WO2022257961A1 - Benzoheterocyclic compound for treating ep2 and ep4 receptor-mediated diseases - Google Patents

Benzoheterocyclic compound for treating ep2 and ep4 receptor-mediated diseases Download PDF

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WO2022257961A1
WO2022257961A1 PCT/CN2022/097611 CN2022097611W WO2022257961A1 WO 2022257961 A1 WO2022257961 A1 WO 2022257961A1 CN 2022097611 W CN2022097611 W CN 2022097611W WO 2022257961 A1 WO2022257961 A1 WO 2022257961A1
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indazole
propane
carboxylic acid
ethyl
compound
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French (fr)
Chinese (zh)
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张学军
臧杨
李群
丁肖华
王俊南
李禹琼
李莉娥
杨俊�
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武汉人福创新药物研发中心有限公司
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    • A61K31/395—Heterocyclic compounds having nitrogen as a ring hetero atom, e.g. guanethidine or rifamycins
    • A61K31/40—Heterocyclic compounds having nitrogen as a ring hetero atom, e.g. guanethidine or rifamycins having five-membered rings with one nitrogen as the only ring hetero atom, e.g. sulpiride, succinimide, tolmetin, buflomedil
    • A61K31/403—Heterocyclic compounds having nitrogen as a ring hetero atom, e.g. guanethidine or rifamycins having five-membered rings with one nitrogen as the only ring hetero atom, e.g. sulpiride, succinimide, tolmetin, buflomedil condensed with carbocyclic rings, e.g. carbazole
    • A61K31/404—Indoles, e.g. pindolol
    • A—HUMAN NECESSITIES
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    • 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
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    • 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
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    • 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
    • A61K31/4965—Non-condensed pyrazines
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    • A61K31/505—Pyrimidines; Hydrogenated pyrimidines, e.g. trimethoprim
    • A61K31/506—Pyrimidines; Hydrogenated pyrimidines, e.g. trimethoprim not condensed and containing further heterocyclic rings
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    • 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
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    • C07D405/10—Heterocyclic compounds containing both one or more hetero rings having oxygen atoms as the only ring hetero atoms, and one or more rings having nitrogen as the only ring hetero atom containing two hetero rings linked by a carbon chain containing aromatic rings

Definitions

  • the invention belongs to the field of medicine, in particular, the invention relates to a benzoheterocyclic compound for treating diseases mediated by EP2 and EP4 receptors, a preparation method and application.
  • Prostaglandin E2 is an endogenous bioactive lipid. PGE2 induces a wide range of upstream and downstream dependent biological responses by activating prostaglandin receptors, and participates in the regulation of inflammation, pain, renal function, and cardiovascular system. , lung function and cancer and many other physiological and pathological processes. It has been reported that PGE2 is highly expressed in cancerous tissues of various cancers, and it has been confirmed that PGE2 is associated with the occurrence, growth and development of cancer and disease conditions in patients. It is generally believed that PGE2 is associated with the activation of cell proliferation and cell death (apoptosis), and plays an important role in the process of cancer cell proliferation, disease progression and cancer metastasis.
  • apoptosis apoptosis
  • EP1 receptor activates the phospholipase C and inositol triphosphate pathways
  • EP2 and EP4 receptors activate adenylate cyclase and cAMP-protein kinase A
  • the activation of the EP3 receptor both inhibits adenylate cyclase and Activates phospholipase C.
  • EP2 and EP4 are expressed in various immune cells (such as macrophages, dendritic cells, NK cells and CTLs), and inhibiting EP2 and EP4 can enhance immune activity and inhibit tumor growth.
  • immune cells such as macrophages, dendritic cells, NK cells and CTLs
  • PGE2 persistently activates EP receptors in the tumor microenvironment (produced in large quantities by tumor cells), which promotes the accumulation and enhances the activity of a variety of immunosuppressive cells, including type 2 tumor-associated macrophages (TAMS) , Treg cells and myeloid-derived suppressor cells (MDSCs).
  • TAMS type 2 tumor-associated macrophages
  • MDSCs myeloid-derived suppressor cells
  • One of the main features of the immunosuppressive tumor microenvironment is the presence of a large number of MDSCs and TAMs, which in turn are associated with gastric cancer, ovarian cancer, breast cancer, bladder cancer, hepatocellular carcinoma (HCC), head and neck cancer, and other types of cancer patients. It is closely related to poor overall survival.
  • PGE2 has been reported to induce immune tolerance by inhibiting the accumulation of antigen-presenting dendritic cells (DCs) in tumors and inhibiting the activation of tumor-infiltrating DCs), thereby helping tumor cells escape immune surveillance.
  • DCs antigen-presenting dendritic cells
  • PGE2 plays a very important role in promoting the occurrence and development of tumors. It has been found that the expression levels of PGE2 and its related receptors EP2 and EP4 are increased in various malignant tumors including colon cancer, lung cancer, breast cancer, and head and neck cancer. Often closely associated with poor prognosis. Therefore, selectively blocking EP2 and EP4 signaling pathways can inhibit tumor development by changing the tumor microenvironment and regulating tumor immune cells.
  • EP4 antagonists have been shown to be effective in alleviating joint inflammation and pain in rodent models of rheumatoid arthritis and osteoarthritis, and have also been shown to be effective in rodent models of autoimmune disease.
  • EP2 antagonists may be shown to be effective in certain chronic inflammatory diseases (especially inflammatory neurodegenerative diseases such as epilepsy, Alzheimer's disease (AD) , Parkinson's disease (PD), amyotrophic lateral sclerosis (ALS) and traumatic brain injury (TBI)).
  • AD Alzheimer's disease
  • PD Parkinson's disease
  • ALS amyotrophic lateral sclerosis
  • TBI traumatic brain injury
  • EP2 and EP4 suppresses the expression of progesterone (P4) signaling machinery proteins in epithelial and mesenchymal cell-specific patterns in endometriotic lesions and inhibits the persistence, invasion, biosynthesis and signaling of PGE2 and estrogen (E2) Transduction, thereby inhibiting the production of pro-inflammatory cytokines, reducing the growth, survival and spread of endometriotic lesions, alleviating pelvic pain, and restoring endometrial receptivity.
  • P4 progesterone
  • E2 and estrogen E2 Transduction
  • the object of the present invention is to provide a benzoheterocyclic compound for treating diseases mediated by EP2 and EP4 receptors and its preparation method and application. and heterocyclic compounds, tautomers, stereoisomers, hydrates, solvates, pharmaceutically acceptable salts or prodrugs thereof.
  • the first aspect of the present invention provides a benzoheterocyclic compound represented by formula I, its tautomers, stereoisomers, hydrates, solvates, pharmaceutically acceptable salts or prodrugs:
  • R 1 and R 2 are respectively hydrogen or -C ⁇ CR 11 , and R 1 and R 2 are different;
  • the R 11 is hydrogen or selected from: C 1 -C 5 alkyl, 3-6 membered cycloalkyl;
  • the R 11 is optionally substituted by one or more substituents selected from the following: hydroxyl, halogen, C 1 -C 5 alkyl; when there are multiple substituents, the substituents are the same or different;
  • L 1 is C 1 -C 5 alkylene
  • L 2 does not exist or is unsubstituted or substituted by Rb C 1 -C 3 alkylene;
  • the L 1 is optionally substituted by Ra 1 and/or Ra 2 ;
  • the Ra 1 and Ra 2 are each independently C 1 -C 5 alkyl, halogen, hydroxyl, amino, C 1 -C 5 alkoxy, C 1 -C 5 haloalkyl, C 1 -C 5 haloalkoxy ;
  • Ra 1 and Ra 2 form a 3-6-membered cycloalkyl group or a 4-6-membered heterocycloalkyl group together with the C atoms they are connected to;
  • the Rb is C 1 -C 3 alkyl or halogen
  • Ring A is phenyl or 5-6 membered heteroaryl
  • Ring B does not exist or is 3-8 membered cycloalkyl, 4-8 membered heterocycloalkyl, phenyl, 5-6 membered heteroaryl;
  • R 3 and R 4 are each independently selected from: hydroxyl, halogen, amino, cyano, C 1 -C 5 alkyl, C 1 -C 5 alkoxy, 3-6 membered cycloalkyl, 4-6 membered hetero Cycloalkyl, 3-6 membered cycloalkoxy; said C 1 -C 5 alkyl, C 1 -C 5 alkoxy, 3-6 membered cycloalkyl, 4-6 membered heterocycloalkyl, 3 -6-membered cycloalkoxy is optionally substituted by one or more substituents selected from the group consisting of hydroxyl, halogen, amino, cyano, 3-6-membered cycloalkyl;
  • n 0, 1, 2 or 3;
  • n 0, 1, 2 or 3.
  • R 1 and R 2 are respectively hydrogen or -C ⁇ CR 11 , and R 1 and R 2 are different;
  • the R 11 is hydrogen or selected from: C 1 -C 5 alkyl, 3-6 membered cycloalkyl;
  • the R 11 is optionally substituted by one or more substituents selected from the following: hydroxyl, halogen, C 1 -C 5 alkyl; when there are multiple substituents, the substituents are the same or different;
  • L 1 is C 1 -C 5 alkylene
  • L 2 does not exist or is unsubstituted or substituted by Rb C 1 -C 3 alkylene;
  • the L 1 is optionally substituted by Ra 1 and/or Ra 2 ;
  • the Ra 1 and Ra 2 are each independently C 1 -C 5 alkyl, halogen, hydroxyl, amino, C 1 -C 5 alkoxy, C 1 -C 5 haloalkyl, C 1 -C 5 haloalkoxy ;
  • Ra 1 and Ra 2 form a 3-6-membered cycloalkyl group or a 4-6-membered heterocycloalkyl group together with the C atoms they are connected to;
  • the Rb is C 1 -C 3 alkyl or halogen
  • Ring A is phenyl or 5-6 membered heteroaryl
  • Ring B does not exist or is 3-6 membered cycloalkyl, 4-6 membered heterocycloalkyl, phenyl, 5-6 membered heteroaryl;
  • R 3 and R 4 are each independently selected from: hydroxyl, halogen, amino, cyano, C 1 -C 5 alkyl, C 1 -C 5 alkoxy, 3-6 membered cycloalkyl, 4-6 membered hetero Cycloalkyl, 3-6 membered cycloalkoxy; said C 1 -C 5 alkyl, C 1 -C 5 alkoxy, 3-6 membered cycloalkyl, 4-6 membered heterocycloalkyl, 3 -6-membered cycloalkoxy is optionally substituted by one or more substituents selected from the group consisting of hydroxyl, halogen, amino, cyano, 3-6-membered cycloalkyl;
  • n 0, 1, 2 or 3;
  • n 0, 1, 2 or 3.
  • the 5-6 membered heteroaryl contains one or more heteroatoms selected from N, O or S; when there are multiple heteroatoms, the The heteroatoms are the same or different; preferably, the 5-6 membered heteroaryl is selected from: thiophene, furan, pyrrole, pyrazole, imidazole, triazole, thiazole, thiadiazole, oxazole, pyridine, pyrimidine , pyridazine, pyrazine.
  • Ra 1 and Ra 2 form oxetane together with their common C atoms.
  • L 1 is -CH 2 -, -CH(CH 3 )-, -CH 2 CH 2 - or -CH 2 CH 2 CH 2 -; preferably, L 1 is -CH 2 - , -CH(CH 3 )-.
  • R 3 and R 4 are each independently selected from: halogen, cyano, C 1 -C 5 alkyl, C 1 -C 5 alkoxy, 3-6 membered cycloalkyl; C 1 -C 5 alkyl, C 1 -C 5 alkoxy, 3-6 membered cycloalkyl are optionally substituted with 1, 2 or 3 substituents selected from the group consisting of hydroxy, fluoro, chloro.
  • ring B does not exist or is 3-6 membered cycloalkyl, 4-6 membered heterocycloalkyl, phenyl, 5-6 membered heteroaryl; said heterocycloalkyl or heteroaryl
  • the heteroatom contained in the group is O, N or S; the heterocycloalkyl group includes a monocyclic ring, a parallel ring, a bridged ring or a spiro ring.
  • ring B is oxetane, Or a saturated or partially saturated heterocycle selected from the following heteroaryl groups: thiophene, furan, pyrrole, pyrazole, imidazole, triazole, thiazole, thiadiazole, oxazole, pyridine, pyrimidine, pyridazine, pyrazine .
  • dihydrothiophene tetrahydrothiophene, dihydrofuran, tetrahydrofuran, dihydropyrrole, tetrahydropyrrole, dihydropyrazole, tetrahydropyrazole, dihydroimidazole, tetrahydroimidazole, dihydrotriazole , tetrahydrotriazole, dihydrothiazole, tetrahydrothiazole, dihydrothiadiazole, tetrahydrothiadiazole, dihydrooxazole, tetrahydrooxazole, dihydropyridine, tetrahydropyridine, dihydropyrimidine, four Hydropyrimidine, dihydropyridazine, tetrahydropyridazine, dihydropyrazine, tetrahydropyrazine.
  • ring B is selected from: oxetane, Cyclopropyl, cyclobutyl, phenyl.
  • formula I has the structure Ia, Ib, Ic, Id,
  • Z 1 , Z 2 , Z 3 , Z 4 , Z 5 are each independently selected from CR 3 or N;
  • Ring B, R 1 , R 2 , R 3 , R 4 , Ra (same as R a1 and R a2 in formula I), n and L 2 are as defined in the first aspect;
  • Z 1 , Z 2 , Z 3 , Z 4 , Z 5 contain at most two Ns;
  • Z 1 , Z 2 , Z 3 , and Z 4 contain at most two Ns.
  • L 2 is absent or selected from -CH 2 -, -CH 2 CH 2 -, -CH 2 CH 2 CH 2 -, -CH(CH 3 )-, -CH(CH 3 )CH 2 -; preferably, L 2 does not exist or is -CH 2 - or -CH(CH 3 )-.
  • R 1 and R 2 are hydrogen or -C ⁇ CR 11 respectively, and R 1 and R 2 are different;
  • R 11 is hydrogen, C 1 -C 5 alkyl or 3-6 membered cycloalkyl ;
  • the R 11 is methyl.
  • Ra is methyl, ethyl, propyl.
  • ring B does not exist or is 3-6 membered cycloalkyl, 4-6 membered heterocycloalkyl, phenyl, 5-6 membered heteroaryl; said heterocycloalkyl or heteroaryl
  • the heteroatom contained in the group is O or N; the heterocycloalkyl group includes a monocyclic ring, a parallel ring, a bridged ring or a spiro ring.
  • Ring B is absent or is cyclopropyl, cyclobutyl, cyclopentyl, cyclohexyl, phenyl, thiophene, furan, pyrrole, pyrazole, imidazole, triazole, thiazole, thiazole Oxadiazole, oxazole, pyridine, pyrimidine, pyridazine, pyrazine.
  • ring B is oxetane, Or a saturated or partially saturated heterocycle selected from the following heteroaryl groups: thiophene, furan, pyrrole, pyrazole, imidazole, triazole, thiazole, thiadiazole, oxazole, pyridine, pyrimidine, pyridazine, pyrazine .
  • dihydrothiophene tetrahydrothiophene, dihydrofuran, tetrahydrofuran, dihydropyrrole, tetrahydropyrrole, dihydropyrazole, tetrahydropyrazole, dihydroimidazole, tetrahydroimidazole, dihydrotriazole , tetrahydrotriazole, dihydrothiazole, tetrahydrothiazole, dihydrothiadiazole, tetrahydrothiadiazole, dihydrooxazole, tetrahydrooxazole, dihydropyridine, tetrahydropyridine, dihydropyrimidine, four Hydropyrimidine, dihydropyridazine, tetrahydropyridazine, dihydropyrazine, tetrahydropyrazine.
  • ring B is selected from: oxetane, Cyclopropyl, cyclobutyl, phenyl.
  • Formula I has the following structures IIa, IIb, IIc, IId, IIe, If, IIg, IIm, IIn
  • the benzoheterocyclic compound represented by formula I its tautomers, stereoisomers, hydrates, solvates, pharmaceutically acceptable salts or prodrugs, the
  • the benzoheterocyclic compounds shown in the formula I include the following structures:
  • benzoheterocyclic compound shown in formula I includes the following structure:
  • the second aspect of the present invention provides an intermediate B-1:
  • R 6 is -OH, -Cl, -OC 1 -C 5 alkyl, -O-benzyl;
  • Ring A, Ring B, L 1 , R 1 , R 2 , R 3 , R 4 , m, n are as defined in the first aspect.
  • the intermediate B-1 has the structure B-1a or B-1b:
  • Z 1 , Z 2 , Z 3 , Z 4 , Z 5 are each independently selected from CR 3 or N;
  • Ring B, R 1 , R 2 , R 3 , R 4 , Ra (same as R a1 and R a2 in formula I), and n are as defined in the first aspect;
  • Z 1 , Z 2 , Z 3 , Z 4 , Z 5 contain at most two Ns;
  • Z 1 , Z 2 , Z 3 , and Z 4 contain at most two Ns.
  • the intermediate B-1 has the structures B-1c, B-1d, B-1f, B-1g:
  • the third aspect of the present invention provides a method for preparing the benzoheterocyclic compound represented by formula I as described in the first aspect, its tautomers, stereoisomers, hydrates, solvates, pharmaceutically acceptable A salt or prodrug method, comprising: 1) reacting intermediate B-1 as described in the second aspect with intermediate B-2 or a salt of intermediate B-2 to obtain the benzo Heterocyclic compounds;
  • the intermediate B-2 has the structure:
  • R 5 is C 1 -C 6 alkyl, benzyl.
  • the salt of intermediate B-2 is hydrochloride.
  • the method also includes:
  • R 6 in the intermediate B-1 is not -OH or -Cl, after converting the group -COR 6 into -COOH or -COCl,
  • the intermediate B-2 has the structure B-2a or B-2b:
  • the fourth aspect of the present invention provides a pharmaceutical composition, which includes: the benzoheterocyclic compound represented by formula I as described in the first aspect, its tautomers, stereoisomers, a hydrate, a solvate, a pharmaceutically acceptable salt or a prodrug; and a pharmaceutically acceptable carrier.
  • the pharmaceutical composition further comprises a second drug.
  • the second drug includes antibodies; preferably, the antibodies include anti-PD-L1 antibodies and anti-PD-1 antibodies.
  • the benzoheterocyclic compound represented by formula I as described in the first aspect its tautomer, stereoisomer, hydrate, solvate, pharmaceutically acceptable salt or
  • the mass ratio of the prodrug to the second drug is 1:100 to 100:1, preferably 10:1 to 100:1, more preferably 10:1 to 50:1, for example 10:1, 15:1 1, 20:1, 25:1, 30:1, 35:1, 40:1, 45:1, 50:1.
  • the fifth aspect of the present invention provides a benzoheterocyclic compound represented by formula I as described in the first aspect, its tautomers, stereoisomers, hydrates, solvates, pharmaceutically acceptable
  • the purposes of salt or prodrug, or the purposes of pharmaceutical composition as described in the fourth aspect, described purposes comprises:
  • the diseases mediated by the EP2 and/or EP4 receptors include inflammatory diseases (such as arthritis and endometriosis), autoimmune diseases (such as multiple sclerosis), neurodegenerative diseases (such as epilepsy, Alzheimer's disease, Parkinson's disease, amyotrophic lateral sclerosis, and traumatic brain injury), cardiovascular disease (such as atherosclerosis), and cancer (such as colon, lung, breast, and head and neck cancers) .
  • inflammatory diseases such as arthritis and endometriosis
  • autoimmune diseases such as multiple sclerosis
  • neurodegenerative diseases such as epilepsy, Alzheimer's disease, Parkinson's disease, amyotrophic lateral sclerosis, and traumatic brain injury
  • cardiovascular disease such as atherosclerosis
  • cancer such as colon, lung, breast, and head and neck cancers
  • the sixth aspect of the present invention provides a method for preventing and/or treating diseases mediated by EP2 and/or EP4 receptors, the method comprising administering an effective amount of the formula I to an individual in need thereof Shown are benzoheterocyclic compounds, tautomers, stereoisomers, hydrates, solvates, pharmaceutically acceptable salts or prodrugs thereof.
  • the method further includes combined use with antibody therapy; preferably, the antibody therapy includes PD-L1 antibody therapy and PD-1 antibody therapy.
  • the mass ratio of prodrug to antibody is 1:100 to 100:1, preferably 10:1 to 100:1, more preferably 10:1 to 50:1, for example 10:1, 15:1, 20 :1, 25:1, 30:1, 35:1, 40:1, 45:1, 50:1.
  • the diseases mediated by the EP2 and/or EP4 receptors include inflammatory diseases (such as arthritis and endometriosis), autoimmune diseases (such as multiple sclerosis), neurological Degenerative diseases (e.g. epilepsy, Alzheimer's disease, Parkinson's disease, amyotrophic lateral sclerosis and traumatic brain injury), cardiovascular diseases (e.g. atherosclerosis) and cancers (e.g. colon, lung, breast and head and neck cancer).
  • inflammatory diseases such as arthritis and endometriosis
  • autoimmune diseases such as multiple sclerosis
  • neurological Degenerative diseases e.g. epilepsy, Alzheimer's disease, Parkinson's disease, amyotrophic lateral sclerosis and traumatic brain injury
  • cardiovascular diseases e.g. atherosclerosis
  • cancers e.g. colon, lung, breast and head and neck cancer.
  • substituents When a substituent is described by a conventional chemical formula written from left to right, the substituent also includes chemically equivalent substituents obtained when the structural formula is written from right to left. For example, CH2O is equivalent to OCH2 .
  • R 1 ", “R1” and “R 1 " have the same meaning and can be substituted for each other. For other symbols such as R 2 , similar definitions have the same meanings.
