WO2021232367A1 - 3-vinyl indazole derivative, preparation method therefor, and use thereof - Google Patents

3-vinyl indazole derivative, preparation method therefor, and use thereof Download PDF

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WO2021232367A1
WO2021232367A1 PCT/CN2020/091613 CN2020091613W WO2021232367A1 WO 2021232367 A1 WO2021232367 A1 WO 2021232367A1 CN 2020091613 W CN2020091613 W CN 2020091613W WO 2021232367 A1 WO2021232367 A1 WO 2021232367A1
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substituted
alkyl
unsubstituted
group
compound
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PCT/CN2020/091613
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French (fr)
Chinese (zh)
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叶庭洪
魏于全
姚于勤
余洛汀
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四川大学
上海医药集团股份有限公司
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Publication of WO2021232367A1 publication Critical patent/WO2021232367A1/en

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    • A—HUMAN NECESSITIES
    • A61—MEDICAL OR VETERINARY SCIENCE; HYGIENE
    • A61K—PREPARATIONS FOR MEDICAL, DENTAL OR TOILETRY PURPOSES
    • A61K31/00—Medicinal preparations containing organic active ingredients
    • A61K31/33—Heterocyclic compounds
    • A61K31/395—Heterocyclic compounds having nitrogen as a ring hetero atom, e.g. guanethidine or rifamycins
    • A61K31/41—Heterocyclic compounds having nitrogen as a ring hetero atom, e.g. guanethidine or rifamycins having five-membered rings with two or more ring hetero atoms, at least one of which being nitrogen, e.g. tetrazole
    • A61K31/415—1,2-Diazoles
    • A61K31/416—1,2-Diazoles condensed with carbocyclic ring systems, e.g. indazole
    • A—HUMAN NECESSITIES
    • A61—MEDICAL OR VETERINARY SCIENCE; HYGIENE
    • A61K—PREPARATIONS FOR MEDICAL, DENTAL OR TOILETRY PURPOSES
    • A61K31/00—Medicinal preparations containing organic active ingredients
    • A61K31/33—Heterocyclic compounds
    • A61K31/395—Heterocyclic compounds having nitrogen as a ring hetero atom, e.g. guanethidine or rifamycins
    • A61K31/435—Heterocyclic compounds having nitrogen as a ring hetero atom, e.g. guanethidine or rifamycins having six-membered rings with one nitrogen as the only ring hetero atom
    • A61K31/44—Non condensed pyridines; Hydrogenated derivatives thereof
    • A61K31/4427—Non condensed pyridines; Hydrogenated derivatives thereof containing further heterocyclic ring systems
    • A61K31/4439—Non condensed pyridines; Hydrogenated derivatives thereof containing further heterocyclic ring systems containing a five-membered ring with nitrogen as a ring hetero atom, e.g. omeprazole
    • A—HUMAN NECESSITIES
    • A61—MEDICAL OR VETERINARY SCIENCE; HYGIENE
    • A61K—PREPARATIONS FOR MEDICAL, DENTAL OR TOILETRY PURPOSES
    • A61K31/00—Medicinal preparations containing organic active ingredients
    • A61K31/33—Heterocyclic compounds
    • A61K31/395—Heterocyclic compounds having nitrogen as a ring hetero atom, e.g. guanethidine or rifamycins
    • A61K31/495—Heterocyclic compounds having nitrogen as a ring hetero atom, e.g. guanethidine or rifamycins having six-membered rings with two or more nitrogen atoms as the only ring heteroatoms, e.g. piperazine or tetrazines
    • A61K31/496—Non-condensed piperazines containing further heterocyclic rings, e.g. rifampin, thiothixene or sparfloxacin
    • A—HUMAN NECESSITIES
    • A61—MEDICAL OR VETERINARY SCIENCE; HYGIENE
    • A61K—PREPARATIONS FOR MEDICAL, DENTAL OR TOILETRY PURPOSES
    • A61K31/00—Medicinal preparations containing organic active ingredients
    • A61K31/33—Heterocyclic compounds
    • A61K31/395—Heterocyclic compounds having nitrogen as a ring hetero atom, e.g. guanethidine or rifamycins
    • A61K31/535—Heterocyclic compounds having nitrogen as a ring hetero atom, e.g. guanethidine or rifamycins having six-membered rings with at least one nitrogen and one oxygen as the ring hetero atoms, e.g. 1,2-oxazines
    • A61K31/5375—1,4-Oxazines, e.g. morpholine
    • A61K31/5377—1,4-Oxazines, e.g. morpholine not condensed and containing further heterocyclic rings, e.g. timolol
    • A—HUMAN NECESSITIES
    • A61—MEDICAL OR VETERINARY SCIENCE; HYGIENE
    • A61P—SPECIFIC THERAPEUTIC ACTIVITY OF CHEMICAL COMPOUNDS OR MEDICINAL PREPARATIONS
    • A61P1/00—Drugs for disorders of the alimentary tract or the digestive system
    • A61P1/16—Drugs for disorders of the alimentary tract or the digestive system for liver or gallbladder disorders, e.g. hepatoprotective agents, cholagogues, litholytics
    • A—HUMAN NECESSITIES
    • A61—MEDICAL OR VETERINARY SCIENCE; HYGIENE
    • A61P—SPECIFIC THERAPEUTIC ACTIVITY OF CHEMICAL COMPOUNDS OR MEDICINAL PREPARATIONS
    • A61P11/00—Drugs for disorders of the respiratory system
    • 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
    • C07D231/00—Heterocyclic compounds containing 1,2-diazole or hydrogenated 1,2-diazole rings
    • C07D231/54—Heterocyclic compounds containing 1,2-diazole or hydrogenated 1,2-diazole rings condensed with carbocyclic rings or ring systems
    • C07D231/56—Benzopyrazoles; Hydrogenated benzopyrazoles
    • C—CHEMISTRY; METALLURGY
    • C07—ORGANIC CHEMISTRY
    • C07D—HETEROCYCLIC COMPOUNDS
    • 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
    • C07D401/06—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 carbon chain containing only aliphatic carbon atoms
    • 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
    • C07D403/06—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 carbon chain containing only aliphatic carbon atoms
    • 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
    • C07D403/10—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 carbon chain containing aromatic rings
    • 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
    • C07D403/14—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 three or more hetero rings
    • C—CHEMISTRY; METALLURGY
    • C07—ORGANIC CHEMISTRY
    • C07D—HETEROCYCLIC COMPOUNDS
    • C07D413/00—Heterocyclic compounds containing two or more hetero rings, at least one ring having nitrogen and oxygen atoms as the only ring hetero atoms
    • C07D413/14—Heterocyclic compounds containing two or more hetero rings, at least one ring having nitrogen and oxygen atoms as the only ring hetero atoms containing three or more hetero rings

