3-(4-氨基-5-(4-氰基-3-氟苯)-7H-吡咯[2,3-d]嘧啶-7-基)吡咯烷-1-甲酸叔丁酯100mg溶于4ml正丁醇中，加入1ml水合肼，氩气保护下回流12小时。浓缩干后柱层析(二氯甲烷：甲醇＝98:2)，可获得62mg固体3-(4-氨基-5-(3-氨基-1H-吲唑-6-基)-7H-吡咯[2,3-d]嘧啶-7-基)吡咯烷-1-甲酸叔丁酯。
1H NMR(300MHz,CDCl3)δ8.33(s,1H),7.65(d,J＝8.2Hz,1H),7.38(s,1H),7.18(d,J＝8.2Hz,1H),7.03(s,1H),5.49(s,1H),5.30(s,2H),4.20(s,2H),3.97–3.87(m,1H),3.70-3.50(m,3H),2.50–2.38(m,1H),2.35-2.23(m,1H),1.47(s,9H).
步骤3：将实施例1步骤11中的3-(4-氨基-5-(7-甲氧基-5-甲基苯并噻吩-2-基)-7H-吡咯[2,3-d]嘧啶-7-基)吡咯烷-1-甲酸叔丁酯替换成上一步获得的3-(4-氨基-5-(3-氨基-1H-吲唑-6-基)-7H-吡咯[2,3-d]嘧啶-7-基)吡咯烷-1-甲酸叔丁酯，按相同步骤可获得标题化合物。
1H NMR(300MHz,CDCl3)δ8.34(s,1H),7.65(d,J＝8.2Hz,1H),7.37(s,1H),7.17(d,J＝9.0Hz,1H),7.00(d,J＝8.2Hz,1H),6.54–6.36(m,2H),5.74(s,1H),5.52(s,1H),5.29(s,2H),4.13(m,2H),3.81(s,2H),2.44(s,2H).
实施例84
1-(3-(4-氨基-3-(7-甲氧基-5-甲基苯并噻吩-2-基)-1H-吡唑[3,4-d]嘧啶-1-基)吡咯烷-1-基)-2-氯乙烷-1-酮
Figure PCTCN2017087222-appb-000204
步骤1：将实施例2步骤7中的丙烯酰氯替换成氯乙酰氯，可制备得到标题化合物。
1H NMR(300MHz,CDCl3)δ8.38(s,1H),7.52(s,1H),7.27(s,1H),6.68(s,1H),5.89(d,J＝7.8Hz,2H),5.58(s,1H),4.20–4.03(m,4H),4.01(s,3H),3.74(s,2H),2.67(s,2H),2.50(s,3H).
实施例85
2-(7-(1-丙烯酰基吡咯烷-3-基)-4-氨基-7H-吡咯[2,3-d]嘧啶-5-基)-7-甲氧基苯并噻吩 -5-甲酸
Figure PCTCN2017087222-appb-000205
步骤1：制备7-羟基苯并噻吩-5-甲酸乙酯
Figure PCTCN2017087222-appb-000206
根据文献J.AM.CHEM.SOC.2007,129,14092-14099类似方法可得到7-羟基苯并噻吩-5-甲酸乙酯。
步骤2：7-甲氧基基苯并噻吩-5-甲酸乙酯
Figure PCTCN2017087222-appb-000207
将上述得到的7-羟基苯并噻吩-5-甲酸乙酯200mg、150mg碳酸钾和155mg碘甲烷溶于乙腈中，封管加热回流20小时。过滤后滤液浓缩柱层析(石油醚100％)可定量获得230mg油状物。
1H NMR(300MHz,CDCl3)δ8.19(d,J＝1.0Hz,1H),7.50(d,J＝5.4Hz,1H),7.43(s,1H),7.40(d,J＝5.4Hz,1H),4.43(q,J＝7.1Hz,2H),4.06(s,3H),1.44(t,J＝7.1Hz,3H).