  • halogen means fluorine, chlorine, bromine, iodine alone or as part of another substituent.
  • alkyl alone or as part of another substituent, means consisting solely of carbon and hydrogen atoms, free of unsaturated bonds, having, for example, 1 to 6 carbon atoms, and is bonded to the molecule by a single bond. The rest are connected straight or branched hydrocarbon chain groups.
  • alkyl groups include, but are not limited to, methyl, ethyl, n-propyl, isopropyl, n-butyl, isobutyl, t-butyl, pentyl, isopentyl, neopentyl, and hexyl.
  • An alkyl group can be unsubstituted or substituted with one or more suitable substituents.
  • alkyl groups may also be isotopomers of naturally abundant alkyl groups that are rich in carbon and/or hydrogen isotopes (ie, deuterium or tritium).
  • alkenyl means an unbranched or branched monovalent hydrocarbon chain containing one or more carbon-carbon double bonds.
  • alkynyl refers to an unbranched or branched monovalent hydrocarbon chain containing one or more carbon-carbon triple bonds. "C 2 -C 5 alkynyl" then contains 2-5 carbon atoms.
  • C 1 -C 5 alkyl alone or as part of another substituent, is understood to mean a linear or branched saturated monovalent hydrocarbon radical having 1, 2, 3, 4 or 5 carbon atoms.
  • the alkyl group is for example methyl, ethyl, propyl, butyl, pentyl, hexyl, isopropyl, isobutyl, sec-butyl, tert-butyl, isopentyl, 2-methylbutyl, 1-methylbutyl, 1-ethylpropyl, 1,2-dimethylpropyl, neopentyl, 1,1-dimethylpropyl, etc. or their isomers.
  • said groups have 1, 2 or 3 carbon atoms (“C 1 -C 3 alkyl”), for example methyl, ethyl, n-propyl or isopropyl.
  • alkylene means a saturated divalent hydrocarbyl group obtained by removing two hydrogen atoms from a saturated straight or branched chain hydrocarbyl group.
  • C 1 -C 5 alkylene refers to an alkylene group containing 1, 2, 3, 4, or 5 carbon atoms, examples of which include methylene (-CH 2 -), ethylene (including -CH 2 CH 2 -or -CH(CH 3 )-), isopropylidene (including -CH(CH 3 )CH 2 - or -C(CH 3 ) 2 -), etc.
  • cycloalkyl or “carbocyclyl” by themselves or as part of another substituent refer to a cyclic alkyl group.
  • mn-membered cycloalkyl or " Cm - Cncycloalkyl” is understood to mean a saturated, unsaturated or partially saturated carbocycle having m to n atoms.
  • 3-15 membered cycloalkyl or “C 3 -C 15 cycloalkyl” refers to a cyclic alkyl group containing 3 to 15, 3 to 9, 3 to 6 or 3 to 5 carbon atoms, which May contain 1 to 4 rings.
  • "3-6 membered cycloalkyl” then contains 3-6 carbon atoms.
  • cycloalkyl groups may be substituted with one or more substituents.
  • a cycloalkyl group can be a cycloalkyl group fused to an aryl or heteroaryl ring group.
  • Halo is used interchangeably with the term “halogen substitution”, alone or as part of another substituent.
  • Haloalkyl or halogen-substituted alkyl refers to both branched and straight-chain saturated aliphatic hydrocarbon groups having the specified number of carbon atoms, substituted with one or more halogens.
  • heterocycloalkyl or “heterocyclyl”, by themselves or as part of another substituent, refer to rings in which one or more (in some embodiments, 1 to 3) carbon atoms are replaced by heteroatoms Alkyl, said heteroatoms such as but not limited to N, O, S and P.
  • mn-membered heterocycloalkyl or " Cm - Cnheterocycloalkyl” is understood to mean a saturated, unsaturated or partially saturated ring having m to n atoms.
  • 4-6 membered heterocycloalkyl is understood to mean a saturated, unsaturated or partially saturated ring having 4 to 6 atoms.
  • the number of carbons is also meant to include heteroatoms.
  • heteroaryl is used interchangeably with the terms “heteroaromatic ring” or “heteroaromatic”, by itself or as part of another substituent, and refers to a monocyclic or polycyclic aromatic ring system, in certain implementations In the scheme, 1 to 3 atoms in the ring system are heteroatoms, that is, elements other than carbon, including but not limited to N, O, S or P.
  • 5-6 membered heteroaryl alone or as part of another substituent, is understood to mean an aromatic ring group having 5 to 6 ring atoms, including heteroatoms. Including but not limited to thiophene, furan, pyrrole, pyrazole, imidazole, triazole, thiazole, thiadiazole, oxazole, pyridine, pyrimidine, pyridazine, pyrazine.
  • the compounds provided herein, including intermediates useful in the preparation of the compounds provided herein, contain reactive functional groups (such as, but not limited to, carboxyl, hydroxyl, and amino moieties), and also include protected derivatives thereof.
  • "Protected derivatives” are those compounds in which one or more reactive sites are blocked by one or more protecting groups (also known as protecting groups).
  • Suitable protecting groups for the carboxyl moiety include benzyl, tert-butyl, etc., and isotopes, etc.
  • Suitable amino and amido protecting groups include acetyl, trifluoroacetyl, t-butoxycarbonyl, benzyloxycarbonyl and the like.
  • Suitable hydroxy protecting groups include benzyl and the like. Other suitable protecting groups are well known to those of ordinary skill in the art.
  • “optional” or “optionally” means that the subsequently described event or circumstance may or may not occur, and that the description includes both the occurrence and non-occurrence of the event or circumstance.
  • “optionally substituted aryl” means that the aryl is substituted or unsubstituted, and the description includes both substituted and unsubstituted aryl.
  • “optional” or “optionally” substitution covers the situation that the compound structure/group is not substituted, and the compound structure/group is substituted by one or more defined substituents.
  • “optionally substituted aryl” means unsubstituted aryl and aryl substituted with one or more substituents selected from the defined substituents.
  • “Multiple” means more than two, including two or more.
  • salt or “pharmaceutically acceptable salt” includes pharmaceutically acceptable acid addition salts and pharmaceutically acceptable base addition salts.
  • pharmaceutically acceptable refers to those compounds, materials, compositions and/or dosage forms which, within the scope of sound medical judgment, are suitable for use in contact with human and animal tissues without excessive Toxicity, irritation, allergic reaction, or other problems or complications, commensurate with a reasonable benefit/risk ratio.
  • “Pharmaceutically acceptable acid addition salt” refers to a salt formed with an inorganic or organic acid that retains the biological effectiveness of the free base without other side effects.
  • “Pharmaceutically acceptable base addition salt” refers to a salt formed with an inorganic base or an organic base that can maintain the biological effectiveness of the free acid without other side effects.
  • other salts are contemplated by the present invention. They may serve as intermediates in the purification of compounds or in the preparation of other pharmaceutically acceptable salts or may be useful in the identification, characterization or purification of compounds of the invention.
  • stereoisomer refers to isomers resulting from differences in the arrangement of atoms in a molecule in space, including cis-trans isomers, enantiomers, diastereoisomers and conformers.
  • the compounds according to the invention may exist as one of the possible isomers or as mixtures thereof, for example as pure optical isomers, or as mixtures of isomers, for example as racemic and non- A mixture of enantiomers, depending on the number of asymmetric carbon atoms.
  • the prefixes D and L or R and S are used to denote the absolute configuration of the molecule with respect to the chiral center (or centers) in the molecule.
  • the prefixes D and L or (+) and (-) are symbols used to designate the rotation of plane polarized light by a compound, where (-) or L indicates that the compound is levorotatory.
  • Compounds prefixed with (+) or D are dextrorotatory.
  • tautomer refers to isomers of functional groups resulting from the rapid movement of an atom in a molecule between two positions.
  • the compounds of the present invention may exhibit tautomerism.
  • Tautomeric compounds can exist in two or more interconvertible species.
  • Prototropic tautomers result from the migration of a covalently bonded hydrogen atom between two atoms.
  • Tautomers generally exist in equilibrium and attempts to isolate a single tautomer usually result in a mixture whose physicochemical properties are consistent with the mixture of compounds. The position of equilibrium depends on the chemical properties within the molecule.
  • the keto form predominates
  • the enol form predominates.
  • the present invention encompasses all tautomeric forms of the compounds.
  • a "pharmaceutical composition” refers to a formulation of a compound of the present invention with a vehicle generally accepted in the art for the delivery of a biologically active compound to a mammal (eg, a human).
  • the medium includes a pharmaceutically acceptable carrier.
  • the purpose of the pharmaceutical composition is to promote the administration of the organism, facilitate the absorption of the active ingredient and thus exert its biological activity.
  • pharmaceutically acceptable carrier includes, but is not limited to, any adjuvant, carrier, excipient, glidant, sweetener approved by the relevant government regulatory agency as acceptable for human or livestock use , diluent, preservative, dye/colorant, flavoring agent, surfactant, wetting agent, dispersing agent, suspending agent, stabilizing agent, isotonic agent, solvent or emulsifying agent.
  • solvate means that the compound of the present invention or its salt includes a stoichiometric or non-stoichiometric solvent bonded by intermolecular non-covalent forces, and when the solvent is water, it is a hydrate.
  • prodrug refers to a compound of the invention that can be converted to biological activity under physiological conditions or by solvolysis.
  • the prodrugs of the present invention are prepared by modifying functional groups in the compounds which can be removed routinely or in vivo to yield the parent compound.
  • Prodrugs include compounds formed by linking a hydroxyl or amino group in the compound of the present invention to any group. When the prodrug of the compound of the present invention is administered to a mammalian individual, the prodrug is split to form free hydroxyl, free of amino.
  • excipient refers to a pharmaceutically acceptable inert ingredient.
  • examples of categories of the term “excipient” include, but are not limited to, binders, disintegrants, lubricants, glidants, stabilizers, fillers, diluents, and the like. Excipients can enhance the handling characteristics of a pharmaceutical formulation, ie make the formulation more suitable for direct compression by increasing flow and/or cohesiveness.
  • patient refers to any animal including mammals, preferably mice, rats, other rodents, rabbits, dogs, cats, pigs, cows, sheep, horses or primates, most preferably humans.
  • terapéuticaally effective amount refers to the amount of an active compound or drug that a researcher, veterinarian, physician, or other clinician is seeking to elicit a biological or medical response in a tissue, system, animal, individual, or human, and includes any of the following or more: (1) Preventing a disease: eg, preventing a disease, disorder or condition in an individual who is susceptible to the disease, disorder or condition but has not yet experienced or developed disease pathology or symptoms. (2) Inhibiting a disease: For example, inhibiting a disease, disorder or condition (ie preventing further development of the pathology and/or symptoms) in an individual experiencing or developing pathology or symptoms of the disease, disorder or condition. (3) Alleviating disease: For example, alleviating a disease, disorder or condition (ie reversing the pathology and/or symptoms) in an individual experiencing or developing the pathology or symptoms of the disease, disorder or condition.
  • treatment and other similar synonyms include the following meanings:
  • antibody includes all classes of immunoglobulins. Antibodies may be monoclonal or polyclonal and may be of any species of origin including, for example, mouse, rat, rabbit, horse or human. Antibodies may be chimeric or humanized, especially when they are used for therapeutic purposes. Antibodies can be obtained or prepared by methods known in the art.
  • P-L1 antibody or "anti-PD-L1” refers to an antibody directed against programmed death ligand 1 (PD-L1).
  • PD-1 antibody or "anti-PD-1” refers to an antibody directed against programmed death protein 1 (PD-1).
  • antibody therapy refers to the medical use of antibodies that bind to target cells or proteins of target cells to treat cancer and/or stimulate an immune response in a subject that results in the recognition, attack and and/or destroy, and in some embodiments of the invention, to activate or stimulate a subject's memory immune response that results in subsequent recognition, attack and/or destruction of cancer cells in the subject.
  • PD-L1 antibody therapy refers to the use of antibodies against programmed death-ligand 1 (anti-PD-L1 ) to modulate the immune response of a subject.
  • the PD-L1 antibody inhibits or blocks the interaction of PD-L1 with programmed cell death protein 1 (PD-1), wherein blockade of the interaction between PD-L1 and PD-1 inhibits PD-1 negatively regulates T cell activation to attack and destroy cancer cells.
  • PD-1 programmed cell death protein 1
  • PD-1 antibody therapy refers to the use of antibodies against the programmed cell death protein 1 PD-1 (anti-PD-1) to modulate the immune response of a subject.
  • the PD1 antibody inhibits or blocks the interaction of PD-1 and PD-L1, wherein the inhibition or blockage of the interaction between PD-L1 and PD-1 inhibits the effect of PD-1 on T cell activation Negative regulation, thereby attacking and destroying cancer cells.
  • the reaction temperature can be appropriately selected according to the solvent, starting material, reagent, etc.
  • the reaction time can also be appropriately selected according to the reaction temperature, solvent, starting material, reagent, etc.
  • the target compound can be separated and purified from the reaction system according to common methods, such as filtration, extraction, recrystallization, washing, silica gel column chromatography and other methods. Without affecting the next step reaction, the target compound can also directly enter the next step reaction without separation and purification.
  • the benzo heterocyclic compound is represented by formula I described in the present invention Show benzoheterocyclic compounds.
  • the benzoheterocyclic compound used for the treatment of diseases mediated by EP2 and EP4 receptors described in the present invention can be used for the treatment of inflammatory diseases (such as arthritis and endometriosis), autoimmune diseases (such as multiple sclerosis), neurodegenerative diseases (such as epilepsy, Alzheimer's disease, Parkinson's disease, amyotrophic lateral sclerosis, and traumatic brain injury), cardiovascular diseases (such as atherosclerosis) and cancers (such as colon cancer , lung, breast and head and neck cancers).
  • inflammatory diseases such as arthritis and endometriosis
  • autoimmune diseases such as multiple sclerosis
  • neurodegenerative diseases such as epilepsy, Alzheimer's disease, Parkinson's disease, amyotrophic lateral sclerosis, and traumatic brain injury
  • cardiovascular diseases such as atherosclerosis
  • cancers such as colon cancer , lung, breast and head and neck cancers.
  • the benzoheterocyclic compound of the present invention has a good antagonism to EP2, a good inhibitory effect on EP4 calcium flow, and a good affinity with EP2 receptors and/or EP4 receptors.
  • the benzoheterocyclic compound of the present invention has low clearance rate of intravenous administration, high oral administration exposure, exhibits excellent pharmacokinetic properties, has a relatively high free fraction in human plasma, and has good thermodynamics Good solubility and druggability.
  • the combination with antibody drugs (such as anti-PD-L1 antibody, anti-PD-1 antibody) showed a significant tumor inhibitory effect.
  • Fig. 1 is the evaluation result of the compound in the present invention to the antigen presenting cell of human PBMC cell differentiation experiment;
  • Fig. 2 is the evaluation result of the compound in the present invention to human PBMC cell differentiation experiment M2 type macrophage;
  • Fig. 3 is the evaluation result of the compound in the present invention to human PBMC cell differentiation experiment M1 type macrophage;
  • Figure 4 is the results of the anti-tumor effect of the compound of the present invention combined with anti-mouse PD-1 antibody in the CT-26 mouse colon cancer tumor model.
  • IC 50 half inhibitory concentration, which refers to the concentration at which half of the maximum inhibitory effect is achieved
  • n-butyllithium 14.56mL, 29.1mmol, 2.5M n-hexane solution
  • HATU O-(7-azabenzotriazol-1-yl)-N,N,N′,N′-tetramethyluronium hexafluorophosphate
  • PE petroleum ether
  • Acidic preparation method A Welch, Ultimate C 18 column, 10 ⁇ m, 21.2 mm ⁇ 250 mm.
  • Mobile phase A was 1 ⁇ pure trifluoroacetic acid aqueous solution
  • mobile phase B was acetonitrile solution.
  • Gradient conditions 0 to 3 minutes, maintain 90% of mobile phase A, gradient elution in 3 to 18 minutes, change from 90% to 5%, and maintain 5% in 18 to 22 minutes.
  • the synthetic route is as follows:
  • the first step the preparation of 4-(propane-1-yn-1-yl)-1H-indazole-7-carboxylic acid methyl ester
  • Step 3 Preparation of 1-((1,1’-diphenyl)-4-ylmethyl)-4-(propane-1-yn-1-yl)-1H-indazole-7-carboxylic acid
  • the starting material 1-((1,1'-diphenyl)-4-ylmethyl)-4-(propan-1-yn-1-yl)-1H-indazole-7-carboxylic acid methyl ester ( 80mg, 0.21mmol) was added to tetrahydrofuran (3mL), methanol (1mL) and water (1mL), lithium hydroxide (15mg, 0.63mmol) was added, and stirred at room temperature for 16h.
  • the synthetic route is as follows:
  • the first step the preparation of 5-(propane-1-yn-1-yl)-1H-indazole-7-carboxylic acid methyl ester
  • Step 3 Preparation of 1-((1,1’-diphenyl)-4-ylmethyl)-5-(propane-1-yn-1-yl)-1H-indazole-7-carboxylic acid
  • the starting material 1-((1,1'-diphenyl)-4-ylmethyl)-5-(propane-1-yn-1-yl)-1H-indazole-7-carboxylic acid methyl ester 50mg, 0.13mmol was added to tetrahydrofuran (3mL), methanol (1mL) and water (1mL), lithium hydroxide (10mg, 0.40mmol) was added, and stirred at room temperature for 4h.
  • the synthetic route is as follows:
  • the first step the preparation of 4-(butane-1-yn-1-yl)-1H-indazole-7-carboxylic acid methyl ester
  • Step 3 Preparation of 1-((1,1’-diphenyl)-4-ylmethyl)-4-(butane-1-yn-1-yl)-1H-indazole-7-carboxylic acid
  • the fifth step 6-(1-((1,1'-diphenyl)-4-ylmethyl)-4-(butane-1-yn-1-yl)-1H-indazole-7- Formamide) spiro[3.3]heptane-2-carboxylic acid (I-3) preparation
  • the synthetic route is as follows:
  • Step 1 Preparation of 1-((1,1’-diphenyl)-4-ylmethyl)-4-bromo-1H-indazole-7-carboxylic acid methyl ester
  • the second step the preparation of 1-((1,1'-diphenyl)-4-ylmethyl)-4-(cyclopropaneethynyl)-1H-indazole-7-carboxylic acid methyl ester
  • Step 3 Preparation of 1-((1,1’-diphenyl)-4-ylmethyl)-4-(cyclopropaneethynyl)-1H-indazole-7-carboxylic acid
  • the synthetic route is as follows:
  • Step 1 Preparation of 1-((1,1’-diphenyl)-4-ylmethyl)-4-bromo-1H-indazole-7-carboxylic acid methyl ester
  • the third step 1-((1,1'-diphenyl)-4-ylmethyl)-4-(3-hydroxy-3-methylbutane-1-yn-1-yl)-1H- Preparation of indazole-7-carboxylic acid
  • the starting material 1-((1,1'-diphenyl)-4-ylmethyl)-4-(3-hydroxy-3-methylbutane-1-yn-1-yl)-1H- Add methyl indazole-7-carboxylate (70mg, 0.16mmol) into tetrahydrofuran (3mL) and water (1mL), add lithium hydroxide (12mg, 0.50mmol), and stir at room temperature for 16h.
  • the fifth step 6-(1-((1,1'-diphenyl)-4-ylmethyl)-4-(3-hydroxy-3-methylbutane-1-yn-1-yl) Preparation of -1H-indazole-7-carboxamide) spiro[3.3]heptane-2-carboxylic acid (I-5)
  • the synthetic route is as follows:
  • the first step the preparation of 4-(propane-1-yn-1-yl)-1H-indazole-7-acetic acid methyl ester
  • Step 3 Preparation of 1-((1,1’-diphenyl)-4-ylmethyl)-4-(propane-1-yn-1-yl)-1H-indazole-7-acetic acid
  • the starting material 1-((1,1'-diphenyl)-4-ylmethyl)-4-(propan-1-yn-1-yl)-1H-indazole-7-acetic acid methyl ester ( 80mg, 0.21mmol) was added to tetrahydrofuran (3mL), methanol (1mL) and water (1mL), lithium hydroxide (15mg, 0.63mmol) was added, and stirred at room temperature for 16h.
  • the synthetic route is as follows:
  • the first step the preparation of 5-(propane-1-yn-1-yl)-1H-indazole-7-acetic acid methyl ester
  • Step 3 Preparation of 1-((1,1’-diphenyl)-4-ylmethyl)-5-(propane-1-yn-1-yl)-1H-indazole-7-acetic acid
  • the synthetic route is as follows:
  • the first step the preparation of 4-(propane-1-yn-1-yl)-1H-indazole-7-carboxylic acid methyl ester
  • the second step the preparation of 4-(propane-1-yn-1-yl)-1-(4-(trifluoromethoxy)benzyl)-1H-indazole-7-carboxylic acid methyl ester
  • the third step the preparation of 4-(propane-1-yn-1-yl)-1-(4-(trifluoromethoxy)benzyl)-1H-indazole-7-carboxylic acid
  • the synthetic route is as follows:
  • the first step the preparation of (4-((7-(methoxycarbonyl)-4-(propane-1-yn-1-yl)-1H-indazol-1-yl)methyl)benzyl)boronic acid
  • the starting material (4-((7-(methoxycarbonyl)-4-(propane-1-yn-1-yl)-1H-indazol-1-yl)methyl)benzyl)boronic acid (80 mg, 0.23mmol) was added to 1,4-dioxane (5mL) and water (1mL), and chloro(2-dicyclohexylphosphino-2,6-dimethoxy-1,1-biphenyl )(2-amino-1,1-biphenyl-2-yl)palladium(II) (17mg, 0.02mmol) and potassium phosphate (196mg, 0.92mmol), 1,1'-bisdiphenylphosphinoferrocene Palladium dichloride (19mg, 0.02mmol), 4-bromo-2-methoxypyrimidine (53mg, 0.28mmol), stirred at 90°C for 1h.