Definitions

  • the invention relates to a 3-vinylindazole derivative, a preparation method and application thereof, and belongs to the field of chemical medicine.
  • the main methods of treating cancer are: traditional therapies such as surgery, radiotherapy, chemotherapy, as well as targeted therapies, including immunotherapies such as PD1/PD-L1, CAR-T therapy, small molecule targeted drugs, monoclonal antibodies, and antibody conjugates Drugs, etc.
  • traditional therapies such as surgery, radiotherapy, chemotherapy, as well as targeted therapies, including immunotherapies such as PD1/PD-L1, CAR-T therapy, small molecule targeted drugs, monoclonal antibodies, and antibody conjugates Drugs, etc.
  • Traditional cancer treatment methods have limited effects, prone to recurrence, metastasis, or toxic side effects, and have a greater impact on the quality of life of patients.
  • Molecular targeted drugs have certain specificity, and small-molecule targeted drugs have always been an important treatment for diseases including cancer.
  • Receptor tyrosine kinases are receptor proteins located on the cell membrane that can transmit extracellular signals into the cell. Most RTKs with carcinogenic effects have low activity or expression levels in normal tissues, but they are over-activated or up-regulated in cancer cells. Receptor tyrosine kinases play an important regulatory role in tumor angiogenesis, tumor cell survival, proliferation, differentiation, and migration.
  • Fibroblast growth factor receptors are receptor tyrosine kinases located on the cell membrane, which can be activated by dimerization and autophosphorylation after binding to the ligand FGF, thereby activating PI3K- AKT, RAS-RAF-MAPK, JAK-STAT and PLC ⁇ four signal pathways.
  • FGFR plays an important role in normal physiological processes such as embryonic development, tissue repair, and maintenance of homeostasis.
  • the abnormality of FGF/FGFR is closely related to cancer and bone diseases.
  • FGFR amplification, mutation and chromosomal translocation make the FGF/FGFR signaling pathway abnormal, and promote tumor occurrence, development, metastasis and drug resistance.
  • FGFR signaling pathway plays a certain regulatory role, affecting the progress and prognosis of cancer.
  • Over-activation and over-expression of FGFR activate downstream signal pathways or autocrine-paracrine signals to produce bypass compensatory effects and make tumors resistant to radiotherapy, chemotherapy, targeted therapy and other therapies. Therefore, targeting FGFR is one of the important strategies for the treatment of cancer.
  • FGFR has also been reported in recent years to prove that it is closely related to pulmonary fibrosis.
  • IPF the proliferation of fibroblasts and the increase of matrix deposition can lead to lung injury and deterioration of lung function.
  • High levels of FGF2 were found in the alveolar lavage fluid and tissues of patients with acute lung injury and pulmonary fibrosis.
  • Mast cells expressing FGF2 accumulate in the extracellular matrix deposition area of IPF and the area where smooth muscle cells/myofibroblast-like cells proliferate.
  • FGF2 plays an important role in the formation of fibrosis, and it directly participates in the cell proliferation and fibrosis after bleomycin-induced lung injury in mice.
  • the increase in angiogenesis in IPF may be mediated by FGF2/FGFR2 in the FGFR signaling pathway.
  • FGF1/FGFR is abundantly expressed in the pathogenesis of IPF patients, suggesting that abnormal FGF1/FGFR signals in IPF patients can promote the migration of fibroblasts and increase the MAPK signal to cause the occurrence of pulmonary fibrosis.
  • TGF- ⁇ 1 induces the expression of ⁇ -SMA in fibroblasts to up-regulate and release FGF2, and FGFR2c can inhibit the proliferation of fibroblasts in pulmonary fibrosis mice through ERK1/2 and Smad3 pathways.
  • FGFR plays an important role in liver fibrosis. Therefore, targeting FGFR may also be a new strategy for the treatment of pulmonary fibrosis and liver fibrosis.
  • the purpose of the present invention is to provide 3-vinylindazole derivatives and preparation methods and uses thereof.
  • the present invention provides the compound represented by formula I, its optical isomer, the compound or its optical isomer pharmaceutically acceptable salt:
  • ring A is selected from substituted or unsubstituted phenyl, substituted or unsubstituted six-membered and five-membered aryl;
  • R 1 , R 2 , and R 3 are independently selected from H, halogen, substituted or unsubstituted alkyl;
  • Ring B is selected from substituted or unsubstituted 5- to 8-membered aryl groups.
  • ring A when ring A is a substituted phenyl group, it contains at least one substituent selected from the group consisting of halogen, unsubstituted alkyl, halogen-substituted alkyl, unsubstituted alkoxy, halogen-substituted alkoxy Group, hydroxyl.
  • the substituted phenyl group contains at least one substituent selected from the group consisting of halogen, unsubstituted C1-C6 alkyl, halogen-substituted C1-C6 alkyl, unsubstituted C1-C6 alkoxy, C1-C6 alkoxy and hydroxy substituted with halogen.
  • the substituted phenyl group contains at least one substituent selected from the group consisting of halogen, unsubstituted C1-C2 alkyl, halogen-substituted C1-C2 alkyl, unsubstituted C1-C2 alkoxy, C1-C2 alkoxy and hydroxy substituted with halogen.
  • the substituted phenyl group contains at least one substituent selected from the group consisting of fluorine, chlorine, trifluoromethyl, methoxy, ethoxy, and hydroxyl.
  • substituted phenyl group is selected from:
  • the substituted phenyl group contains at least one substituent selected from the group consisting of fluorine, chlorine, and methoxy.
  • substituted phenyl group is selected from:
  • ring A is a substituted or unsubstituted six-membered five-membered aryl group
  • the six-membered aryl group contains 0 to 2 heteroatoms.
  • the six-membered aryl group is phenyl.
  • the five-membered aryl group contains at least one heteroatom nitrogen.
  • the five-membered aryl group is selected from substituted or unsubstituted imidazolyl, oxazolyl or pyrrolyl.
  • the five-membered aryl group is selected from Wherein, R 4 to R 9 are independently selected from H, substituted or unsubstituted alkyl.
  • R 4 to R 9 are independently selected from H, unsubstituted C1-C6 alkyl or halogen-substituted C1-C6 alkyl.
  • R 4 to R 9 are independently selected from H, methyl or ethyl.
  • R 5 is selected from H, methyl or ethyl, and R 4 , R 6 , R 7 , R 8 , and R 9 are all H.
  • the six- and five-membered aryl group is selected from:
  • R 1 , R 2 , and R 3 are independently selected from H, halogen, unsubstituted C1-C6 alkyl or halogen-substituted C1-C6 alkyl.
  • R 1 , R 2 , and R 3 are independently selected from H, halogen or unsubstituted C1-C3 alkyl.
  • R 1 , R 2 , R 3 are independently selected from H, fluorine, chlorine or methyl.
  • R 2 is selected from H, fluorine, chlorine or methyl, and R 1 and R 3 are both H.
  • R 2 is selected from H or methyl, and both R 1 and R 3 are H.
  • ring B is selected from substituted or unsubstituted 5- to 6-membered aryl groups.
  • the aryl group contains 0 to 2 heteroatoms.
  • the heteroatom is nitrogen.
  • the aryl group is selected from phenyl, pyridyl, pyrimidinyl or pyrazolyl.
  • substituent selected from the group consisting of halogen, unsubstituted alkyl, halogen-substituted alkyl, unsubstituted
  • substituent selected from the group consisting
  • R 10 is selected from Substituted C1-C6 alkyl or halogen-substit
  • R 10 is an unsubstituted C1-C6 alkyl group.
  • R 10 is a methyl group.
  • n is 0 or 1.
  • R 11 and R 12 are independently selected from unsubstituted C1-C6 alkyl groups, or R 11 and R 12 are connected to form a substituted or unsubstituted morpholinyl or piperazinyl group.
  • R 11 and R 12 are both methyl groups.
  • the substituted morpholinyl and piperazinyl groups contain at least one substituent selected from the following group: substituted or unsubstituted C1-C6 alkyl, C1-C6 alkoxycarbonyl, benzyloxycarbonyl, fluorenyloxy Carbonyl.
  • the substituted morpholinyl and piperazinyl groups contain at least one substituent selected from the group consisting of methyl and tert-butoxycarbonyl.
  • R 11 and R 12 are connected to form Wherein, R 13 and R 15 are independently selected from H or methyl, and R 14 is selected from H, methyl or tert-butoxycarbonyl.
  • ring B is a substituted 6-membered aryl group, it is selected from:
  • ring B is an unsubstituted 6-membered aryl group, it is selected from
  • ring B is a substituted pyrazolyl group, and contains at least one substituent selected from the group consisting of unsubstituted alkyl, halogen-substituted alkyl, and hydroxy-substituted alkyl.
  • the substituted pyrazolyl group contains at least one substituent selected from the group consisting of unsubstituted C1-C6 alkyl, halogen-substituted C1-C6 alkyl, and hydroxy-substituted C1-C6 alkyl.
  • the substituted pyrazolyl contains only one substituent, and the substituent is a C1-C6 alkyl substituted with a hydroxy group.
  • the substituted pyrazolyl contains only one substituent, and the substituent is Further preferably, ring B is
  • ring B is selected from:
  • the compound is selected from:
  • the present invention provides a method for preparing the compound, its optical isomer, the compound or its optical isomer pharmaceutically acceptable salt, which comprises the following steps:
  • Y is halogen
  • R 16 and R 17 are independently selected from H, substituted or unsubstituted alkyl, or R 16 and R 17 are connected to form an alicyclic ring;
  • the nitrite is R 18 is selected from substituted or unsubstituted alkyl, and R 21 is selected from substituted or unsubstituted alkyl;
  • R 19 and R 20 are independently selected from H, substituted or unsubstituted alkyl, or R 19 and R 20 are connected to form an alicyclic ring.
  • Y is bromine
  • R 16 and R 17 are independently selected from H or an unsubstituted C1-C6 alkyl group, or R 16 and R 17 are connected to form a 5-membered alicyclic ring.
  • compound 2 is
  • R 18 is an unsubstituted C1-C6 alkyl group.
  • R 18 is isopentyl.
  • R 21 is an unsubstituted C1-C6 alkyl group.
  • R 21 is a methyl group.
  • R 19 and R 20 are independently selected from H or an unsubstituted C1-C6 alkyl group, or R 19 and R 20 are connected to form a 5-membered alicyclic ring.
  • compound 3 is
  • preparation method meets at least one of the following:
  • the palladium catalyst in step a is palladium acetate, [1,1'-bis(diphenylphosphino)ferrocene]palladium dichloride,
  • Step a Adding a base to the reaction system, the base being one or more of sodium carbonate, sodium bicarbonate, potassium carbonate, potassium phosphate, and cesium carbonate;
  • step a the molar ratio of compound 1: compound 2: base: palladium catalyst is 1: (1.1 ⁇ 1.5): (2.0 ⁇ 3.0): (0.003 ⁇ 0.010);
  • the reaction solvent in step a is one or more of dioxane, water, toluene, DMF, n-butanol, isopropanol, and sec-butanol;
  • the reaction solvent in step a is a mixed solvent of 1,4-dioxane:water volume ratio (4-8):1;
  • step a The reaction temperature in step a is 90-110°C;
  • step a The reaction time of step a is 5-10h;
  • Step a is reacted under N 2 protection
  • Step b Dissolve Intermediate I in the reaction solvent, add alkali and acid anhydride and stir thoroughly, and add nitrite for reaction to obtain Intermediate II;
  • the base in step b is potassium acetate
  • step b the molar ratio of intermediate I: base: acid anhydride: nitrite is 1: (1.1 ⁇ 1.5): (1.8 ⁇ 2.5): (3 ⁇ 5);
  • the reaction solvent in step b is toluene
  • Step b adding nitrite and refluxing reaction for 4-8h;
  • Step c reacts under acidic conditions
  • step c hydrochloric acid is added to the reaction system
  • step c 6N HCl is added to the reaction system;
  • the reaction solvent in step c is an alcohol solvent
  • the reaction solvent in step c is methanol
  • step d the intermediate III is dissolved in the reaction solvent, a base is added, and I 2 is dissolved in the reaction solvent and added dropwise to the reaction solution to obtain the intermediate IV through the reaction;
  • the alkali is one or more of sodium bicarbonate, sodium carbonate, potassium carbonate, sodium hydroxide, and potassium hydroxide;
  • step d the molar ratio of Intermediate III: I 2 : Base is 1: (1.5-2.0): 2.0;
  • step d is DMF
  • step d The reaction temperature in step d is 25 to 80°C;
  • step d The reaction time of step d is 2-10h;
  • the palladium catalyst in step e is palladium acetate, [1,1'-bis(diphenylphosphino)ferrocene]palladium dichloride, [1,1'-bis(diphenylphosphino)ferrocene] One or more of palladium dichloride dichloromethane complex and tris(dibenzylidene indeneacetone) dipalladium;
  • Step e adding a base to the reaction system, the base being one or more of DIEA, sodium carbonate, sodium bicarbonate, potassium carbonate, potassium phosphate, and cesium carbonate;
  • step e the molar ratio of intermediate IV: compound 3: base: palladium catalyst is 1: (1.1 ⁇ 1.5): (2.0 ⁇ 3.0): (0.003 ⁇ 0.010);
  • the reaction solvent in step e is one or more of dioxane, water, toluene, DMF, n-butanol, isopropanol, and sec-butanol;
  • the reaction solvent in step e is a mixed solvent of 1,4-dioxane:water volume ratio (4-8):1;
  • step e The reaction temperature in step e is 90-110°C;
  • step e The reaction time of step e is 5-10h;
  • Step e is reacted under N 2 protection.
  • the present invention provides the use of the compound, its optical isomer, the compound or its optical isomer pharmaceutically acceptable salt in the preparation of FGFR kinase inhibitor drugs.
  • the drug is an FGFR1, FGFR2 and/or FGFR3 kinase inhibitor.
  • the present invention provides the use of the compound, its optical isomer, the compound or its optical isomer pharmaceutically acceptable salt in the preparation of a medicine for treating and/or preventing cancer.
  • the cancer is breast cancer, lung cancer, gastric cancer, kidney cancer, colorectal cancer, liver cancer, melanoma, bladder cancer, urothelial cancer and/or cholangiocarcinoma.
  • the lung cancer is non-small cell lung cancer.
  • the present invention provides the use of the compound, its optical isomer, the compound or its optical isomer pharmaceutically acceptable salt in the preparation of a medicine for treating and/or preventing organ fibrosis.
  • the organ fibrosis is pulmonary fibrosis or liver fibrosis.
  • the present invention provides a pharmaceutical composition, which uses the compound, its optical isomer, the compound or its pharmaceutically acceptable salt as the active ingredient, and pharmaceutically acceptable excipients or auxiliary materials are added.
  • the compounds and derivatives provided by the present invention can be named according to the IUPAC (International Union of Pure and Applied Chemistry) or CAS (Chemical Abstracts Service, Columbus, OH) naming system.
  • alkyl is a linear or branched saturated hydrocarbon group.
  • Examples of C 1 -C 6 alkyl groups include but are not limited to methyl (C 1 ), ethyl (C 2 ), n-propyl (C 3 ), isopropyl (C 3 ), n-butyl (C 4 ) , Tert-butyl (C 4 ), sec-butyl (C 4 ), isobutyl (C 4 ), n-pentyl (C 5 ), 3-pentyl (C 5 ), pentyl (C 5 ), new Pentyl (C 5 ), 3-methyl-2-butyl (C 5 ), tert-pentyl (C 5 ) and n-hexyl (C 6 ).
  • alkoxy refers to the group -OR, where R is alkyl as defined above.
  • Examples of C 1 ⁇ C 6 alkoxy include but are not limited to methoxy, ethoxy, n-propoxy, isopropoxy, n-butoxy, tert-butoxy, sec-butoxy, n-pentoxy Group, n-hexyloxy and 1,2-dimethylbutoxy.
  • aryl refers to a group of a 4n+2 aromatic ring system containing or not containing heteroatoms in the aromatic ring system, wherein the heteroatoms are selected from nitrogen, oxygen and/or sulfur.
  • alicyclic refers to a saturated or partially unsaturated cyclic hydrocarbon group.
  • alkoxycarbonyl refers to the group ROC (O) -, wherein R is alkyl as defined above, preferably R is a C 1 ⁇ C 6 alkyl (i.e., C1 ⁇ C6 of the present invention, the alkoxycarbonyl group) .
  • R is alkyl as defined above, preferably R is a C 1 ⁇ C 6 alkyl (i.e., C1 ⁇ C6 of the present invention, the alkoxycarbonyl group) .
  • Examples include, but are not limited to, methoxycarbonyl, ethoxycarbonyl, propoxycarbonyl, isopropoxycarbonyl, butoxycarbonyl, isobutoxycarbonyl, tert-butoxycarbonyl.
  • pharmaceutically acceptable means that a certain carrier, carrier, diluent, excipient, and/or the formed salt is usually chemically or physically compatible with other ingredients constituting a certain pharmaceutical dosage form, and physiologically Compatible with the receptor.
  • pharmaceutically acceptable salt refers to the acid and/or basic salt formed by the compound of the present invention with inorganic and/or organic acids and bases, and also includes zwitterionic salts (internal salts), and also includes quaternary ammonium salts, For example, alkyl ammonium salts.
  • zwitterionic salts internal salts
  • quaternary ammonium salts For example, alkyl ammonium salts.
  • These salts can be obtained directly in the final isolation and purification of the compound. It can also be obtained by appropriately mixing the above-mentioned compound with a certain amount of acid or base (e.g., equivalent).
  • These salts may form a precipitate in the solution and be collected by filtration, or recovered after evaporation of the solvent, or prepared by freeze-drying after reaction in an aqueous medium.
  • the salt in the present invention can be the hydrochloride, sulfate, citrate, benzenesulfonate, hydrobromide, hydrofluoride, phosphate, acetate, propionate, butane Acid salt, oxalate, malate, succinate, fumarate, maleate, tartrate or trifluoroacetate.
  • the method of administration of the compound or pharmaceutical composition of the present invention is not particularly limited, and representative administration methods include (but are not limited to): oral, parenteral (intravenous, intramuscular, or subcutaneous), and topical administration.
  • the pharmaceutically acceptable excipients in the present invention refer to substances contained in the dosage form in addition to the active ingredients.
  • the pharmaceutically acceptable auxiliary component of the present invention has certain physiological activity, but the addition of the component will not change the dominant position of the above-mentioned pharmaceutical composition in the course of disease treatment, but only exerts auxiliary functions. These auxiliary functions only It is the utilization of the known activity of the ingredient, and it is a commonly used adjuvant therapy in the medical field. If the aforementioned auxiliary components are used in combination with the pharmaceutical composition of the present invention, they should still fall within the protection scope of the present invention.
  • the present invention provides a class of 3-vinylindazole derivatives with novel structures.
  • Biological experiments have proved that the compound provided by the present invention has a significant inhibitory effect on the activity of FGFR kinase, and can effectively inhibit breast cancer, lung cancer, gastric cancer, kidney cancer, colorectal cancer, liver cancer, melanoma, bladder cancer, urothelial cancer and
  • the proliferation of a variety of cancer cells such as cholangiocarcinoma has a broad-spectrum anti-cancer effect; in addition, it also shows a significant inhibitory effect on the proliferation of fibroblasts.
  • the cloth is comparable, the anti-fibrosis effect is significant, and it also has a good inhibitory effect on hepatic stellate cells.
  • the invention provides new options for the development and application of anti-cancer and anti-fibrosis drugs.
  • Figure 1 is a drug-time curve diagram of compound 4-20 of the present invention in biological experiments
  • Figure 2 is a graph of HE and Masson staining of lung tissue 14 days after administration in an animal experiment
  • Figure 3 shows the HE and Masson staining of lung tissue 28 days after administration in an animal experiment.
  • the raw materials and equipment used in the specific embodiments of the present invention are all known products, which are obtained by purchasing commercially available products.
  • the main biological experiment instruments and equipment are as follows. Ultra-clean workbench BHC-1000IIA/B3: Sujing Antai Biotechnology Company; Constant temperature water bath PolyScience 9505: PolyScience Company; Sterilizer MLS-3780: SANYO Company; Oven: Binder Company; Ultra-pure water meter Milli-Q Integral 10 :Millipore company; microplate reader Multiscan MK3, cell incubator, low-speed centrifuge Sorvall ST1: Thermofisher company; Centrifuge 5415C ultracentrifuge: Germany Eppendorf company; NUAIRE NU-425-600E biological safety cabinet: American Nuaire company; BCD-215YD Type ordinary refrigerator: China Haier company; SANYO (-80°C) ultra-low temperature refrigerator: Japan Sanyo Electric Group; Rocker 51702 shaker: American Cole Parmer company; 96-well cell culture plate: Costa Corning company; ordinary optical microscope: Olympus company; Liquid gun: Thermo Company; PH meter
  • the cell line used in the experiment was purchased from ATCC, USA.
  • Various necessities for cell culture were purchased from Gibco BRL, including DMEM medium, RPMI 1640 medium, fetal bovine serum (FBS) and pancreatin.
  • Tetramethylazazole blue (MTT) and dimethyl sulfoxide (DMSO) were purchased from Sigma Company in the United States.
  • the compound was diluted with DMSO to 50 times the final highest inhibitor concentration required in the reaction. Transfer 100 ⁇ L of compound dilution to a 96-well plate. Add 100 ⁇ L of DMSO to the two blank wells of the same 96-well plate. This 96-well plate serves as the source plate. Transfer 10 ⁇ L of compound from the source plate to the 96-well plate as the intermediate plate. Add 90 ⁇ L of l1x kinase buffer to each well of the middle plate. Mix the compounds in the middle plate on a shaker for 10 minutes. Transfer 5 ⁇ L from each well of the 96-well middle plate to a 384-well plate, and set up secondary wells. Add kinase to 1x Kinase Alkaline Buffer.
  • the assay plate already contains 5 ⁇ L of compound 10% DMSO solution. Add 10 ⁇ L of l2.5x enzyme solution to each well of the 384-well assay plate. Incubate for 10 minutes at room temperature. Add 10 ⁇ L of 2.5x peptide solution to each well of the 384-well assay plate. After incubating at 28°C for a specific time, add 25 ⁇ L of stop buffer to stop the reaction. Collect data on Caliper and convert the data to IC 50 .
  • Passage is generally 1 time in 3 to 4 days; Passage of adherent growth cells: The cells adhere to the wall and grow to about 80% of the bottom of the bottle, remove the culture bottle from the incubator, aspirate the medium, and wash once with 0.25% pancreatin Then add 0.25% trypsin digestion solution for digestion. After observing the cell shrinkage and rounding, add complete medium to stop the digestion, and pipette to disperse and fall off the cells. Collect the cell suspension, centrifuge at 1500rpm/min for 3min, and pour the supernatant. , The cell pellet is resuspended in complete medium and pipetted evenly, and then divided into 3 to 5 bottles for culture. Generally, it is passaged once every 3 to 4 days.
  • Collect the cells in the logarithmic growth phase (4T1 mouse breast cancer cell line; MDA-MB-231 human breast cancer cell line; A549 human non-small cell lung cancer line; SUN-16 human poorly differentiated gastric cancer cell; NIH3T3 mouse adult Fibroblasts; human lung fibroblasts HPF (Catalog#3300)), seeded in a 96-well plate at a rate of 2.5 ⁇ 10 3 to 1 ⁇ 10 4 per well, in a cell incubator at 37°C and 5% CO 2 Incubate in medium overnight for 24 hours, dilute the drug to be tested with DMEM medium and add it to a 96-well plate. Each drug has 8 gradients, and each gradient contains 3 replicate wells.
  • the dosing group add 100 ⁇ L of the compound medium solution to each well according to the gradient (final concentrations are 1000, 333, 127, 42.3, 14.1, 4.7, 1.56, 0.53 nM), and each concentration has 3 replicate wells; negative control
  • 100 ⁇ L of blank medium containing 1 ⁇ DMSO was added to each well for a total of 6 multiple wells; in the blank control group, only 100 ⁇ L of medium was added to each well. Place the plate in a 37°C, 5% CO 2 cell culture incubator for 72 hours.
  • the invisible control group and the blank group add 20 ⁇ L of MTT solution (5mg/mL) to each well, continue to incubate for 2-4 hours.
  • the pharmacokinetic analysis of the compound 4-20 of the present invention is completed by the testing service provided by Medicipuya Pharmaceutical Technology (Shanghai) Co., Ltd. 8 SPF-grade male SD rats (6 experimental rats, and the rest as blank controls) used in this experiment were transferred from the experimental institution animal reserve (999M-017), Shanghai Xipuer-Bikai Experimental Animal Co., Ltd. The experiment was divided into two groups, each group of 3 mice, respectively: a single intravenous injection with a dose of 2 mg/kg, a dose of 0.4 mg/mL, a dose of 5 mL/kg, and a dose of 20 mg/kg. The drug concentration was 2 mg/mL and the administration volume was 10 mL/kg.
  • the animals in the oral administration group were fasted overnight for 10-14 hours before administration and 4 hours after administration. And set the blood sampling time points as follows: before administration, after administration 0.083h, 0.25h, 0.5h, 1h, 2h, 4h, 6h, 8h, 24h, 10 times, after blood collection through the jugular vein, each sample is collected about 0.20 mL, heparin sodium is anticoagulated and placed on ice after collection.
  • Plasma sample processing place the blood sample on ice after collection, and centrifuge to separate the plasma within 1 hour (centrifugation conditions: 6800g, 6mins, 2-8°C). Plasma samples were stored in a refrigerator at -80°C before analysis.
  • mice Male C57BL/6 mice (7-9 weeks old, weighing 18-22 g) were purchased from Hua Fukang (Beijing, China). The mice are housed and maintained in the facility under SPF conditions. On the 0th day of the experiment, the mice were anesthetized with 10% chloral hydrate, and then a single intratracheal instillation of bleomycin sulfate dissolved in normal saline (2 mg/kg body weight) was given to the mice, and an equal volume was injected at the same time To the rats in the control group.
  • mice were randomly divided into groups on the second day, each with 10 mice, and the compound 4-20 (YTH-17; 30mg/kg, 60mg/kg) and 4-22 (YTH-18; 30mg/kg) of the present invention were orally administered orally every day 60mg/kg), the positive control is Nintedanib (BIBF1120) (30mg/Kg, the solvent is (5% DMSO + 40% PEG400 + 55% normal saline)) medicine and an equal volume of solvent as a control. After 14 or 28 days of administration, the mice were sacrificed.
  • mice were sacrificed on the 14th or 28th day of the experiment.
  • the lung tissue samples were placed in 4% (m/v) PBS-buffered paraformaldehyde solution. Three days later, part of the tissue was rinsed in water for 2 hours, then dehydrated with gradient ethanol and embedded in paraffin.
  • the tissues wrapped in paraffin were cut into serial sections (3 ⁇ m) and stained with hematoxylin and eosin or Masson trichrome to evaluate the histopathological changes and the degree of accumulated collagen fibers.
  • Table 2 lists the inhibition of FGFR1 kinase by some of the compounds synthesized in the present invention.
  • the letter A represents an IC 50 of 50 nM or less
  • the letter B represents an IC 50 of 50 nM to 100 nM
  • the letter C represents an IC 50 of 100 nM to 500 nM
  • the letter D represents an IC 50 of 500 nM or more.
  • Table 2 The IC 50 value of some compounds of the present invention for inhibiting FGFR1 kinase
  • Table 3 lists the inhibitory effects of some of the compounds synthesized in the present invention on various kinases.
  • the letter A represents an IC 50 of 50 nM or less
  • the letter B represents an IC 50 of 50 nM to 100 nM
  • the letter C represents an IC 50 of 100 nM to 500 nM
  • the letter D represents an IC 50 of 500 nM or more.
  • Table 3 The IC 50 values of some compounds of the present invention for inhibiting related kinases
  • Table 4 lists the inhibition of the proliferation of SNU16 cells by some of the compounds synthesized by the present invention under 72 hours of action.
  • the letter A represents an IC 50 of 50 nM or less
  • the letter B represents an IC 50 of 50 nM to 100 nM
  • the letter C represents an IC 50 of 100 nM to 500 nM
  • the letter D represents an IC 50 of 500 nM or more.
  • Table 4 The IC 50 value of some compounds of the present invention in inhibiting the proliferation of SUN16 cells
  • Table 5 The IC 50 value of some compounds of the present invention in inhibiting tumor cell proliferation
  • Table 6 lists the inhibition of the proliferation of NIH-3T3 cells and human lung fibroblasts of some compounds synthesized by the present invention after 72 hours of action.
  • Table 6 The IC 50 values of some compounds of the present invention in inhibiting the proliferation of NIH-3T3 cells and human lung fibroblasts
  • Figure 1 shows the blood concentration and time curve.
  • a single intravenous injection (1-A) or oral gavage (1-B) dose: 2mg/kg, 20mg/kg
  • SD rats three male rats in each group, respectively numbered : 101/102/103, 201/202/203
  • the study found that the average half-life after intravenous administration was 5.81h, and the average AUC(0-t) was 2771.74h*ng/mL.
  • the average half-life was 3.44h
  • the average AUC(0-t) was 20917.54h*ng/mL
  • the average oral bioavailability was calculated to be 75.47%.

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Abstract

The present invention relates to the field of chemical medicine, and relates to a 3-vinyl indazole derivative, a preparation method therefor, and use thereof. The present invention provides a compound represented by formula I, an optical isomer thereof, and a pharmaceutically acceptable salt of the compound or the optical isomer thereof. Biological experiments show that the 3-vinyl indazole derivative provided in the present invention has a significant inhibitory effect on the activity of FGFR kinase, can effectively inhibit the proliferation of multiple types of cancer cells in breast cancer, lung cancer, gastric cancer, kidney cancer, colorectal cancer, liver cancer, melanoma, bladder cancer, urothelial carcinoma, cholangiocarcinoma, and the like, and has a broad-spectrum anti-cancer effect; in addition, the 3-vinyl indazole derivative also shows a significant inhibitory effect on the proliferation of fibroblasts, has an effect equivalent to the drug Nintedanib currently clinically used for treating pulmonary fibrosis, and has a significant anti-fibrosis therapeutic effect; moreover, the 3-vinyl indazole derivative also has a good inhibitory effect on hepatic stellate cells. The present invention provides a new option for the development and application of anti-cancer and anti-fibrosis drugs.