步骤3：制备7-甲氧基基苯并噻吩-5-甲酸
Figure PCTCN2017087222-appb-000208
将步骤2获得的200mg7-甲氧基基苯并噻吩-5-甲酸乙酯溶于4ml四氢呋喃、1ml甲醇和1ml水的混合溶液中，加入36mg氢氧化锂搅拌24小时。用2N稀盐酸调节PH至 2，乙酸乙酯萃取。有机相浓缩干柱层析(二氯甲烷：甲醇＝98:2),可获得165mg白色固体，收率93％。
1H NMR(300MHz,CDCl3)δ8.30(s,1H),7.53(d,J＝5.4Hz,1H),7.49(s,1H),7.44(d,J＝5.3Hz,1H),4.09(s,3H).
步骤4：将实施例1步骤6的7-甲氧基-5-甲基苯并噻吩和正丁基锂分别替换成7-甲氧基基苯并噻吩-5-甲酸和二异丙基氨基锂，按照实施例1相同步骤可制备得到标题化合物。
1H NMR(300MHz,DMSO-d6)δ12.96(s,1H),8.21(s,1H),8.12(s,1H),7.65(d,J＝17.5Hz,1H),7.55(s,1H),7.40(s,1H),6.70–6.47(m,3H),6.17(d,J＝16.4Hz,1H),5.67(d,J＝12.6Hz,1H),5.37(m,1H),4.12(m,1H),4.03(s,3H),3.90(m,1H),3.79–3.68(m,1H),3.54(m,1H),2.39(m,2H).
实施例86
(S)-1-(3-(4-氨基-5-(7-甲氧基苯并噻吩-2-基)-7H-吡咯[2,3-d]嘧啶-7-基)吡咯烷-1-基)丙-2-烯-1-酮
Figure PCTCN2017087222-appb-000209
将实施例12步骤3中的(7-甲氧基-5-甲基苯并噻吩-2-基)硼酸替换成实施例22中得到的(7-甲氧基苯并噻吩-2-基)硼酸，可制备得到标题化合物。
1H NMR(300MHz,CDCl3)δ8.34(s,1H),7.45–7.31(m,2H),7.29(d,J＝1.7Hz,1H),7.12(d,J＝10.7Hz,1H),6.81(d,J＝7.8Hz,1H),6.51–6.45(m,1H),6.45–6.39(m,1H),5.81–5.68(m,1H),5.58(s,2H),5.51(dd,J＝14.3,6.9Hz,1H),4.25–4.04(m,2H),4.02(s,3H),3.75-3.93(m,2H),2.40-2.60(m,2H).
对照化合物
(S)-1-(3-(4-氨基-5-(喹啉-3-基)-7H-吡咯[2,3-d]嘧啶-7-基)吡咯烷-1-基)丙-2-烯-1-酮
Figure PCTCN2017087222-appb-000210
制备方法参照实施例1。
1H NMR(300MHz,DMSO-d)δ9.05(s,1H),8.35(s,1H),8.21(s,1H),8.11–7.96(m,2H),7.76(d,J＝7.2Hz,1H),7.69(d,J＝16.2Hz,1H),7.66–7.59(m,1H),6.72–6.53(m,1H),6.39(s,2H),6.17(d,J＝16.5Hz,1H),5.69(t,J＝11.4Hz,1H),5.51–5.31(m,1H),4.22–4.09(m,0.5H),4.05–3.86(m,1.5H),3.76(dd,J＝12.4,6.2Hz,1.5H),3.55(dd,J＝20.0,7.9Hz,0.5H),2.41(dd,J＝16.2,8.9Hz,2H).