  • Step 3 Preparation of 1-(4-(2-methoxypyrimidin-4-yl)benzyl)-4-(propane-1-yn-1-yl)-1H-indazole-7-carboxylic acid
  • the starting material 1-(4-(2-methoxypyrimidin-4-yl)benzyl)-4-(propane-1-yn-1-yl)-1H-indazole-7-carboxylic acid methyl ester ( 70mg, 0.17mmol) was added to tetrahydrofuran (3mL), methanol (1mL) and water (1mL), lithium hydroxide (13mg, 0.51mmol) was added, and stirred at room temperature for 16h.
  • the synthetic route is as follows:
  • the first step the preparation of (4-((7-(methoxycarbonyl)-4-(propane-1-yn-1-yl)-1H-indazol-1-yl)methyl)benzyl)boronic acid
  • the second step the preparation of 4-(propane-1-yn-1-yl)-1-(4-(thiazol-2-yl)benzyl)-1H-indazole-7-carboxylic acid methyl ester
  • the third step the preparation of 4-(propane-1-yn-1-yl)-1-(4-(thiazol-2-yl)benzyl)-1H-indazole-7-carboxylic acid
  • the synthetic route is as follows:
  • Step 1 Preparation of 1-(4-iodophenyl)-4-(propane-1-yn-1-yl)-1H-indazole-7-carboxylic acid methyl ester
  • the second step the preparation of 1-(4-(1H-pyrazol-1-yl)benzyl)-4-(propane-1-yn-1-yl)-1H-indazole-7-carboxylic acid methyl ester
  • Step 4 Preparation of 1-(4-(1H-pyrazol-1-yl)benzyl)-4-(propane-1-yn-1-yl)-1H-indazole-7-carboxylic acid
  • the synthetic route is as follows:
  • the first step the preparation of 2-amino-4-bromo-3-methylbenzoic acid methyl ester
  • the second step the preparation of (3-bromo-6-(methoxycarbonyl)-2-methylphenyl)-1-tetrafluoroborate diazonium salt
  • the compound 2-amino-4-bromo-3-methylbenzoate (5.0 g, 20.5 mmol) was added to 50% HBF 4 aqueous solution (5 mL) at room temperature, cooled to 0 ° C, and sodium nitrite ( 1.42g, 20.6mmol) in 5mL aqueous solution, stirred at 0°C for 1h. After filtration and drying, the crude compound (3-bromo-6-(methoxycarbonyl)-2-methylphenyl)-1-tetrafluoroborate diazonium salt (6.6 g, yield 94%) was obtained.
  • the third step the preparation of 4-bromo-1H-indazole-7-carboxylic acid methyl ester
  • Step 4 Preparation of 4-(propane-1-yn-1-yl)-1H-indazole-7-carboxylic acid methyl ester
  • Step 5 Preparation of 1-(4-cyclopropylbenzyl)-4-(propane-1-yn-1-yl)-1H-indazole-7-carboxylic acid methyl ester
  • Step 6 Preparation of 1-(4-cyclopropylbenzyl)-4-(propane-1-yn-1-yl)-1H-indazole-7-carboxylic acid
  • the seventh step 6-(1-(4-cyclopropylbenzyl)-4-(propane-1-yn-1-yl)-1H-indazole-7-carboxamide)spiro[3.3]heptane - Preparation of methyl 2-formate
  • the synthetic route is as follows:
  • Step 1 Preparation of 1-(4-iodophenyl)-4-(propane-1-yn-1-yl)-1H-indazole-7-carboxylic acid methyl ester
  • the third step the preparation of 1-(4-(2-methoxypyridin-4-yl)phenyl)-4-(propane-1-yn-1-yl)-1H-indazole-7-carboxylic acid
  • Methyl 1-(4-(2-methoxypyridin-4-yl)phenyl)-4-(propane-1-yn-1-yl)-1H-indazole-7-carboxylate (100mg , 0.24mmol) was added to tetrahydrofuran (3mL) and water (1mL), lithium hydroxide (18mg, 0.73mmol) was added, and stirred at room temperature for 16h.
  • the synthetic route is as follows:
  • the first step the preparation of 2-(4-(bromomethyl)benzyl)propan-2-ol
  • the second step the preparation of 1-(4-(2-hydroxypropan-2-yl)benzyl)-4-(propane-1-yn-1-yl)-1H-indazole-7-carboxylic acid methyl ester
  • Step 3 Preparation of 1-(4-(2-hydroxypropan-2-yl)benzyl)-4-(propane-1-yn-1-yl)-1H-indazole-7-carboxylic acid
  • the synthetic route is as follows:
  • the first step the preparation of 4-(propane-1-yn-1-yl)-1H-indazole-7-carboxylic acid methyl ester
  • the second step the preparation of 4-(propane-1-yn-1-yl)-1-(4-(trifluoromethyl)benzyl)-1H-indazole-7-carboxylic acid methyl ester
  • the third step the preparation of 4-(propane-1-yn-1-yl)-1-(4-(trifluoromethyl)benzyl)-1H-indazole-7-carboxylic acid
  • the first step the preparation of 1-(bromomethyl)-3-cyclopropylbenzene
  • the second step the preparation of 1-(3-cyclopropylbenzyl)-4-(propane-1-yn-1-yl)-1H-indazole-7-carboxylic acid methyl ester
  • Step 3 Preparation of 1-(3-cyclopropylbenzyl)-4-(propane-1-yn-1-yl)-1H-indazole-7-carboxylic acid
  • the synthetic route is as follows:
  • the first step the preparation of 4-(propane-1-yn-1-yl)-1H-indazole-7-carboxylic acid methyl ester
  • the second step the preparation of 4-(propane-1-yn-1-yl)-1-(3-(trifluoromethoxy)benzyl)-1H-indazole-7-carboxylic acid methyl ester
  • the third step the preparation of 4-(propane-1-yn-1-yl)-1-(3-(trifluoromethoxy)benzyl)-1H-indazole-7-carboxylic acid
  • the synthetic route is as follows:
  • the first step the preparation of 4-(propane-1-yn-1-yl)-1H-indazole-7-carboxylic acid methyl ester
  • the second step the preparation of 1-(1-(4-cyclopropylphenyl)ethyl)-4-(propane-1-yn-1-yl)-1H-indazole-7-carboxylic acid methyl ester
  • Step 3 Preparation of 1-(1-(4-cyclopropylphenyl)ethyl)-4-(propane-1-yn-1-yl)-1H-indazole-7-carboxylic acid
  • the synthetic route is as follows:
  • the first step the preparation of 4-(propane-1-yn-1-yl)-1H-indazole-7-carboxylic acid methyl ester
  • the second step the preparation of (R)-1-(1-(4-bromophenyl)ethyl)-4-(propane-1-yn-1-yl)-1H-indazole-7-carboxylic acid methyl ester
  • Step 3 Preparation of (R)-1-(1-(4-cyclopropylphenyl)ethyl)-(propane-1-yn-1-yl)-1H-indazole-7-carboxylic acid methyl ester
  • Step 4 Preparation of (R)-1-(1-(4-cyclopropylphenyl)ethyl)-4-(propane-1-yn-1-yl)-1H-indazole-7-carboxylic acid
  • the synthetic route is as follows:
  • the synthetic route is as follows:
  • the first step the preparation of 4-(propane-1-yn-1-yl)-1H-indazole-7-carboxylic acid methyl ester
  • the second step the preparation of 4-(propane-1-yn-1-yl)-1-(4-(trifluoromethoxy)phenyl)ethyl)-1H-indazole-7-carboxylic acid methyl ester
  • the third step the preparation of 4-(propane-1-yn-1-yl)-1-(4-(trifluoromethoxy)phenyl)ethyl)-1H-indazole-7-carboxylic acid
  • the synthetic route is as follows:
  • Step 1 Preparation of (R)-1-(4-(4,4,5,5-tetramethyl-1,3,2-dioxaborolan-2-yl)phenyl)ethanol
  • the second step the preparation of (R)-1-(4-(2-methoxypyrimidin-4-yl)phenyl)ethanol
  • the synthetic route is as follows:
  • the synthetic method of compound I-20b refers to the synthetic method of its isomer I-20a, changing (R)-1-(4-bromophenyl)ethanol into (s)-1-(4-bromophenyl)ethanol , through the same six-step reaction to obtain the compound (Sa, R)-6-(1-(1-(4-(2-methoxypyrimidin-4-yl)phenyl)ethyl)-4-(propane- 1-yn-1-yl)-1H-indazole-7-carboxamido)spiro[3.3]heptane-2-carboxylic acid (34.5 mg, 59% yield).
  • the synthetic route is as follows:
  • the first step the preparation of 4-(propane-1-yn-1-yl)-1H-indazole-7-carboxylic acid methyl ester
  • the third step 1-(3-(4-cyclopropylphenyl)oxetan-3-yl)-4-(propane-1-yn-1-yl)-1H-indazole-7-formic acid Preparation of esters
  • Methyl 1-(3-(4-bromophenyl)oxetan-3-yl)-4-(propane-1-yn-1-yl)-1H-indazole-7-carboxylic acid methyl ester 300mg , 0.71mmol
  • cyclopropylboronic acid 300mg, 3.53mmol
  • tetrakis (triphenylphosphine) palladium 80mg, 0.071mmol
  • potassium fluoride 123mg, 2.12mmol was added to toluene (3ml) and water (0.5 ml), heated to 100°C and stirred for 12h.
  • the synthetic route is as follows:
  • Methyltriphenylphosphine bromide (9.86g, 27.6mmol) was dissolved in tetrahydrofuran (100mL), potassium tert-butoxide (1mol/L solution in tetrahydrofuran, 27.6mL) was slowly added dropwise under ice cooling, and stirred for 1 hour.
  • 1-(4-Bromophenyl)-3-oxetanyl-1-carbonitrile (6.9g, 27.6mmol) was dissolved in tetrahydrofuran (10mL) and slowly added to the above solution, gradually raised to room temperature and stirred overnight for 16 Hour.
  • Step 8 Preparation of 1-(4-(1-methyl-2-oxabicyclo[2.2.1]hexane-4-yl)phenyl)ethan-1-one
  • Step 9 Preparation of 1-(4-(1-methyl-2-oxabicyclo[2.2.1]hexane-4-yl)phenyl)ethan-1-ol
  • reaction solution was concentrated and purified by column chromatography to obtain 1-(4-(1-methyl-2-oxabicyclo[2.2.1]hexane-4-yl)phenyl)ethyl)-4-(propane-1 -Alkyn-1-yl)-1H-indazole-7-carboxylic acid methyl ester (500 mg, 86% yield).
  • the synthetic route is as follows:
  • reaction solution was poured into water (30 mL) and extracted with ethyl acetate (20 mL*3). The organic phases were combined, washed with saturated brine (10 mL*2), dried over anhydrous sodium sulfate, filtered and concentrated to obtain a crude product.
  • the reaction solution was concentrated to obtain a crude product.
  • the synthetic route is as follows:
  • the first step the preparation of N-methoxy-N-methyl-6-(trifluoromethoxy)pyridinamide
  • the second step the preparation of 1-(6-(trifluoromethoxy)pyridin-3-yl)ethan-1-one
  • the third step the preparation of 1-(6-(trifluoromethoxy)pyridin-3-yl)ethan-1-ol
  • the fifth step 4-(propane-1-yn-1-yl)-1-(1-(6-(trifluoromethoxy)pyridin-3-yl)ethane)-1H-indazole-7- Preparation of formic acid
  • the synthetic route is as follows:
  • Step 1 Preparation of 1-(1-(5-bromopyridin-2-yl)ethyl)-4-(propane-1-yn-1-yl)-1H-indazole-7-carboxylic acid methyl ester
  • Methyl 1-(1-(5-bromopyridin-2-yl)ethyl)-4-(propane-1-yn-1-yl)-1H-indazole-7-carboxylate 400 mg, 1.00 mmol
  • tetrakistriphenylphosphopalladium 116mg, 0.1mmol
  • cyclopropylboronic acid 431mg, 5.02mmol
  • potassium fluoride 321mg, 5.52mmol
  • reaction solution was concentrated and purified by column chromatography to obtain 1-(1-(5-cyclopropylpyridin-2-yl)ethyl)-4-(propane-1-yn-1-yl)-1H-indazole-7 - Methyl formate (250 mg, 69% yield).
  • Step 3 Preparation of 1-(1-(5-cyclopropylpyridin-2-yl)ethyl)-4-(propane-1-yn-1-yl)-1H-indazole-7-carboxylic acid
  • the synthetic route is as follows:
  • the first step the preparation of 1-(5-chloropyrazin-2-yl)-ethan-1-ol
  • the second step the preparation of 1-(5-cyclopropylpyrazin-2-yl)-ethan-1-ol
  • 1-(5-cyclopropylpyrazin-2-yl)-ethan-1-ol (613 mg, 3.73 mmol), methyl 4-(propan-1-yn-1-yl)-1H-indazole- 7-Carboxylate (400mg, 1.87mmol) and triphenylphosphine (1.47g, 5.60mmol) were dissolved in tetrahydrofuran (10mL), and azobisisopropyl ester (1.13g, 5.60mmol) was slowly added dropwise under ice-cooling , followed by warming to room temperature and stirring for 16 hours.
  • reaction solution was concentrated and purified by column chromatography to obtain 1-(1-(5-cyclopropylpyrazin-2-yl)ethyl)-4-(propane-1-yn-1-yl)-1H-indazole- 7-Methyl carboxylate (400 mg, 30% yield).
  • Step 4 Preparation of 1-(1-(5-cyclopropylpyrazin-2-yl)ethyl)-4-(propane-1-yn-1-yl)-1H-indazole-7-carboxylic acid
  • the synthetic route is as follows:
  • the synthetic route is as follows:
  • Step 1 Preparation of 1-(1-(6-bromopyridin-3-yl)ethyl)-4-(propane-1-yn-1-yl)-1H-indazole-7-carboxylic acid methyl ester
  • reaction solution was poured into water (50 mL), extracted with ethyl acetate (20 mL*3). The organic phases were combined, washed with saturated brine (20 mL*2), dried over anhydrous sodium sulfate, filtered and concentrated to obtain a brown crude product.
  • Step 3 Preparation of 1-(1-(6-cyclopropylpyridin-3-yl)ethyl)-4-(propane-1-yn-1-yl)-1H-indazole-7-carboxylic acid
  • reaction solution was added to water (50 mL), and extracted with ethyl acetate (10 mL*3). The combined organic phases were washed with saturated brine (10 mL*3), dried over anhydrous sodium sulfate, filtered, and concentrated to obtain a crude product.
  • Resolution method Use Column: Chiralpak AD-3 type chiral resolution column (specification: 50 ⁇ 4.6mm, 3um) for chiral resolution, mobile phase A is supercritical fluid CO 2 , mobile phase B is isopropyl Alcohol (containing 0.05% diethanolamine); gradient conditions: mobile phase B is 40%, flow rate is 3mL/min; column temperature is maintained at 35°C, and column pressure is maintained at 100bar.
  • (R)-6-(1-(1-(6-cyclopropylpyridin-3-yl)ethyl)-4-(propane-1-yn-1-yl)-1H-indazole-7 -Carboxamido)spiro[3.3]heptane-2-carboxylic acid methyl ester retention time is 0.754min.
  • the retention time of amido)spiro[3.3]heptane-2-carboxylic acid methyl ester was 1.448min.
  • Step 7 (Sa,S)-6-(1-(1-(6-cyclopropylpyridin-3-yl)ethyl)-4-(propane-1-yn-1-yl)-1H- Preparation of indazole-7-carboxamido)spiro[3.3]heptane-2-carboxylic acid (I-30b)
  • the synthetic route is as follows:
  • reaction solution was concentrated and purified by column chromatography to obtain 1-(1-(2-cyclopropylpyrimidin-5-yl)ethyl)-4-(propane-1-yn-1-yl)-1H-indazole-7 - Methyl formate (200 mg, 43% yield).
  • Step 3 Preparation of 1-(1-(2-cyclopropylpyrimidin-5-yl)ethyl)-4-(propane-1-yn-1-yl)-1H-indazole-7-carboxylic acid
  • the synthetic route is as follows:
  • the first step 1-(1-(5-bromopyrimidin-2-yl)ethyl)-4-(propane-1-yn-1-yl)-1H-7-indazole-7-carboxylic acid methyl ester preparation
  • reaction solution was poured into water (50 mL), extracted with ethyl acetate (20 mL*3). The organic phases were combined, washed with saturated brine (20 mL*2), dried over anhydrous sodium sulfate, filtered and concentrated to obtain a crude product.
  • Step 3 Preparation of 1-(1-(5-cyclopropylpyrimidin-2-yl)ethyl)-4-(propane-1-yn-1-yl)-1H-indazole-7-carboxylic acid
  • reaction solution was added to water (50 mL), and extracted with ethyl acetate (10 mL*3). The combined organic phases were washed with saturated brine (10 mL*3), dried over anhydrous sodium sulfate, filtered, and concentrated to obtain a crude product.
  • SFC resolution method use Kromasil (S, S) Whelk-O1 type chiral resolution column (specification: 50 ⁇ 4.6mm, 3.5um) for chiral resolution, mobile phase A is supercritical fluid CO 2 , flow Phase B is ethanol (containing 0.05% diethanolamine); Gradient conditions: mobile phase B is from 5% to 80%, flow rate is 10mL/min; 40% gradient keeps 10min, 50% gradient keeps 10min, column temperature keeps at At 35°C, the column pressure was maintained at 100 bar.
  • the seventh step (Sa,S)-6-(1-(1-(5-cyclopropylpyrimidin-2-yl)ethyl)-4-(propane-1-yn-1-yl)-1H- Preparation of indazole-7-carboxamido)spiro[3.3]heptane-2-carboxylic acid (I-32b)
  • the synthetic route is as follows:
  • the first step the preparation of 1-(4-(difluoromethyl)phenyl)ethan-1-one
  • the second step the preparation of 1-(4-(difluoromethyl)phenyl)ethan-1-ol
  • reaction solution was concentrated and purified by column chromatography to obtain 1-(1-(4-(difluoromethyl)phenyl)ethyl)-4-(propane-1-yn-1-yl)-1H-indazole-7 - Methyl formate (450 mg, 60% yield).
  • Step 4 Preparation of 1-(1-(4-(difluoromethyl)phenyl)ethyl)-4-(propane-1-yn-1-yl)-1H-indazole-7-carboxylic acid
  • SFC resolution method use Chiralpak AD-3 chiral resolution column (specification: 50 ⁇ 4.6mm, 3um) for chiral resolution, mobile phase A is supercritical fluid CO 2 , mobile phase B is isopropanol (containing 0.05% diethanolamine); Gradient conditions: the mobile phase B is 40% of the gradient to maintain 10min, the column temperature is maintained at 35 ° C, the column pressure is maintained at 100bar, the peak time: 0.581min.
  • the synthetic route is as follows:
  • SFC resolution method use Chiralpak AD-3 chiral resolution column (specification: 50 ⁇ 4.6mm, 3um) for chiral resolution, mobile phase A is supercritical fluid CO 2 , mobile phase B is isopropanol (Containing 0.05% diethanolamine); Gradient conditions: mobile phase B is 40% gradient and kept for 10min, column temperature was kept at 35°C, column pressure was kept at 100bar, peak eluting time: 2.245min.
  • reaction solution was concentrated, water (3mL) was added, and then dilute hydrochloric acid (1mol/L aqueous solution) was added to adjust the pH value of the solution to about 2.
  • (Sa,S)-6-(1-(1-(4-(difluoro Methyl)phenyl)ethyl)-4-(propane-1-yn-1-yl)-1H-indazole-7-carboxamide)spiro[3.3]heptane-2-carboxylic acid (I-34b) (62.5 mg, yield 80%).
  • the synthetic route is as follows:
  • the first step 1-((1R)-1-(4-(3-azabicyclo[3.1.0]hexan-3-yl)phenyl)ethyl)-4-(propane-1-yne- Preparation of 1-yl)-1H-indazole-7-carboxylic acid methyl ester
  • the synthetic route is as follows:
  • the first step 1-((1S)-1-(4-(3-azabicyclo[3.1.0]hexan-3-yl)phenyl)ethyl)-4-(propane-1-yne- Preparation of 1-yl)-1H-indazole-7-carboxylic acid methyl ester

Abstract

A benzoheterocyclic compound as represented by formula (I), a tautomer, a stereoisomer, a hydrate, a solvate, a pharmaceutically acceptable salt or a prodrug thereof for treating EP2 and EP4 receptor-mediated diseases, which can be used to treat EP2 and EP4 receptor-mediated diseases.

Description

用于治疗EP2、EP4受体介导的疾病的苯并杂环化合物Benzoheterocyclic compounds for treating diseases mediated by EP2 and EP4 receptors
本申请要求2021年6月8日向中国国家知识产权局提交的，专利申请号为202110638805.9，发明名称为“用于治疗EP2、EP4受体介导的疾病的苯并杂环化合物”以及2021年12月9日向中国国家知识产权局提交的，专利申请号为202111499017.2，发明名称为“用于治疗EP2、EP4受体介导的疾病的苯并杂环化合物”的两件在先申请的优先权。所述两件申请的全文通过引用的方式结合于本申请中。This application requires submission to the State Intellectual Property Office of China on June 8, 2021. The patent application number is 202110638805.9, and the title of the invention is "benzoheterocyclic compounds for the treatment of diseases mediated by EP2 and EP4 receptors" and on December 2021 The patent application number is 202111499017.2, submitted to the State Intellectual Property Office of China on September 9, and the invention title is "Benzo heterocyclic compounds for the treatment of EP2, EP4 receptor-mediated diseases". Both applications are hereby incorporated by reference in their entirety.