Description

3-乙烯基吲唑类衍生物及其制备方法和用途3-vinyl indazole derivatives and preparation method and use thereof 技术领域Technical field
本发明涉及3-乙烯基吲唑类衍生物及其制备方法和用途，属于化学医药领域。The invention relates to a 3-vinylindazole derivative, a preparation method and application thereof, and belongs to the field of chemical medicine.
背景技术Background technique
目前治疗癌症的主要方法有：手术、放疗、化疗等传统疗法，以及靶向疗法，包括PD1/PD-L1、CAR-T疗法等免疫疗法，小分子靶向药物、单克隆抗体和抗体偶联药物等。传统癌症治疗方法效果有限，容易出现复发、转移或毒副作用，且对患者生活质量有较大影响。分子靶向药物具有一定特异性，其中小分子靶向药物一直是包括癌症等在内的疾病治疗的重要治疗方式。At present, the main methods of treating cancer are: traditional therapies such as surgery, radiotherapy, chemotherapy, as well as targeted therapies, including immunotherapies such as PD1/PD-L1, CAR-T therapy, small molecule targeted drugs, monoclonal antibodies, and antibody conjugates Drugs, etc. Traditional cancer treatment methods have limited effects, prone to recurrence, metastasis, or toxic side effects, and have a greater impact on the quality of life of patients. Molecular targeted drugs have certain specificity, and small-molecule targeted drugs have always been an important treatment for diseases including cancer.
受体酪氨酸激酶(Receptor tyrosine kinases,RTKs)作为分子靶向药物的一类靶点，是位于细胞膜上的受体蛋白，可以将胞外信号传导至细胞内。大多数具有致癌作用的RTKs在正常组织中的活性或表达水平很低，但在癌细胞中过度活化或表达上调。受体酪氨酸激酶对肿瘤血管生成、肿瘤细胞存活、增殖、分化、迁移等发挥重要调控作用。Receptor tyrosine kinases (Receptor tyrosine kinases, RTKs), as a class of molecular targeted drugs, are receptor proteins located on the cell membrane that can transmit extracellular signals into the cell. Most RTKs with carcinogenic effects have low activity or expression levels in normal tissues, but they are over-activated or up-regulated in cancer cells. Receptor tyrosine kinases play an important regulatory role in tumor angiogenesis, tumor cell survival, proliferation, differentiation, and migration.
成纤维生长因子受体(Fibroblast growth factor receptors,FGFRs)是一种位于细胞膜上的受体酪氨酸激酶，能与配体FGF结合后发生二聚化和自磷酸化而激活，进而激活PI3K-AKT、RAS-RAF-MAPK、JAK-STAT和PLCγ四条信号通路。FGFR在胚胎发育、组织修复、维持体内稳态等正常生理过程中发挥重要作用。FGF/FGFR的异常与癌症和骨骼疾病等有密切联系。FGFR扩增、突变和染色体易位使FGF/FGFR信号通路发生异常，促进肿瘤的发生、发展、转移和耐药。在肿瘤形成、发展、转移、复发等各个阶段，FGFR信号通路都发挥着一定的调控作用，影响癌症的进展和预后。FGFR的过度活化和过表达激活下游信号通路或者自分泌-旁分泌信号，产生旁路代偿作用，使肿瘤对放疗、化疗、靶向治疗等疗法耐药。因此，靶向FGFR是治疗癌症的重要策略之一。Fibroblast growth factor receptors (FGFRs) are receptor tyrosine kinases located on the cell membrane, which can be activated by dimerization and autophosphorylation after binding to the ligand FGF, thereby activating PI3K- AKT, RAS-RAF-MAPK, JAK-STAT and PLCγ four signal pathways. FGFR plays an important role in normal physiological processes such as embryonic development, tissue repair, and maintenance of homeostasis. The abnormality of FGF/FGFR is closely related to cancer and bone diseases. FGFR amplification, mutation and chromosomal translocation make the FGF/FGFR signaling pathway abnormal, and promote tumor occurrence, development, metastasis and drug resistance. In the various stages of tumor formation, development, metastasis, and recurrence, FGFR signaling pathway plays a certain regulatory role, affecting the progress and prognosis of cancer. Over-activation and over-expression of FGFR activate downstream signal pathways or autocrine-paracrine signals to produce bypass compensatory effects and make tumors resistant to radiotherapy, chemotherapy, targeted therapy and other therapies. Therefore, targeting FGFR is one of the important strategies for the treatment of cancer.
FGFR除了与癌症密切相关，近年来也有报道证明它与肺纤维化有着密切的关系。在IPF中，成纤维细胞的增殖和基质沉积的增加会导致肺损伤和肺功能的恶化。急性肺损伤和肺纤维化患者的肺泡灌洗液和组织中均发现了高水平的FGF2。表达FGF2的肥大细胞积聚在IPF的细胞外基质沉积区和平滑肌细胞/肌成纤维细胞样细胞增殖的区域。FGF2在纤维化形成中具有重要作用，其直接参与了博莱霉素诱导的小鼠肺损伤后的细胞增殖和纤维增生。IPF中血管新生的增加可能是由FGFR信号通路中的FGF2/FGFR2 介导。FGF1/FGFR在IPF患者的发病部位大量表达，提示IPF患者出现异常FGF1/FGFR信号后，可以通过促进成纤维细胞的迁移和増加MAPK信号导致肺纤维化的发生。在肺纤维化过程中，TGF-β1诱导成纤维细胞中的α-SMA的表达上调和释放FGF2，而FGFR2c可通过ERK1/2及Smad3通路抑制肺纤维化小鼠的成纤维细胞增殖。同时，FGFR在肝纤维化中具有重要作用也被报道，所以，靶向FGFR也可能是治疗肺纤维化和肝纤维化的新的策略。In addition to being closely related to cancer, FGFR has also been reported in recent years to prove that it is closely related to pulmonary fibrosis. In IPF, the proliferation of fibroblasts and the increase of matrix deposition can lead to lung injury and deterioration of lung function. High levels of FGF2 were found in the alveolar lavage fluid and tissues of patients with acute lung injury and pulmonary fibrosis. Mast cells expressing FGF2 accumulate in the extracellular matrix deposition area of IPF and the area where smooth muscle cells/myofibroblast-like cells proliferate. FGF2 plays an important role in the formation of fibrosis, and it directly participates in the cell proliferation and fibrosis after bleomycin-induced lung injury in mice. The increase in angiogenesis in IPF may be mediated by FGF2/FGFR2 in the FGFR signaling pathway. FGF1/FGFR is abundantly expressed in the pathogenesis of IPF patients, suggesting that abnormal FGF1/FGFR signals in IPF patients can promote the migration of fibroblasts and increase the MAPK signal to cause the occurrence of pulmonary fibrosis. In the process of pulmonary fibrosis, TGF-β1 induces the expression of α-SMA in fibroblasts to up-regulate and release FGF2, and FGFR2c can inhibit the proliferation of fibroblasts in pulmonary fibrosis mice through ERK1/2 and Smad3 pathways. At the same time, it has also been reported that FGFR plays an important role in liver fibrosis. Therefore, targeting FGFR may also be a new strategy for the treatment of pulmonary fibrosis and liver fibrosis.
发明内容Summary of the invention
本发明的目的在于提供3-乙烯基吲唑类衍生物及其制备方法和用途。The purpose of the present invention is to provide 3-vinylindazole derivatives and preparation methods and uses thereof.
本发明提供了式Ⅰ所示的化合物、其光学异构体、化合物或其光学异构体在药学上可接受的盐：The present invention provides the compound represented by formula I, its optical isomer, the compound or its optical isomer pharmaceutically acceptable salt:
Figure PCTCN2020091613-appb-000001
Figure PCTCN2020091613-appb-000001
其中，环A选自取代或未取代的苯基、取代或未取代的六元并五元芳基；Wherein, ring A is selected from substituted or unsubstituted phenyl, substituted or unsubstituted six-membered and five-membered aryl;
R 1、R 2、R 3独立选自H、卤素、取代或未取代的烷基； R 1 , R 2 , and R 3 are independently selected from H, halogen, substituted or unsubstituted alkyl;
环B选自取代或未取代的5～8元芳基。Ring B is selected from substituted or unsubstituted 5- to 8-membered aryl groups.
进一步地，当环A为取代的苯基时，含有至少一个选自下组的取代基：卤素、未取代的烷基、卤素取代的烷基、未取代的烷氧基、卤素取代的烷氧基、羟基。Further, when ring A is a substituted phenyl group, it contains at least one substituent selected from the group consisting of halogen, unsubstituted alkyl, halogen-substituted alkyl, unsubstituted alkoxy, halogen-substituted alkoxy Group, hydroxyl.
优选地，所述取代的苯基含有至少一个选自下组的取代基：卤素、未取代的C1～C6烷基、卤素取代的C1～C6烷基、未取代的C1～C6烷氧基、卤素取代的C1～C6烷氧基、羟基。Preferably, the substituted phenyl group contains at least one substituent selected from the group consisting of halogen, unsubstituted C1-C6 alkyl, halogen-substituted C1-C6 alkyl, unsubstituted C1-C6 alkoxy, C1-C6 alkoxy and hydroxy substituted with halogen.
优选地，所述取代的苯基含有至少一个选自下组的取代基：卤素、未取代的C1～C2烷基、卤素取代的C1～C2烷基、未取代的C1～C2烷氧基、卤素取代的C1～C2烷氧基、羟基。Preferably, the substituted phenyl group contains at least one substituent selected from the group consisting of halogen, unsubstituted C1-C2 alkyl, halogen-substituted C1-C2 alkyl, unsubstituted C1-C2 alkoxy, C1-C2 alkoxy and hydroxy substituted with halogen.
优选地，所述取代的苯基含有至少一个选自下组的取代基：氟、氯、三氟甲基、甲氧基、乙氧基、羟基。Preferably, the substituted phenyl group contains at least one substituent selected from the group consisting of fluorine, chlorine, trifluoromethyl, methoxy, ethoxy, and hydroxyl.
进一步优选地，所述取代的苯基选自：Further preferably, the substituted phenyl group is selected from:
Figure PCTCN2020091613-appb-000002
Figure PCTCN2020091613-appb-000002
优选地，所述取代的苯基含有至少一个选自下组的取代基：氟、氯、甲氧基。Preferably, the substituted phenyl group contains at least one substituent selected from the group consisting of fluorine, chlorine, and methoxy.
进一步优选地，所述取代的苯基选自：Further preferably, the substituted phenyl group is selected from:
Figure PCTCN2020091613-appb-000003
Figure PCTCN2020091613-appb-000003
进一步地，当环A为取代或未取代的六元并五元芳基时，所述六元并五元芳基中，六元芳基含有0～2个杂原子。Further, when ring A is a substituted or unsubstituted six-membered five-membered aryl group, in the six-membered five-membered aryl group, the six-membered aryl group contains 0 to 2 heteroatoms.
优选地，六元芳基为苯基。Preferably, the six-membered aryl group is phenyl.
所述六元并五元芳基中，五元芳基含有至少一个杂原子氮。In the six-membered five-membered aryl group, the five-membered aryl group contains at least one heteroatom nitrogen.
优选地，五元芳基选自取代或未取代的咪唑基、恶唑基或吡咯基。Preferably, the five-membered aryl group is selected from substituted or unsubstituted imidazolyl, oxazolyl or pyrrolyl.
优选地，五元芳基选自
Figure PCTCN2020091613-appb-000004
其中，R 4～R 9独立选自H、取代或未取代的烷基。 Preferably, the five-membered aryl group is selected from
Figure PCTCN2020091613-appb-000004
Wherein, R 4 to R 9 are independently selected from H, substituted or unsubstituted alkyl.
优选地，R 4～R 9独立选自H、未取代的C1～C6烷基或卤素取代的C1～C6烷基。 Preferably, R 4 to R 9 are independently selected from H, unsubstituted C1-C6 alkyl or halogen-substituted C1-C6 alkyl.
优选地，R 4～R 9独立选自H、甲基或乙基。 Preferably, R 4 to R 9 are independently selected from H, methyl or ethyl.
优选地，R 5选自H、甲基或乙基，R 4、R 6、R 7、R 8、R 9均为H。 Preferably, R 5 is selected from H, methyl or ethyl, and R 4 , R 6 , R 7 , R 8 , and R 9 are all H.
进一步优选地，所述六元并五元芳基选自：Further preferably, the six- and five-membered aryl group is selected from:
Figure PCTCN2020091613-appb-000005
Figure PCTCN2020091613-appb-000005
进一步地，R 1、R 2、R 3独立选自H、卤素、未取代的C1～C6烷基或卤素取代的C1～C6 烷基。 Further, R 1 , R 2 , and R 3 are independently selected from H, halogen, unsubstituted C1-C6 alkyl or halogen-substituted C1-C6 alkyl.
优选地，R 1、R 2、R 3独立选自H、卤素或未取代的C1～C3烷基。 Preferably, R 1 , R 2 , and R 3 are independently selected from H, halogen or unsubstituted C1-C3 alkyl.
优选地，R 1、R 2、R 3独立选自H、氟、氯或甲基。 Preferably, R 1 , R 2 , R 3 are independently selected from H, fluorine, chlorine or methyl.
进一步优选地，R 2选自H、氟、氯或甲基，R 1、R 3均为H。 More preferably, R 2 is selected from H, fluorine, chlorine or methyl, and R 1 and R 3 are both H.
进一步优选地，R 2选自H或甲基，R 1、R 3均为H。 More preferably, R 2 is selected from H or methyl, and both R 1 and R 3 are H.
进一步地，环B选自取代或未取代的5～6元芳基。Further, ring B is selected from substituted or unsubstituted 5- to 6-membered aryl groups.
优选地，所述芳基含有0～2个杂原子。Preferably, the aryl group contains 0 to 2 heteroatoms.
优选地，所述杂原子为氮。Preferably, the heteroatom is nitrogen.
优选地，所述芳基选自苯基、吡啶基、嘧啶基或吡唑基。Preferably, the aryl group is selected from phenyl, pyridyl, pyrimidinyl or pyrazolyl.
进一步地，当环B为取代的6元芳基时，含有至少一个选自下组的取代基：卤素、未取代的烷基、卤素取代的烷基、未取代的烷氧基、卤素取代的烷氧基、-C(＝O)OR 10、-X-NR 11R 12，其中，R 10选自取代或未取代的烷基，X选自-(CH 2) n-、-C(＝O)-或-SO 2-，n为0～6的整数，R 11、R 12独立选自取代或未取代的烷基，或者，R 11与R 12相连形成脂环。 Further, when ring B is a substituted 6-membered aryl group, it contains at least one substituent selected from the group consisting of halogen, unsubstituted alkyl, halogen-substituted alkyl, unsubstituted alkoxy, halogen-substituted Alkoxy, -C(=O)OR 10 , -X-NR 11 R 12 , wherein R 10 is selected from substituted or unsubstituted alkyl, and X is selected from -(CH 2 ) n -, -C(= O)- or -SO 2 -, n is an integer of 0-6, R 11 and R 12 are independently selected from substituted or unsubstituted alkyl groups, or R 11 and R 12 are connected to form an alicyclic ring.
优选地，当环B为取代的6元芳基时，含有至少一个选自下组的取代基：卤素、未取代的C1～C6烷基、卤素取代的C1～C6烷基、未取代的C1～C6烷氧基、卤素取代的C1～C6烷氧基、-C(＝O)OR 10、-X-NR 11R 12，其中，R 10选自取代或未取代的C1～C6烷基，X选自-(CH 2) n-、-C(＝O)-或-SO 2-，n为0～6的整数，R 11、R 12独立选自取代或未取代的C1～C6烷基，或者，R 11与R 12相连形成6元脂环。 Preferably, when ring B is a substituted 6-membered aryl group, it contains at least one substituent selected from the group consisting of halogen, unsubstituted C1-C6 alkyl, halogen-substituted C1-C6 alkyl, unsubstituted C1 -C6 alkoxy, halogen-substituted C1-C6 alkoxy, -C(=O)OR 10 , -X-NR 11 R 12 , wherein R 10 is selected from substituted or unsubstituted C1-C6 alkyl, X is selected from -(CH 2 ) n -, -C(=O)- or -SO 2 -, n is an integer of 0-6, R 11 and R 12 are independently selected from substituted or unsubstituted C1-C6 alkyl Or, R 11 and R 12 are connected to form a 6-membered alicyclic ring.
优选地，当环B为取代的6元芳基时，含有至少一个选自下组的取代基：卤素、甲基、三氟甲基、甲氧基、三氟甲氧基、-C(＝O)OR 10、-X-NR 11R 12，其中，R 10选自取代或未取代的C1～C6烷基，X选自-(CH 2) n-、-C(＝O)-或-SO 2-，n为0～3的整数，R 11、R 12独立选自取代或未取代的C1～C6烷基，或者，R 11与R 12相连形成6元脂环，所述6元脂环含有2个杂原子。 Preferably, when ring B is a substituted 6-membered aryl group, it contains at least one substituent selected from the group consisting of halogen, methyl, trifluoromethyl, methoxy, trifluoromethoxy, -C(= O) OR 10 , -X-NR 11 R 12 , wherein R 10 is selected from substituted or unsubstituted C1-C6 alkyl, and X is selected from -(CH 2 ) n -, -C(=O)- or- SO 2 -, n is an integer from 0 to 3, R 11 and R 12 are independently selected from substituted or unsubstituted C1-C6 alkyl groups, or R 11 and R 12 are connected to form a 6-membered alicyclic ring, the 6-membered aliphatic The ring contains 2 heteroatoms.
优选地，当环B为取代的6元芳基时，仅含有一个选自下组的取代基：-C(＝O)OR 10、-X-NR 11R 12，其中，R 10选自未取代的C1～C6烷基或卤素取代的C1～C6烷基，X选自-(CH 2) n-或-C(＝O)，n为0～3的整数，R 11、R 12独立选自未取代的C1～C6烷基或卤素取代的C1～C6烷基，或者，R 11与R 12相连形成6元脂环，所述6元脂环含有2个杂原子，所述杂原子为氮或氧。 Preferably, when ring B is a substituted 6-membered aryl group, it contains only one substituent selected from the following group: -C(=O)OR 10 , -X-NR 11 R 12 , wherein R 10 is selected from Substituted C1-C6 alkyl or halogen-substituted C1-C6 alkyl, X is selected from -(CH 2 ) n -or -C(=O), n is an integer of 0 to 3, R 11 and R 12 are independently selected From an unsubstituted C1-C6 alkyl group or a halogen-substituted C1-C6 alkyl group, or, R 11 and R 12 are connected to form a 6-membered alicyclic ring, the 6-membered alicyclic ring contains 2 heteroatoms, and the heteroatoms are Nitrogen or oxygen.
进一步优选地，R 10为未取代的C1～C6烷基。 More preferably, R 10 is an unsubstituted C1-C6 alkyl group.
进一步优选地，R 10为甲基。 More preferably, R 10 is a methyl group.
进一步优选地，n为0或1。More preferably, n is 0 or 1.
进一步优选地，R 11、R 12独立选自未取代的C1～C6烷基，或者，R 11与R 12相连形成取代或未取代的吗啉基、哌嗪基。 More preferably, R 11 and R 12 are independently selected from unsubstituted C1-C6 alkyl groups, or R 11 and R 12 are connected to form a substituted or unsubstituted morpholinyl or piperazinyl group.
进一步优选地，R 11、R 12均为甲基。 More preferably, R 11 and R 12 are both methyl groups.
进一步优选地，所述取代的吗啉基、哌嗪基含有至少一个选自下组的取代基：取代或未取代的C1～C6烷基、C1～C6烷氧羰基、苄氧羰基、芴氧羰基。Further preferably, the substituted morpholinyl and piperazinyl groups contain at least one substituent selected from the following group: substituted or unsubstituted C1-C6 alkyl, C1-C6 alkoxycarbonyl, benzyloxycarbonyl, fluorenyloxy Carbonyl.
进一步优选地，所述取代的吗啉基、哌嗪基含有至少一个选自下组的取代基：甲基、叔丁氧羰基。More preferably, the substituted morpholinyl and piperazinyl groups contain at least one substituent selected from the group consisting of methyl and tert-butoxycarbonyl.
进一步优选地，R 11与R 12相连形成
Figure PCTCN2020091613-appb-000006
其中，R 13、R 15独立选自H或甲基，R 14选自H、甲基或叔丁氧羰基。 Further preferably, R 11 and R 12 are connected to form
Figure PCTCN2020091613-appb-000006
Wherein, R 13 and R 15 are independently selected from H or methyl, and R 14 is selected from H, methyl or tert-butoxycarbonyl.
进一步优选地，当环B为取代的6元芳基时，选自：More preferably, when ring B is a substituted 6-membered aryl group, it is selected from:
Figure PCTCN2020091613-appb-000007
Figure PCTCN2020091613-appb-000007
进一步地，当环B为未取代的6元芳基时，选自
Figure PCTCN2020091613-appb-000008
Further, when ring B is an unsubstituted 6-membered aryl group, it is selected from
Figure PCTCN2020091613-appb-000008
进一步地，环B为取代的吡唑基，含有至少一个选自下组的取代基：未取代的烷基、卤素取代的烷基、羟基取代的烷基。Further, ring B is a substituted pyrazolyl group, and contains at least one substituent selected from the group consisting of unsubstituted alkyl, halogen-substituted alkyl, and hydroxy-substituted alkyl.
优选地，所述取代的吡唑基含有至少一个选自下组的取代基：未取代的C1～C6烷基、卤素取代的C1～C6烷基、羟基取代的C1～C6烷基。Preferably, the substituted pyrazolyl group contains at least one substituent selected from the group consisting of unsubstituted C1-C6 alkyl, halogen-substituted C1-C6 alkyl, and hydroxy-substituted C1-C6 alkyl.
优选地，所述取代的吡唑基仅含有一个取代基，所述取代基为羟基取代的C1～C6烷基。Preferably, the substituted pyrazolyl contains only one substituent, and the substituent is a C1-C6 alkyl substituted with a hydroxy group.
优选地，所述取代的吡唑基仅含有一个取代基，所述取代基为
Figure PCTCN2020091613-appb-000009
进一步优选地，环B为
Figure PCTCN2020091613-appb-000010
Preferably, the substituted pyrazolyl contains only one substituent, and the substituent is
Figure PCTCN2020091613-appb-000009