化合物分子水平对FGFR1、FGFR4酶活的影响
1、试验方法
将酶反应底物Poly(Glu,Tyr)4:1用无钾离子的PBS(10mM磷酸钠缓冲液，150mM NaCl,pH 7.2-7.4)稀释成20μg/mL，125μL/孔包被酶标板，置37℃反应12-16小时。弃去孔中液体后洗板，用200μL/孔的T-PBS(含0.1％Tween-20的PBS)洗板三次，每次5分钟。于37℃烘箱中干燥酶标板1-2小时。
每孔加入用反应缓冲液(50mM HEPES pH 7.4,50mM MgCl2,0.5mM MnCl2,0.2mM Na3VO4,1mM DTT)稀释的ATP溶液50μL，终浓度5μM。化合物用DMSO稀释成合适的浓度，1μL/孔或者或含相应浓度的DMSO(阴性对照孔)，再加入用49μL反应缓冲液稀释的FGFR1、FGFR4激酶域重组蛋白启动反应，每次实验需设无ATP对照孔两孔。置37℃摇床(100rpm)反应1小时。T-PBS洗板三次。加入一抗PY99稀释液100μL/孔，37℃摇床反应0.5小时。T-PBS洗板三次。加入二抗辣根过氧化物酶标记羊抗鼠的IgG稀释液100μL/孔，37℃摇床反应0.5小时。T-PBS洗板三次。加入2mg/ml的OPD显色液100μL/孔(用含有0.03％H2O2的0.1M柠檬酸-柠檬酸钠缓冲液(pH＝5.4)稀释)，25℃避光反应1-10分钟。(OPD溶解时需用超声，显色液需现配现用)。加入2M H2SO4 50μL/孔中止反应，用可调波长式微孔板酶标仪SPECTRA MAX 190读数，波长为490nm。
样品的抑制率通过下列公式求得：
Figure PCTCN2017087222-appb-000211
IC50值采用酶标仪随机附带软件以四参数法回归求得。
2、实验结果
表2.化合物对FGFR1和FGFR4酶活抑制活性
Figure PCTCN2017087222-appb-000212
Figure PCTCN2017087222-appb-000213
Figure PCTCN2017087222-appb-000214
表3.化合物对EGFR酶活抑制活性
实施例 EGFR
NO.12 >1000nM
其中：A表示IC50小于(≤)10nM
B表示IC50小于(≤)100nM且大于(＞)10nM
C表示IC50大于(＞)100nM
从表2可以看出：本发明化合物对FGFR1和FGFR4酶在nM水平具有明显的抑制作用，由于FGFR1和FGFR2、3具有很高的同源性，可以预见本发明化合物对于FGFR1-4都具有显著的抑制作用。
此外，从表2还可以得出如下结论：
i)对比化合物NO.1-8(含取代的苯丙噻吩基团且Y-E-Z结构较复杂)和化合物NO.9-10以及NO.50(含取代的苯丙噻吩基团，但是Y-E-Z结构较简单)可知，当Y-E-Z结构较为复杂时，所述化合物具有更高的FGFR抑制活性；类似的结论也可从化合物NO.2和化合物NO.50的测试结果对比得出；
ii)对比化合物NO.13(含取代的苯丙噻吩基)和化合物NO.23(含取代的苯丙呋喃基)可知，当所述化合物包含取代的苯丙噻吩基时，其将具有更高的FGFR抑制活性；
iii)对比化合物NO.43和化合物NO.22可知，苯丙噻吩上甲氧基的取代是利于增强所述化合物的FGFR抑制活性的；
iv)对比化合物NO.43和化合物NO.13可知，苯丙噻吩上甲氧基和甲基的取代是利于增强所述化合物的FGFR抑制活性的；
v)对比化合物NO.12(R1为取代的苯丙噻吩基)和对照化合物(R1为未取代的喹啉基)可知，相比于R1为取代的苯丙噻吩基的化合物，当R1为未取代的喹啉基时，所得化合物对FGFR的抑制活性将显著降低；换言之，当R1为未取代的喹啉基时，所得化合物基本不具有FGFR抑制活性。
从表3可以看出：化合物NO.12作为本发明的代表性的化合物，其基本不具有EGFR抑制活性。
化合物对SNU16细胞增殖的影响
1、试验方法