技术领域technical field
本发明属于医药领域，具体地，本发明涉及到一种用于治疗EP2、EP4受体介导的疾病的苯并杂环化合物及制备方法和用途。The invention belongs to the field of medicine, in particular, the invention relates to a benzoheterocyclic compound for treating diseases mediated by EP2 and EP4 receptors, a preparation method and application.
背景技术Background technique
前列腺素E2(Prostaglandin E2，PGE2)是一种内源性生物活性脂质，PGE2通过激活前列腺素受体引起广泛的上下游依赖性生物应答，参与调控包括炎症、疼痛、肾功能、心血管系统、肺功能以及癌症等诸多生理和病理过程。据报道，PGE2在各种癌症的癌变组织中高度表达，并且已证实PGE2与患者的癌症和疾病状况的发生、成长和发展相关。普遍认为，PGE2与细胞增殖和细胞死亡(凋亡)的激活相关，并且在癌细胞增殖、疾病进展和癌症转移的过程中起重要作用。Prostaglandin E2 (Prostaglandin E2, PGE2) is an endogenous bioactive lipid. PGE2 induces a wide range of upstream and downstream dependent biological responses by activating prostaglandin receptors, and participates in the regulation of inflammation, pain, renal function, and cardiovascular system. , lung function and cancer and many other physiological and pathological processes. It has been reported that PGE2 is highly expressed in cancerous tissues of various cancers, and it has been confirmed that PGE2 is associated with the occurrence, growth and development of cancer and disease conditions in patients. It is generally believed that PGE2 is associated with the activation of cell proliferation and cell death (apoptosis), and plays an important role in the process of cancer cell proliferation, disease progression and cancer metastasis.
PGE2的受体存在EP1、EP2、EP3和EP4共4种亚型，广泛表达于各种组织中。EP1受体激活磷脂酶C和三磷酸肌醇途径，EP2和EP4受体激活腺苷酸环化酶和cAMP-蛋白激酶A，EP3受体的激活既能抑制腺苷酸环化酶，又能激活磷脂酶C。There are four subtypes of PGE2 receptors, EP1, EP2, EP3 and EP4, which are widely expressed in various tissues. The EP1 receptor activates the phospholipase C and inositol triphosphate pathways, the EP2 and EP4 receptors activate adenylate cyclase and cAMP-protein kinase A, and the activation of the EP3 receptor both inhibits adenylate cyclase and Activates phospholipase C.
其中EP2和EP4表达于多种免疫细胞(如巨噬细胞、树突状细胞、NK细胞和CTL)，抑制EP2和EP4可增强免疫活性，抑制肿瘤生长。Among them, EP2 and EP4 are expressed in various immune cells (such as macrophages, dendritic cells, NK cells and CTLs), and inhibiting EP2 and EP4 can enhance immune activity and inhibit tumor growth.
PGE2在肿瘤微环境中持续地激活肿瘤微环境中的EP受体(由肿瘤细胞大量产生)，会促进多种免疫抑制细胞的积累并增强其活性，包括2型肿瘤相关巨噬细胞(TAMS)、Treg细胞和髓系来源的抑制细胞(MDSCs)。免疫抑制肿瘤微环境的主要特征之一是存在大量的MDSCs和TAM，它们反过来又与胃癌、卵巢癌、乳腺癌、膀胱癌、肝细胞癌(HCC)、头颈癌和其他类型的癌症患者的总体生存率低密切相关。此外，据报道PGE2通过抑制抗原提呈树突状细胞(DC)在肿瘤中的积聚以及抑制肿瘤浸润性DC的激活来诱导免疫耐受)，从而帮助肿瘤细胞逃避免疫监视。PGE2在促进肿瘤发生发展中发挥十分重要的作用，在包括结肠癌、肺癌、乳腺癌、和头颈癌等各类恶性肿瘤中都发现PGE2及其相关受体EP2、EP4的表达水平升高，并往往和不良预后密切相关。因此，选择性阻断EP2和EP4信号通路可以通过改变肿瘤微环境、调控肿瘤免疫细胞抑制肿瘤发生发展。PGE2 persistently activates EP receptors in the tumor microenvironment (produced in large quantities by tumor cells), which promotes the accumulation and enhances the activity of a variety of immunosuppressive cells, including type 2 tumor-associated macrophages (TAMS) , Treg cells and myeloid-derived suppressor cells (MDSCs). One of the main features of the immunosuppressive tumor microenvironment is the presence of a large number of MDSCs and TAMs, which in turn are associated with gastric cancer, ovarian cancer, breast cancer, bladder cancer, hepatocellular carcinoma (HCC), head and neck cancer, and other types of cancer patients. It is closely related to poor overall survival. In addition, PGE2 has been reported to induce immune tolerance by inhibiting the accumulation of antigen-presenting dendritic cells (DCs) in tumors and inhibiting the activation of tumor-infiltrating DCs), thereby helping tumor cells escape immune surveillance. PGE2 plays a very important role in promoting the occurrence and development of tumors. It has been found that the expression levels of PGE2 and its related receptors EP2 and EP4 are increased in various malignant tumors including colon cancer, lung cancer, breast cancer, and head and neck cancer. Often closely associated with poor prognosis. Therefore, selectively blocking EP2 and EP4 signaling pathways can inhibit tumor development by changing the tumor microenvironment and regulating tumor immune cells.
EP2和/或EP4的选择性和双重拮抗剂可以用于治疗其他疾病和病症。在类风湿性关节炎 和骨关节炎的啮齿动物模型中，已证明EP4拮抗剂可有效缓解关节炎症和疼痛，也已证明EP4拮抗剂在自身免疫疾病的啮齿动物模型中有效。Selective and dual antagonists of EP2 and/or EP4 can be used to treat other diseases and conditions. EP4 antagonists have been shown to be effective in alleviating joint inflammation and pain in rodent models of rheumatoid arthritis and osteoarthritis, and have also been shown to be effective in rodent models of autoimmune disease.
PGE2是通过EP2受体介导促炎功能的主要前列腺素，因此EP2拮抗剂可显示出作为某些慢性炎性疾病(尤其是炎性神经变性疾病，如癫痫、阿尔茨海默病(AD)、帕金森病(PD)、肌萎缩侧索硬化(ALS)和创伤性脑损伤(TBI))的治疗剂的效用。在阿尔茨海默病鼠模型中，EP4拮抗剂ONO-AE3-208减少了淀粉样蛋白-β并且改善了行为表现。PGE2 is the main prostaglandin that mediates pro-inflammatory functions through the EP2 receptor, thus EP2 antagonists may be shown to be effective in certain chronic inflammatory diseases (especially inflammatory neurodegenerative diseases such as epilepsy, Alzheimer's disease (AD) , Parkinson's disease (PD), amyotrophic lateral sclerosis (ALS) and traumatic brain injury (TBI)). In a mouse model of Alzheimer's disease, the EP4 antagonist ONO-AE3-208 reduced amyloid-β and improved behavioral performance.
已有研究证实，子宫内膜异位症(内异症)患者腹腔液中的致痛物质PGE2表达明显增高，而PGE2增高可进一步刺激雌激素合成限速酶(芳香酶)的表达，使雌激素的合成增加，从而促进内异症的发生和发展。抑制EP2和EP4可抑制内异症病变中上皮和间质细胞特异性模式中孕激素(P4)信号转导机制蛋白的表达，抑制PGE2和雌激素(E2)的留存、侵袭、生物合成和信号转导，从而抑制促炎细胞因子的产生，减少腹膜内异症病变组织的生长、存活和扩散，减轻盆腔疼痛，恢复子宫内膜的容受性。Studies have confirmed that the expression of the pain-causing substance PGE2 in the peritoneal fluid of patients with endometriosis (endometriosis) is significantly increased, and the increase of PGE2 can further stimulate the expression of the rate-limiting enzyme (aromatase) for estrogen synthesis, making estrogen The synthesis of hormones increases, thereby promoting the occurrence and development of endometriosis. Inhibition of EP2 and EP4 suppresses the expression of progesterone (P4) signaling machinery proteins in epithelial and mesenchymal cell-specific patterns in endometriotic lesions and inhibits the persistence, invasion, biosynthesis and signaling of PGE2 and estrogen (E2) Transduction, thereby inhibiting the production of pro-inflammatory cytokines, reducing the growth, survival and spread of endometriotic lesions, alleviating pelvic pain, and restoring endometrial receptivity.
因此开发可用于治疗EP2和/或EP4受体介导的疾病的新型化合物具有重要意义。这类化合物具有可用于治疗炎性疾病、自身免疫疾病、神经变性疾病、心血管疾病和癌症的潜力。Therefore, it is of great significance to develop new compounds that can be used to treat diseases mediated by EP2 and/or EP4 receptors. Such compounds have the potential to be useful in the treatment of inflammatory diseases, autoimmune diseases, neurodegenerative diseases, cardiovascular diseases and cancer.
发明内容Contents of the invention
本发明的目的是提供一种用于治疗EP2、EP4受体介导的疾病的苯并杂环化合物及制备方法和用途，所述苯并杂环化合物为本发明所述的式I所示苯并杂环化合物、其互变异构体、立体异构体、水合物、溶剂化物、药学上可接受的盐或前药。The object of the present invention is to provide a benzoheterocyclic compound for treating diseases mediated by EP2 and EP4 receptors and its preparation method and application. and heterocyclic compounds, tautomers, stereoisomers, hydrates, solvates, pharmaceutically acceptable salts or prodrugs thereof.
本发明第一方面，提供了一种式I所示苯并杂环化合物、其互变异构体、立体异构体、水合物、溶剂化物、药学上可接受的盐或前药：The first aspect of the present invention provides a benzoheterocyclic compound represented by formula I, its tautomers, stereoisomers, hydrates, solvates, pharmaceutically acceptable salts or prodrugs:
Figure PCTCN2022097611-appb-000001
Figure PCTCN2022097611-appb-000001
其中，R 1、R 2分别为氢或-C≡C-R 11，且R 1、R 2不同； Wherein, R 1 and R 2 are respectively hydrogen or -C≡CR 11 , and R 1 and R 2 are different;
所述R 11为氢或选自：C 1-C 5烷基、3-6元环烷基； The R 11 is hydrogen or selected from: C 1 -C 5 alkyl, 3-6 membered cycloalkyl;
所述R 11任选地被一个或多个选自下列的取代基取代：羟基、卤素、C 1-C 5烷基；当取代基为多个时，所述取代基相同或不同； The R 11 is optionally substituted by one or more substituents selected from the following: hydroxyl, halogen, C 1 -C 5 alkyl; when there are multiple substituents, the substituents are the same or different;
L 1为C 1-C 5亚烷基； L 1 is C 1 -C 5 alkylene;
L 2不存在或为未取代或被Rb取代的C 1-C 3亚烷基； L 2 does not exist or is unsubstituted or substituted by Rb C 1 -C 3 alkylene;
所述L 1任选地被Ra 1和/或Ra 2取代； The L 1 is optionally substituted by Ra 1 and/or Ra 2 ;
所述Ra 1、Ra 2各自独立地为C 1-C 5烷基、卤素、羟基、氨基、C 1-C 5烷氧基、C 1-C 5卤代 烷基、C 1-C 5卤代烷氧基； The Ra 1 and Ra 2 are each independently C 1 -C 5 alkyl, halogen, hydroxyl, amino, C 1 -C 5 alkoxy, C 1 -C 5 haloalkyl, C 1 -C 5 haloalkoxy ;
或所述Ra 1、Ra 2与它们共同连接的C原子一起形成3-6元环烷基或4-6元杂环烷基； Or the Ra 1 and Ra 2 form a 3-6-membered cycloalkyl group or a 4-6-membered heterocycloalkyl group together with the C atoms they are connected to;
所述Rb为C 1-C 3烷基或卤素； The Rb is C 1 -C 3 alkyl or halogen;
环A为苯基或5-6元杂芳基；Ring A is phenyl or 5-6 membered heteroaryl;
环B不存在或为3-8元环烷基、4-8元杂环烷基、苯基、5-6元杂芳基；Ring B does not exist or is 3-8 membered cycloalkyl, 4-8 membered heterocycloalkyl, phenyl, 5-6 membered heteroaryl;
R 3、R 4各自独立地选自：羟基、卤素、氨基、氰基、C 1-C 5烷基、C 1-C 5烷氧基、3-6元环烷基、4-6元杂环烷基、3-6元环烷氧基；所述C 1-C 5烷基、C 1-C 5烷氧基、3-6元环烷基、4-6元杂环烷基、3-6元环烷氧基任选地被一个或多个选自下列的取代基取代：羟基、卤素、氨基、氰基、3-6元环烷基； R 3 and R 4 are each independently selected from: hydroxyl, halogen, amino, cyano, C 1 -C 5 alkyl, C 1 -C 5 alkoxy, 3-6 membered cycloalkyl, 4-6 membered hetero Cycloalkyl, 3-6 membered cycloalkoxy; said C 1 -C 5 alkyl, C 1 -C 5 alkoxy, 3-6 membered cycloalkyl, 4-6 membered heterocycloalkyl, 3 -6-membered cycloalkoxy is optionally substituted by one or more substituents selected from the group consisting of hydroxyl, halogen, amino, cyano, 3-6-membered cycloalkyl;
m为0、1、2或3；m is 0, 1, 2 or 3;
n为0、1、2或3。n is 0, 1, 2 or 3.
在本发明的一些实施方式中，提供了一种式I所示苯并杂环化合物、其互变异构体、立体异构体、水合物、溶剂化物、药学上可接受的盐或前药：In some embodiments of the present invention, there is provided a benzoheterocyclic compound represented by formula I, its tautomer, stereoisomer, hydrate, solvate, pharmaceutically acceptable salt or prodrug :
Figure PCTCN2022097611-appb-000002
Figure PCTCN2022097611-appb-000002
其中，R 1、R 2分别为氢或-C≡C-R 11，且R 1、R 2不同； Wherein, R 1 and R 2 are respectively hydrogen or -C≡CR 11 , and R 1 and R 2 are different;
所述R 11为氢或选自：C 1-C 5烷基、3-6元环烷基； The R 11 is hydrogen or selected from: C 1 -C 5 alkyl, 3-6 membered cycloalkyl;
所述R 11任选地被一个或多个选自下列的取代基取代：羟基、卤素、C 1-C 5烷基；当取代基为多个时，所述取代基相同或不同； The R 11 is optionally substituted by one or more substituents selected from the following: hydroxyl, halogen, C 1 -C 5 alkyl; when there are multiple substituents, the substituents are the same or different;
L 1为C 1-C 5亚烷基； L 1 is C 1 -C 5 alkylene;
L 2不存在或为未取代或被Rb取代的C 1-C 3亚烷基； L 2 does not exist or is unsubstituted or substituted by Rb C 1 -C 3 alkylene;
所述L 1任选地被Ra 1和/或Ra 2取代； The L 1 is optionally substituted by Ra 1 and/or Ra 2 ;
所述Ra 1、Ra 2各自独立地为C 1-C 5烷基、卤素、羟基、氨基、C 1-C 5烷氧基、C 1-C 5卤代烷基、C 1-C 5卤代烷氧基； The Ra 1 and Ra 2 are each independently C 1 -C 5 alkyl, halogen, hydroxyl, amino, C 1 -C 5 alkoxy, C 1 -C 5 haloalkyl, C 1 -C 5 haloalkoxy ;
或所述Ra 1、Ra 2与它们共同连接的C原子一起形成3-6元环烷基或4-6元杂环烷基； Or the Ra 1 and Ra 2 form a 3-6-membered cycloalkyl group or a 4-6-membered heterocycloalkyl group together with the C atoms they are connected to;
所述Rb为C 1-C 3烷基或卤素； The Rb is C 1 -C 3 alkyl or halogen;
环A为苯基或5-6元杂芳基；Ring A is phenyl or 5-6 membered heteroaryl;
环B不存在或为3-6元环烷基、4-6元杂环烷基、苯基、5-6元杂芳基；Ring B does not exist or is 3-6 membered cycloalkyl, 4-6 membered heterocycloalkyl, phenyl, 5-6 membered heteroaryl;
R 3、R 4各自独立地选自：羟基、卤素、氨基、氰基、C 1-C 5烷基、C 1-C 5烷氧基、3-6元环烷基、4-6元杂环烷基、3-6元环烷氧基；所述C 1-C 5烷基、C 1-C 5烷氧基、3-6元环烷基、4-6元杂环烷基、3-6元环烷氧基任选地被一个或多个选自下列的取代基取代：羟基、卤素、氨基、氰基、3-6元环烷基； R 3 and R 4 are each independently selected from: hydroxyl, halogen, amino, cyano, C 1 -C 5 alkyl, C 1 -C 5 alkoxy, 3-6 membered cycloalkyl, 4-6 membered hetero Cycloalkyl, 3-6 membered cycloalkoxy; said C 1 -C 5 alkyl, C 1 -C 5 alkoxy, 3-6 membered cycloalkyl, 4-6 membered heterocycloalkyl, 3 -6-membered cycloalkoxy is optionally substituted by one or more substituents selected from the group consisting of hydroxyl, halogen, amino, cyano, 3-6-membered cycloalkyl;
m为0、1、2或3；m is 0, 1, 2 or 3;
n为0、1、2或3。n is 0, 1, 2 or 3.
在一优选实施方式中，在所述环A中，所述5-6元杂芳基含有一个或多个选自N、O或S的杂原子；当所述杂原子为多个时，所述杂原子相同或不同；较佳地，所述5-6元杂芳基选自：噻吩、呋喃、吡咯、吡唑、咪唑、三氮唑、噻唑、噻二唑、噁唑、吡啶、嘧啶、哒嗪、吡嗪。In a preferred embodiment, in the ring A, the 5-6 membered heteroaryl contains one or more heteroatoms selected from N, O or S; when there are multiple heteroatoms, the The heteroatoms are the same or different; preferably, the 5-6 membered heteroaryl is selected from: thiophene, furan, pyrrole, pyrazole, imidazole, triazole, thiazole, thiadiazole, oxazole, pyridine, pyrimidine , pyridazine, pyrazine.
在一优选实施方式中，Ra 1、Ra 2与它们共同连接的C原子一起形成氧杂环丁烷。 In a preferred embodiment, Ra 1 and Ra 2 form oxetane together with their common C atoms.
在一优选实施方式中，L 1为-CH 2-、-CH(CH 3)-、-CH 2CH 2-或-CH 2CH 2CH 2-；较佳地，L 1为-CH 2-、-CH(CH 3)-。 In a preferred embodiment, L 1 is -CH 2 -, -CH(CH 3 )-, -CH 2 CH 2 - or -CH 2 CH 2 CH 2 -; preferably, L 1 is -CH 2 - , -CH(CH 3 )-.
在一优选实施方式中，R 3、R 4各自独立地选自：卤素、氰基、C 1-C 5烷基、C 1-C 5烷氧基、3-6元环烷基；所述C 1-C 5烷基、C 1-C 5烷氧基、3-6元环烷基任选地被1、2或3个选自下列的取代基取代：羟基、氟、氯。 In a preferred embodiment, R 3 and R 4 are each independently selected from: halogen, cyano, C 1 -C 5 alkyl, C 1 -C 5 alkoxy, 3-6 membered cycloalkyl; C 1 -C 5 alkyl, C 1 -C 5 alkoxy, 3-6 membered cycloalkyl are optionally substituted with 1, 2 or 3 substituents selected from the group consisting of hydroxy, fluoro, chloro.
在一优选实施方式中，环B不存在或为3-6元环烷基、4-6元杂环烷基、苯基、5-6元杂芳基；所述杂环烷基或杂芳基中含有杂原子为O、N或S；所述杂环烷基包括单环、并环、桥环或螺环。In a preferred embodiment, ring B does not exist or is 3-6 membered cycloalkyl, 4-6 membered heterocycloalkyl, phenyl, 5-6 membered heteroaryl; said heterocycloalkyl or heteroaryl The heteroatom contained in the group is O, N or S; the heterocycloalkyl group includes a monocyclic ring, a parallel ring, a bridged ring or a spiro ring.
在一优选实施方式中，环B为氧杂环丁烷、
Figure PCTCN2022097611-appb-000003
或为选自下列杂芳基的饱和或部分饱和的杂环：噻吩、呋喃、吡咯、吡唑、咪唑、三氮唑、噻唑、噻二唑、噁唑、吡啶、嘧啶、哒嗪、吡嗪。例如，可选自二氢噻吩、四氢噻吩、二氢呋喃、四氢呋喃、二氢吡咯、四氢吡咯、二氢吡唑、四氢吡唑、二氢咪唑、四氢咪唑、二氢三氮唑、四氢三氮唑、二氢噻唑、四氢噻唑、二氢噻二唑、四氢噻二唑、二氢噁唑、四氢噁唑、二氢吡啶、四氢吡啶、二氢嘧啶、四氢嘧啶、二氢哒嗪、四氢哒嗪、二氢吡嗪、四氢吡嗪。 In a preferred embodiment, ring B is oxetane,
Figure PCTCN2022097611-appb-000003
Or a saturated or partially saturated heterocycle selected from the following heteroaryl groups: thiophene, furan, pyrrole, pyrazole, imidazole, triazole, thiazole, thiadiazole, oxazole, pyridine, pyrimidine, pyridazine, pyrazine . For example, may be selected from dihydrothiophene, tetrahydrothiophene, dihydrofuran, tetrahydrofuran, dihydropyrrole, tetrahydropyrrole, dihydropyrazole, tetrahydropyrazole, dihydroimidazole, tetrahydroimidazole, dihydrotriazole , tetrahydrotriazole, dihydrothiazole, tetrahydrothiazole, dihydrothiadiazole, tetrahydrothiadiazole, dihydrooxazole, tetrahydrooxazole, dihydropyridine, tetrahydropyridine, dihydropyrimidine, four Hydropyrimidine, dihydropyridazine, tetrahydropyridazine, dihydropyrazine, tetrahydropyrazine.