Further preferably, ring B is
Figure PCTCN2020091613-appb-000010
进一步地，环B选自：Further, ring B is selected from:
Figure PCTCN2020091613-appb-000011
Figure PCTCN2020091613-appb-000011
进一步地，所述化合物选自：Further, the compound is selected from:
Figure PCTCN2020091613-appb-000012
Figure PCTCN2020091613-appb-000012
Figure PCTCN2020091613-appb-000013
Figure PCTCN2020091613-appb-000013
Figure PCTCN2020091613-appb-000014
Figure PCTCN2020091613-appb-000014
Figure PCTCN2020091613-appb-000015
Figure PCTCN2020091613-appb-000015
Figure PCTCN2020091613-appb-000016
Figure PCTCN2020091613-appb-000016
Figure PCTCN2020091613-appb-000017
Figure PCTCN2020091613-appb-000017
Figure PCTCN2020091613-appb-000018
Figure PCTCN2020091613-appb-000018
本发明提供了所述的化合物、其光学异构体、化合物或其光学异构体药学上可接受的盐的制备方法，包括如下步骤：The present invention provides a method for preparing the compound, its optical isomer, the compound or its optical isomer pharmaceutically acceptable salt, which comprises the following steps:
a、化合物1与化合物2在钯催化剂作用下偶联，得到中间体Ⅰ：a. Compound 1 and compound 2 are coupled under the action of palladium catalyst to obtain Intermediate I:
Figure PCTCN2020091613-appb-000019
Figure PCTCN2020091613-appb-000019
其中，Y为卤素，R 16、R 17独立选自H、取代或未取代的烷基，或者，R 16与R 17相连形成脂环； Wherein, Y is halogen, R 16 and R 17 are independently selected from H, substituted or unsubstituted alkyl, or R 16 and R 17 are connected to form an alicyclic ring;
b、中间体Ⅰ在碱存在的条件下与酸酐和亚硝酸酯反应，得到中间体Ⅱ：b. Intermediate I reacts with acid anhydride and nitrite in the presence of a base to obtain Intermediate II:
Figure PCTCN2020091613-appb-000020
Figure PCTCN2020091613-appb-000020
所述亚硝酸酯为
Figure PCTCN2020091613-appb-000021
R 18选自取代或未取代的烷基，R 21选自取代或未取代的烷基； The nitrite is
Figure PCTCN2020091613-appb-000021
R 18 is selected from substituted or unsubstituted alkyl, and R 21 is selected from substituted or unsubstituted alkyl;
c、中间体Ⅱ脱除酰基，得到中间体Ⅲ：c. Intermediate II removes the acyl group to obtain Intermediate III:
Figure PCTCN2020091613-appb-000022
Figure PCTCN2020091613-appb-000022
d、中间体Ⅲ经卤代，得到中间体Ⅳ：d. Intermediate III is halogenated to obtain Intermediate IV:
Figure PCTCN2020091613-appb-000023
Figure PCTCN2020091613-appb-000023
e、中间体Ⅳ与化合物3在钯催化剂作用下偶联，即得：e. Intermediate IV and compound 3 are coupled under the action of palladium catalyst to obtain:
Figure PCTCN2020091613-appb-000024
Figure PCTCN2020091613-appb-000024
其中，R 19、R 20独立选自H、取代或未取代的烷基，或者，R 19与R 20相连形成脂环。 Wherein, R 19 and R 20 are independently selected from H, substituted or unsubstituted alkyl, or R 19 and R 20 are connected to form an alicyclic ring.
优选地，Y为溴。Preferably, Y is bromine.
优选地，R 16、R 17独立选自H或未取代的C1～C6烷基，或者，R 16与R 17相连形成5元脂环。 Preferably, R 16 and R 17 are independently selected from H or an unsubstituted C1-C6 alkyl group, or R 16 and R 17 are connected to form a 5-membered alicyclic ring.
进一步优选地，化合物2为
Figure PCTCN2020091613-appb-000025
Further preferably, compound 2 is
Figure PCTCN2020091613-appb-000025
优选地，R 18为未取代的C1～C6烷基。 Preferably, R 18 is an unsubstituted C1-C6 alkyl group.
进一步优选地，R 18为异戊基。 More preferably, R 18 is isopentyl.
优选地，R 21为未取代的C1～C6烷基。 Preferably, R 21 is an unsubstituted C1-C6 alkyl group.
进一步优选地，R 21为甲基。 More preferably, R 21 is a methyl group.
优选地，R 19、R 20独立选自H或未取代的C1～C6烷基，或者，R 19与R 20相连形成5元脂环。 Preferably, R 19 and R 20 are independently selected from H or an unsubstituted C1-C6 alkyl group, or R 19 and R 20 are connected to form a 5-membered alicyclic ring.
进一步优选地，化合物3为
Figure PCTCN2020091613-appb-000026
More preferably, compound 3 is
Figure PCTCN2020091613-appb-000026
进一步地，所述的制备方法满足以下至少一项：Further, the preparation method meets at least one of the following:
步骤a所述钯催化剂为醋酸钯、[1,1'-双(二苯基膦基)二茂铁]二氯化钯、The palladium catalyst in step a is palladium acetate, [1,1'-bis(diphenylphosphino)ferrocene]palladium dichloride,
[1,1'-双(二苯基膦)二茂铁]二氯化钯二氯甲烷络合物、三(二亚苄基茚丙酮)二钯中一种或两种以上；[1,1'-Bis(diphenylphosphine)ferrocene]dichloropalladium dichloromethane complex, one or more of tris(dibenzylidene indeneacetone)dipalladium;
步骤a向反应体系中加入碱，所述碱为碳酸钠、碳酸氢钠、碳酸钾、磷酸钾、碳酸铯中一种或两种以上；Step a: Adding a base to the reaction system, the base being one or more of sodium carbonate, sodium bicarbonate, potassium carbonate, potassium phosphate, and cesium carbonate;
步骤a中化合物1：化合物2：碱：钯催化剂的摩尔配比为1：(1.1～1.5)：(2.0～3.0)：(0.003～0.010)；In step a, the molar ratio of compound 1: compound 2: base: palladium catalyst is 1: (1.1～1.5): (2.0～3.0): (0.003～0.010);
步骤a的反应溶剂为二氧六环、水、甲苯、DMF、正丁醇、异丙醇、仲丁醇中一种或两种以上；The reaction solvent in step a is one or more of dioxane, water, toluene, DMF, n-butanol, isopropanol, and sec-butanol;
优选地，步骤a的反应溶剂为1,4-二氧六环：水体积比为(4～8)：1的混合溶剂；Preferably, the reaction solvent in step a is a mixed solvent of 1,4-dioxane:water volume ratio (4-8):1;
步骤a的反应温度为90～110℃；The reaction temperature in step a is 90-110°C;
步骤a的反应时间为5～10h；The reaction time of step a is 5-10h;
步骤a于N 2保护下反应； Step a is reacted under N 2 protection;
步骤b将中间体I溶于反应溶剂中，加入碱和酸酐充分搅拌，加入亚硝酸酯反应，得到中间体Ⅱ；Step b: Dissolve Intermediate I in the reaction solvent, add alkali and acid anhydride and stir thoroughly, and add nitrite for reaction to obtain Intermediate II;
步骤b所述碱为乙酸钾；The base in step b is potassium acetate;
步骤b中中间体I：碱：酸酐：亚硝酸酯的摩尔配比为1：(1.1～1.5)：(1.8～2.5)：(3～5)；In step b, the molar ratio of intermediate I: base: acid anhydride: nitrite is 1: (1.1～1.5): (1.8～2.5): (3～5);
步骤b的反应溶剂为甲苯；The reaction solvent in step b is toluene;
步骤b加入亚硝酸酯回流反应4～8h；Step b: adding nitrite and refluxing reaction for 4-8h;
步骤c在酸性条件下反应；Step c reacts under acidic conditions;
优选地，步骤c向反应体系中加入盐酸；Preferably, in step c, hydrochloric acid is added to the reaction system;
优选地，步骤c向反应体系中加入6NHCl；Preferably, in step c, 6N HCl is added to the reaction system;
步骤c的反应溶剂为醇系溶剂；The reaction solvent in step c is an alcohol solvent;
优选地，步骤c的反应溶剂为甲醇；Preferably, the reaction solvent in step c is methanol;
步骤c回流反应1～2h；Step c reflux reaction for 1～2h;
当Z为碘时，步骤d将中间体Ⅲ溶于反应溶剂中，加入碱，将I 2溶于反应溶剂中滴加至反应液，反应得到中间体Ⅳ； When Z is iodine, in step d, the intermediate III is dissolved in the reaction solvent, a base is added, and I 2 is dissolved in the reaction solvent and added dropwise to the reaction solution to obtain the intermediate IV through the reaction;
优选地，所述碱为碳酸氢钠、碳酸钠、碳酸钾、氢氧化钠、氢氧化钾中一种或两种以上；Preferably, the alkali is one or more of sodium bicarbonate, sodium carbonate, potassium carbonate, sodium hydroxide, and potassium hydroxide;
步骤d中中间体Ⅲ：I 2：碱的摩尔配比为1：(1.5-2.0)：2.0； In step d, the molar ratio of Intermediate III: I 2 : Base is 1: (1.5-2.0): 2.0;
步骤d的反应溶剂为DMF；The reaction solvent in step d is DMF;
步骤d的反应温度为25～80℃；The reaction temperature in step d is 25 to 80°C;
步骤d的反应时间为2～10h；The reaction time of step d is 2-10h;
步骤e所述钯催化剂为醋酸钯、[1,1'-双(二苯基膦基)二茂铁]二氯化钯、[1,1'-双(二苯基膦)二茂铁]二氯化钯二氯甲烷络合物、三(二亚苄基茚丙酮)二钯中一种或两种以上；The palladium catalyst in step e is palladium acetate, [1,1'-bis(diphenylphosphino)ferrocene]palladium dichloride, [1,1'-bis(diphenylphosphino)ferrocene] One or more of palladium dichloride dichloromethane complex and tris(dibenzylidene indeneacetone) dipalladium;
步骤e向反应体系中加入碱，所述碱为DIEA、碳酸钠、碳酸氢钠、碳酸钾、磷酸钾、碳酸铯中一种或两种以上；Step e: adding a base to the reaction system, the base being one or more of DIEA, sodium carbonate, sodium bicarbonate, potassium carbonate, potassium phosphate, and cesium carbonate;
步骤e中中间体Ⅳ：化合物3：碱：钯催化剂的摩尔配比为1：(1.1～1.5)：(2.0～3.0)：(0.003～0.010)；In step e, the molar ratio of intermediate IV: compound 3: base: palladium catalyst is 1: (1.1～1.5): (2.0～3.0): (0.003～0.010);
步骤e的反应溶剂为二氧六环、水、甲苯、DMF、正丁醇、异丙醇、仲丁醇中一种或两种以上；The reaction solvent in step e is one or more of dioxane, water, toluene, DMF, n-butanol, isopropanol, and sec-butanol;
优选地，步骤e的反应溶剂为1,4-二氧六环：水体积比为(4～8)：1的混合溶剂；Preferably, the reaction solvent in step e is a mixed solvent of 1,4-dioxane:water volume ratio (4-8):1;
步骤e的反应温度为90～110℃；The reaction temperature in step e is 90-110°C;
步骤e的反应时间为5～10h；The reaction time of step e is 5-10h;
步骤e于N 2保护下反应。 Step e is reacted under N 2 protection.
本发明提供了所述的化合物、其光学异构体、化合物或其光学异构体药学上可接受的盐在制备FGFR激酶抑制剂类药物中的用途。The present invention provides the use of the compound, its optical isomer, the compound or its optical isomer pharmaceutically acceptable salt in the preparation of FGFR kinase inhibitor drugs.
优选地，所述药物是FGFR1、FGFR2和/或FGFR3激酶抑制剂。Preferably, the drug is an FGFR1, FGFR2 and/or FGFR3 kinase inhibitor.
本发明提供了所述的化合物、其光学异构体、化合物或其光学异构体药学上可接受的盐在制备治疗和/或预防癌症的药物中的用途。The present invention provides the use of the compound, its optical isomer, the compound or its optical isomer pharmaceutically acceptable salt in the preparation of a medicine for treating and/or preventing cancer.
优选地，所述癌症为乳腺癌、肺癌、胃癌、肾癌、结直肠癌、肝癌、黑色素瘤、膀胱癌、尿路上皮癌和/或胆管癌。Preferably, the cancer is breast cancer, lung cancer, gastric cancer, kidney cancer, colorectal cancer, liver cancer, melanoma, bladder cancer, urothelial cancer and/or cholangiocarcinoma.
进一步优选地，所述肺癌为非小细胞肺癌。Further preferably, the lung cancer is non-small cell lung cancer.
本发明提供了所述的化合物、其光学异构体、化合物或其光学异构体药学上可接受的盐在制备治疗和/或预防器官纤维化的药物中的用途。The present invention provides the use of the compound, its optical isomer, the compound or its optical isomer pharmaceutically acceptable salt in the preparation of a medicine for treating and/or preventing organ fibrosis.
优选地，所述器官纤维化为肺纤维化或肝纤维化。Preferably, the organ fibrosis is pulmonary fibrosis or liver fibrosis.
本发明提供了药物组合物，它是以所述的化合物、其光学异构体、化合物或其光学异构体在药学上可接受的盐为活性成分，加入药学上可接受的辅料或者辅助性成分制备而成的制剂；优选地，所述制剂为口服制剂或注射制剂。The present invention provides a pharmaceutical composition, which uses the compound, its optical isomer, the compound or its pharmaceutically acceptable salt as the active ingredient, and pharmaceutically acceptable excipients or auxiliary materials are added. A preparation prepared from ingredients; preferably, the preparation is an oral preparation or an injection preparation.
术语定义：Definition of Terms:
本发明提供的化合物和衍生物可以根据IUPAC(国际纯粹与应用化学联合会)或CAS(化学文摘服务社，Columbus，OH)命名系统命名。The compounds and derivatives provided by the present invention can be named according to the IUPAC (International Union of Pure and Applied Chemistry) or CAS (Chemical Abstracts Service, Columbus, OH) naming system.
术语“烷基”是直链或支链的饱和烃基的基团。C 1～C 6烷基的实例包括但不限于甲 基(C 1)、乙基(C 2)、正丙基(C 3)、异丙基(C 3)、正丁基(C 4)、叔丁基(C 4)、仲丁基(C 4)、异丁基(C 4)、正戊基(C 5)、3-戊基(C 5)、戊基(C 5)、新戊基(C 5)、3-甲基-2-丁基(C 5)、叔戊基(C 5)和正己基(C 6)。 The term "alkyl" is a linear or branched saturated hydrocarbon group. Examples of C 1 -C 6 alkyl groups include but are not limited to methyl (C 1 ), ethyl (C 2 ), n-propyl (C 3 ), isopropyl (C 3 ), n-butyl (C 4 ) , Tert-butyl (C 4 ), sec-butyl (C 4 ), isobutyl (C 4 ), n-pentyl (C 5 ), 3-pentyl (C 5 ), pentyl (C 5 ), new Pentyl (C 5 ), 3-methyl-2-butyl (C 5 ), tert-pentyl (C 5 ) and n-hexyl (C 6 ).
术语“烷氧基”是指基团-OR，其中R是上文所定义的烷基。C 1～C 6烷氧基的实例包括但不限于甲氧基、乙氧基、正丙氧基、异丙氧基、正丁氧基、叔丁氧基、仲丁氧基、正戊氧基、正己氧基和1,2-二甲基丁氧基。 The term "alkoxy" refers to the group -OR, where R is alkyl as defined above. Examples of C 1 ～C 6 alkoxy include but are not limited to methoxy, ethoxy, n-propoxy, isopropoxy, n-butoxy, tert-butoxy, sec-butoxy, n-pentoxy Group, n-hexyloxy and 1,2-dimethylbutoxy.
术语“芳基”是指在芳族环系中包含或不包含杂原子的4n+2芳族环系的基团，其中，杂原子选自氮、氧和/或硫。The term "aryl" refers to a group of a 4n+2 aromatic ring system containing or not containing heteroatoms in the aromatic ring system, wherein the heteroatoms are selected from nitrogen, oxygen and/or sulfur.
术语“脂环”是指饱和或部分不饱和的环状烃基基团。The term "alicyclic" refers to a saturated or partially unsaturated cyclic hydrocarbon group.
术语“烷氧羰基”是指基团R-O-C(O)-，其中R是上文所定义的烷基，优选R是C 1～C 6烷基(即本发明所述C1～C6烷氧羰基)。其例子包括但不限于甲氧羰基、乙氧羰基、丙氧羰基、异丙氧羰基、丁氧羰基、异丁氧羰基、叔丁氧羰基。 The term "alkoxycarbonyl" refers to the group ROC (O) -, wherein R is alkyl as defined above, preferably R is a C 1 ~ C 6 alkyl (i.e., C1 ~ C6 of the present invention, the alkoxycarbonyl group) . Examples include, but are not limited to, methoxycarbonyl, ethoxycarbonyl, propoxycarbonyl, isopropoxycarbonyl, butoxycarbonyl, isobutoxycarbonyl, tert-butoxycarbonyl.
术语“药学上可接受的”是指某载体、运载物、稀释剂、辅料，和/或所形成的盐通常在化学上或物理上与构成某药物剂型的其它成分相兼容，并在生理上与受体相兼容。The term "pharmaceutically acceptable" means that a certain carrier, carrier, diluent, excipient, and/or the formed salt is usually chemically or physically compatible with other ingredients constituting a certain pharmaceutical dosage form, and physiologically Compatible with the receptor.
术语“药学上可接受的盐”是指本发明化合物与无机和/或有机酸和碱形成的酸式和/或碱式盐，也包括两性离子盐(内盐)，还包括季铵盐，例如烷基铵盐。这些盐可以是在化合物的最后分离和纯化中直接得到。也可以是通过将上述化合物与一定数量的酸或碱适当(例如等当量)进行混合而得到。这些盐可能在溶液中形成沉淀而以过滤方法收集，或在溶剂蒸发后回收而得到，或在水介质中反应后冷冻干燥制得。本发明中所述盐可以是化合物的盐酸盐、硫酸盐、枸橼酸盐、苯磺酸盐、氢溴酸盐、氢氟酸盐、磷酸盐、乙酸盐、丙酸盐、丁二酸盐、草酸盐、苹果酸盐、琥珀酸盐、富马酸盐、马来酸盐、酒石酸盐或三氟乙酸盐。The term "pharmaceutically acceptable salt" refers to the acid and/or basic salt formed by the compound of the present invention with inorganic and/or organic acids and bases, and also includes zwitterionic salts (internal salts), and also includes quaternary ammonium salts, For example, alkyl ammonium salts. These salts can be obtained directly in the final isolation and purification of the compound. It can also be obtained by appropriately mixing the above-mentioned compound with a certain amount of acid or base (e.g., equivalent). These salts may form a precipitate in the solution and be collected by filtration, or recovered after evaporation of the solvent, or prepared by freeze-drying after reaction in an aqueous medium. The salt in the present invention can be the hydrochloride, sulfate, citrate, benzenesulfonate, hydrobromide, hydrofluoride, phosphate, acetate, propionate, butane Acid salt, oxalate, malate, succinate, fumarate, maleate, tartrate or trifluoroacetate.
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、肠胃外(静脉内、肌肉内或皮下)、和局部给药。The method of administration of the compound or pharmaceutical composition of the present invention is not particularly limited, and representative administration methods include (but are not limited to): oral, parenteral (intravenous, intramuscular, or subcutaneous), and topical administration.
本发明所述药学上可接受的辅料，是指除活性成分以外包含在剂型中的物质。The pharmaceutically acceptable excipients in the present invention refer to substances contained in the dosage form in addition to the active ingredients.
本发明所述药学上可接受的辅助性成分，它具有一定生理活性，但该成分的加入不会改变上述药物组合物在疾病治疗过程中的主导地位，而仅仅发挥辅助功效，这些辅助功效仅仅是对该成分已知活性的利用，是医药领域惯用的辅助治疗方式。若将上述辅助性成分与本发明药物组合物配合使用，仍然应属于本发明保护的范围。The pharmaceutically acceptable auxiliary component of the present invention has certain physiological activity, but the addition of the component will not change the dominant position of the above-mentioned pharmaceutical composition in the course of disease treatment, but only exerts auxiliary functions. These auxiliary functions only It is the utilization of the known activity of the ingredient, and it is a commonly used adjuvant therapy in the medical field. If the aforementioned auxiliary components are used in combination with the pharmaceutical composition of the present invention, they should still fall within the protection scope of the present invention.
本发明提供了一类结构新颖的3-乙烯基吲唑类衍生物。生物学实验证明，本发明提 供的化合物对FGFR激酶的活性具有显著抑制作用，能有效抑制乳腺癌、肺癌、胃癌、肾癌、结直肠癌、肝癌、黑色素瘤、膀胱癌、尿路上皮癌及胆管癌等多种癌细胞的增殖，具有广谱的抗癌作用；此外，对成纤维细胞的增殖也表现出明显的抑制作用，效果与目前临床上用于治疗肺纤维化的药物尼达尼布相当，抗纤维化疗效显著，同时对肝星状细胞也具有良好的抑制作用。本发明为抗癌、抗纤维化药物的开发和应用提供了新的选择。The present invention provides a class of 3-vinylindazole derivatives with novel structures. Biological experiments have proved that the compound provided by the present invention has a significant inhibitory effect on the activity of FGFR kinase, and can effectively inhibit breast cancer, lung cancer, gastric cancer, kidney cancer, colorectal cancer, liver cancer, melanoma, bladder cancer, urothelial cancer and The proliferation of a variety of cancer cells such as cholangiocarcinoma has a broad-spectrum anti-cancer effect; in addition, it also shows a significant inhibitory effect on the proliferation of fibroblasts. The cloth is comparable, the anti-fibrosis effect is significant, and it also has a good inhibitory effect on hepatic stellate cells. The invention provides new options for the development and application of anti-cancer and anti-fibrosis drugs.
附图说明Description of the drawings
图1为生物学实验中本发明化合物4-20的药-时曲线图；Figure 1 is a drug-time curve diagram of compound 4-20 of the present invention in biological experiments;
图2为动物体内实验中给药14天后肺组织的HE和Masson染色图；Figure 2 is a graph of HE and Masson staining of lung tissue 14 days after administration in an animal experiment;
图3为动物体内实验中给药28天后肺组织的HE和Masson染色图。Figure 3 shows the HE and Masson staining of lung tissue 28 days after administration in an animal experiment.
具体实施方式Detailed ways