化合物对SNU16细胞的增殖抑制作用以CCK-8细胞计数试剂盒(Dojindo)检测。具体步骤如下：处于对数生长期的SNU16细胞按合适密度接种至96孔培养板中，每孔90μL，培养过夜后，加入不同浓度的化合物作用72hr，并设定溶剂对照组(阴性对照)。待化合物作用细胞72h后，化合物对细胞增殖的影响采用CCK-8细胞计数试剂盒(Dojindo)检测，每孔加入10μL CCK-8试剂，置于37℃培养箱中放置2-4小时后，用全波长式微孔板酶标仪SpectraMax 190读数，测定波长为450nm。采用以下列公式计算化合物对肿瘤细胞生长的抑制率(％)：抑制率(％)＝(OD阴性对照孔-OD给药孔)/OD阴性对照孔×100％。IC50值采用酶标仪随机附带软件以四参数法回归求得。
2、实验结果
表4.化合物对SNU16细胞增殖的影响
Figure PCTCN2017087222-appb-000215
其中：A表示IC50小于(≤)10nM
B表示IC50小于(≤)100nM且大于(＞)10nM
C表示IC50大于(＞)100nM
从表4可以看出：本发明化合物对于FGFR依赖性的细胞株的生殖活性具有显著的抑制作用。
化合物对非小细胞肺癌细胞NCI-H1581裸小鼠移植瘤生长的影响
实验方法
取生长旺盛期的瘤组织剪切成1.5mm3左右，在无菌条件下，接种于裸小鼠右侧腋窝皮下。裸小鼠皮下移植瘤用游标卡尺测量移植瘤直径，待肿瘤生长至平均体积约为90mm3组左右后将动物随机分组。将实施例12所得化合物NO.12分别按照100mg/kg和20mg/kg的剂量，每天口服给药一次，连续给药14天，测量化合物NO.12对人肺癌NCI-H1581裸小鼠皮下移植瘤的生长抑制作用。
溶剂对照组则给以等量生理盐水。整个实验过程中，每周2次测量移植瘤直径，同时称量小鼠体重。肿瘤体积(tumor volume,TV)的计算公式为：TV＝1/2×a×b2，其中a、b分别表示长、宽。根据测量的结果计算出相对肿瘤体积(relative tumor volume，RTV)，计算公式为：RTV＝Vt/V0，其中V0为分笼给药时(即d0)测量所得肿瘤体积，Vt为每一次测量时的肿瘤体积。
抗肿瘤活性的评价指标为：1)相对肿瘤增殖率T/C(％)，计算公式如下：T/C(％)＝(TRTV/CRTV)×100％，TRTV：治疗组RTV；CRTV：阴性对照组RTV；2)肿瘤体积增长抑制率GI％，计算公式如下：GI％＝[1-(TVt-TV0)/(CVt-CV0)]×100％，TVt为治疗组每次测量的瘤体积；TV0为治疗组分笼给药时所得瘤体积；CVt为对照组每次测量的瘤体积；CV0为对照组分笼给药时所得瘤体积；3)瘤重抑制率，计算公式如下：瘤重抑制率％＝(Wc-WT)/Wc×100％，Wc：对照组瘤重，WT：治疗组瘤重。
实验结束当天给药2小时后，各组收集血液样本及肿瘤组织保存。
实验结果
表5.化合物NO.12对人肺癌NCI-H1581裸小鼠移植瘤的实验治疗作用
Figure PCTCN2017087222-appb-000216
从表5可以得出如下结论：
i)相比于阴性对照组，施用化合物NO.12 14天后，动物的体重变化分别为-5.9％(100mg/kg，po)和24.2％(20mg/kg，po)，远小于阴性对照组的36.8％；
ii)相比于阴性对照组，施用化合物NO.12 14天后，动物的肿瘤体积变化分别为54.8 ％(100mg/kg，po)和1500％(20mg/kg，po)，远小于阴性对照组的5223％；
iii)相比于阴性对照组，施用化合物NO.12 14天后，动物的相对肿瘤体积分别为1.51(100mg/kg，po)和16.33(20mg/kg，po)，远小于阴性对照组的52.22；
iv)相比于阴性对照组，施用化合物NO.12 14天后，动物的相对肿瘤增殖率分别为2.9％(100mg/kg，po)和31.28％(20mg/kg，po)。
在本发明提及的所有文献都在本申请中引用作为参考，就如同每一篇文献被单独引用作为参考那样。此外应理解，在阅读了本发明的上述讲授内容之后，本领域技术人员可以对本发明作各种改动或修改，这些等价形式同样落于本申请所附权利要求书所限定的范围。

Claims (10)

  1. 一种式I所示化合物或其立体异构体、几何异构体、互变异构体、其药用盐、其前药及其水合物或溶剂合物，