在一优选实施方式中，环B选自：氧杂环丁烷、
Figure PCTCN2022097611-appb-000004
环丙基、环丁基、苯基。 In a preferred embodiment, ring B is selected from: oxetane,
Figure PCTCN2022097611-appb-000004
Cyclopropyl, cyclobutyl, phenyl.
在一优选实施方式中，式I具有结构Ia、Ib、Ic、Id，In a preferred embodiment, formula I has the structure Ia, Ib, Ic, Id,
Figure PCTCN2022097611-appb-000005
Figure PCTCN2022097611-appb-000005
Figure PCTCN2022097611-appb-000006
Figure PCTCN2022097611-appb-000006
其中，Z 1、Z 2、Z 3、Z 4、Z 5各自独立地选自CR 3或N； Wherein, Z 1 , Z 2 , Z 3 , Z 4 , Z 5 are each independently selected from CR 3 or N;
环B、R 1、R 2、R 3、R 4、Ra(同式I中R a1、R a2)、n、L 2的定义如第一方面中所述； Ring B, R 1 , R 2 , R 3 , R 4 , Ra (same as R a1 and R a2 in formula I), n and L 2 are as defined in the first aspect;
较佳地，在结构Ia中，Z 1、Z 2、Z 3、Z 4、Z 5中至多含有两个N； Preferably, in structure Ia, Z 1 , Z 2 , Z 3 , Z 4 , Z 5 contain at most two Ns;
较佳地，在结构Ib中，Z 1、Z 2、Z 3、Z 4中至多含有两个N。 Preferably, in structure Ib, Z 1 , Z 2 , Z 3 , and Z 4 contain at most two Ns.
在一优选实施方式中，L 2不存在或选自-CH 2-、-CH 2CH 2-、-CH 2CH 2CH 2-、-CH(CH 3)-、-CH(CH 3)CH 2-；较佳地，L 2不存在或为-CH 2-或-CH(CH 3)-。 In a preferred embodiment, L 2 is absent or selected from -CH 2 -, -CH 2 CH 2 -, -CH 2 CH 2 CH 2 -, -CH(CH 3 )-, -CH(CH 3 )CH 2 -; preferably, L 2 does not exist or is -CH 2 - or -CH(CH 3 )-.
在一优选实施方式中，R 1、R 2分别为氢或-C≡C-R 11，且R 1、R 2不同；R 11为氢、C 1-C 5烷基或3-6元环烷基；较佳地，所述R 11为甲基。 In a preferred embodiment, R 1 and R 2 are hydrogen or -C≡CR 11 respectively, and R 1 and R 2 are different; R 11 is hydrogen, C 1 -C 5 alkyl or 3-6 membered cycloalkyl ; Preferably, the R 11 is methyl.
在一优选实施方式中，Ra为甲基、乙基、丙基。In a preferred embodiment, Ra is methyl, ethyl, propyl.
在一优选实施方式中，环B不存在或为3-6元环烷基、4-6元杂环烷基、苯基、5-6元杂芳基；所述杂环烷基或杂芳基中含有杂原子为O或N；所述杂环烷基包括单环、并环、桥环或螺环。In a preferred embodiment, ring B does not exist or is 3-6 membered cycloalkyl, 4-6 membered heterocycloalkyl, phenyl, 5-6 membered heteroaryl; said heterocycloalkyl or heteroaryl The heteroatom contained in the group is O or N; the heterocycloalkyl group includes a monocyclic ring, a parallel ring, a bridged ring or a spiro ring.
在一优选实施方式中，环B为不存在或为环丙基、环丁基、环戊基、环己基、苯基、噻吩、呋喃、吡咯、吡唑、咪唑、三氮唑、噻唑、噻二唑、噁唑、吡啶、嘧啶、哒嗪、吡嗪。In a preferred embodiment, Ring B is absent or is cyclopropyl, cyclobutyl, cyclopentyl, cyclohexyl, phenyl, thiophene, furan, pyrrole, pyrazole, imidazole, triazole, thiazole, thiazole Oxadiazole, oxazole, pyridine, pyrimidine, pyridazine, pyrazine.
在一优选实施方式中，环B为氧杂环丁烷、
Figure PCTCN2022097611-appb-000007
或为选自下列杂芳基的饱和或部分饱和的杂环：噻吩、呋喃、吡咯、吡唑、咪唑、三氮唑、噻唑、噻二唑、噁唑、吡啶、嘧啶、哒嗪、吡嗪。例如，可选自二氢噻吩、四氢噻吩、二氢呋喃、四氢呋喃、二氢吡咯、四氢吡咯、二氢吡唑、四氢吡唑、二氢咪唑、四氢咪唑、二氢三氮唑、四氢三氮唑、二氢噻唑、四氢噻唑、二氢噻二唑、四氢噻二唑、二氢噁唑、四氢噁唑、二氢吡啶、四氢吡啶、二氢嘧啶、四氢嘧啶、二氢哒嗪、四氢哒嗪、二氢吡嗪、四氢吡嗪。 In a preferred embodiment, ring B is oxetane,
Figure PCTCN2022097611-appb-000007
Or a saturated or partially saturated heterocycle selected from the following heteroaryl groups: thiophene, furan, pyrrole, pyrazole, imidazole, triazole, thiazole, thiadiazole, oxazole, pyridine, pyrimidine, pyridazine, pyrazine . For example, may be selected from dihydrothiophene, tetrahydrothiophene, dihydrofuran, tetrahydrofuran, dihydropyrrole, tetrahydropyrrole, dihydropyrazole, tetrahydropyrazole, dihydroimidazole, tetrahydroimidazole, dihydrotriazole , tetrahydrotriazole, dihydrothiazole, tetrahydrothiazole, dihydrothiadiazole, tetrahydrothiadiazole, dihydrooxazole, tetrahydrooxazole, dihydropyridine, tetrahydropyridine, dihydropyrimidine, four Hydropyrimidine, dihydropyridazine, tetrahydropyridazine, dihydropyrazine, tetrahydropyrazine.
在一优选实施方式中，环B选自：氧杂环丁烷、
Figure PCTCN2022097611-appb-000008
环丙基、环丁基、苯基。 In a preferred embodiment, ring B is selected from: oxetane,
Figure PCTCN2022097611-appb-000008
Cyclopropyl, cyclobutyl, phenyl.
在一优选实施方式中，式I具有如下结构IIa、IIb、IIc、IId、IIe、IIf、IIg、IIm、IInIn a preferred embodiment, Formula I has the following structures IIa, IIb, IIc, IId, IIe, If, IIg, IIm, IIn
Figure PCTCN2022097611-appb-000009
Figure PCTCN2022097611-appb-000009
Figure PCTCN2022097611-appb-000010
Figure PCTCN2022097611-appb-000010
在一优选实施方式中，所述的式I所示苯并杂环化合物、其互变异构体、立体异构体、水合物、溶剂化物、药学上可接受的盐或前药中，所述的式I所示苯并杂环化合物包括如下结构：In a preferred embodiment, in the benzoheterocyclic compound represented by formula I, its tautomers, stereoisomers, hydrates, solvates, pharmaceutically acceptable salts or prodrugs, the The benzoheterocyclic compounds shown in the formula I include the following structures:
Figure PCTCN2022097611-appb-000011
Figure PCTCN2022097611-appb-000011
Figure PCTCN2022097611-appb-000012
Figure PCTCN2022097611-appb-000012
Figure PCTCN2022097611-appb-000013
Figure PCTCN2022097611-appb-000013
Figure PCTCN2022097611-appb-000014
Figure PCTCN2022097611-appb-000014
在一优选实施方式中，所述的式I所示苯并杂环化合物包括如下结构：In a preferred embodiment, the benzoheterocyclic compound shown in formula I includes the following structure:
Figure PCTCN2022097611-appb-000015
Figure PCTCN2022097611-appb-000015
Figure PCTCN2022097611-appb-000016
Figure PCTCN2022097611-appb-000016
Figure PCTCN2022097611-appb-000017
Figure PCTCN2022097611-appb-000017
本发明第二方面，提供了一种中间体B-1：The second aspect of the present invention provides an intermediate B-1:
Figure PCTCN2022097611-appb-000018
Figure PCTCN2022097611-appb-000018
其中，R 6为-OH、-Cl、-O-C 1-C 5烷基、-O-苄基； Wherein, R 6 is -OH, -Cl, -OC 1 -C 5 alkyl, -O-benzyl;
环A、环B、L 1、R 1、R 2、R 3、R 4、m、n的定义如第一方面中所述。 Ring A, Ring B, L 1 , R 1 , R 2 , R 3 , R 4 , m, n are as defined in the first aspect.
在一优选实施方式中，所述中间体B-1具有结构B-1a或B-1b：In a preferred embodiment, the intermediate B-1 has the structure B-1a or B-1b:
Figure PCTCN2022097611-appb-000019
Figure PCTCN2022097611-appb-000019
其中，Z 1、Z 2、Z 3、Z 4、Z 5各自独立地选自CR 3或N； Wherein, Z 1 , Z 2 , Z 3 , Z 4 , Z 5 are each independently selected from CR 3 or N;
环B、R 1、R 2、R 3、R 4、Ra(同式I中R a1、R a2)、n的定义如第一方面中所述； Ring B, R 1 , R 2 , R 3 , R 4 , Ra (same as R a1 and R a2 in formula I), and n are as defined in the first aspect;
较佳地，在结构Ia中，Z 1、Z 2、Z 3、Z 4、Z 5中至多含有两个N； Preferably, in structure Ia, Z 1 , Z 2 , Z 3 , Z 4 , Z 5 contain at most two Ns;
较佳地，在结构Ib中，Z 1、Z 2、Z 3、Z 4中至多含有两个N。 Preferably, in structure Ib, Z 1 , Z 2 , Z 3 , and Z 4 contain at most two Ns.
较佳地，所述中间体B-1具有结构B-1c、B-1d、B-1f、B-1g：Preferably, the intermediate B-1 has the structures B-1c, B-1d, B-1f, B-1g:
Figure PCTCN2022097611-appb-000020
Figure PCTCN2022097611-appb-000020
本发明第三方面，提供了一种制备如第一方面所述的式I所示苯并杂环化合物、其互变异构体、立体异构体、水合物、溶剂化物、药学上可接受的盐或前药的方法，包括：1)将如第二方面所述的中间体B-1与中间体B-2或中间体B-2的盐反应，得到所述式I所示苯并杂环化合物；The third aspect of the present invention provides a method for preparing the benzoheterocyclic compound represented by formula I as described in the first aspect, its tautomers, stereoisomers, hydrates, solvates, pharmaceutically acceptable A salt or prodrug method, comprising: 1) reacting intermediate B-1 as described in the second aspect with intermediate B-2 or a salt of intermediate B-2 to obtain the benzo Heterocyclic compounds;
所述中间体B-2具有结构：The intermediate B-2 has the structure:
Figure PCTCN2022097611-appb-000021
Figure PCTCN2022097611-appb-000021
其中，R 5为C 1-C 6烷基、苄基。 Wherein, R 5 is C 1 -C 6 alkyl, benzyl.
在一优选实施方式中，所述中间体B-2的盐为盐酸盐。In a preferred embodiment, the salt of intermediate B-2 is hydrochloride.
在一优选实施方式中，所述方法还包括：In a preferred embodiment, the method also includes:
2)当所述中间体B-1中R 6不为-OH或-Cl时，将基团-COR 6转化为-COOH或-COCl后， 2) When R 6 in the intermediate B-1 is not -OH or -Cl, after converting the group -COR 6 into -COOH or -COCl,
再与中间体B-2反应；和/或Reaction with intermediate B-2; and/or
3)在所述中间体B-1与中间体B-2反应后，将基团-COOR 5水解为-COOH。 3) After the reaction of intermediate B-1 with intermediate B-2, the group -COOR 5 is hydrolyzed to -COOH.
较佳地，所述中间体B-2具有结构B-2a或B-2b：Preferably, the intermediate B-2 has the structure B-2a or B-2b:
Figure PCTCN2022097611-appb-000022
Figure PCTCN2022097611-appb-000022
本发明第四方面，提供了一种药物组合物，所述药物组合物包括：如第一方面所述的式I所示苯并杂环化合物、其互变异构体、立体异构体、水合物、溶剂化物、药学上可接受的盐或前药；和药学上可接受的载体。The fourth aspect of the present invention provides a pharmaceutical composition, which includes: the benzoheterocyclic compound represented by formula I as described in the first aspect, its tautomers, stereoisomers, a hydrate, a solvate, a pharmaceutically acceptable salt or a prodrug; and a pharmaceutically acceptable carrier.
根据本发明的实施方案，所述药物组合物还包括第二种药物。According to an embodiment of the present invention, the pharmaceutical composition further comprises a second drug.
在一优选实施方式中，所述第二种药物包括抗体；较佳地，所述抗体包括抗PD-L1抗体、抗PD-1抗体。In a preferred embodiment, the second drug includes antibodies; preferably, the antibodies include anti-PD-L1 antibodies and anti-PD-1 antibodies.
在一些实施方式中，所述如第一方面所述的式I所示苯并杂环化合物、其互变异构体、立体异构体、水合物、溶剂化物、药学上可接受的盐或前药与第二种药物的质量比例为1：100至100:1，优选的为10:1至100:1，更优选的为10:1至50:1，例如为10:1，15:1，20:1，25:1，30:1，35:1，40:1，45:1，50:1。In some embodiments, the benzoheterocyclic compound represented by formula I as described in the first aspect, its tautomer, stereoisomer, hydrate, solvate, pharmaceutically acceptable salt or The mass ratio of the prodrug to the second drug is 1:100 to 100:1, preferably 10:1 to 100:1, more preferably 10:1 to 50:1, for example 10:1, 15:1 1, 20:1, 25:1, 30:1, 35:1, 40:1, 45:1, 50:1.
本发明第五方面，提供了一种如第一方面所述的式I所示苯并杂环化合物、其互变异构体、立体异构体、水合物、溶剂化物、药学上可接受的盐或前药的用途，或如第四方面所述的药物组合物的用途，所述用途包括：The fifth aspect of the present invention provides a benzoheterocyclic compound represented by formula I as described in the first aspect, its tautomers, stereoisomers, hydrates, solvates, pharmaceutically acceptable The purposes of salt or prodrug, or the purposes of pharmaceutical composition as described in the fourth aspect, described purposes comprises:
1)对EP2和/或EP4产生拮抗作用；1) produce antagonistic effect on EP2 and/or EP4;
2)与EP2和/或EP4受体结合；2) Binding to EP2 and/or EP4 receptors;
3)预防、治疗EP2和/或EP4受体介导的疾病疾病、3) Prevention and treatment of diseases mediated by EP2 and/or EP4 receptors,
4)制备EP2和/或EP4拮抗剂、4) preparing EP2 and/or EP4 antagonists,
5)制备预防、治疗与EP2和/或EP4受体介导的疾病的药物、药物组合物或制剂。5) Preparation of medicines, pharmaceutical compositions or preparations for preventing and treating diseases mediated by EP2 and/or EP4 receptors.
较佳地，其中所述EP2和/或EP4受体介导的疾病包括炎性疾病(例如关节炎和子宫内膜异位症)、自身免疫疾病(例如多发性硬化症)、神经变性疾病(例如癫痫、阿尔茨海默病、帕金森病、肌萎缩侧索硬化和创伤性脑损伤)、心血管疾病(例如动脉粥样硬化)和癌症(例如结肠癌、肺癌、乳腺癌和头颈癌)。Preferably, the diseases mediated by the EP2 and/or EP4 receptors include inflammatory diseases (such as arthritis and endometriosis), autoimmune diseases (such as multiple sclerosis), neurodegenerative diseases ( such as epilepsy, Alzheimer's disease, Parkinson's disease, amyotrophic lateral sclerosis, and traumatic brain injury), cardiovascular disease (such as atherosclerosis), and cancer (such as colon, lung, breast, and head and neck cancers) .
本发明第六方面，提供了一种用于预防和/或治疗EP2和/或EP4受体介导的疾病的方法，所述方法包括向有此需要的个体给予有效量的所述式I所示苯并杂环化合物、其互变异构体、立体异构体、水合物、溶剂化物、药学上可接受的盐或前药。The sixth aspect of the present invention provides a method for preventing and/or treating diseases mediated by EP2 and/or EP4 receptors, the method comprising administering an effective amount of the formula I to an individual in need thereof Shown are benzoheterocyclic compounds, tautomers, stereoisomers, hydrates, solvates, pharmaceutically acceptable salts or prodrugs thereof.
在一优选实施方式中，所述方法还包括与抗体疗法联合使用；较佳地，所述抗体疗法包括PD-L1抗体疗法、PD-1抗体疗法。在一些实施方式中，所述如第一方面所述的式I所示苯并杂环化合物、其互变异构体、立体异构体、水合物、溶剂化物、药学上可接受的盐或前药与抗体的质量比为1：100至100:1，优选的为10:1至100:1，更优选的为10:1至50:1，例如为10:1，15:1，20:1，25:1，30:1，35:1，40:1，45:1，50:1。In a preferred embodiment, the method further includes combined use with antibody therapy; preferably, the antibody therapy includes PD-L1 antibody therapy and PD-1 antibody therapy. In some embodiments, the benzoheterocyclic compound represented by formula I as described in the first aspect, its tautomer, stereoisomer, hydrate, solvate, pharmaceutically acceptable salt or The mass ratio of prodrug to antibody is 1:100 to 100:1, preferably 10:1 to 100:1, more preferably 10:1 to 50:1, for example 10:1, 15:1, 20 :1, 25:1, 30:1, 35:1, 40:1, 45:1, 50:1.
在一优选实施方式中，其中所述EP2和/或EP4受体介导的疾病包括炎性疾病(例如关节炎和子宫内膜异位症)、自身免疫疾病(例如多发性硬化症)、神经变性疾病(例如癫痫、阿尔茨海默病、帕金森病、肌萎缩侧索硬化和创伤性脑损伤)、心血管疾病(例如动脉粥样硬化)和癌症(例如结肠癌、肺癌、乳腺癌和头颈癌)。In a preferred embodiment, the diseases mediated by the EP2 and/or EP4 receptors include inflammatory diseases (such as arthritis and endometriosis), autoimmune diseases (such as multiple sclerosis), neurological Degenerative diseases (e.g. epilepsy, Alzheimer's disease, Parkinson's disease, amyotrophic lateral sclerosis and traumatic brain injury), cardiovascular diseases (e.g. atherosclerosis) and cancers (e.g. colon, lung, breast and head and neck cancer).
本发明的附加方面和优点将在下面的描述中部分给出，部分将从下面的描述中变得明显，或通过本发明的实践了解到。Additional aspects and advantages of the invention will be set forth in the description which follows, and in part will be obvious from the description, or may be learned by practice of the invention.
术语和定义Terms and Definitions
除非另有说明，本申请说明书和权利要求书中记载的基团和术语定义，包括其作为实例的定义、示例性的定义、优选的定义、表格中记载的定义、实施例中具体化合物的定义等，可以彼此之间任意组合和结合。这样的组合和结合后的基团定义及化合物结构，应当属于本申请说明书记载的范围内。Unless otherwise stated, the definitions of groups and terms recorded in the specification and claims of this application include their definitions as examples, exemplary definitions, preferred definitions, definitions recorded in tables, and definitions of specific compounds in the examples etc., can be arbitrarily combined and combined with each other. Such combinations and combined group definitions and compound structures should fall within the scope of the description of the present application.
除非另有定义，否则本文所有科技术语具有的涵义与权利要求主题所属领域技术人员通常理解的涵义相同。除非另有说明，本文全文引用的所有专利、专利申请、公开材料通过引用方式整体并入本文。如果本文对术语有多个定义，以本章的定义为准。Unless defined otherwise, all technical and scientific terms used herein have the same meaning as commonly understood by one of ordinary skill in the art to which the claimed subject matter belongs. All patents, patent applications, and publications cited in their entirety herein are hereby incorporated by reference in their entirety unless otherwise indicated. If there is more than one definition of a term herein, the definition in this chapter shall prevail.
应理解，上述简述和下文的详述为示例性且仅用于解释，而不对本发明主题作任何限制。在本申请中，除非另有具体说明，否则使用单数时也包括复数。必须注意，除非文中另有清楚的说明，否则在本说明书和权利要求书中所用的单数形式包括所指事物的复数形式。还应注意，除非另有说明，否则所用“或”、“或者”表示“和/或”。此外，所用术语“包括”以及其它形式，例如“包含”、“含”和“含有”并非限制性。It is to be understood that both the foregoing brief description and the following detailed description are exemplary and explanatory only and are not restrictive of the inventive subject matter. In this application, the use of the singular also includes the plural unless specifically stated otherwise. It must be noted that, unless the context clearly dictates otherwise, as used in the specification and claims, the singular includes the plural of the referents. It should also be noted that the use of "or", "or" means "and/or" unless stated otherwise. Furthermore, the use of the term "comprises" as well as other forms, such as "comprises", "comprises" and "comprises" is not limiting.