本发明具体实施方式中使用的原料、设备均为已知产品，通过购买市售产品获得。The raw materials and equipment used in the specific embodiments of the present invention are all known products, which are obtained by purchasing commercially available products.
实施例1 本发明化合物的制备Example 1 Preparation of the compound of the present invention
Figure PCTCN2020091613-appb-000027
Figure PCTCN2020091613-appb-000027
取化合物1-1(1.86g,10mmol)，化合物2-1(3.17g,12mmol)，Pd(dppf)Cl 2(5mmol％)，碳酸钾(2.76g,20mmol)，加入1,4-二氧六环与水(体积比为4:1)的混合溶剂50mL中，N 2置换后于100℃下反应6小时。TLC监测反应，反应完全后将反应液减压蒸除，剩余物用二氯甲烷与甲醇混合溶剂溶解，经硅藻土过滤，滤液浓缩后，经柱层析(PE:EA＝2:1)得到中间体Ⅰ-1(浅黄色固体，73％)，MSm/z(ESI)：244.1[M+H] +。 Take compound 1-1 (1.86g, 10mmol), compound 2-1 (3.17g, 12mmol), Pd(dppf)Cl 2 (5mmol%), potassium carbonate (2.76g, 20mmol), add 1,4-diox In 50 mL of a mixed solvent of hexacyclic ring and water (volume ratio 4:1), the mixture was replaced with N 2 and reacted at 100° C. for 6 hours. The reaction was monitored by TLC. After the reaction was completed, the reaction solution was evaporated under reduced pressure. The residue was dissolved in a mixed solvent of dichloromethane and methanol, filtered through Celite, and the filtrate was concentrated and subjected to column chromatography (PE:EA=2:1) Intermediate I-1 (light yellow solid, 73%) was obtained, MS m/z (ESI): 244.1 [M+H] + .
Figure PCTCN2020091613-appb-000028
Figure PCTCN2020091613-appb-000028
将中间体Ⅰ-1(1.21g,5mmol)溶于甲苯30mL中，加入乙酸钾(0.59g,6mmol)，乙酸酐(0.94mL,10mmol)，室温搅拌1h后加入亚硝酸异戊酯(2mL,15mmol)，回流5h后TLC监测反应。反应完全后将溶剂减压浓缩，剩余物加水搅拌后用EA萃取，有机相用无水硫酸钠干燥后浓缩，柱层析(PE：EA＝4:1)得中间体Ⅱ-1(浅黄色固体，57％)，MSm/z(ESI)：297.2[M+H] +。 Intermediate I-1 (1.21g, 5mmol) was dissolved in 30mL of toluene, potassium acetate (0.59g, 6mmol), acetic anhydride (0.94mL, 10mmol) were added, stirred at room temperature for 1h, and then isoamyl nitrite (2mL, 15mmol), and TLC monitored the reaction after refluxing for 5h. After the reaction was completed, the solvent was concentrated under reduced pressure, the residue was stirred with water and extracted with EA, the organic phase was dried over anhydrous sodium sulfate and concentrated, and column chromatography (PE:EA=4:1) gave Intermediate II-1 (light yellow) Solid, 57%), MS m/z (ESI): 297.2 [M+H] + .
Figure PCTCN2020091613-appb-000029
Figure PCTCN2020091613-appb-000029
将中间体Ⅱ-1(1.48g,5mmol)溶于甲醇50mL中，加入6NHCl(20mL)，回流1h后TLC监测。反应完毕后减压浓缩溶剂，剩余物用碳酸氢钠水溶液调节pH至8，将析出的固体过滤干燥，得到中间体Ⅲ-1(浅棕色固体，69％)，MSm/z(ESI)：255.1[M+H] +。 Intermediate II-1 (1.48 g, 5 mmol) was dissolved in 50 mL of methanol, 6N HCl (20 mL) was added, and TLC was monitored after refluxing for 1 h. After the reaction, the solvent was concentrated under reduced pressure. The residue was adjusted to pH 8 with aqueous sodium bicarbonate solution. The precipitated solid was filtered and dried to obtain Intermediate III-1 (light brown solid, 69%), MSm/z(ESI): 255.1 [M+H] + .
Figure PCTCN2020091613-appb-000030
Figure PCTCN2020091613-appb-000030
将中间体Ⅲ-1(1.27g,5mmol)溶于DMF10mL中，加入碳酸钾(1.38g,10mmol)，室温搅拌下加入I 2(1.9g,7.5mmol)，65℃下反应8h后TLC监测。反应完毕后用保险粉的饱和水溶液淬灭反应，将反应液倒入水中，有白色固体析出，将固体过滤后干燥，得中间体Ⅳ-1(浅黄色固体，92％)，MSm/z(ESI)：381.0[M+H] +。 Intermediate III-1 (1.27g, 5mmol) was dissolved in DMF10mL, potassium carbonate (1.38g, 10mmol) was added, I 2 (1.9g, 7.5mmol) was added with stirring at room temperature, and the reaction was carried out at 65°C for 8 hours, followed by TLC monitoring. After the completion of the reaction, the reaction was quenched with a saturated aqueous solution of sodium hydroxide. The reaction solution was poured into water. A white solid was precipitated. The solid was filtered and dried to obtain Intermediate IV-1 (light yellow solid, 92%), MSm/z( ESI): 381.0[M+H] + .
Figure PCTCN2020091613-appb-000031
Figure PCTCN2020091613-appb-000031
取4-溴-N,N-二甲基苄胺(642mg,3mmol)，乙烯基硼酸频哪醇酯(612μL,3.6mmol)，DIEA(884μL,5.4mmol)，加入干燥甲苯20mL，N 2置换后于95℃下反应6小时.TLC监测反应。反应完全后减压蒸除溶剂，剩余物用二氯甲烷与甲醇混合溶剂溶解，经硅藻土过滤，滤液浓缩后，经薄层色谱层析(DCM:MeOH＝15:1)得到化合物3-1(浅黄色固体，77％)，MSm/z(ESI)：288.2[M+H] +。 Take 4-bromo-N,N-dimethylbenzylamine (642mg, 3mmol), vinyl boronic acid pinacol ester (612μL, 3.6mmol), DIEA (884μL, 5.4mmol), add 20mL of dry toluene, replace with N 2 The reaction was carried out at 95°C for 6 hours. The reaction was monitored by TLC. After the reaction was complete, the solvent was evaporated under reduced pressure, the residue was dissolved in a mixed solvent of dichloromethane and methanol, filtered through Celite, and the filtrate was concentrated, and then the compound was obtained by thin layer chromatography (DCM:MeOH=15:1). 1 (light yellow solid, 77%), MS m/z (ESI): 288.2 [M+H] + .
Figure PCTCN2020091613-appb-000032
Figure PCTCN2020091613-appb-000032
取中间体Ⅳ-1(190mg,0.5mmol)，化合物3-1(172mg,0.6mmol)，Pd(dppf)Cl 2(5mmol％)，碳酸钾(138mg,1mmol)，加入1,4-二氧六环与水(体积比4:1)的混合溶剂15mL中，N 2置换后于100℃下反应8小时。TLC监测反应，反应完全后将反应液减压蒸除，剩余物用二氯甲烷与甲醇混合溶剂溶解，经硅藻土过滤，滤液浓缩后，经薄层色谱层析(DCM:MeOH＝10:1)得到化合物4-1(浅黄色固体，17％)。 1H NMR(400MHz,DMSO-d 6)δ13.20(s,1H),8.23(d,J＝8.4Hz,1H),7.77–7.72(m,1H),7.67(d,J＝8.0Hz,2H),7.57–7.47(m,3H),7.32(d,J＝8.0Hz,2H),6.87(d,J＝2.3Hz,2H),6.55(t,J＝2.2Hz,1H),3.84(s,6H),3.40(s,2H),2.16(s,6H). Take Intermediate IV-1 (190mg, 0.5mmol), Compound 3-1 (172mg, 0.6mmol), Pd(dppf)Cl 2 (5mmol%), potassium carbonate (138mg, 1mmol), add 1,4-diox In 15 mL of a mixed solvent of hexacyclic ring and water (volume ratio 4:1), the mixture was replaced with N 2 and reacted at 100° C. for 8 hours. The reaction was monitored by TLC. After the reaction was completed, the reaction solution was evaporated under reduced pressure. The residue was dissolved in a mixed solvent of dichloromethane and methanol, filtered through celite, and the filtrate was concentrated and subjected to thin layer chromatography (DCM:MeOH=10: 1) Compound 4-1 (light yellow solid, 17%) was obtained. 1 H NMR(400MHz,DMSO-d 6 )δ13.20(s,1H), 8.23(d,J=8.4Hz,1H), 7.77–7.72(m,1H), 7.67(d,J=8.0Hz, 2H), 7.57–7.47 (m, 3H), 7.32 (d, J = 8.0 Hz, 2H), 6.87 (d, J = 2.3 Hz, 2H), 6.55 (t, J = 2.2 Hz, 1H), 3.84 ( s,6H), 3.40(s,2H), 2.16(s,6H).
使用类似方法可得到以下化合物：Using similar methods, the following compounds can be obtained:
化合物4-2Compound 4-2
Figure PCTCN2020091613-appb-000033
Figure PCTCN2020091613-appb-000033
1H NMR(400MHz,DMSO-d 6)δ13.08(s,1H),8.19(d,J＝8.5Hz,1H),7.72(s,1H),7.57(d,J＝8.5Hz,2H),7.51–7.39(m,2H),7.34(d,J＝16.7Hz,1H),6.96(d,J＝8.5Hz,2H),6.86(d,J＝2.2Hz,2H),6.54(t,J＝2.2Hz,1H),3.84(s,6H),3.20(t,J＝5.0Hz,4H),2.49–2.41(m,4H),2.24(s,3H). 1 H NMR(400MHz,DMSO-d 6 )δ13.08(s,1H), 8.19(d,J=8.5Hz,1H), 7.72(s,1H), 7.57(d,J=8.5Hz,2H) ,7.51–7.39(m,2H),7.34(d,J=16.7Hz,1H), 6.96(d,J=8.5Hz,2H), 6.86(d,J=2.2Hz,2H), 6.54(t, J = 2.2Hz, 1H), 3.84 (s, 6H), 3.20 (t, J = 5.0 Hz, 4H), 2.49-2.41 (m, 4H), 2.24 (s, 3H).
化合物4-3Compound 4-3
Figure PCTCN2020091613-appb-000034
Figure PCTCN2020091613-appb-000034
1H NMR(400MHz,DMSO-d 6)δ13.35(s,1H),8.28(d,J＝8.5Hz,1H),7.99(d,J＝8.1Hz,2H),7.88(d,J＝8.2Hz,2H),7.75(d,J＝17.2Hz,2H),7.62(d,J＝16.7Hz,1H),7.53(dd,J＝8.5,1.5Hz,1H),6.88(d,J＝2.2Hz,2H),6.56(d,J＝2.3Hz,1H),3.87(s,3H),3.85(s,6H). 1 H NMR (400MHz, DMSO-d 6 ) δ 13.35 (s, 1H), 8.28 (d, J = 8.5 Hz, 1H), 7.99 (d, J = 8.1 Hz, 2H), 7.88 (d, J = 8.2Hz, 2H), 7.75 (d, J = 17.2 Hz, 2H), 7.62 (d, J = 16.7 Hz, 1H), 7.53 (dd, J = 8.5, 1.5 Hz, 1H), 6.88 (d, J = 2.2Hz, 2H), 6.56 (d, J = 2.3Hz, 1H), 3.87 (s, 3H), 3.85 (s, 6H).
化合物4-4Compound 4-4
Figure PCTCN2020091613-appb-000035
Figure PCTCN2020091613-appb-000035
1H NMR(400MHz,DMSO-d 6)δ13.28(s,1H),8.26(d,J＝8.4Hz,1H),7.81(d,J＝8.0Hz,2H),7.75(s,1H),7.66(d,J＝16.8Hz,1H),7.57(d,J＝16.7Hz,1H),7.52(dd,J＝8.5,1.5Hz,1H),7.45(d,J＝8.2Hz,2H),6.87(d,J＝2.2Hz,2H),6.55(t,J＝2.2Hz,1H),3.84(s,6H),3.40(s,8H),1.42(s,9H). 1 H NMR (400MHz, DMSO-d 6 ) δ 13.28 (s, 1H), 8.26 (d, J = 8.4 Hz, 1H), 7.81 (d, J = 8.0 Hz, 2H), 7.75 (s, 1H) ,7.66(d,J=16.8Hz,1H),7.57(d,J=16.7Hz,1H),7.52(dd,J=8.5,1.5Hz,1H),7.45(d,J=8.2Hz,2H) ,6.87(d,J=2.2Hz,2H), 6.55(t,J=2.2Hz,1H), 3.84(s,6H), 3.40(s,8H),1.42(s,9H).
化合物4-6Compound 4-6
Figure PCTCN2020091613-appb-000036
Figure PCTCN2020091613-appb-000036
1H NMR(400MHz,DMSO-d 6)δ13.33(s,1H),8.28(d,J＝8.5Hz,1H),7.86–7.76(m,3H),7.67(d,J＝16.7Hz,1H),7.62–7.51(m,2H),7.45(d,J＝7.9Hz,2H),7.23–7.13(m,2H),6.91–6.83(m,1H),4.08(s,4H),3.87(s,6H),3.09(s,2H),1.41(s,9H),1.10(d,J＝19.9Hz,6H). 1H NMR(400MHz,DMSO-d 6 )δ13.33(s,1H), 8.28(d,J=8.5Hz,1H),7.86-7.76(m,3H),7.67(d,J=16.7Hz,1H ), 7.62–7.51(m, 2H), 7.45(d, J=7.9Hz, 2H), 7.23–7.13(m, 2H), 6.91–6.83(m, 1H), 4.08(s, 4H), 3.87( s, 6H), 3.09 (s, 2H), 1.41 (s, 9H), 1.10 (d, J = 19.9 Hz, 6H).
化合物4-8Compound 4-8
Figure PCTCN2020091613-appb-000037
Figure PCTCN2020091613-appb-000037
1H NMR(400MHz,DMSO-d 6)δ13.10(s,1H),8.15(d,J＝8.5Hz,1H),8.01(s,1H),7.84(s,1H),7.76(s,1H),7.49(d,J＝8.5Hz,1H),7.38(d,J＝16.7Hz,1H),7.28–7.11(m,3H),6.86(dt,J＝10.9,2.3Hz,1H),4.93(s,1H),4.16(t,J＝5.6Hz,2H),3.87(s,6H),3.77(q,J＝5.5Hz,2H). 1H NMR (400MHz, DMSO-d 6 ) δ 13.10 (s, 1H), 8.15 (d, J = 8.5 Hz, 1H), 8.01 (s, 1H), 7.84 (s, 1H), 7.76 (s, 1H) ), 7.49 (d, J = 8.5Hz, 1H), 7.38 (d, J = 16.7Hz, 1H), 7.28-7.11 (m, 3H), 6.86 (dt, J = 10.9, 2.3 Hz, 1H), 4.93 (s, 1H), 4.16 (t, J = 5.6 Hz, 2H), 3.87 (s, 6H), 3.77 (q, J = 5.5 Hz, 2H).
化合物4-9Compound 4-9
Figure PCTCN2020091613-appb-000038
Figure PCTCN2020091613-appb-000038
1H NMR(400MHz,DMSO-d 6)δ13.40(s,1H),8.28(d,J＝8.5Hz,1H),7.82(d,J＝8.1Hz,3H),7.65(t,J＝17.6Hz,2H),7.58–7.51(m,1H),7.47(d,J＝7.9Hz,2H),7.24–7.11(m,2H),6.87(d,J＝10.9Hz,1H),3.87(s,6H),3.63(s,4H),3.00(s,4H)2.20(s,3H). 1 H NMR (400MHz, DMSO-d 6 ) δ 13.40 (s, 1H), 8.28 (d, J = 8.5 Hz, 1H), 7.82 (d, J = 8.1 Hz, 3H), 7.65 (t, J = 17.6Hz, 2H), 7.58–7.51 (m, 1H), 7.47 (d, J = 7.9 Hz, 2H), 7.24–7.11 (m, 2H), 6.87 (d, J = 10.9 Hz, 1H), 3.87 ( s, 6H), 3.63 (s, 4H), 3.00 (s, 4H) 2.20 (s, 3H).
实施例2 本发明化合物的制备Example 2 Preparation of the compound of the present invention
Figure PCTCN2020091613-appb-000039
Figure PCTCN2020091613-appb-000039
取化合物4-4(50mg)，加入HCl/1,4-二氧六环溶液10ml，室温下搅拌5h，浓缩溶剂，剩余物用甲醇溶解后经薄层色谱层析(DCM:MeOH＝8:1)得化合物4-5(浅棕色固体，52％)。 1H NMR(400MHz,DMSO-d 6)δ9.58(s,2H),8.26(d,J＝8.5Hz,1H),7.83(d,J＝7.9Hz,2H),7.77(s,1H),7.68(d,J＝16.7Hz,1H),7.58(d,J＝16.7Hz,1H),7.51(dd,J＝7.9,4.4Hz,3H),6.87(d,J＝2.2Hz,2H),6.55(t,J＝2.2Hz,1H),3.84(s,6H),3.78–3.63 (m,4H),3.16(s,4H). Take compound 4-4 (50mg), add 10ml of HCl/1,4-dioxane solution, stir at room temperature for 5h, concentrate the solvent, the residue is dissolved in methanol and then subjected to thin layer chromatography (DCM:MeOH=8: 1) Compound 4-5 (light brown solid, 52%) was obtained. 1 H NMR(400MHz,DMSO-d 6 )δ9.58(s,2H), 8.26(d,J=8.5Hz,1H), 7.83(d,J=7.9Hz,2H), 7.77(s,1H) ,7.68(d,J=16.7Hz,1H),7.58(d,J=16.7Hz,1H),7.51(dd,J=7.9,4.4Hz,3H), 6.87(d,J=2.2Hz,2H) ,6.55(t,J=2.2Hz,1H),3.84(s,6H),3.78–3.63 (m,4H),3.16(s,4H).
使用类似方法可以得到以下化合物：Using similar methods, the following compounds can be obtained:
化合物4-7Compound 4-7
Figure PCTCN2020091613-appb-000040
Figure PCTCN2020091613-appb-000040
1H NMR(400MHz,DMSO-d6)δ13.35(s,1H),8.28(d,J＝8.5Hz,1H),7.84–7.78(m,3H),7.66(d,J＝16.8Hz,1H),7.61–7.51(m,2H),7.40(d,J＝8.0Hz,2H),7.23–7.13(m,2H),6.87(dt,J＝10.9,2.3Hz,1H),3.87(s,6H),3.63–3.35(m,3H),2.69(d,J＝8.5Hz,3H),1.10–0.80(m,7H).1H NMR(400MHz,DMSO-d6)δ13.35(s,1H), 8.28(d,J=8.5Hz,1H),7.84-7.78(m,3H),7.66(d,J=16.8Hz,1H) ,7.61-7.51(m,2H),7.40(d,J=8.0Hz,2H),7.23-7.13(m,2H),6.87(dt,J=10.9,2.3Hz,1H),3.87(s,6H ), 3.63–3.35(m,3H), 2.69(d,J=8.5Hz,3H), 1.10–0.80(m,7H).
下表1中化合物的制备方法与上述化合物类似。The preparation methods of the compounds in Table 1 below are similar to the above compounds.
表1 本发明部分化合物的质谱数据Table 1 Mass spectrometry data of some compounds of the present invention
Figure PCTCN2020091613-appb-000041
Figure PCTCN2020091613-appb-000041
Figure PCTCN2020091613-appb-000042
Figure PCTCN2020091613-appb-000042
Figure PCTCN2020091613-appb-000043
Figure PCTCN2020091613-appb-000043
Figure PCTCN2020091613-appb-000044
Figure PCTCN2020091613-appb-000044
Figure PCTCN2020091613-appb-000045
Figure PCTCN2020091613-appb-000045
Figure PCTCN2020091613-appb-000046
Figure PCTCN2020091613-appb-000046
Figure PCTCN2020091613-appb-000047
Figure PCTCN2020091613-appb-000047
Figure PCTCN2020091613-appb-000048
Figure PCTCN2020091613-appb-000048
Figure PCTCN2020091613-appb-000049
Figure PCTCN2020091613-appb-000049
Figure PCTCN2020091613-appb-000050
Figure PCTCN2020091613-appb-000050
Figure PCTCN2020091613-appb-000051
Figure PCTCN2020091613-appb-000051
Figure PCTCN2020091613-appb-000052
Figure PCTCN2020091613-appb-000052
以下通过生物学实验证明本发明的有益效果。The following biological experiments prove the beneficial effects of the present invention.
一、实验仪器及材料1. Experimental equipment and materials
主要生物实验仪器和设备如下所述。超净工作台BHC-1000IIA/B3：苏净安泰生物技术公司；恒温水浴箱PolyScience 9505：PolyScience公司；灭菌锅MLS-3780：SANYO公司；烘箱：Binder公司；超纯水仪Milli-Q Integral 10：Millipore公司；酶标仪Multiscan MK3、细胞培养箱、低速离心机Sorvall ST1：Thermofisher公司；Centrifuge 5415C超速离心机：德国Eppendorf公司；NUAIRE NU-425-600E生物安全柜：美国Nuaire公司；BCD-215YD型普通冰箱：中国Haier公司；SANYO(-80℃)超低温冰箱：日本三洋电器集团；Rocker 51702摇床：美国Cole Parmer公司；96孔细胞培养板：Costa Corning公司；普通光学显微镜：Olympus公司；移液枪：Thermo公司；PH计：Coring公司；高压灭菌锅：SANYO公司。The main biological experiment instruments and equipment are as follows. Ultra-clean workbench BHC-1000IIA/B3: Sujing Antai Biotechnology Company; Constant temperature water bath PolyScience 9505: PolyScience Company; Sterilizer MLS-3780: SANYO Company; Oven: Binder Company; Ultra-pure water meter Milli-Q Integral 10 ：Millipore company; microplate reader Multiscan MK3, cell incubator, low-speed centrifuge Sorvall ST1: Thermofisher company; Centrifuge 5415C ultracentrifuge: Germany Eppendorf company; NUAIRE NU-425-600E biological safety cabinet: American Nuaire company; BCD-215YD Type ordinary refrigerator: China Haier company; SANYO (-80℃) ultra-low temperature refrigerator: Japan Sanyo Electric Group; Rocker 51702 shaker: American Cole Parmer company; 96-well cell culture plate: Costa Corning company; ordinary optical microscope: Olympus company; Liquid gun: Thermo Company; PH meter: Coring Company; Autoclave: SANYO Company.
实验中采用的细胞系购自美国ATCC公司。培养细胞所用各种必需品购自Gibco BRL公司，包括DMEM培养基、RPMI 1640培养基、胎牛血清(FBS)和胰酶。四甲基偶氮唑蓝(MTT)、二甲基亚砜(DMSO)购自美国Sigma公司。The cell line used in the experiment was purchased from ATCC, USA. Various necessities for cell culture were purchased from Gibco BRL, including DMEM medium, RPMI 1640 medium, fetal bovine serum (FBS) and pancreatin. Tetramethylazazole blue (MTT) and dimethyl sulfoxide (DMSO) were purchased from Sigma Company in the United States.
二、实验方法2. Experimental method
1、激酶测试1. Kinase test
用DMSO将化合物稀释至反应中最终所需最高抑制剂浓度的50倍。将100μL化合物稀释液转移到96孔板中。在同一96孔板的两个空白孔中加入100μL DMSO。该96孔板作为源板。将10μL化合物从源板转移到的96孔板中，作为中间板。向中间板的每个孔中加入90μL l1x激酶缓冲液。在振荡器上将中间板中的化合物混合10分钟。将96孔中间板的每孔取5μL转移至384孔板，设置副孔。在1x激酶碱缓冲液中加入激酶。在1x激酶碱缓冲液中加入FAM标记的肽和ATP。测定板中已含有5μL化合物10％DMSO溶液。向384孔测定板的每个孔中加入10μL l2.5x酶溶液。在室温下孵育10分钟。向384孔测定板的每个孔中加入10μL的2.5x肽溶液。在28℃下孵育特定时间后，加入25μL终止缓冲液以停止反应。在Caliper上收集数据，将数据进一步换算为IC 50。 The compound was diluted with DMSO to 50 times the final highest inhibitor concentration required in the reaction. Transfer 100 μL of compound dilution to a 96-well plate. Add 100μL of DMSO to the two blank wells of the same 96-well plate. This 96-well plate serves as the source plate. Transfer 10 μL of compound from the source plate to the 96-well plate as the intermediate plate. Add 90 μL of l1x kinase buffer to each well of the middle plate. Mix the compounds in the middle plate on a shaker for 10 minutes. Transfer 5μL from each well of the 96-well middle plate to a 384-well plate, and set up secondary wells. Add kinase to 1x Kinase Alkaline Buffer. Add FAM-labeled peptide and ATP to 1x Kinase Alkaline Buffer. The assay plate already contains 5 μL of compound 10% DMSO solution. Add 10 μL of l2.5x enzyme solution to each well of the 384-well assay plate. Incubate for 10 minutes at room temperature. Add 10 μL of 2.5x peptide solution to each well of the 384-well assay plate. After incubating at 28°C for a specific time, add 25 μL of stop buffer to stop the reaction. Collect data on Caliper and convert the data to IC 50 .
2、细胞培养2. Cell culture