    Figure PCTCN2017087222-appb-100001
    式中，R1选自下组：取代或未取代的含有1-3个选自S、O、N和Se杂原子的5-14元杂芳基和取代或未取代的6-14元芳基，并且所述取代指被选自下组的1个或多个基团取代：C1-C6烷基、卤代的C1-C6烷基、C3-C8环烷基、卤代的C3-C8环烷基、羟基、C1-C6烷氧基、卤代的C1-C6烷氧基、-O-(C3-C8环烷基)、-O-(C3-C8的卤代环烷基)、-NR6R7、卤素、-(C1-C6亚烷基)-L1、C(＝O)R8；并且当R1为含N的5-14元杂芳基时，R1不是选自下组的基团：未取代的喹啉基、未取代的异喹啉基；
    R2和R3可相同或不同，分别独立地选自：H、取代或未取代的C1-C6烷基、取代或未取代的C3-C8环烷基、C(＝O)R8和S(＝O)2R9；
    R4选自H、取代或未取代的C1-C6烷基、取代或未取代的C3-C8环烷基、NR6R7、卤素、羟基、氰基、取代或未取代的C1-C6烷氧基和取代或未取代的C1-C6烷硫基；
    X1和X2可相同或不同，分别独立地选自N和CR10；R10选自H、取代或未取代的C1-C6烷基、取代或未取代的C3-C8环烷基、卤素、羟基、氰基、取代或未取代的C1-C6烷氧基和取代或未取代的C1-C6烷硫基；
    n为0、1、2、3、4或5；
    Y选自取代或未取代的含有1-3个选自N、O和S杂原子的3-10元杂环基、-NR6R7、取代或未取代的C3-C8环烷基和-L2-(取代或未取代的C6-C10芳基)-，并且所述取代指被选自下组的1个或多个基团独立地取代：C1-C6烷基、卤代的C1-C6烷基、C3-C8环烷基、卤代的C3-C8环烷基、羟基、C1-C6烷氧基、卤代的C1-C6烷氧基、-O-(C3-C8环烷基)、-O-(C3-C8的卤代环烷基)、NR11R12、卤素、含有1-3个选自N、O和S杂原子的4-10元杂环基、-(C1-C6亚烷基)-L1、C(＝O)R8和-Boc,
    E选自下组：无、C(＝O)、S(＝O)2、C(＝S)和S(＝O)；
    Z选自下组：无、取代或未取代的C2-C6烯基、取代或未取代的C2-C6炔基、取代或未取代的C1-C6烷基、取代或未取代的C3-C8环烷基、-(R13)-N(R11)-(R14)-(取代或未取代的C1-C6烷氧基)；
    R6和R7可相同或不同，分别独立地选自：H、取代或未取代的C1-C6烷基、取代或未取代的C3-C8环烷基、C(＝O)R8、-(C1-C6亚烷基)-L1、-(C1-C6亚烷基)-(取代或未取代的含1-3 个选自N、O和S杂原子的4-8元杂环基)、-CN、卤素、-OH、-(C1-C6亚烷基)-(取代或未取代的C4-C8环烷基)、或-(C1-C6亚烷基)-L2-(C1-C6亚烷基)-(取代或未取代的C1-C6烷氧基)；
    R8和R9可相同或不同，分别独立地选自：H、羟基、取代或未取代的C1-C6烷基、取代或未取代的C3-C8环烷基、取代或未取代的C1-C6烷氧基、取代或未取代的C2-C4烯基、取代或未取代的C2-C4炔基、-L2-(C1-C6亚烷基)-L1和-NR11R12；
    L1选自-OH、C1-C4烷氧基、-NR11R12、或含1或2个N原子的4-7元杂环；和
    L2选自下组：-NR11-、或-N(取代或未取代的C3-C6环烷基)；
    对于R2至R10、X1、X2、E、Z和L2，所述取代指被选自下组的1个或多个基团独立地取代：C1-C6烷基、卤代的C1-C6烷基、C3-C8环烷基、卤代的C3-C8环烷基、羟基、C1-C6烷氧基、卤代的C1-C6烷氧基、-O-(C3-C8环烷基)、-O-(C3-C8的卤代环烷基)、NR11R12、卤素、-CN、含有1-3个选自N、O和S杂原子的4-10元杂环基、-(C1-C6亚烷基)-L1和C(＝O)R8；其中，所述的4-10元杂环基可任选地具有1-3个选自下组的取代基：C1-C3烷基、卤素、-OH、C1-C3卤代烷基、C3-C4环烷基；