除非另有说明，否则采用本领域技术范围内的常规方法，如质谱、NMR、IR和UV/VIS光谱法和药理学方法。除非提出具体定义，否则本文在分析化学、有机合成化学以及药物和药物化学的有关描述中采用的术语是本领域已知的。可在化学合成、化学分析、药物制备、制剂和递送，以及对患者的治疗中使用标准技术。例如，可利用厂商对试剂盒的使用说明，或者按照本领域公知的方式或本发明的说明来实施反应和进行纯化。通常可根据本说明书中引用和讨论的多个概要性和较具体的文献中的描述，按照本领域熟知的常规方法实施上述技术和方法。在本说明书中，可由本领域技术人员选择基团及其取代基以提供稳定的结构部分和化合物。Unless otherwise indicated, conventional methods within the skill of the art, such as mass spectroscopy, NMR, IR and UV/VIS spectroscopy and pharmacological methods are employed. Unless specific definitions are set forth, terms employed herein in the relevant descriptions of analytical chemistry, synthetic organic chemistry, and pharmaceutical and medicinal chemistry are those that are known in the art. Standard techniques can be used in chemical syntheses, chemical analyses, pharmaceutical preparation, formulation and delivery, and treatment of patients. For example, reactions and purifications can be carried out using the manufacturer's instructions for the kit, or by methods known in the art or as described herein. The techniques and methods described above can generally be performed according to conventional methods well known in the art as described in various general and more specific documents that are cited and discussed in this specification. In this specification, groups and substituents thereof can be selected by those skilled in the art to provide stable moieties and compounds.
当通过从左向右书写的常规化学式描述取代基时，该取代基也同样包括从右向左书写结构式时所得到的在化学上等同的取代基。举例而言，CH 2O等同于OCH 2。如本文所用，
Figure PCTCN2022097611-appb-000023
Figure PCTCN2022097611-appb-000024
表示基团的连接位点。如本文所用，“R 1”、“R1”和“R 1”的含义相同，可相互替换。对于R 2等其它其他符号，类似定义的含义相同。 When a substituent is described by a conventional chemical formula written from left to right, the substituent also includes chemically equivalent substituents obtained when the structural formula is written from right to left. For example, CH2O is equivalent to OCH2 . As used herein,
Figure PCTCN2022097611-appb-000023
Figure PCTCN2022097611-appb-000024
Indicates the attachment site of the group. As used herein, "R 1 ", "R1" and "R 1 " have the same meaning and can be substituted for each other. For other symbols such as R 2 , similar definitions have the same meanings.
本文所用的章节标题仅用于组织文章的目的，而不应被解释为对所述主题的限制。本申请中引用的所有文献或文献部分包括但不限于专利、专利申请、文章、书籍、操作手册和论文，均通过引用方式整体并入本文。The section headings used herein are for the purpose of organizing the article only and should not be construed as limitations on the subject matter described. All documents or portions of documents cited in this application, including but not limited to patents, patent applications, articles, books, manuals, and treatises, are hereby incorporated by reference in their entirety.
除前述以外，当用于本申请的说明书及权利要求书中时，除非另外特别指明，否则以下术语具有如下所示的含义。In addition to the foregoing, when used in the specification and claims of the present application, the following terms have the meanings shown below unless otherwise specified.
在本申请中，在单独或作为其他取代基一部分时，术语“卤素”是指氟、氯、溴、碘。In this application, the term "halogen" means fluorine, chlorine, bromine, iodine alone or as part of another substituent.
如本文所用，在单独或作为其他取代基一部分时，术语“烷基”意指仅由碳原子和氢原 子组成、不含不饱和键、具有例如1至6个碳原子且通过单键与分子的其余部分连接的直链或支链的烃链基团。烷基的实例包括但不限于甲基、乙基、正丙基、异丙基、正丁基、异丁基，叔丁基，戊基，异戊基，新戊基和己基。烷基可以是未取代的或被一个或多个合适的取代基取代。烷基也可以是富含碳和/或氢的同位素(即氘或氚)的天然丰度烷基的同位素异构体。如本文所用，术语“烯基”表示无支链或支链的单价烃链，其含有一个或多个碳-碳双键。如本文所用，术语“炔基”是指无支链或支链的一价烃链，其含有一个或多个碳-碳三键。“C 2-C 5炔基”则含有2-5个碳原子。 As used herein, the term "alkyl", alone or as part of another substituent, means consisting solely of carbon and hydrogen atoms, free of unsaturated bonds, having, for example, 1 to 6 carbon atoms, and is bonded to the molecule by a single bond. The rest are connected straight or branched hydrocarbon chain groups. Examples of alkyl groups include, but are not limited to, methyl, ethyl, n-propyl, isopropyl, n-butyl, isobutyl, t-butyl, pentyl, isopentyl, neopentyl, and hexyl. An alkyl group can be unsubstituted or substituted with one or more suitable substituents. The alkyl groups may also be isotopomers of naturally abundant alkyl groups that are rich in carbon and/or hydrogen isotopes (ie, deuterium or tritium). As used herein, the term "alkenyl" means an unbranched or branched monovalent hydrocarbon chain containing one or more carbon-carbon double bonds. As used herein, the term "alkynyl" refers to an unbranched or branched monovalent hydrocarbon chain containing one or more carbon-carbon triple bonds. "C 2 -C 5 alkynyl" then contains 2-5 carbon atoms.
在单独或作为其他取代基一部分时，术语“C 1-C 5烷基”应理解为表示具有1、2、3、4或5个碳原子的直链或支链饱和一价烃基。所述烷基是例如甲基、乙基、丙基、丁基、戊基、己基、异丙基、异丁基、仲丁基、叔丁基、异戊基、2-甲基丁基、1-甲基丁基、1-乙基丙基、1,2-二甲基丙基、新戊基、1,1-二甲基丙基等或它们的异构体。特别地，所述基团具有1、2或3个碳原子(“C 1-C 3烷基”)，例如甲基、乙基、正丙基或异丙基。 The term "C 1 -C 5 alkyl", alone or as part of another substituent, is understood to mean a linear or branched saturated monovalent hydrocarbon radical having 1, 2, 3, 4 or 5 carbon atoms. The alkyl group is for example methyl, ethyl, propyl, butyl, pentyl, hexyl, isopropyl, isobutyl, sec-butyl, tert-butyl, isopentyl, 2-methylbutyl, 1-methylbutyl, 1-ethylpropyl, 1,2-dimethylpropyl, neopentyl, 1,1-dimethylpropyl, etc. or their isomers. In particular, said groups have 1, 2 or 3 carbon atoms (“C 1 -C 3 alkyl”), for example methyl, ethyl, n-propyl or isopropyl.
术语“亚烷基”表示从饱和的直链或支链烃基中去掉两个氢原子所得到的饱和的二价烃基基团。“C 1-C 5亚烷基”指含有1、2、3、4、5个碳原子的亚烷基，其实例包括亚甲基(-CH 2-)，亚乙基(包括-CH 2CH 2-或-CH(CH 3)-)，亚异丙基(包括-CH(CH 3)CH 2-或-C(CH 3) 2-)等。 The term "alkylene" means a saturated divalent hydrocarbyl group obtained by removing two hydrogen atoms from a saturated straight or branched chain hydrocarbyl group. "C 1 -C 5 alkylene" refers to an alkylene group containing 1, 2, 3, 4, or 5 carbon atoms, examples of which include methylene (-CH 2 -), ethylene (including -CH 2 CH 2 -or -CH(CH 3 )-), isopropylidene (including -CH(CH 3 )CH 2 - or -C(CH 3 ) 2 -), etc.
在单独或作为其他取代基一部分时，术语“环烷基”或“碳环基”是指一种环状烷基。术语“m-n元环烷基”或者“C m-C n环烷基”应理解为表示具有m至n个原子的饱和、不饱和或部分饱和的碳环。例如，“3-15元环烷基”或者“C 3-C 15环烷基”是指含有3至15，3至9，3至6或3至5个碳原子的环状烷基，它可能包含1至4个环。“3-6元环烷基”则含有3-6个碳原子。包括单环、二环、三环、螺环或桥环。未取代的环烷基的实例包括但不限于环丙基，环丁基，环戊基，环己基。环烷基可以被一个或多个取代基取代。在一些实施方案中，环烷基可以是与芳基或杂芳环基稠合的环烷基。 The terms "cycloalkyl" or "carbocyclyl" by themselves or as part of another substituent refer to a cyclic alkyl group. The term "mn-membered cycloalkyl" or " Cm - Cncycloalkyl " is understood to mean a saturated, unsaturated or partially saturated carbocycle having m to n atoms. For example, "3-15 membered cycloalkyl" or "C 3 -C 15 cycloalkyl" refers to a cyclic alkyl group containing 3 to 15, 3 to 9, 3 to 6 or 3 to 5 carbon atoms, which May contain 1 to 4 rings. "3-6 membered cycloalkyl" then contains 3-6 carbon atoms. Including monocyclic, bicyclic, tricyclic, spiro or bridged rings. Examples of unsubstituted cycloalkyl include, but are not limited to, cyclopropyl, cyclobutyl, cyclopentyl, cyclohexyl. Cycloalkyl groups may be substituted with one or more substituents. In some embodiments, a cycloalkyl group can be a cycloalkyl group fused to an aryl or heteroaryl ring group.
在单独或作为其他取代基一部分时，术语“卤代”可与术语“卤素取代”互换使用。“卤代烷基”或“卤素取代的烷基”指包括具有特定数目的碳原子、被一或多个卤素取代的支链和直链的饱和脂族烃基。The term "halo" is used interchangeably with the term "halogen substitution", alone or as part of another substituent. "Haloalkyl" or "halogen-substituted alkyl" refers to both branched and straight-chain saturated aliphatic hydrocarbon groups having the specified number of carbon atoms, substituted with one or more halogens.
在单独或作为其他取代基一部分时，术语“杂环烷基”或“杂环基”是指其中一个或多个(在一些实施方案中为1至3个)碳原子被杂原子取代的环烷基，所述杂原子例如但不限于N、O、S和P。术语“m-n元杂环烷基”或者“C m-C n杂环烷基”应理解为表示具有m至n个原子的饱和、不饱和或部分饱和的环。例如，术语“4-6元杂环烷基”应理解为表示具有4至6个原子的饱和、不饱和或部分饱和的环。当诸如4-6元的前缀用于表示杂环烷基时，碳的数目也意味着包括杂原子。 The terms "heterocycloalkyl" or "heterocyclyl", by themselves or as part of another substituent, refer to rings in which one or more (in some embodiments, 1 to 3) carbon atoms are replaced by heteroatoms Alkyl, said heteroatoms such as but not limited to N, O, S and P. The term "mn-membered heterocycloalkyl" or " Cm - Cnheterocycloalkyl " is understood to mean a saturated, unsaturated or partially saturated ring having m to n atoms. For example, the term "4-6 membered heterocycloalkyl" is understood to mean a saturated, unsaturated or partially saturated ring having 4 to 6 atoms. When a prefix such as 4-6 membered is used to denote heterocycloalkyl, the number of carbons is also meant to include heteroatoms.
在单独或作为其他取代基一部分时，术语“杂芳基”可与术语“杂芳环”或“杂芳族基”互换使用，是指单环或多环芳环系统，在某些实施方案中，环系统中1至3个原子是杂原子，即除碳以外的元素，包括但不限于N，O、S或P。The term "heteroaryl" is used interchangeably with the terms "heteroaromatic ring" or "heteroaromatic", by itself or as part of another substituent, and refers to a monocyclic or polycyclic aromatic ring system, in certain implementations In the scheme, 1 to 3 atoms in the ring system are heteroatoms, that is, elements other than carbon, including but not limited to N, O, S or P.
在单独或作为其他取代基一部分时，术语“5-6元杂芳基”应理解为具有5～6个环原子且包含杂原子的芳族环基团。包括但不限于噻吩、呋喃、吡咯、吡唑、咪唑、三氮唑、噻唑、噻二唑、噁唑、吡啶、嘧啶、哒嗪、吡嗪。The term "5-6 membered heteroaryl", alone or as part of another substituent, is understood to mean an aromatic ring group having 5 to 6 ring atoms, including heteroatoms. Including but not limited to thiophene, furan, pyrrole, pyrazole, imidazole, triazole, thiazole, thiadiazole, oxazole, pyridine, pyrimidine, pyridazine, pyrazine.
本文提供的化合物，包括可用于制备本文提供的化合物的中间体，其含有反应性官能团 (例如但不限于羧基，羟基和氨基部分)，还包括其保护的衍生物。“受保护的衍生物”是其中一个或多个反应性位点被一个或多个保护基团(也称为保护基团)封闭的那些化合物。合适的羧基部分保护基包括苄基，叔丁基等，以及同位素等。合适的氨基和酰氨基保护基包括乙酰基，三氟乙酰基，叔丁氧基羰基，苄氧基羰基等。合适的羟基保护基包括苄基等。其他合适的保护基团是本领域普通技术人员所熟知的。The compounds provided herein, including intermediates useful in the preparation of the compounds provided herein, contain reactive functional groups (such as, but not limited to, carboxyl, hydroxyl, and amino moieties), and also include protected derivatives thereof. "Protected derivatives" are those compounds in which one or more reactive sites are blocked by one or more protecting groups (also known as protecting groups). Suitable protecting groups for the carboxyl moiety include benzyl, tert-butyl, etc., and isotopes, etc. Suitable amino and amido protecting groups include acetyl, trifluoroacetyl, t-butoxycarbonyl, benzyloxycarbonyl and the like. Suitable hydroxy protecting groups include benzyl and the like. Other suitable protecting groups are well known to those of ordinary skill in the art.
在本申请中，“任选的”或“任选地”表示随后描述的事件或状况可能发生也可能不发生，且该描述同时包括该事件或状况发生和不发生的情况。例如，“任选地被取代的芳基”表示芳基被取代或未被取代，且该描述同时包括被取代的芳基与未被取代的芳基。进一步的，“任选的”或“任选地”取代情形涵盖了化合物结构/基团无取代，以及化合物结构/基团被选自一个或多个所定义的取代基取代的情形。例如，“任选地被取代的芳基”表示未被取代的芳基与被选自一个或多个所定义的取代基取代的芳基。“多个”意为两个以上，即包含两个、三个以上。In this application, "optional" or "optionally" means that the subsequently described event or circumstance may or may not occur, and that the description includes both the occurrence and non-occurrence of the event or circumstance. For example, "optionally substituted aryl" means that the aryl is substituted or unsubstituted, and the description includes both substituted and unsubstituted aryl. Further, "optional" or "optionally" substitution covers the situation that the compound structure/group is not substituted, and the compound structure/group is substituted by one or more defined substituents. For example, "optionally substituted aryl" means unsubstituted aryl and aryl substituted with one or more substituents selected from the defined substituents. "Multiple" means more than two, including two or more.
在本申请中，术语“盐”或“药学上可接受的盐”，包括药学上可接受的酸加成盐和药学上可接受的碱加成盐。术语“药学上可接受的”，是针对那些化合物、材料、组合物和/或剂型而言，它们在可靠的医学判断的范围之内，适用于与人类和动物的组织接触使用，而没有过多的毒性、刺激性、过敏性反应或其它问题或并发症，与合理的利益/风险比相称。In this application, the term "salt" or "pharmaceutically acceptable salt" includes pharmaceutically acceptable acid addition salts and pharmaceutically acceptable base addition salts. The term "pharmaceutically acceptable" refers to those compounds, materials, compositions and/or dosage forms which, within the scope of sound medical judgment, are suitable for use in contact with human and animal tissues without excessive Toxicity, irritation, allergic reaction, or other problems or complications, commensurate with a reasonable benefit/risk ratio.
“药学上可接受的酸加成盐”是指能够保留游离碱的生物有效性而无其它副作用的，与无机酸或有机酸所形成的盐。“药学上可接受的碱加成盐”是指能够保持游离酸的生物有效性而无其它副作用的、与无机碱或有机碱所形成的盐。除了药学可接受的盐外，本发明还考虑其他盐。它们可以在化合物纯化中或在制备其它药学上课接受的盐中充当中间体或可用于本发明化合物的鉴别、表征或纯化。"Pharmaceutically acceptable acid addition salt" refers to a salt formed with an inorganic or organic acid that retains the biological effectiveness of the free base without other side effects. "Pharmaceutically acceptable base addition salt" refers to a salt formed with an inorganic base or an organic base that can maintain the biological effectiveness of the free acid without other side effects. In addition to pharmaceutically acceptable salts, other salts are contemplated by the present invention. They may serve as intermediates in the purification of compounds or in the preparation of other pharmaceutically acceptable salts or may be useful in the identification, characterization or purification of compounds of the invention.
术语“立体异构体”是指由分子中原子在空间上排列方式不同所产生的异构体，包括顺反异构体、对映异构体、非对应异构体和构象异构体。The term "stereoisomer" refers to isomers resulting from differences in the arrangement of atoms in a molecule in space, including cis-trans isomers, enantiomers, diastereoisomers and conformers.
依据原料和方法的选择，本发明化合物可以以可能的异构体中的一个或它们的混合物的形式存在，例如作为纯旋光异构体，或作为异构体混合物，如作为外消旋和非对映异构体混合物，这取决于不对称碳原子的数量。当描述具有光学活性的化合物时，使用前缀D和L或R和S来表示就分子中的手性中心(或多个手性中心)而言分子的绝对构型。前缀D和L或(+)和(–)是用于指定化合物所致平面偏振光旋转的符号，其中(–)或L表示化合物是左旋的。前缀为(+)或D的化合物是右旋的。Depending on the choice of starting materials and processes, the compounds according to the invention may exist as one of the possible isomers or as mixtures thereof, for example as pure optical isomers, or as mixtures of isomers, for example as racemic and non- A mixture of enantiomers, depending on the number of asymmetric carbon atoms. When describing optically active compounds, the prefixes D and L or R and S are used to denote the absolute configuration of the molecule with respect to the chiral center (or centers) in the molecule. The prefixes D and L or (+) and (-) are symbols used to designate the rotation of plane polarized light by a compound, where (-) or L indicates that the compound is levorotatory. Compounds prefixed with (+) or D are dextrorotatory.
当将本发明式中与手性碳的键描写直成线时，应当理解为，手性碳的(R)和(S)两种构型和由此产生的其对映体纯的化合物和混合物两者包括在该通式范围内。本文中消旋体或者对映体纯的化合物的图示法来自Maehr，J.Chem.Ed.1985，62:114-120。用楔形键和虚线键表示一个立体中心的绝对构型。When the bond with the chiral carbon in the formula of the present invention is described as a straight line, it should be understood that the (R) and (S) two configurations of the chiral carbon and the resulting enantiomerically pure compounds and Mixtures of both are included within the scope of the general formula. Graphical representations of racemic or enantiomerically pure compounds herein are from Maehr, J. Chem. Ed. 1985, 62:114-120. The absolute configuration of a stereocenter is indicated by wedge-shaped bonds and dashed-line bonds.
术语“互变异构体”是指因分子中某一原子在两个位置迅速移动而产生的官能团异构体。本发明化合物可表现出互变异构现象。互变异构的化合物可以存在两种或多种可相互转化的种类。质子移变互变异构体来自两个原子之间共价键合的氢原子的迁移。互变异构体一般以平衡形式存在，尝试分离单一互变异构体时通常产生一种混合物，其理化性质与化合物的混合物是一致的。平衡的位置取决于分子内的化学特性。例如，在很多脂族醛和酮如乙醛中，酮型占优势；而在酚中，烯醇型占优势。本发明包含化合物的所有互变异构形式。The term "tautomer" refers to isomers of functional groups resulting from the rapid movement of an atom in a molecule between two positions. The compounds of the present invention may exhibit tautomerism. Tautomeric compounds can exist in two or more interconvertible species. Prototropic tautomers result from the migration of a covalently bonded hydrogen atom between two atoms. Tautomers generally exist in equilibrium and attempts to isolate a single tautomer usually result in a mixture whose physicochemical properties are consistent with the mixture of compounds. The position of equilibrium depends on the chemical properties within the molecule. For example, in many aliphatic aldehydes and ketones such as acetaldehyde, the keto form predominates; in phenols, the enol form predominates. The present invention encompasses all tautomeric forms of the compounds.
在本申请中，“药物组合物”是指本发明化合物与本领域通常接受的用于将生物活性化合物输送至哺乳动物(例如人)的介质的制剂。该介质包括药学上可接受的载体。药物组合物的目的是促进生物体的给药，利于活性成分的吸收进而发挥生物活性。In this application, a "pharmaceutical composition" refers to a formulation of a compound of the present invention with a vehicle generally accepted in the art for the delivery of a biologically active compound to a mammal (eg, a human). The medium includes a pharmaceutically acceptable carrier. The purpose of the pharmaceutical composition is to promote the administration of the organism, facilitate the absorption of the active ingredient and thus exert its biological activity.
在本申请中，“药学上可接受的载体”包括但不限于任何被相关的政府管理部门许可为可接受供人类或家畜使用的佐剂、载体、赋形剂、助流剂、增甜剂、稀释剂、防腐剂、染料/着色剂、矫味剂、表面活性剂、润湿剂、分散剂、助悬剂、稳定剂、等渗剂、溶剂或乳化剂。In this application, "pharmaceutically acceptable carrier" includes, but is not limited to, any adjuvant, carrier, excipient, glidant, sweetener approved by the relevant government regulatory agency as acceptable for human or livestock use , diluent, preservative, dye/colorant, flavoring agent, surfactant, wetting agent, dispersing agent, suspending agent, stabilizing agent, isotonic agent, solvent or emulsifying agent.
术语“溶剂化物”指本发明化合物或其盐包括以分子间非共价力结合的化学计量或非化学计量的溶剂，当溶剂为水时，则为水合物。The term "solvate" means that the compound of the present invention or its salt includes a stoichiometric or non-stoichiometric solvent bonded by intermolecular non-covalent forces, and when the solvent is water, it is a hydrate.