从液氮中取出冻存保种的肿瘤细胞，迅速置于37℃恒温水浴复温融化，在无菌条件下用培养基洗涤1次。然后用完全培养基接种于培养瓶内，在37℃，5％CO 2培养箱中培养，第二天置换新鲜细胞培养液。悬浮生长细胞的传代：细胞培养2～3天后，从培养箱中取出培养瓶，收集细胞悬液于离心管中，1500rpm/min，离心3min，倒去上清液，细胞沉淀用完全培养基重悬并吹打均匀，然后分至3～5瓶培养。一般3～4天传代1次； 贴壁生长细胞的传代：细胞贴壁长满至瓶底的80％左右，从培养箱中取出培养瓶，吸去培养基，用0.25％胰酶洗涤1次，然后加入0.25％的胰酶消化液消化，观察细胞收缩变圆后，加入完全培养基终止消化，并吹打使细胞分散脱落，收集细胞悬液，1500rpm/min，离心3min，倒去上清液，细胞沉淀用完全培养基重悬并吹打均匀，然后分至3～5瓶瓶培养。一般3～4天传代1次。 Take out the cryopreserved tumor cells from the liquid nitrogen, quickly place them in a 37°C constant temperature water bath to rewarm and thaw, and wash them once with the culture medium under aseptic conditions. Then inoculate the culture flask with complete medium, culture in a 37°C, 5% CO 2 incubator, and replace with fresh cell culture medium the next day. Subculture of suspended growth cells: After cell culture for 2 to 3 days, remove the culture flask from the incubator, collect the cell suspension in a centrifuge tube, centrifuge at 1500 rpm/min for 3 minutes, pour the supernatant, and regenerate the cell pellet with complete medium Suspend and pipette evenly, and then divide into 3 to 5 bottles for culture. Passage is generally 1 time in 3 to 4 days; Passage of adherent growth cells: The cells adhere to the wall and grow to about 80% of the bottom of the bottle, remove the culture bottle from the incubator, aspirate the medium, and wash once with 0.25% pancreatin Then add 0.25% trypsin digestion solution for digestion. After observing the cell shrinkage and rounding, add complete medium to stop the digestion, and pipette to disperse and fall off the cells. Collect the cell suspension, centrifuge at 1500rpm/min for 3min, and pour the supernatant. , The cell pellet is resuspended in complete medium and pipetted evenly, and then divided into 3 to 5 bottles for culture. Generally, it is passaged once every 3 to 4 days.
3、细胞增殖抑制实验(MTT法)3. Cell proliferation inhibition test (MTT method)
收集对数生长期的细胞(4T1鼠源乳腺癌细胞株；MDA-MB-231人源乳腺癌细胞株；A549人源非小细胞肺癌株；SUN-16人低分化胃癌细胞；NIH3T3小鼠成纤维细胞；人肺成纤维细胞HPF(Catalog#3300))，以每孔2.5×10 3～1×10 4个的数量接种于96孔板中，在37℃、5％CO 2的细胞培养箱中培养过夜24小时，采用DMEM培养基稀释待测药物并加入到96孔板中，每种药物8个梯度，每个梯度含3个复孔。加药组中，按梯度(终浓度分别为1000、333、127、42.3、14.1、4.7、1.56、0.53nM)每孔加入100μL化合物的培养基溶液，每个浓度设3个复孔；阴性对照组每孔中加入含1‰DMSO的空白培养基100μL，共6个复孔；空白对照组每孔中只加入100μL培养基。将板置于37℃、5％CO 2细胞培养孵箱内培养72小时。药物处理组、隐形对照组和空白组每孔加入20μL MTT溶液(5mg/mL)，继续培养2-4小时，待甲瓒形成后，终止培养，倾去上清液后每孔加150μL DMSO(对于悬浮细胞则直接加入50μL 20％SDS溶液)，在摇床上摇15～20分钟。用酶标仪在570nm波长下检测每孔细胞吸光度(OD 570)，取其平均值记录结果。细胞增殖抑制率＝(对照组OD 570-实验组OD 570)/(对照组OD 570-空白OD 570)×100％。最后，使用Graphpad Prism软件拟合半数抑制浓度。 Collect the cells in the logarithmic growth phase (4T1 mouse breast cancer cell line; MDA-MB-231 human breast cancer cell line; A549 human non-small cell lung cancer line; SUN-16 human poorly differentiated gastric cancer cell; NIH3T3 mouse adult Fibroblasts; human lung fibroblasts HPF (Catalog#3300)), seeded in a 96-well plate at a rate of 2.5×10 3 to 1×10 4 per well, in a cell incubator at 37°C and 5% CO 2 Incubate in medium overnight for 24 hours, dilute the drug to be tested with DMEM medium and add it to a 96-well plate. Each drug has 8 gradients, and each gradient contains 3 replicate wells. In the dosing group, add 100 μL of the compound medium solution to each well according to the gradient (final concentrations are 1000, 333, 127, 42.3, 14.1, 4.7, 1.56, 0.53 nM), and each concentration has 3 replicate wells; negative control In the group, 100 μL of blank medium containing 1‰ DMSO was added to each well for a total of 6 multiple wells; in the blank control group, only 100 μL of medium was added to each well. Place the plate in a 37°C, 5% CO 2 cell culture incubator for 72 hours. In the drug treatment group, the invisible control group and the blank group, add 20μL of MTT solution (5mg/mL) to each well, continue to incubate for 2-4 hours. For suspended cells, add 50 μL of 20% SDS solution directly, and shake on a shaker for 15-20 minutes. Detect the cell absorbance (OD 570 ) of each well with a microplate reader at a wavelength of 570 nm, and take the average value and record the result. Cell proliferation inhibition rate=(control group OD570 -experimental group OD570 )/(control group OD570 -blank OD570 )×100%. Finally, use Graphpad Prism software to fit the half inhibitory concentration.
4、药代动力学研究4. Pharmacokinetic research
本发明化合物4-20药代动力学分析采用美迪西普亚医药科技(上海)有限公司提供的测试服务完成。该实验采用的SPF级SD雄性大鼠8只(6只为实验鼠，其余为空白对照)转移自实验机构动物储备库(999M-017)，上海西普尔-必凯实验动物有限公司。实验分为两组，每组3只鼠，分别为：给药剂量2mg/kg，给药浓度0.4mg/mL，给药体积5mL/kg的单次静脉注射和给药剂量20mg/kg，给药浓度2mg/mL，给药体积10mL/kg的灌胃口服(口服给药组动物给药前禁食过夜10-14h，给药4h后给食)。并设置采血时间点为：给药前，给药后0.083h、0.25h、0.5h、1h、2h、4h、6h、8h、24h共十次，经颈静脉采血后，每个样品采集约0.20mL，肝素钠抗凝，采集后放置冰上。血浆样品处理：血液样本采集后置于冰上，并于1小时之内离心分离血浆(离心条件：6800g，6mins， 2-8℃)。血浆样本在分析前存放时则放于-80℃冰箱内。结果分析：通过不同时间点的血药浓度数据，运用Phoenix WinNonlin7.0计算药代动力学参数，提供AUC 0-t、AUC 0- ∞、MRT 0-∞、C max、T max和T 1/2等参数及其平均值和标准差。 The pharmacokinetic analysis of the compound 4-20 of the present invention is completed by the testing service provided by Medicipuya Pharmaceutical Technology (Shanghai) Co., Ltd. 8 SPF-grade male SD rats (6 experimental rats, and the rest as blank controls) used in this experiment were transferred from the experimental institution animal reserve (999M-017), Shanghai Xipuer-Bikai Experimental Animal Co., Ltd. The experiment was divided into two groups, each group of 3 mice, respectively: a single intravenous injection with a dose of 2 mg/kg, a dose of 0.4 mg/mL, a dose of 5 mL/kg, and a dose of 20 mg/kg. The drug concentration was 2 mg/mL and the administration volume was 10 mL/kg. The animals in the oral administration group were fasted overnight for 10-14 hours before administration and 4 hours after administration. And set the blood sampling time points as follows: before administration, after administration 0.083h, 0.25h, 0.5h, 1h, 2h, 4h, 6h, 8h, 24h, 10 times, after blood collection through the jugular vein, each sample is collected about 0.20 mL, heparin sodium is anticoagulated and placed on ice after collection. Plasma sample processing: place the blood sample on ice after collection, and centrifuge to separate the plasma within 1 hour (centrifugation conditions: 6800g, 6mins, 2-8°C). Plasma samples were stored in a refrigerator at -80°C before analysis. Result analysis: According to the blood drug concentration data at different time points, Phoenix WinNonlin7.0 is used to calculate the pharmacokinetic parameters, providing AUC 0-t , AUC 0- ∞ , MRT 0-∞ , C max , T max and T 1/ 2 and other parameters and their mean and standard deviation.
5、动物体内抑制肺纤维化实验5. Experiments on inhibiting pulmonary fibrosis in animals
1.动物模型及给药1. Animal model and administration
雄性C57BL/6小鼠(7-9周龄，体重18-22g)购自华阜康(中国北京)。将小鼠圈养并在SPF条件下维持在设施中。在实验的第0天，先用10％水合氯醛将小鼠麻醉，然后给小鼠单次气管内滴注溶于生理盐水(2mg/kg体重)的硫酸博来霉素，同时注射等体积的生理盐水到对照组的大鼠中。第二天随机将小鼠分组，每组10只，每天口服灌胃本发明化合物4-20(YTH-17；30mg/kg，60mg/kg)，4-22(YTH-18；30mg/kg，60mg/kg)，阳性对照为尼达尼布(Nintedanib，BIBF1120)(30mg/Kg，溶剂为(5％DMSO+40％PEG400+55％生理盐水))药和等体积的溶剂做对照。给药14或28天后，处死小鼠。Male C57BL/6 mice (7-9 weeks old, weighing 18-22 g) were purchased from Hua Fukang (Beijing, China). The mice are housed and maintained in the facility under SPF conditions. On the 0th day of the experiment, the mice were anesthetized with 10% chloral hydrate, and then a single intratracheal instillation of bleomycin sulfate dissolved in normal saline (2 mg/kg body weight) was given to the mice, and an equal volume was injected at the same time To the rats in the control group. The mice were randomly divided into groups on the second day, each with 10 mice, and the compound 4-20 (YTH-17; 30mg/kg, 60mg/kg) and 4-22 (YTH-18; 30mg/kg) of the present invention were orally administered orally every day 60mg/kg), the positive control is Nintedanib (BIBF1120) (30mg/Kg, the solvent is (5% DMSO + 40% PEG400 + 55% normal saline)) medicine and an equal volume of solvent as a control. After 14 or 28 days of administration, the mice were sacrificed.
2.HE和Masson染色2. HE and Masson staining
在实验的第14天或28天处死小鼠。将肺组织样品置于4％(m/v)PBS-缓冲的多聚甲醛溶液中，三天后，取部分组织冲水2h，后使用梯度乙醇脱水并包埋在石蜡中。将包裹在石蜡中的组织切割连续切片(3μm)并用苏木精和伊红或Masson三色染色以评估组织病理学变化和累积的胶原蛋白纤维的程度。The mice were sacrificed on the 14th or 28th day of the experiment. The lung tissue samples were placed in 4% (m/v) PBS-buffered paraformaldehyde solution. Three days later, part of the tissue was rinsed in water for 2 hours, then dehydrated with gradient ethanol and embedded in paraffin. The tissues wrapped in paraffin were cut into serial sections (3 μm) and stained with hematoxylin and eosin or Masson trichrome to evaluate the histopathological changes and the degree of accumulated collagen fibers.
三、体外实验结果3. Results of in vitro experiments
1、下表2列举了本发明合成的部分化合物对FGFR1激酶的抑制情况。字母A代表IC 50为50nM以下，字母B代表IC 50为50nM至100nM，字母C代表IC 50为100nM至500nM，字母D代表IC 50为500nM以上。 1. Table 2 below lists the inhibition of FGFR1 kinase by some of the compounds synthesized in the present invention. The letter A represents an IC 50 of 50 nM or less, the letter B represents an IC 50 of 50 nM to 100 nM, the letter C represents an IC 50 of 100 nM to 500 nM, and the letter D represents an IC 50 of 500 nM or more.
表2 本发明部分化合物抑制FGFR1激酶的IC 50值 Table 2 The IC 50 value of some compounds of the present invention for inhibiting FGFR1 kinase
Figure PCTCN2020091613-appb-000053
Figure PCTCN2020091613-appb-000053
Figure PCTCN2020091613-appb-000054
Figure PCTCN2020091613-appb-000054
2、下表3列举了本发明合成的部分化合物对多种激酶的抑制情况。字母A代表IC 50为50nM以下，字母B代表IC 50为50nM至100nM，字母C代表IC 50为100nM至500nM，字母D代表IC 50为500nM以上。 2. Table 3 below lists the inhibitory effects of some of the compounds synthesized in the present invention on various kinases. The letter A represents an IC 50 of 50 nM or less, the letter B represents an IC 50 of 50 nM to 100 nM, the letter C represents an IC 50 of 100 nM to 500 nM, and the letter D represents an IC 50 of 500 nM or more.
表3 本发明部分化合物抑制相关激酶的IC 50值 Table 3 The IC 50 values of some compounds of the present invention for inhibiting related kinases
Figure PCTCN2020091613-appb-000055
Figure PCTCN2020091613-appb-000055
Figure PCTCN2020091613-appb-000056
Figure PCTCN2020091613-appb-000056
3、下表4列举了本发明合成的部分化合物在72小时作用下对SNU16细胞的增殖抑制情况。字母A代表IC 50为50nM以下，字母B代表IC 50为50nM至100nM，字母C代表IC 50为100nM至500nM，字母D代表IC 50为500nM以上。 3. Table 4 below lists the inhibition of the proliferation of SNU16 cells by some of the compounds synthesized by the present invention under 72 hours of action. The letter A represents an IC 50 of 50 nM or less, the letter B represents an IC 50 of 50 nM to 100 nM, the letter C represents an IC 50 of 100 nM to 500 nM, and the letter D represents an IC 50 of 500 nM or more.
表4 本发明部分化合物抑制SUN16细胞增殖的IC 50值 Table 4 The IC 50 value of some compounds of the present invention in inhibiting the proliferation of SUN16 cells
Figure PCTCN2020091613-appb-000057
Figure PCTCN2020091613-appb-000057
4、下表5列举了本发明合成的部分化合物在72小时对部分肿瘤细胞的增殖抑制情况。4. Table 5 below lists the inhibitory effects of some compounds synthesized by the present invention on the proliferation of some tumor cells at 72 hours.
表5 本发明部分化合物抑制肿瘤细胞增殖的IC 50值 Table 5 The IC 50 value of some compounds of the present invention in inhibiting tumor cell proliferation
Figure PCTCN2020091613-appb-000058
Figure PCTCN2020091613-appb-000058
NT表示未测试。NT means not tested.
5、下表6列举了本发明合成的部分化合物在作用72h后对NIH-3T3细胞和人肺成纤维细胞的增殖抑制情况。5. Table 6 below lists the inhibition of the proliferation of NIH-3T3 cells and human lung fibroblasts of some compounds synthesized by the present invention after 72 hours of action.
表6 本发明部分化合物抑制NIH-3T3细胞和人肺成纤维细胞增殖的IC 50值 Table 6 The IC 50 values of some compounds of the present invention in inhibiting the proliferation of NIH-3T3 cells and human lung fibroblasts
Figure PCTCN2020091613-appb-000059
Figure PCTCN2020091613-appb-000059
NT表示未测试。NT means not tested.
四、药代动力学实验结果4. Results of pharmacokinetic experiments
图1显示血药浓度和时间曲线。经单次静脉注射(1-A)或灌胃口服(1-B)(给药剂量分别为：2mg/kg、20mg/kg)给予SD大鼠(每组各三只雄性鼠，分别编号为：101/102/103、201/202/203)受试物4-20后，研究发现：静脉给药后平均半衰期为5.81h，平均AUC(0-t)为2771.74h*ng/mL。口服给药后，平均半衰期为3.44h，平均AUC(0-t)为20917.54h*ng/mL，算出平均口服生物利用度为75.47％。Figure 1 shows the blood concentration and time curve. A single intravenous injection (1-A) or oral gavage (1-B) (dose: 2mg/kg, 20mg/kg) were given to SD rats (three male rats in each group, respectively numbered : 101/102/103, 201/202/203) After 4-20 of the test substance, the study found that the average half-life after intravenous administration was 5.81h, and the average AUC(0-t) was 2771.74h*ng/mL. After oral administration, the average half-life was 3.44h, the average AUC(0-t) was 20917.54h*ng/mL, and the average oral bioavailability was calculated to be 75.47%.
五、动物体内实验结果5. Results of in vivo animal experiments
实验结果见图2、图3。图2，给药14天后，HE染色结果显示：假手术组，小鼠肺组织结构完整清晰，无炎性细胞侵入；溶剂对照组，小鼠肺泡结构略有破坏，肺泡间隔稍有增宽，较为明显炎性细胞浸润及成纤维细胞增生；YTH-17(4-20)高低剂量组，YTH-18(4-22)高低剂量组以及尼达尼布阳性对照组，与溶剂对照组相比，情况明显好转，病变范围明显减少。马氏染色结果显示：假手术组，小鼠肺组织结构正常，肺泡壁未见增厚；溶剂对照组，肺泡、肺间隔以及终末端支气管等周围稍有有增生的胶原纤维，呈少量现纤维化状态；YTH-17(4-20)高低剂量组，YTH-18(4-22)高低剂量组以及尼达尼布阳性对照组，与溶剂对照组相比，情况明显好转，胶原总体含量减少。The experimental results are shown in Figure 2 and Figure 3. Figure 2. After 14 days of administration, the HE staining results showed that in the sham operation group, the lung tissue structure of the mice was intact and clear, without inflammatory cell invasion; in the solvent control group, the alveolar structure of the mice was slightly damaged, and the alveolar interval was slightly widened. More obvious inflammatory cell infiltration and fibroblast proliferation; YTH-17 (4-20) high and low dose groups, YTH-18 (4-22) high and low dose groups and nintedanib positive control group, compared with the solvent control group , The situation has improved significantly, and the lesion area has been significantly reduced. The results of Markov's staining showed that in the sham operation group, the structure of the lung tissue of the mice was normal, and the alveolar wall was not thickened; in the solvent control group, there were slightly proliferated collagen fibers around the alveoli, lung septum, and terminal bronchus, showing a small amount of existing fibers. Chemical status; YTH-17 (4-20) high and low dose groups, YTH-18 (4-22) high and low dose groups and nintedanib positive control group, compared with the solvent control group, the situation is significantly improved, and the overall collagen content is reduced .
图3，药物治疗28天后，HE染色结果显示：假手术组，小鼠肺组织结构完整清晰，肺泡壁未见增厚，无炎性细胞侵入，无成纤维细胞增生；溶剂对照组，小鼠肺泡结构破坏严重，肺泡间隔明显增宽，大量的炎性细胞浸润及成纤维细胞增生，呈现明显纤维化状态；YTH-17(4-20)高低剂量组，YTH-18(4-22)高低剂量组以及尼达尼布阳性对照组，与溶剂对照组相比，小鼠肺组织较为结构完整，清晰情况明显好转，病变程度明显减少。马氏染色结果显示：假手术组，小鼠肺组织结构正常，肺泡壁未见增厚；无胶原沉积；溶剂对照组，肺泡、肺间隔以及终末端支气管等周围有增生的胶原纤维，呈现纤维化状态；YTH-17(4-20)高低剂量组，YTH-18(4-22)高低剂量组以及尼达尼布阳性对照组，较溶剂组相比，小鼠肺组织结构完整，胶原沉积现象明显减少，呈明显的好转，胶原总体含量减少，减轻胶原增生等，改善纤维化程度。Figure 3. After 28 days of drug treatment, HE staining results showed that in the sham operation group, the lung tissue structure of the mice was intact and clear, the alveolar wall was not thickened, there was no inflammatory cell invasion, and no fibroblast proliferation; solvent control group, mice The alveolar structure is severely damaged, the alveolar space is significantly widened, a large number of inflammatory cells infiltrate and fibroblast proliferation, showing an obvious fibrotic state; YTH-17 (4-20) high and low dose group, YTH-18 (4-22) high and low Compared with the solvent control group, in the dose group and the nintedanib positive control group, the lung tissue of the mice was more complete, the clear condition was improved significantly, and the degree of lesions was significantly reduced. The results of Markov staining showed that in the sham operation group, the lung tissue structure of the mice was normal, and the alveolar wall was not thickened; there was no collagen deposition; in the solvent control group, there were proliferating collagen fibers around the alveoli, lung septum, and terminal bronchus. Chemical status; YTH-17 (4-20) high and low dose groups, YTH-18 (4-22) high and low dose groups and nintedanib positive control group, compared with the solvent group, the mouse lung tissue structure is complete, collagen deposition The phenomenon was significantly reduced, showing a significant improvement, the overall collagen content was reduced, collagen proliferation was reduced, and the degree of fibrosis was improved.
以上实验结果表明，本发明化合物4-20(YTH-17)和4-22(YTH-18)均具有抗肺纤维化的作用。The above experimental results show that the compounds 4-20 (YTH-17) and 4-22 (YTH-18) of the present invention have anti-pulmonary fibrosis effects.
需要说明的是，本说明书中描述的具体特征、结构、材料或者特点可以在任一个或多个实施例中以合适的方式结合。此外，在不相互矛盾的情况下，本领域的技术人员可以将本说明书中描述的不同实施例以及不同实施例的特征进行结合和组合。It should be noted that the specific features, structures, materials or characteristics described in this specification can be combined in any one or more embodiments in a suitable manner. In addition, those skilled in the art can combine and combine the different embodiments and the features of the different embodiments described in this specification without contradicting each other.