    R11和R12独立地选自H、C1-C6烷基、C1-C6卤代烷基、C3-C6环烷基、C3-C6卤代环烷基、-CO(C2-C4烯基)、或-CO(C2-C4炔基)；或R11和R12与相邻的N构成含1-2个N原子和0-2个O或S原子的4-7元杂环；
    R13和R14独立地选自取代或未取代的C1-C6亚烷基、或取代或未取代的C2-C6亚烯基。
  2. 如权利要求1所述的式I所示化合物或其立体异构体、几何异构体、互变异构体、其药用盐、其前药及其水合物或溶剂合物，其特征在于，所述的R1为下式的二环基团：
    Figure PCTCN2017087222-appb-100002
    式中，环A为取代或未取代的5元杂芳环；而环B为取代或未取代的6元杂芳环或取代或未取代的苯基。
  3. 如权利要求1所述的式I所示化合物或其立体异构体、几何异构体、互变异构体、其药用盐、其前药及其水合物或溶剂合物，其特征在于，R1为取代的5-14元杂芳基或取代的6-14元芳基。
  4. 如权利要求1所述的式I所示化合物或其立体异构体、几何异构体、互变异构体、其药用盐、其前药及其水合物或溶剂合物，其特征在于，所述的式I所示化合物选自下组：
    Figure PCTCN2017087222-appb-100003
    Figure PCTCN2017087222-appb-100004
    Figure PCTCN2017087222-appb-100005
    Figure PCTCN2017087222-appb-100006
    Figure PCTCN2017087222-appb-100007
    Figure PCTCN2017087222-appb-100008
    Figure PCTCN2017087222-appb-100009
    Figure PCTCN2017087222-appb-100010
    Figure PCTCN2017087222-appb-100011
    Figure PCTCN2017087222-appb-100012
    Figure PCTCN2017087222-appb-100013
  5. 一种药物组合物，其特征在于，所述药物组合物包含治疗有效量的权利要求1所述的化合物或其立体异构体、几何异构体、互变异构体、其药用盐、其前药及其水合物或溶剂合物中的一种或多种以及任选的药学上可接受的载体。
  6. 一种权利要求1所述的化合物或其立体异构体、几何异构体、互变异构体、其药用盐、其前药及其水合物或溶剂合物或权利要求5所述药物组合物的用途，其特征在于，用于制备预防和/或治疗选自下组的疾病的药物：
    a)肿瘤相关疾病；
    b)蛋白酪氨酸激酶活性相关疾病。
  7. 一种FGFR抑制剂，其特征在于，所述FGFR抑制剂包含抑制有效量的权利要求1所述的化合物或其立体异构体、几何异构体、互变异构体、其药用盐、其前药及其水合物或溶剂合物中的一种或多种。
  8. 一种权利要求1所述的化合物或其立体异构体、几何异构体、互变异构体、其药用盐、其前药及其水合物或溶剂合物的制备方法，其特征在于，所述方法包括如下步骤：
    (i)在惰性溶剂中，将式4化合物或其盐与式15所示的酰卤化合物或者酸进行缩合反应，从而形成式I化合物；
    Figure PCTCN2017087222-appb-100014
    各式中，R1、R2、R3、R4、n、X1、X2、Y、E和Z的定义同上；
    X选自下组：卤素、羟基。
  9. 一种权利要求5所述药物组合物的制备方法，其特征在于，包括步骤：将药学上可接受的载体与权利要求1所述的化合物或其立体异构体、几何异构体、互变异构体、其药用盐、其前药及其水合物或溶剂合物进行混合，从而形成药物组合物。
  10. 一种非诊断性、非治疗性抑制FGFR活性的方法，其特征在于，所述方法包括向所需患者施用抑制有效量的权利要求1所述的化合物或其立体异构体、几何异构体、互变异构体、其药用盐、其前药及其水合物或溶剂合物或权利要求5所述药物组合物。
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