术语“前药”是指可以在生理条件下或者通过溶剂解转化为具有生物活性的本发明化合物。本发明的前药通过修饰在该化合物中的功能基团来制备，该修饰可以按常规的操作或者在体内被除去，而得到母体化合物。前药包括本发明化合物中的一个羟基或者氨基连接到任何基团上所形成的化合物，当本发明化合物的前药被施予哺乳动物个体时，前药被割裂而分别形成游离的羟基、游离的氨基。The term "prodrug" refers to a compound of the invention that can be converted to biological activity under physiological conditions or by solvolysis. The prodrugs of the present invention are prepared by modifying functional groups in the compounds which can be removed routinely or in vivo to yield the parent compound. Prodrugs include compounds formed by linking a hydroxyl or amino group in the compound of the present invention to any group. When the prodrug of the compound of the present invention is administered to a mammalian individual, the prodrug is split to form free hydroxyl, free of amino.
术语“辅料”是指可药用惰性成分。术语“赋形剂”的种类实例非限制性地包括粘合剂、崩解剂、润滑剂、助流剂、稳定剂、填充剂和稀释剂等。赋形剂能增强药物制剂的操作特性，即通过增加流动性和/或粘着性使制剂更适于直接压缩。The term "excipient" refers to a pharmaceutically acceptable inert ingredient. Examples of categories of the term "excipient" include, but are not limited to, binders, disintegrants, lubricants, glidants, stabilizers, fillers, diluents, and the like. Excipients can enhance the handling characteristics of a pharmaceutical formulation, ie make the formulation more suitable for direct compression by increasing flow and/or cohesiveness.
术语“患者”是指包括哺乳动物在内的任何动物，优选小鼠、大鼠、其它啮齿类动物、兔、狗、猫、猪、牛、羊、马或灵长类动物，最优选人。The term "patient" refers to any animal including mammals, preferably mice, rats, other rodents, rabbits, dogs, cats, pigs, cows, sheep, horses or primates, most preferably humans.
术语“治疗有效量”是指研究人员、兽医、医师或其它临床医师正在组织、系统、动物、个体或人中寻找的引起生物学或医学反应的活性化合物或药物的量，它包括以下一项或多项：(1)预防疾病：例如在易感染疾病、紊乱或病症但尚未经历或出现疾病病理或症状的个体中预防疾病、紊乱或病症。(2)抑制疾病：例如在正经历或出现疾病、紊乱或病症的病理或症状的个体中抑制疾病、紊乱或病症(即阻止病理和/或症状的进一步发展)。(3)缓解疾病：例如在正经历或出现疾病、紊乱或病症的病理或症状的个体中缓解疾病、紊乱或病症(即逆转病理和/或症状)。The term "therapeutically effective amount" refers to the amount of an active compound or drug that a researcher, veterinarian, physician, or other clinician is seeking to elicit a biological or medical response in a tissue, system, animal, individual, or human, and includes any of the following or more: (1) Preventing a disease: eg, preventing a disease, disorder or condition in an individual who is susceptible to the disease, disorder or condition but has not yet experienced or developed disease pathology or symptoms. (2) Inhibiting a disease: For example, inhibiting a disease, disorder or condition (ie preventing further development of the pathology and/or symptoms) in an individual experiencing or developing pathology or symptoms of the disease, disorder or condition. (3) Alleviating disease: For example, alleviating a disease, disorder or condition (ie reversing the pathology and/or symptoms) in an individual experiencing or developing the pathology or symptoms of the disease, disorder or condition.
本文所用的术语“治疗”和其它类似的同义词包括以下含义：As used herein, the term "treatment" and other similar synonyms include the following meanings:
(i)预防疾病或病症在哺乳动物中出现，特别是当这类哺乳动物易患有该疾病或病症，但尚未被诊断为已患有该疾病或病症时；(i) preventing the occurrence of a disease or condition in a mammal, especially when such mammal is susceptible to the disease or condition but has not been diagnosed as having the disease or condition;
(ii)抑制疾病或病症，即遏制其发展；(ii) inhibiting a disease or condition, i.e. arresting its development;
(iii)缓解疾病或病症，即，使该疾病或病症的状态消退；或者(iii) ameliorating a disease or condition, i.e., causing regression of the state of the disease or condition; or
(iv)减轻该疾病或病症所造成的症状。(iv) Alleviating the symptoms caused by the disease or condition.
术语“抗体”包括所有类型的免疫球蛋白。抗体可以是单克隆或多克隆的，并且可以属于任何物种的来源，包括如小鼠、大鼠、兔、马或人。尤其当其用于治疗的目的时，抗体可以是嵌合的或人源化的。抗体可以通过所属领域中已知的方法获得或制备。The term "antibody" includes all classes of immunoglobulins. Antibodies may be monoclonal or polyclonal and may be of any species of origin including, for example, mouse, rat, rabbit, horse or human. Antibodies may be chimeric or humanized, especially when they are used for therapeutic purposes. Antibodies can be obtained or prepared by methods known in the art.
术语PD-L1抗体”或“抗PD-L1”是指针对程序性死亡配体1(PD-L1)的抗体。The term "PD-L1 antibody" or "anti-PD-L1" refers to an antibody directed against programmed death ligand 1 (PD-L1).
术语“PD-1抗体”或“抗PD-1”是指针对程序性死亡蛋白1(PD-1)的抗体。The term "PD-1 antibody" or "anti-PD-1" refers to an antibody directed against programmed death protein 1 (PD-1).
术语“抗体疗法”是指医学使用结合靶细胞或靶细胞蛋白的抗体，以治疗癌症和/或刺激受试对象的免疫应答，所述免疫应答导致受试对象中的癌细胞的识别、攻击和/或破坏，并且 在本发明的一些实施方案中，以激活或刺激受试对象的记忆免疫应答，所述记忆免疫应答导致受试对象中的癌细胞的随后识别、攻击和/或破坏。The term "antibody therapy" refers to the medical use of antibodies that bind to target cells or proteins of target cells to treat cancer and/or stimulate an immune response in a subject that results in the recognition, attack and and/or destroy, and in some embodiments of the invention, to activate or stimulate a subject's memory immune response that results in subsequent recognition, attack and/or destruction of cancer cells in the subject.
术语“PD-L1抗体疗法”是指使用针对程序性死亡配体1的抗体(抗PD-L1)调节受试对象的免疫应答。在一些实施方案中，PD-L1抗体抑制或阻断PD-L1与程序性细胞死亡蛋白1(PD-1)的相互作用，其中PD-L1与PD-1之间的相互作用的阻断抑制PD-1对T细胞激活的负向调节，从而攻击和破坏癌细胞。The term "PD-L1 antibody therapy" refers to the use of antibodies against programmed death-ligand 1 (anti-PD-L1 ) to modulate the immune response of a subject. In some embodiments, the PD-L1 antibody inhibits or blocks the interaction of PD-L1 with programmed cell death protein 1 (PD-1), wherein blockade of the interaction between PD-L1 and PD-1 inhibits PD-1 negatively regulates T cell activation to attack and destroy cancer cells.
术语“PD-1抗体疗法”是指使用针对程序性细胞死亡蛋白1PD-1的抗体(抗PD-1)调节受试对象的免疫应答。在一些实施方案中，PD1抗体抑制或阻断PD-1与PD-L1的相互作用，其中PD-L1与PD-1之间的相互作用的抑制或阻断抑制PD-1对T细胞激活的负向调节，从而攻击和破坏癌细胞。The term "PD-1 antibody therapy" refers to the use of antibodies against the programmed cell death protein 1 PD-1 (anti-PD-1) to modulate the immune response of a subject. In some embodiments, the PD1 antibody inhibits or blocks the interaction of PD-1 and PD-L1, wherein the inhibition or blockage of the interaction between PD-L1 and PD-1 inhibits the effect of PD-1 on T cell activation Negative regulation, thereby attacking and destroying cancer cells.
各步骤的反应，反应温度可因溶剂、起始原料、试剂等适宜选择，反应时间也可因反应温度、溶剂、起始原料、试剂等适宜选择。各步骤反应结束后，目标化合物可按常用方法自反应体系中进行分离、提纯等步骤，如过滤、萃取、重结晶、洗涤、硅胶柱层析等方法。在不影响下一步反应的情况下，目标化合物也可不经过分离、纯化直接进入下一步反应。In the reaction of each step, the reaction temperature can be appropriately selected according to the solvent, starting material, reagent, etc., and the reaction time can also be appropriately selected according to the reaction temperature, solvent, starting material, reagent, etc. After the reaction of each step, the target compound can be separated and purified from the reaction system according to common methods, such as filtration, extraction, recrystallization, washing, silica gel column chromatography and other methods. Without affecting the next step reaction, the target compound can also directly enter the next step reaction without separation and purification.
有益效果Beneficial effect
本发明人经过广泛而深入地研究，意外地开发了用于治疗EP2、EP4受体介导的疾病的苯并杂环化合物，所述苯并杂环化合物为本发明中所述的式I所示苯并杂环化合物。本发明所述的用于治疗EP2、EP4受体介导的疾病的苯并杂环化合物，可以用于治疗炎性疾病(例如关节炎和子宫内膜异位症)、自身免疫疾病(例如多发性硬化症)、神经变性疾病(例如癫痫、阿尔茨海默病、帕金森病、肌萎缩侧索硬化和创伤性脑损伤)、心血管疾病(例如动脉粥样硬化)和癌症(例如结肠癌、肺癌、乳腺癌和头颈癌)。After extensive and in-depth research, the inventors unexpectedly developed a benzo heterocyclic compound for treating diseases mediated by EP2 and EP4 receptors. The benzo heterocyclic compound is represented by formula I described in the present invention Show benzoheterocyclic compounds. The benzoheterocyclic compound used for the treatment of diseases mediated by EP2 and EP4 receptors described in the present invention can be used for the treatment of inflammatory diseases (such as arthritis and endometriosis), autoimmune diseases (such as multiple sclerosis), neurodegenerative diseases (such as epilepsy, Alzheimer's disease, Parkinson's disease, amyotrophic lateral sclerosis, and traumatic brain injury), cardiovascular diseases (such as atherosclerosis) and cancers (such as colon cancer , lung, breast and head and neck cancers).
实验证明，本发明所述苯并杂环化合物对EP2具有很好的拮抗作用，对EP4钙流显示较好抑制作用，与EP2受体和/或EP4受体具有很好的亲和力。本发明所述苯并杂环化合物的静脉给药清除率低，口服给药暴露量高，表现出优良的药代动力学性质，在人血浆中有较高的游离分数，具有很好的热力学溶解度，成药性好。另外，与抗体药物(例如抗PD-L1抗体、抗PD-1抗体)联用表现出显著的抑瘤作用。Experiments have proved that the benzoheterocyclic compound of the present invention has a good antagonism to EP2, a good inhibitory effect on EP4 calcium flow, and a good affinity with EP2 receptors and/or EP4 receptors. The benzoheterocyclic compound of the present invention has low clearance rate of intravenous administration, high oral administration exposure, exhibits excellent pharmacokinetic properties, has a relatively high free fraction in human plasma, and has good thermodynamics Good solubility and druggability. In addition, the combination with antibody drugs (such as anti-PD-L1 antibody, anti-PD-1 antibody) showed a significant tumor inhibitory effect.
附图说明Description of drawings
图1为本发明中的化合物对人PBMC细胞分化实验抗原呈递细胞的评价结果；Fig. 1 is the evaluation result of the compound in the present invention to the antigen presenting cell of human PBMC cell differentiation experiment;
图2为本发明中的化合物对人PBMC细胞分化实验M2型巨噬细胞的评价结果；Fig. 2 is the evaluation result of the compound in the present invention to human PBMC cell differentiation experiment M2 type macrophage;
图3为本发明中的化合物对人PBMC细胞分化实验M1型巨噬细胞的评价结果；Fig. 3 is the evaluation result of the compound in the present invention to human PBMC cell differentiation experiment M1 type macrophage;
图4为本发明中的化合物联合anti-mouse PD-1抗体法在CT-26鼠源结肠癌肿瘤模型中的抗肿瘤作用结果。Figure 4 is the results of the anti-tumor effect of the compound of the present invention combined with anti-mouse PD-1 antibody in the CT-26 mouse colon cancer tumor model.
具体实施方式Detailed ways
以下结合具体实施例，进一步说明本发明。需理解，以下的描述仅为本发明的最优选实 施方式，而不应当被认为是对于本发明保护范围的限制。在充分理解本发明的基础上，下列实施例中未注明具体条件的实验方法，通常按照常规条件，或按照制造厂商所建议的条件，本领域技术人员可以对本发明的技术方案作出非本质的改动，这样的改动应当被视为包括于本发明的保护范围之中的。Below in conjunction with specific embodiment, further illustrate the present invention. It should be understood that the following description is only the most preferred embodiment of the present invention, and should not be considered as limiting the protection scope of the present invention. On the basis of fully understanding the present invention, the experimental methods that do not indicate specific conditions in the following examples usually follow conventional conditions, or according to the conditions suggested by the manufacturer, those skilled in the art can make unessential changes to the technical solution of the present invention Changes, such changes should be considered included in the protection scope of the present invention.
本申请具有如下定义：This application has the following definitions:
符号或单位：Symbol or unit:
IC 50：半数抑制浓度，指达到最大抑制效果一半时的浓度 IC 50 : half inhibitory concentration, which refers to the concentration at which half of the maximum inhibitory effect is achieved
M：mol/L，例如正丁基锂(14.56mL，29.1mmol，2.5M的正己烷溶液)表示摩尔浓度为2.5mol/L的正丁基锂的正己烷溶液M: mol/L, such as n-butyllithium (14.56mL, 29.1mmol, 2.5M n-hexane solution) means n-hexane solution of n-butyllithium with a molar concentration of 2.5mol/L
N：当量浓度，例如2N盐酸表示2mol/L盐酸溶液N: Equivalent concentration, for example, 2N hydrochloric acid means 2mol/L hydrochloric acid solution
HATU：O-(7-氮杂苯并三唑-1-基)-N,N,N′,N′-四甲基脲六氟磷酸酯HATU: O-(7-azabenzotriazol-1-yl)-N,N,N′,N′-tetramethyluronium hexafluorophosphate
PE：石油醚PE: petroleum ether
EA：乙酸乙酯EA: ethyl acetate
试验或检测方法：Test or detection method:
酸性制备方法A：Welch，Ultimate C 18柱，10μm，21.2mm×250mm。流动相A为1‰的三氟醋酸纯水溶液，流动相B为乙腈溶液。梯度条件：0～3分钟，流动相A保持90％，3～18分钟梯度洗脱，由90％变为5％，18～22分钟保持5％。 Acidic preparation method A: Welch, Ultimate C 18 column, 10 μm, 21.2 mm×250 mm. Mobile phase A was 1‰ pure trifluoroacetic acid aqueous solution, and mobile phase B was acetonitrile solution. Gradient conditions: 0 to 3 minutes, maintain 90% of mobile phase A, gradient elution in 3 to 18 minutes, change from 90% to 5%, and maintain 5% in 18 to 22 minutes.
实施例1化合物I-1的制备The preparation of embodiment 1 compound I-1
合成路线如下所示：The synthetic route is as follows:
Figure PCTCN2022097611-appb-000025
Figure PCTCN2022097611-appb-000025
第一步：4-(丙烷-1-炔-1-基)-1H-吲唑-7-甲酸甲酯的制备The first step: the preparation of 4-(propane-1-yn-1-yl)-1H-indazole-7-carboxylic acid methyl ester
室温下将化合物4-溴-1H-吲唑-7-甲酸甲酯(1.0g,3.94mmol)加入到1,4-二氧六环(10mL)中，加入二三苯基膦二氯化钯(274mg,0.39mmol)，1-丙炔-三正丁基锡(1.52g,4.61mmol)，氮气保护下加热至90℃，搅拌16h。冷却至室温，加入水(50mL)稀释，用乙酸乙酯(50mL×3)萃取，分液，合并有机相，有机相用无水硫酸钠干燥，过滤，浓缩，残留物用薄层硅胶板分 离纯化(石油醚:乙酸乙酯(V/V)＝5:1)得化合物4-(丙烷-1-炔-1-基)-1H-吲唑-7-甲酸甲酯(500mg,产率42％)。Add the compound 4-bromo-1H-indazole-7-carboxylic acid methyl ester (1.0g, 3.94mmol) to 1,4-dioxane (10mL) at room temperature, add ditriphenylphosphine palladium dichloride (274mg, 0.39mmol), 1-propyne-tri-n-butyltin (1.52g, 4.61mmol), heated to 90°C under nitrogen protection, and stirred for 16h. Cool to room temperature, add water (50mL) to dilute, extract with ethyl acetate (50mL×3), separate the liquids, combine the organic phases, dry the organic phases with anhydrous sodium sulfate, filter, concentrate, and separate the residue with a thin-layer silica gel plate Purification (petroleum ether: ethyl acetate (V/V) = 5:1) gave compound 4-(propane-1-yn-1-yl)-1H-indazole-7-carboxylic acid methyl ester (500mg, yield 42 %).
第二步：1-((1,1’-二苯基)-4-基甲基)-4-(丙烷-1-炔-1-基)-1H-吲唑-7-甲酸甲酯的制备The second step: 1-((1,1'-diphenyl)-4-ylmethyl)-4-(propane-1-yn-1-yl)-1H-indazole-7-carboxylic acid methyl ester preparation
室温下将化合物4-(丙烷-1-炔-1-基)-1H-吲唑-7-甲酸甲酯(100mg,0.47mmol)加入到N,N-二甲基甲酰胺(5mL)中，加入4-苯基苄溴(174mg,0.71mmol)，碳酸铯(460mg,1.40mmol)，室温搅拌16h。加入水(20mL)稀释，用乙酸乙酯(20mL×3)萃取，分液，合并有机相，有机相用无水硫酸钠干燥，过滤，浓缩，残留物用薄层硅胶板分离纯化(石油醚:乙酸乙酯(V/V)＝10:1)得化合物1-((1,1’-二苯基)-4-基甲基)-4-(丙烷-1-炔-1-基)-1H-吲唑-7-甲酸甲酯(80mg,产率45％)。Compound 4-(propane-1-yn-1-yl)-1H-indazole-7-carboxylic acid methyl ester (100 mg, 0.47 mmol) was added into N,N-dimethylformamide (5 mL) at room temperature, Add 4-phenylbenzyl bromide (174mg, 0.71mmol), cesium carbonate (460mg, 1.40mmol), and stir at room temperature for 16h. Add water (20mL) to dilute, extract with ethyl acetate (20mL×3), separate the layers, combine the organic phases, dry the organic phases with anhydrous sodium sulfate, filter, concentrate, and separate and purify the residue with a thin-layer silica gel plate (petroleum ether : Ethyl acetate (V/V)=10:1) to obtain compound 1-((1,1'-diphenyl)-4-ylmethyl)-4-(propane-1-yn-1-yl) - 1H-indazole-7-carboxylic acid methyl ester (80 mg, yield 45%).
第三步：1-((1,1’-二苯基)-4-基甲基)-4-(丙烷-1-炔-1-基)-1H-吲唑-7-甲酸的制备Step 3: Preparation of 1-((1,1’-diphenyl)-4-ylmethyl)-4-(propane-1-yn-1-yl)-1H-indazole-7-carboxylic acid
室温下将原料1-((1,1’-二苯基)-4-基甲基)-4-(丙烷-1-炔-1-基)-1H-吲唑-7-甲酸甲酯(80mg,0.21mmol)加入到四氢呋喃(3mL)，甲醇(1mL)和水(1mL)中，加入氢氧化锂(15mg,0.63mmol)，室温搅拌16h。用1N盐酸调节pH＝4，用乙酸乙酯(20mL×3)萃取，分液，合并有机相，有机相用无水硫酸钠干燥，过滤，浓缩，得化合物粗品1-((1,1’-二苯基)-4-基甲基)-4-(丙烷-1-炔-1-基)-1H-吲唑-7-甲酸(50mg,产率65％)。The starting material 1-((1,1'-diphenyl)-4-ylmethyl)-4-(propan-1-yn-1-yl)-1H-indazole-7-carboxylic acid methyl ester ( 80mg, 0.21mmol) was added to tetrahydrofuran (3mL), methanol (1mL) and water (1mL), lithium hydroxide (15mg, 0.63mmol) was added, and stirred at room temperature for 16h. Use 1N hydrochloric acid to adjust pH=4, extract with ethyl acetate (20mL×3), separate the layers, combine the organic phases, dry the organic phases with anhydrous sodium sulfate, filter, and concentrate to obtain the crude compound 1-((1,1' -diphenyl)-4-ylmethyl)-4-(propan-1-yn-1-yl)-1H-indazole-7-carboxylic acid (50 mg, 65% yield).