Claims (16)

  1. 式Ⅰ所示的化合物、其光学异构体、化合物或其光学异构体在药学上可接受的盐：The compound represented by formula I, its optical isomer, the compound or its optical isomer pharmaceutically acceptable salt:
    Figure PCTCN2020091613-appb-100001
    Figure PCTCN2020091613-appb-100001
    其中，环A选自取代或未取代的苯基、取代或未取代的六元并五元芳基；Wherein, ring A is selected from substituted or unsubstituted phenyl, substituted or unsubstituted six-membered and five-membered aryl;
    R 1、R 2、R 3独立选自H、卤素、取代或未取代的烷基； R 1 , R 2 , and R 3 are independently selected from H, halogen, substituted or unsubstituted alkyl;
    环B选自取代或未取代的5～8元芳基。Ring B is selected from substituted or unsubstituted 5- to 8-membered aryl groups.
  2. 如权利要求1所述的化合物，其特征是：当环A为取代的苯基时，含有至少一个选自下组的取代基：卤素、未取代的烷基、卤素取代的烷基、未取代的烷氧基、卤素取代的烷氧基、羟基；The compound of claim 1, wherein when ring A is substituted phenyl, it contains at least one substituent selected from the group consisting of halogen, unsubstituted alkyl, halogen-substituted alkyl, unsubstituted The alkoxy group, halogen-substituted alkoxy group, hydroxyl group;
    优选地，所述取代的苯基含有至少一个选自下组的取代基：卤素、未取代的C1～C6烷基、卤素取代的C1～C6烷基、未取代的C1～C6烷氧基、卤素取代的C1～C6烷氧基、羟基；Preferably, the substituted phenyl group contains at least one substituent selected from the group consisting of halogen, unsubstituted C1-C6 alkyl, halogen-substituted C1-C6 alkyl, unsubstituted C1-C6 alkoxy, C1-C6 alkoxy and hydroxy substituted by halogen;
    优选地，所述取代的苯基含有至少一个选自下组的取代基：卤素、未取代的C1～C2烷基、卤素取代的C1～C2烷基、未取代的C1～C2烷氧基、卤素取代的C1～C2烷氧基、羟基；Preferably, the substituted phenyl group contains at least one substituent selected from the group consisting of halogen, unsubstituted C1-C2 alkyl, halogen-substituted C1-C2 alkyl, unsubstituted C1-C2 alkoxy, C1-C2 alkoxy and hydroxyl substituted by halogen;
    优选地，所述取代的苯基含有至少一个选自下组的取代基：氟、氯、三氟甲基、甲氧基、乙氧基、羟基；Preferably, the substituted phenyl group contains at least one substituent selected from the group consisting of fluorine, chlorine, trifluoromethyl, methoxy, ethoxy, and hydroxyl;
    进一步优选地，所述取代的苯基选自：Further preferably, the substituted phenyl group is selected from:
    Figure PCTCN2020091613-appb-100002
    Figure PCTCN2020091613-appb-100002
    Figure PCTCN2020091613-appb-100003
    Figure PCTCN2020091613-appb-100003
    优选地，所述取代的苯基含有至少一个选自下组的取代基：氟、氯、甲氧基；Preferably, the substituted phenyl group contains at least one substituent selected from the group consisting of fluorine, chlorine, and methoxy;
    进一步优选地，所述取代的苯基选自：Further preferably, the substituted phenyl group is selected from:
    Figure PCTCN2020091613-appb-100004
    Figure PCTCN2020091613-appb-100004
  3. 如权利要求1或2所述的化合物，其特征是：当环A为取代或未取代的六元并五元芳基时，所述六元并五元芳基中，六元芳基含有0～2个杂原子；The compound of claim 1 or 2, wherein when ring A is a substituted or unsubstituted six-membered five-membered aryl group, in the six-membered five-membered aryl group, the six-membered aryl group contains 0 ～2 heteroatoms;
    优选地，六元芳基为苯基；Preferably, the six-membered aryl group is phenyl;
    所述六元并五元芳基中，五元芳基含有至少一个杂原子氮；In the six-membered five-membered aryl group, the five-membered aryl group contains at least one heteroatom nitrogen;
    优选地，五元芳基选自取代或未取代的咪唑基、恶唑基或吡咯基；Preferably, the five-membered aryl group is selected from substituted or unsubstituted imidazolyl, oxazolyl or pyrrolyl;
    优选地，五元芳基选自
    Figure PCTCN2020091613-appb-100005
    其中，R 4～R 9独立选自H、取代或未取代的烷基；     Preferably, the five-membered aryl group is selected from
    Figure PCTCN2020091613-appb-100005
    Wherein, R 4 to R 9 are independently selected from H, substituted or unsubstituted alkyl;
    优选地，R 4～R 9独立选自H、未取代的C1～C6烷基或卤素取代的C1～C6烷基； Preferably, R 4 to R 9 are independently selected from H, unsubstituted C1 to C6 alkyl or halogen substituted C1 to C6 alkyl;
    优选地，R 4～R 9独立选自H、甲基或乙基； Preferably, R 4 to R 9 are independently selected from H, methyl or ethyl;
    优选地，R 5选自H、甲基或乙基，R 4、R 6、R 7、R 8、R 9均为H； Preferably, R 5 is selected from H, methyl or ethyl, and R 4 , R 6 , R 7 , R 8 , and R 9 are all H;
    进一步优选地，所述六元并五元芳基选自：Further preferably, the six- and five-membered aryl group is selected from:
    Figure PCTCN2020091613-appb-100006
    Figure PCTCN2020091613-appb-100006
  4. 如权利要求1～3任意一项所述的化合物，其特征是：R 1、R 2、R 3独立选自H、卤素、未取代的C1～C6烷基或卤素取代的C1～C6烷基； The compound according to any one of claims 1 to 3, wherein R 1 , R 2 , and R 3 are independently selected from H, halogen, unsubstituted C1-C6 alkyl or halogen-substituted C1-C6 alkyl ；
    优选地，R 1、R 2、R 3独立选自H、卤素或未取代的C1～C3烷基； Preferably, R 1 , R 2 , R 3 are independently selected from H, halogen or unsubstituted C1-C3 alkyl;
    优选地，R 1、R 2、R 3独立选自H、氟、氯或甲基； Preferably, R 1 , R 2 , R 3 are independently selected from H, fluorine, chlorine or methyl;
    进一步优选地，R 2选自H、氟、氯或甲基，R 1、R 3均为H； More preferably, R 2 is selected from H, fluorine, chlorine or methyl, and R 1 and R 3 are both H;
    进一步优选地，R 2选自H或甲基，R 1、R 3均为H。 More preferably, R 2 is selected from H or methyl, and both R 1 and R 3 are H.
  5. 如权利要求1～4任意一项所述的化合物，其特征是：环B选自取代或未取代的5～6元芳基；The compound according to any one of claims 1 to 4, wherein the ring B is selected from substituted or unsubstituted 5- to 6-membered aryl groups;
    优选地，所述芳基含有0～2个杂原子；Preferably, the aryl group contains 0 to 2 heteroatoms;
    优选地，所述杂原子为氮；Preferably, the heteroatom is nitrogen;
    优选地，所述芳基选自苯基、吡啶基、嘧啶基或吡唑基。Preferably, the aryl group is selected from phenyl, pyridyl, pyrimidinyl or pyrazolyl.
  6. 如权利要求5所述的化合物，其特征是：当环B为取代的6元芳基时，含有至少一个选自下组的取代基：卤素、未取代的烷基、卤素取代的烷基、未取代的烷氧基、卤素取代的烷氧基、-C(＝O)OR 10、-X-NR 11R 12，其中，R 10选自取代或未取代的烷基，X选自-(CH 2) n-、-C(＝O)-或-SO 2-，n为0～6的整数，R 11、R 12独立选自取代或未取代的烷基，或者，R 11与R 12相连形成脂环； The compound of claim 5, wherein when ring B is a substituted 6-membered aryl group, it contains at least one substituent selected from the group consisting of halogen, unsubstituted alkyl, halogen-substituted alkyl, Unsubstituted alkoxy, halogen-substituted alkoxy, -C(=O)OR 10 , -X-NR 11 R 12 , wherein R 10 is selected from substituted or unsubstituted alkyl, and X is selected from -( CH 2 ) n -, -C(=O)- or -SO 2 -, n is an integer of 0-6, R 11 and R 12 are independently selected from substituted or unsubstituted alkyl groups, or, R 11 and R 12 Connected to form an alicyclic ring;
    优选地，当环B为取代的6元芳基时，含有至少一个选自下组的取代基：卤素、未取代的C1～C6烷基、卤素取代的C1～C6烷基、未取代的C1～C6烷氧基、卤素取代的C1～C6烷氧基、-C(＝O)OR 10、-X-NR 11R 12，其中，R 10选自取代或未取代的C1～C6烷基，X选自-(CH 2) n-、-C(＝O)-或-SO 2-，n为0～6的整数，R 11、R 12独立选自取代或未取代的C1～C6烷基，或者，R 11与R 12相连形成6元脂环； Preferably, when ring B is a substituted 6-membered aryl group, it contains at least one substituent selected from the group consisting of halogen, unsubstituted C1-C6 alkyl, halogen-substituted C1-C6 alkyl, unsubstituted C1 -C6 alkoxy, halogen-substituted C1-C6 alkoxy, -C(=O)OR 10 , -X-NR 11 R 12 , wherein R 10 is selected from substituted or unsubstituted C1-C6 alkyl, X is selected from -(CH 2 ) n -, -C(=O)- or -SO 2 -, n is an integer of 0-6, R 11 and R 12 are independently selected from substituted or unsubstituted C1-C6 alkyl , Or, R 11 and R 12 are connected to form a 6-membered alicyclic ring;
    优选地，当环B为取代的6元芳基时，含有至少一个选自下组的取代基：卤素、甲基、三氟甲基、甲氧基、三氟甲氧基、-C(＝O)OR 10、-X-NR 11R 12，其中，R 10选自取代或未取代的C1～C6烷基，X选自-(CH 2) n-、-C(＝O)-或-SO 2-，n为0～3的整数，R 11、R 12独立选自取代或未取代的C1～C6烷基，或者，R 11与R 12相连形成6元脂环，所述6元脂环含有2个杂原子； Preferably, when ring B is a substituted 6-membered aryl group, it contains at least one substituent selected from the group consisting of halogen, methyl, trifluoromethyl, methoxy, trifluoromethoxy, -C(= O) OR 10 , -X-NR 11 R 12 , wherein R 10 is selected from substituted or unsubstituted C1-C6 alkyl, and X is selected from -(CH 2 ) n -, -C(=O)- or- SO 2 -, n is an integer from 0 to 3, R 11 and R 12 are independently selected from substituted or unsubstituted C1-C6 alkyl groups, or R 11 and R 12 are connected to form a 6-membered alicyclic ring, the 6-membered aliphatic The ring contains 2 heteroatoms;
    优选地，当环B为取代的6元芳基时，仅含有一个选自下组的取代基：-C(＝O)OR 10、-X-NR 11R 12，其中，R 10选自未取代的C1～C6烷基或卤素取代的C1～C6烷基，X选自-(CH 2) n-或-C(＝O)，n为0～3的整数，R 11、R 12独立选自未取代的C1～C6烷基或卤素取代的C1～C6烷基，或者，R 11与R 12相连形成6元脂环，所述6元脂环含有2个杂原子，所述杂原子为氮或氧； Preferably, when ring B is a substituted 6-membered aryl group, it contains only one substituent selected from the following group: -C(=O)OR 10 , -X-NR 11 R 12 , wherein R 10 is selected from Substituted C1-C6 alkyl or halogen-substituted C1-C6 alkyl, X is selected from -(CH 2 ) n -or -C(=O), n is an integer of 0 to 3, R 11 and R 12 are independently selected From an unsubstituted C1-C6 alkyl group or a halogen-substituted C1-C6 alkyl group, or, R 11 and R 12 are connected to form a 6-membered alicyclic ring, the 6-membered alicyclic ring contains 2 heteroatoms, and the heteroatoms are Nitrogen or oxygen;
    进一步优选地，R 10为未取代的C1～C6烷基； More preferably, R 10 is an unsubstituted C1-C6 alkyl group;
    进一步优选地，R 10为甲基； More preferably, R 10 is methyl;
    进一步优选地，n为0或1；Further preferably, n is 0 or 1;
    进一步优选地，R 11、R 12独立选自未取代的C1～C6烷基，或者，R 11与R 12 相连形成取代或未取代的吗啉基、哌嗪基； Further preferably, R 11 and R 12 are independently selected from unsubstituted C1-C6 alkyl groups, or R 11 and R 12 are connected to form a substituted or unsubstituted morpholinyl or piperazinyl group;
    进一步优选地，R 11、R 12均为甲基； More preferably, R 11 and R 12 are both methyl groups;
    进一步优选地，所述取代的吗啉基、哌嗪基含有至少一个选自下组的取代基：取代或未取代的C1～C6烷基、C1～C6烷氧羰基、苄氧羰基、芴氧羰基；Further preferably, the substituted morpholinyl and piperazinyl groups contain at least one substituent selected from the following group: substituted or unsubstituted C1-C6 alkyl, C1-C6 alkoxycarbonyl, benzyloxycarbonyl, fluorenyloxy Carbonyl
    进一步优选地，所述取代的吗啉基、哌嗪基含有至少一个选自下组的取代基：甲基、叔丁氧羰基；Further preferably, the substituted morpholinyl and piperazinyl groups contain at least one substituent selected from the group consisting of methyl and tert-butoxycarbonyl;
    进一步优选地，R 11与R 12相连形成
    Figure PCTCN2020091613-appb-100007
    其中，R 13、R 15独立选自H或甲基，R 14选自H、甲基或叔丁氧羰基；     Further preferably, R 11 and R 12 are connected to form
    Figure PCTCN2020091613-appb-100007
    Wherein, R 13 and R 15 are independently selected from H or methyl, and R 14 is selected from H, methyl or tert-butoxycarbonyl;
    进一步优选地，当环B为取代的6元芳基时，选自：More preferably, when ring B is a substituted 6-membered aryl group, it is selected from:
    Figure PCTCN2020091613-appb-100008
    Figure PCTCN2020091613-appb-100008
  7. 如权利要求5所述的化合物，其特征是：当环B为未取代的6元芳基时，选自
    Figure PCTCN2020091613-appb-100009
    The compound of claim 5, wherein when ring B is an unsubstituted 6-membered aryl group, it is selected from
    Figure PCTCN2020091613-appb-100009
  8. 如权利要求5所述的化合物，其特征是：环B为取代的吡唑基，含有至少一个选自下组的取代基：未取代的烷基、卤素取代的烷基、羟基取代的烷基；The compound according to claim 5, characterized in that: ring B is a substituted pyrazolyl and contains at least one substituent selected from the group consisting of unsubstituted alkyl, halogen-substituted alkyl, and hydroxy-substituted alkyl ；
    优选地，所述取代的吡唑基含有至少一个选自下组的取代基：未取代的C1～C6烷基、卤素取代的C1～C6烷基、羟基取代的C1～C6烷基；Preferably, the substituted pyrazolyl contains at least one substituent selected from the group consisting of unsubstituted C1-C6 alkyl, halogen-substituted C1-C6 alkyl, and hydroxy-substituted C1-C6 alkyl;
    优选地，所述取代的吡唑基仅含有一个取代基，所述取代基为羟基取代的C1～C6烷基；Preferably, the substituted pyrazolyl contains only one substituent, and the substituent is a C1-C6 alkyl substituted by a hydroxy group;
    优选地，所述取代的吡唑基仅含有一个取代基，所述取代基为
    Figure PCTCN2020091613-appb-100010
    Preferably, the substituted pyrazolyl contains only one substituent, and the substituent is
    Figure PCTCN2020091613-appb-100010
    进一步优选地，环B为
    Figure PCTCN2020091613-appb-100011
    Further preferably, ring B is
    Figure PCTCN2020091613-appb-100011
  9. 如权利要求1、5～8任意一项所述的化合物，其特征是：环B选自：The compound according to any one of claims 1, 5 to 8, wherein the ring B is selected from:
    Figure PCTCN2020091613-appb-100012
    Figure PCTCN2020091613-appb-100012
  10. 如权利要求1～9任意一项所述的化合物，其特征是：所述化合物选自：The compound according to any one of claims 1-9, wherein the compound is selected from:
    Figure PCTCN2020091613-appb-100013
    Figure PCTCN2020091613-appb-100013
    Figure PCTCN2020091613-appb-100014
    Figure PCTCN2020091613-appb-100014
    Figure PCTCN2020091613-appb-100015
    Figure PCTCN2020091613-appb-100015
    Figure PCTCN2020091613-appb-100016
    Figure PCTCN2020091613-appb-100016
    Figure PCTCN2020091613-appb-100017
    Figure PCTCN2020091613-appb-100017
    Figure PCTCN2020091613-appb-100018
    Figure PCTCN2020091613-appb-100018
    Figure PCTCN2020091613-appb-100019
    Figure PCTCN2020091613-appb-100019
  11. 权利要求1～10任意一项所述的化合物、其光学异构体、化合物或其光学异构体药学上可接受的盐的制备方法，其特征是：包括如下步骤：The method for preparing the compound, its optical isomer, the compound or its optical isomer pharmaceutically acceptable salt according to any one of claims 1 to 10, characterized in that it comprises the following steps:
    a、化合物1与化合物2在钯催化剂作用下偶联，得到中间体Ⅰ：a. Compound 1 and compound 2 are coupled under the action of palladium catalyst to obtain Intermediate I:
    Figure PCTCN2020091613-appb-100020
    Figure PCTCN2020091613-appb-100020
    其中，Y为卤素，R 16、R 17独立选自H、取代或未取代的烷基，或者，R 16与R 17相连形成脂环； Wherein, Y is halogen, R 16 and R 17 are independently selected from H, substituted or unsubstituted alkyl, or R 16 and R 17 are connected to form an alicyclic ring;
    优选地，Y为溴；Preferably, Y is bromine;
    优选地，R 16、R 17独立选自H或未取代的C1～C6烷基，或者，R 16与R 17相连形成5元脂环； Preferably, R 16 and R 17 are independently selected from H or an unsubstituted C1-C6 alkyl group, or R 16 and R 17 are connected to form a 5-membered alicyclic ring;
    进一步优选地，化合物2为
    Figure PCTCN2020091613-appb-100021
    Further preferably, compound 2 is
    Figure PCTCN2020091613-appb-100021
    b、中间体Ⅰ在碱存在的条件下与酸酐和亚硝酸酯反应，得到中间体Ⅱ：b. Intermediate I reacts with acid anhydride and nitrite in the presence of a base to obtain Intermediate II:
    Figure PCTCN2020091613-appb-100022
    Figure PCTCN2020091613-appb-100022
    所述亚硝酸酯为
    Figure PCTCN2020091613-appb-100023
    R 18选自取代或未取代的烷基，R 21选自取代或未取代的烷基；     The nitrite is
    Figure PCTCN2020091613-appb-100023
    R 18 is selected from substituted or unsubstituted alkyl, and R 21 is selected from substituted or unsubstituted alkyl;
    优选地，R 18为未取代的C1～C6烷基； Preferably, R 18 is an unsubstituted C1-C6 alkyl group;
    进一步优选地，R 18为异戊基； More preferably, R 18 is isopentyl;
    优选地，R 21为未取代的C1～C6烷基； Preferably, R 21 is an unsubstituted C1-C6 alkyl group;
    进一步优选地，R 21为甲基； More preferably, R 21 is a methyl group;
    c、中间体Ⅱ脱除酰基，得到中间体Ⅲ：c. Intermediate II removes the acyl group to obtain Intermediate III:
    Figure PCTCN2020091613-appb-100024
    Figure PCTCN2020091613-appb-100024
    d、中间体Ⅲ经卤代，得到中间体Ⅳ：d. Intermediate III is halogenated to obtain Intermediate IV:
    Figure PCTCN2020091613-appb-100025
    Figure PCTCN2020091613-appb-100025
    优选地，Z为碘；Preferably, Z is iodine;
    e、中间体Ⅳ与化合物3在钯催化剂作用下偶联，即得：e. Intermediate IV and compound 3 are coupled under the action of palladium catalyst to obtain:
    Figure PCTCN2020091613-appb-100026
    Figure PCTCN2020091613-appb-100026
    其中，R 19、R 20独立选自H、取代或未取代的烷基，或者，R 19与R 20相连形成脂环； Wherein, R 19 and R 20 are independently selected from H, substituted or unsubstituted alkyl, or R 19 and R 20 are connected to form an alicyclic ring;
    优选地，R 19、R 20独立选自H或未取代的C1～C6烷基，或者，R 19与R 20相连形成5元脂环； Preferably, R 19 and R 20 are independently selected from H or an unsubstituted C1-C6 alkyl group, or R 19 and R 20 are connected to form a 5-membered alicyclic ring;
    进一步优选地，化合物3为
    Figure PCTCN2020091613-appb-100027
    More preferably, compound 3 is
    Figure PCTCN2020091613-appb-100027
  12. 如权利要求11所述的制备方法，其特征是：满足以下至少一项：The preparation method according to claim 11, characterized in that: at least one of the following is satisfied:
    步骤a所述钯催化剂为醋酸钯、[1,1'-双(二苯基膦基)二茂铁]二氯化钯、The palladium catalyst in step a is palladium acetate, [1,1'-bis(diphenylphosphino)ferrocene]palladium dichloride,
    [1,1'-双(二苯基膦)二茂铁]二氯化钯二氯甲烷络合物、三(二亚苄基茚丙酮)二钯中一种或两种以上；[1,1'-Bis(diphenylphosphine)ferrocene]dichloropalladium dichloromethane complex, one or more of tris(dibenzylidene indeneacetone)dipalladium;
    步骤a向反应体系中加入碱，所述碱为碳酸钠、碳酸氢钠、碳酸钾、磷酸钾、碳酸铯中一种或两种以上；Step a: Adding a base to the reaction system, the base being one or more of sodium carbonate, sodium bicarbonate, potassium carbonate, potassium phosphate, and cesium carbonate;
    步骤a中化合物1：化合物2：碱：钯催化剂的摩尔配比为1：(1.1～1.5)：In step a, the molar ratio of compound 1: compound 2: base: palladium catalyst is 1: (1.1～1.5):
    (2.0～3.0)：(0.003～0.010)；(2.0～3.0): (0.003～0.010);
    步骤a的反应溶剂为二氧六环、水、甲苯、DMF、正丁醇、异丙醇、仲丁醇中一种或两种以上；The reaction solvent in step a is one or more of dioxane, water, toluene, DMF, n-butanol, isopropanol, and sec-butanol;
    优选地，步骤a的反应溶剂为1,4-二氧六环：水体积比为(4～8)：1的混合溶剂；Preferably, the reaction solvent in step a is a mixed solvent of 1,4-dioxane:water volume ratio (4-8):1;
    步骤a的反应温度为90～110℃；The reaction temperature in step a is 90-110°C;
    步骤a的反应时间为5～10h；The reaction time of step a is 5-10h;
    步骤a于N 2保护下反应； Step a is reacted under N 2 protection;
    步骤b将中间体I溶于反应溶剂中，加入碱和酸酐充分搅拌，加入亚硝酸酯反应，得到中间体Ⅱ；Step b: Dissolve Intermediate I in the reaction solvent, add alkali and acid anhydride and stir thoroughly, and add nitrite for reaction to obtain Intermediate II;
    步骤b所述碱为乙酸钾；The base in step b is potassium acetate;
    步骤b中中间体I：碱：酸酐：亚硝酸酯的摩尔配比为1：(1.1～1.5)：(1.8～2.5)：(3～5)；In step b, the molar ratio of intermediate I: base: acid anhydride: nitrite is 1: (1.1～1.5): (1.8～2.5): (3～5);
    步骤b的反应溶剂为甲苯；The reaction solvent in step b is toluene;
    步骤b加入亚硝酸酯回流反应4～8h；Step b: adding nitrite and refluxing reaction for 4-8h;
    步骤c在酸性条件下反应；Step c reacts under acidic conditions;
    优选地，步骤c向反应体系中加入盐酸；Preferably, in step c, hydrochloric acid is added to the reaction system;
    优选地，步骤c向反应体系中加入6N HCl；Preferably, step c adds 6N HCl to the reaction system;
    步骤c的反应溶剂为醇系溶剂；The reaction solvent in step c is an alcohol solvent;
    优选地，步骤c的反应溶剂为甲醇；Preferably, the reaction solvent in step c is methanol;
    步骤c回流反应1～2h；Step c reflux reaction for 1～2h;
    当Z为碘时，步骤d将中间体Ⅲ溶于反应溶剂中，加入碱，将I 2溶于反应溶剂中滴加至反应液，反应得到中间体Ⅳ； When Z is iodine, in step d, the intermediate III is dissolved in the reaction solvent, a base is added, and I 2 is dissolved in the reaction solvent and added dropwise to the reaction solution to obtain the intermediate IV through the reaction;
    优选地，所述碱为碳酸氢钠、碳酸钠、碳酸钾、氢氧化钠、氢氧化钾中一种或两种以上；Preferably, the alkali is one or more of sodium bicarbonate, sodium carbonate, potassium carbonate, sodium hydroxide, and potassium hydroxide;
    步骤d中中间体Ⅲ：I 2：碱的摩尔配比为1：(1.5-2.0)：2.0； In step d, the molar ratio of Intermediate III: I 2 : Base is 1: (1.5-2.0): 2.0;
    步骤d的反应溶剂为DMF；The reaction solvent in step d is DMF;
    步骤d的反应温度为25～80℃；The reaction temperature in step d is 25 to 80°C;
    步骤d的反应时间为2～10h；The reaction time of step d is 2-10h;
    步骤e所述钯催化剂为醋酸钯、[1,1'-双(二苯基膦基)二茂铁]二氯化钯、[1,1'-双(二苯基膦)二茂铁]二氯化钯二氯甲烷络合物、三(二亚苄基茚丙酮)二钯中一种 或两种以上；The palladium catalyst in step e is palladium acetate, [1,1'-bis(diphenylphosphino)ferrocene]palladium dichloride, [1,1'-bis(diphenylphosphino)ferrocene] One or more of palladium dichloride dichloromethane complex and tris(dibenzylidene indeneacetone) dipalladium;
    步骤e向反应体系中加入碱，所述碱为DIEA、碳酸钠、碳酸氢钠、碳酸钾、磷酸钾、碳酸铯中一种或两种以上；Step e: adding a base to the reaction system, the base being one or more of DIEA, sodium carbonate, sodium bicarbonate, potassium carbonate, potassium phosphate, and cesium carbonate;
    步骤e中中间体Ⅳ：化合物3：碱：钯催化剂的摩尔配比为1：(1.1～1.5)：(2.0～3.0)：(0.003～0.010)；In step e, the molar ratio of intermediate IV: compound 3: base: palladium catalyst is 1: (1.1～1.5): (2.0～3.0): (0.003～0.010);
    步骤e的反应溶剂为二氧六环、水、甲苯、DMF、正丁醇、异丙醇、仲丁醇中一种或两种以上；The reaction solvent in step e is one or more of dioxane, water, toluene, DMF, n-butanol, isopropanol, and sec-butanol;
    优选地，步骤e的反应溶剂为1,4-二氧六环：水体积比为(4～8)：1的混合溶剂；Preferably, the reaction solvent in step e is a mixed solvent of 1,4-dioxane:water volume ratio (4-8):1;
    步骤e的反应温度为90～110℃；The reaction temperature in step e is 90-110°C;
    步骤e的反应时间为5～10h；The reaction time of step e is 5-10h;
    步骤e于N 2保护下反应。 Step e is reacted under N 2 protection.
  13. 权利要求1～10任意一项所述的化合物、其光学异构体、化合物或其光学异构体药学上可接受的盐在制备FGFR激酶抑制剂类药物中的用途；优选地，所述药物是FGFR1、FGFR2和/或FGFR3激酶抑制剂。Use of the compound, optical isomer, compound or optical isomer pharmaceutically acceptable salt thereof according to any one of claims 1 to 10 in the preparation of FGFR kinase inhibitor drugs; preferably, the drug It is an FGFR1, FGFR2 and/or FGFR3 kinase inhibitor.
  14. 权利要求1～10任意一项所述的化合物、其光学异构体、化合物或其光学异构体药学上可接受的盐在制备治疗和/或预防癌症的药物中的用途；优选地，所述癌症为乳腺癌、肺癌、胃癌、肾癌、结直肠癌、肝癌、黑色素瘤、膀胱癌、尿路上皮癌和/或胆管癌；进一步优选地，所述肺癌为非小细胞肺癌。Use of the compound, its optical isomer, the compound or its optical isomer pharmaceutically acceptable salt according to any one of claims 1 to 10 in the preparation of a medicine for the treatment and/or prevention of cancer; preferably, The cancer is breast cancer, lung cancer, gastric cancer, kidney cancer, colorectal cancer, liver cancer, melanoma, bladder cancer, urothelial cancer and/or cholangiocarcinoma; further preferably, the lung cancer is non-small cell lung cancer.
  15. 权利要求1～10任意一项所述的化合物、其光学异构体、化合物或其光学异构体药学上可接受的盐在制备治疗和/或预防器官纤维化的药物中的用途；优选地，所述器官纤维化为肺纤维化或肝纤维化。Use of the compound, its optical isomer, the compound or its optical isomer pharmaceutically acceptable salt according to any one of claims 1 to 10 in the preparation of a medicament for the treatment and/or prevention of organ fibrosis; preferably , Said organ fibrosis is pulmonary fibrosis or liver fibrosis.
  16. 药物组合物，其特征是：它是以权利要求1～10任意一项所述的化合物、其光学异构体、化合物或其光学异构体在药学上可接受的盐为活性成分，加入药学上可接受的辅料或者辅助性成分制备而成的制剂；优选地，所述制剂为口服制剂或注射制剂。The pharmaceutical composition is characterized in that it is the active ingredient of the compound, its optical isomer, the compound or its pharmaceutically acceptable salt of the optical isomer of any one of claims 1-10, and added to the pharmaceutical A preparation prepared from above-acceptable adjuvants or auxiliary components; preferably, the preparation is an oral preparation or an injection preparation.
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Citations (4)

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Publication number Priority date Publication date Assignee Title
CN1374950A (en) * 1999-07-02 2002-10-16 阿古龙制药公司 Indazole compounds and pharmaceutical compositions for inhibiting protein kinases, and methods for their use
WO2018136010A1 (en) * 2017-01-20 2018-07-26 Aslan Pharmaceuticals Pte Ltd Combination therapy
CN110452176A (en) * 2018-05-07 2019-11-15 四川大学 Indazole analog derivative and its preparation method and application
CN111205227A (en) * 2018-11-22 2020-05-29 四川大学 3-vinyl indazole derivative and preparation method and application thereof

Patent Citations (4)

* Cited by examiner, † Cited by third party
Publication number Priority date Publication date Assignee Title
CN1374950A (en) * 1999-07-02 2002-10-16 阿古龙制药公司 Indazole compounds and pharmaceutical compositions for inhibiting protein kinases, and methods for their use
WO2018136010A1 (en) * 2017-01-20 2018-07-26 Aslan Pharmaceuticals Pte Ltd Combination therapy
CN110452176A (en) * 2018-05-07 2019-11-15 四川大学 Indazole analog derivative and its preparation method and application
CN111205227A (en) * 2018-11-22 2020-05-29 四川大学 3-vinyl indazole derivative and preparation method and application thereof

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