第四步：6-(1-((1,1’-二苯基)-4-基甲基)-4-(丙烷-1-炔-1-基)-1H-吲唑-7-甲酰胺)螺环[3.3]庚烷-2-甲酸甲酯的制备The fourth step: 6-(1-((1,1'-diphenyl)-4-ylmethyl)-4-(propane-1-yn-1-yl)-1H-indazole-7-methan Preparation of amido)spiro[3.3]heptane-2-carboxylic acid methyl ester
室温下将化合物1-((1,1’-二苯基)-4-基甲基)-4-(丙烷-1-炔-1-基)-1H-吲唑-7-甲酸(50mg,0.14mmol)加入到N,N-二甲基甲酰胺(3mL)中，加入2-(6-氨基螺环[3.3]庚烷-2-基)甲酸甲酯盐酸盐(43mg,0.21mmol)，二异丙基乙胺(36mg,0.28mmol)，HATU(160mg,0.42mmol)，搅拌16h。加入水(20mL)稀释，用乙酸乙酯(20mL×3)萃取，分液，合并有机相，有机相用无水硫酸钠干燥，过滤，浓缩，残留物用薄层硅胶板分离纯化(石油醚:乙酸乙酯(V/V)＝1:1)得化合物6-(1-((1,1’-二苯基)-4-基甲基)-4-(丙烷-1-炔-1-基)-1H-吲唑-7-甲酰胺)螺环[3.3]庚烷-2-甲酸甲酯(12mg,产率17％)。Compound 1-((1,1'-diphenyl)-4-ylmethyl)-4-(propane-1-yn-1-yl)-1H-indazole-7-carboxylic acid (50mg, 0.14mmol) was added to N,N-dimethylformamide (3mL), and methyl 2-(6-aminospiro[3.3]heptane-2-yl)formate hydrochloride (43mg, 0.21mmol) was added , diisopropylethylamine (36mg, 0.28mmol), HATU (160mg, 0.42mmol), stirred for 16h. Add water (20mL) to dilute, extract with ethyl acetate (20mL×3), separate the layers, combine the organic phases, dry the organic phases with anhydrous sodium sulfate, filter, concentrate, and separate and purify the residue with a thin-layer silica gel plate (petroleum ether : Ethyl acetate (V/V)=1:1) to obtain compound 6-(1-((1,1'-diphenyl)-4-ylmethyl)-4-(propane-1-yne-1 -yl)-lH-indazole-7-carboxamide)spiro[3.3]heptane-2-carboxylic acid methyl ester (12 mg, yield 17%).
第五步：6-(1-((1,1’-二苯基)-4-基甲基)-4-(丙烷-1-炔-1-基)-1H-吲唑-7-甲酰胺)螺环[3.3]庚烷-2-甲酸(I-1)的制备The fifth step: 6-(1-((1,1'-diphenyl)-4-ylmethyl)-4-(propane-1-yn-1-yl)-1H-indazole-7-methan Preparation of amide) spiro[3.3]heptane-2-carboxylic acid (I-1)
Figure PCTCN2022097611-appb-000026
Figure PCTCN2022097611-appb-000026
室温下将化合物6-(1-((1,1’-二苯基)-4-基甲基)-4-(丙烷-1-炔-1-基)-1H-吲唑-7-甲酰胺)螺环[3.3]庚烷-2-甲酸甲酯(12mg,0.02mmol)加入到四氢呋喃(3mL)和水(1mL)中，加入氢氧化锂(2.0mg,0.08mmol)，室温搅拌6h。用1N盐酸调节pH＝4，用乙酸乙酯(20mL×3)萃取，分液，合并有机相，有机相用无水硫酸钠干燥，过滤，浓缩，残留物经酸性制备方法A制备得 化合物6-(1-((1,1’-二苯基)-4-基甲基)-4-(丙烷-1-炔-1-基)-1H-吲唑-7-甲酰胺)螺环[3.3]庚烷-2-甲酸(I-1)(10mg,产率86％)。Compound 6-(1-((1,1'-diphenyl)-4-ylmethyl)-4-(propane-1-yn-1-yl)-1H-indazole-7-methan Amide) spiro[3.3]heptane-2-carboxylic acid methyl ester (12mg, 0.02mmol) was added to tetrahydrofuran (3mL) and water (1mL), lithium hydroxide (2.0mg, 0.08mmol) was added, and stirred at room temperature for 6h. Use 1N hydrochloric acid to adjust pH=4, extract with ethyl acetate (20mL×3), separate the layers, combine the organic phases, dry the organic phases with anhydrous sodium sulfate, filter, concentrate, and prepare the residue by acidic preparation method A to obtain compound 6 -(1-((1,1'-diphenyl)-4-ylmethyl)-4-(propane-1-yn-1-yl)-1H-indazole-7-carboxamide) spiro[ 3.3] Heptane-2-carboxylic acid (I-1) (10 mg, yield 86%).
1H NMR(400mHz,DMSO-d6)δ12.0(s,1H),8.77(d,1H),8.25(s,1H),7.58(d,2H),7.52(d,2H),7.45(t,2H),7.36(d,2H),7.22(d,1H),7.05(d,2H),5.81(s,2H),4.31-4.25(m,1H),2.94(t,1H),2.43-2.37(m,1H),2.32-2.21(m,3H),2.18(s,3H),2.10-1.99(m,2H),1.93-1.83(m,2H)。 1 H NMR (400mHz, DMSO-d6) δ12.0(s,1H),8.77(d,1H),8.25(s,1H),7.58(d,2H),7.52(d,2H),7.45(t ,2H),7.36(d,2H),7.22(d,1H),7.05(d,2H),5.81(s,2H),4.31-4.25(m,1H),2.94(t,1H),2.43- 2.37 (m, 1H), 2.32-2.21 (m, 3H), 2.18 (s, 3H), 2.10-1.99 (m, 2H), 1.93-1.83 (m, 2H).
LC-MS,M/Z(ESI):504.7[M+H] +。 LC-MS, M/Z (ESI): 504.7 [M+H] + .
实施例2化合物I-2的制备The preparation of embodiment 2 compound 1-2
合成路线如下所示：The synthetic route is as follows:
Figure PCTCN2022097611-appb-000027
Figure PCTCN2022097611-appb-000027
第一步：5-(丙烷-1-炔-1-基)-1H-吲唑-7-甲酸甲酯的制备The first step: the preparation of 5-(propane-1-yn-1-yl)-1H-indazole-7-carboxylic acid methyl ester
室温下将化合物5-溴-1H-吲唑-7-甲酸甲酯(1.0g,3.92mmol)加入到1,4-二氧六环(10mL)中，加入二三苯基膦二氯化钯(274mg,0.39mmol)，1-丙炔-三正丁基锡(1.55g,4.70mmol)，氮气保护下加热至90℃，搅拌16h。冷却至室温，加入水(50mL)稀释，用乙酸乙酯(50mL×3)萃取，分液，合并有机相，有机相用无水硫酸钠干燥，过滤，浓缩，残留物用薄层硅胶板分离纯化(石油醚:乙酸乙酯(V/V)＝5:1)得化合物5-(丙烷-1-炔-1-基)-1H-吲唑-7-甲酸甲酯(300mg，产率36％)。Add the compound 5-bromo-1H-indazole-7-carboxylic acid methyl ester (1.0g, 3.92mmol) to 1,4-dioxane (10mL) at room temperature, add ditriphenylphosphine palladium dichloride (274mg, 0.39mmol), 1-propyne-tri-n-butyltin (1.55g, 4.70mmol), heated to 90°C under nitrogen protection, and stirred for 16h. Cool to room temperature, add water (50mL) to dilute, extract with ethyl acetate (50mL×3), separate the liquids, combine the organic phases, dry the organic phases with anhydrous sodium sulfate, filter, concentrate, and separate the residue with a thin-layer silica gel plate Purification (petroleum ether: ethyl acetate (V/V) = 5:1) gave compound 5-(propane-1-yn-1-yl)-1H-indazole-7-carboxylic acid methyl ester (300mg, yield 36 %).
第二步：1-((1,1’-二苯基)-4-基甲基)-5-(丙烷-1-炔-1-基)-1H-吲唑-7-甲酸甲酯的制备The second step: 1-((1,1'-diphenyl)-4-ylmethyl)-5-(propane-1-yn-1-yl)-1H-indazole-7-carboxylic acid methyl ester preparation
室温下将化合物5-(丙烷-1-炔-1-基)-1H-吲唑-7-甲酸甲酯(100mg,0.47mmol)加入到N,N-二甲基甲酰胺(5mL)中，加入4-苯基苄溴(150mg,0.61mmol)，碳酸铯(460mg,1.40mmol)，室温搅拌16h。加入水(20mL)稀释，用乙酸乙酯(20mL×3)萃取，分液，合并有机相，有机相用无水硫酸钠干燥，过滤，浓缩，残留物用薄层硅胶板分离纯化(石油醚:乙酸乙酯(V/V)＝10:1)得化合物1-((1,1’-二苯基)-4-基甲基)-5-(丙烷-1-炔-1-基)-1H-吲唑-7-甲酸甲酯(50mg,产率28％)。The compound 5-(propane-1-yn-1-yl)-1H-indazole-7-carboxylic acid methyl ester (100 mg, 0.47 mmol) was added into N,N-dimethylformamide (5 mL) at room temperature, Add 4-phenylbenzyl bromide (150mg, 0.61mmol), cesium carbonate (460mg, 1.40mmol), and stir at room temperature for 16h. Add water (20mL) to dilute, extract with ethyl acetate (20mL×3), separate the layers, combine the organic phases, dry the organic phases with anhydrous sodium sulfate, filter, concentrate, and separate and purify the residue with a thin-layer silica gel plate (petroleum ether : Ethyl acetate (V/V)=10:1) to obtain compound 1-((1,1'-diphenyl)-4-ylmethyl)-5-(propane-1-yn-1-yl) - 1H-indazole-7-carboxylic acid methyl ester (50 mg, yield 28%).
第三步：1-((1,1’-二苯基)-4-基甲基)-5-(丙烷-1-炔-1-基)-1H-吲唑-7-甲酸的制备Step 3: Preparation of 1-((1,1’-diphenyl)-4-ylmethyl)-5-(propane-1-yn-1-yl)-1H-indazole-7-carboxylic acid
室温下将原料1-((1,1’-二苯基)-4-基甲基)-5-(丙烷-1-炔-1-基)-1H-吲唑-7-甲酸甲酯(50mg,0.13mmol)加入到四氢呋喃(3mL)，甲醇(1mL)和水(1mL)中，加入氢氧化锂(10mg,0.40mmol)，室温搅拌4h。用1N盐酸调节pH＝4，用乙酸乙酯(20mL×3)萃取，分液，合并有机相，有机相用无水硫酸钠干燥，过滤，浓缩，得化合物粗品1-((1,1’-二苯基)-4-基甲基)-5-(丙烷-1-炔-1-基)-1H-吲唑-7-甲酸(40mg,产率83％).The starting material 1-((1,1'-diphenyl)-4-ylmethyl)-5-(propane-1-yn-1-yl)-1H-indazole-7-carboxylic acid methyl ester ( 50mg, 0.13mmol) was added to tetrahydrofuran (3mL), methanol (1mL) and water (1mL), lithium hydroxide (10mg, 0.40mmol) was added, and stirred at room temperature for 4h. Use 1N hydrochloric acid to adjust pH=4, extract with ethyl acetate (20mL×3), separate the layers, combine the organic phases, dry the organic phases with anhydrous sodium sulfate, filter, and concentrate to obtain the crude compound 1-((1,1' -Diphenyl)-4-ylmethyl)-5-(propane-1-yn-1-yl)-1H-indazole-7-carboxylic acid (40mg, yield 83%).
第四步：6-(1-((1,1’-二苯基)-4-基甲基)-5-(丙烷-1-炔-1-基)-1H-吲唑-7-甲酰胺)螺环[3.3]庚烷-2-甲酸甲酯的制备The fourth step: 6-(1-((1,1'-diphenyl)-4-ylmethyl)-5-(propane-1-yn-1-yl)-1H-indazole-7-methan Preparation of amido)spiro[3.3]heptane-2-carboxylic acid methyl ester
室温下将化合物1-((1,1’-二苯基)-4-基甲基)-5-(丙烷-1-炔-1-基)-1H-吲唑-7-甲酸(40mg,0.11mmol)加入到N,N-二甲基甲酰胺(3mL)中，加入2-(6-氨基螺环[3.3]庚烷-2-基)甲酸甲酯盐酸盐(34mg,0.16mmol)，二异丙基乙胺(43mg,0.33mmol)，HATU(63mg,0.16mmol)，搅拌16h。加入水(20mL)稀释，用乙酸乙酯(20mL×3)萃取，分液，合并有机相，有机相用无水硫酸钠干燥，过滤，浓缩，残留物用薄层硅胶板分离纯化(石油醚:乙酸乙酯(V/V)＝1:1)得化合物6-(1-((1,1’-二苯基)-4-基甲基)-5-(丙烷-1-炔-1-基)-1H-吲唑-7-甲酰胺)螺环[3.3]庚烷-2-甲酸甲酯(50mg,产率88％)。Compound 1-((1,1'-diphenyl)-4-ylmethyl)-5-(propane-1-yn-1-yl)-1H-indazole-7-carboxylic acid (40mg, 0.11mmol) was added to N,N-dimethylformamide (3mL), and methyl 2-(6-aminospiro[3.3]heptane-2-yl)formate hydrochloride (34mg, 0.16mmol) was added , diisopropylethylamine (43mg, 0.33mmol), HATU (63mg, 0.16mmol), stirred for 16h. Add water (20mL) to dilute, extract with ethyl acetate (20mL×3), separate the layers, combine the organic phases, dry the organic phases with anhydrous sodium sulfate, filter, concentrate, and separate and purify the residue with a thin-layer silica gel plate (petroleum ether : Ethyl acetate (V/V)=1:1) to obtain compound 6-(1-((1,1'-diphenyl)-4-ylmethyl)-5-(propane-1-yne-1 -yl)-1H-indazole-7-carboxamide)spiro[3.3]heptane-2-carboxylic acid methyl ester (50 mg, yield 88%).
第五步：6-(1-((1,1’-二苯基)-4-基甲基)-5-(丙烷-1-炔-1-基)-1H-吲唑-7-甲酰胺)螺环[3.3]庚烷-2-甲酸(I-2)的制备The fifth step: 6-(1-((1,1'-diphenyl)-4-ylmethyl)-5-(propane-1-yn-1-yl)-1H-indazole-7-methan Preparation of amide) spiro[3.3]heptane-2-carboxylic acid (I-2)
Figure PCTCN2022097611-appb-000028
Figure PCTCN2022097611-appb-000028
室温下将化合物6-(1-((1,1’-二苯基)-4-基甲基)-5-(丙烷-1-炔-1-基)-1H-吲唑-7-甲酰胺)螺环[3.3]庚烷-2-甲酸甲酯(50mg,0.10mmol)加入到四氢呋喃(3mL)，甲醇(1mL)和水(1mL)中，加入氢氧化锂(8.0mg,0.33mmol)，室温搅拌16h。用1N盐酸调节pH＝4，用乙酸乙酯(20mL×3)萃取，分液，合并有机相，有机相用无水硫酸钠干燥，过滤，浓缩，残留物经酸性制备方法A制备得化合物白色固体6-(1-((1,1’-二苯基)-4-基甲基)-5-(丙烷-1-炔-1-基)-1H-吲唑-7-甲酰胺)螺环[3.3]庚烷-2-甲酸(I-2)(12mg,产率25％)。Compound 6-(1-((1,1'-diphenyl)-4-ylmethyl)-5-(propane-1-yn-1-yl)-1H-indazole-7-meth Amide) spiro[3.3]heptane-2-carboxylate (50mg, 0.10mmol) was added to tetrahydrofuran (3mL), methanol (1mL) and water (1mL), lithium hydroxide (8.0mg, 0.33mmol) was added , stirred at room temperature for 16h. Use 1N hydrochloric acid to adjust pH=4, extract with ethyl acetate (20mL×3), separate the layers, combine the organic phases, dry the organic phases with anhydrous sodium sulfate, filter, concentrate, and the residue is prepared by acidic preparation method A to obtain compound white Solid 6-(1-((1,1'-diphenyl)-4-ylmethyl)-5-(propane-1-yn-1-yl)-1H-indazole-7-carboxamide)spiro Cyclo[3.3]heptane-2-carboxylic acid (I-2) (12 mg, 25% yield).
1H NMR(400mHz,DMSO-d6)δ11.9(s,1H),8.79(d,1H),8.26(s,1H),7.93(s,1H),7.58(d,2H),7.52(d,2H),7.45(t,2H),7.37(t,2H),7.05(d,2H),5.79(s,2H),4.29-4.23(m,1H),2.96(t,1H),2.43-2.37(m,1H),2.32-2.21(m,3H),2.18(s,3H),2.11-1.98(m,2H),1.99-1.84(m,2H)。 1 H NMR(400mHz,DMSO-d6)δ11.9(s,1H),8.79(d,1H),8.26(s,1H),7.93(s,1H),7.58(d,2H),7.52(d ,2H),7.45(t,2H),7.37(t,2H),7.05(d,2H),5.79(s,2H),4.29-4.23(m,1H),2.96(t,1H),2.43- 2.37 (m, 1H), 2.32-2.21 (m, 3H), 2.18 (s, 3H), 2.11-1.98 (m, 2H), 1.99-1.84 (m, 2H).
LC-MS,M/Z(ESI):504.6[M+H] +。 LC-MS, M/Z (ESI): 504.6 [M+H] + .
实施例3化合物I-3的制备The preparation of embodiment 3 compound 1-3
合成路线如下所示：The synthetic route is as follows:
Figure PCTCN2022097611-appb-000029
Figure PCTCN2022097611-appb-000029
第一步：4-(丁烷-1-炔-1-基)-1H-吲唑-7-甲酸甲酯的制备The first step: the preparation of 4-(butane-1-yn-1-yl)-1H-indazole-7-carboxylic acid methyl ester
室温下将化合物4-溴-1H-吲唑-7-甲酸甲酯(600mg,2.35mmol)加入到N,N-二甲基乙酰胺(6mL)中，加入醋酸钯(70mg,0.31mmol)，(丁-1-炔-1-基)三甲基硅烷(595mg,4.71mmol)，碘化亚铜(138mg,0.72mmol)，碳酸铯(1.53g,4.71mmol)，氮气保护下加热至80℃，搅拌16h。冷却至室温，加入水(50mL)稀释，用乙酸乙酯(50mL×3)萃取，分液，合并有机相，有机相用无水硫酸钠干燥，过滤，浓缩，残留物用薄层硅胶板分离纯化(石油醚:乙酸乙酯(V/V)＝5:1)得化合物4-(丁烷-1-炔-1-基)-1H-吲唑-7-甲酸甲酯(200mg,产率37％)。Compound 4-bromo-1H-indazole-7-carboxylic acid methyl ester (600mg, 2.35mmol) was added to N,N-dimethylacetamide (6mL) at room temperature, palladium acetate (70mg, 0.31mmol) was added, (But-1-yn-1-yl)trimethylsilane (595mg, 4.71mmol), cuprous iodide (138mg, 0.72mmol), cesium carbonate (1.53g, 4.71mmol), heated to 80°C under nitrogen protection , stirred for 16h. Cool to room temperature, add water (50mL) to dilute, extract with ethyl acetate (50mL×3), separate the liquids, combine the organic phases, dry the organic phases with anhydrous sodium sulfate, filter, concentrate, and separate the residue with a thin-layer silica gel plate Purification (petroleum ether: ethyl acetate (V/V) = 5:1) gave compound 4-(butane-1-yn-1-yl)-1H-indazole-7-carboxylic acid methyl ester (200mg, yield 37%).
第二步：1-((1,1’-二苯基)-4-基甲基)-4-(丁烷-1-炔-1-基)-1H-吲唑-7-甲酸甲酯的制备The second step: 1-((1,1'-diphenyl)-4-ylmethyl)-4-(butane-1-yn-1-yl)-1H-indazole-7-carboxylic acid methyl ester preparation of
室温下将化合物4-(丁烷-1-炔-1-基)-1H-吲唑-7-甲酸甲酯(150mg,0.66mmol)加入到N,N-二甲基甲酰胺(2mL)中，加入4-苯基苄溴(245mg,0.99mmol)，碳酸铯(645mg,1.98mmol)，室温搅拌16h。加入水(20mL)稀释，用乙酸乙酯(20mL×3)萃取，分液，合并有机相，有机相用无水硫酸钠干燥，过滤，浓缩，残留物用薄层硅胶板分离纯化(石油醚:乙酸乙酯(V/V)＝1:1)得化合物1-((1,1’-二苯基)-4-基甲基)-4-(丁烷-1-炔-1-基)-1H-吲唑-7-甲酸甲酯(130mg,产率50％)。Compound 4-(butane-1-yn-1-yl)-1H-indazole-7-carboxylic acid methyl ester (150 mg, 0.66 mmol) was added to N,N-dimethylformamide (2 mL) at room temperature , Add 4-phenylbenzyl bromide (245mg, 0.99mmol), cesium carbonate (645mg, 1.98mmol), and stir at room temperature for 16h. Add water (20mL) to dilute, extract with ethyl acetate (20mL×3), separate the layers, combine the organic phases, dry the organic phases with anhydrous sodium sulfate, filter, concentrate, and separate and purify the residue with a thin-layer silica gel plate (petroleum ether : Ethyl acetate (V/V)=1:1) to obtain compound 1-((1,1'-diphenyl)-4-ylmethyl)-4-(butane-1-yn-1-yl )-1H-indazole-7-carboxylic acid methyl ester (130 mg, yield 50%).
第三步：1-((1,1’-二苯基)-4-基甲基)-4-(丁烷-1-炔-1-基)-1H-吲唑-7-甲酸的制备Step 3: Preparation of 1-((1,1’-diphenyl)-4-ylmethyl)-4-(butane-1-yn-1-yl)-1H-indazole-7-carboxylic acid
室温下将原料1-((1,1’-二苯基)-4-基甲基)-4-(丁烷-1-炔-1-基)-1H-吲唑-7-甲酸甲酯(130mg,0.33mmol)加入到四氢呋喃(3mL)，甲醇(1mL)和水(1mL)中，加入氢氧化锂(24mg,1.0mmol)，室温搅拌16h。用1N盐酸调节pH＝4，用乙酸乙酯(20mL×3)萃取，分液，合并有机相，有机相用无水硫酸钠干燥，过滤，浓缩，得化合物粗品1-((1,1’-二苯基)-4-基甲基)-4-(丁烷-1-炔-1-基)-1H-吲唑-7-甲酸(120mg,产率96％).Starting material 1-((1,1'-diphenyl)-4-ylmethyl)-4-(butane-1-yn-1-yl)-1H-indazole-7-carboxylic acid methyl ester at room temperature (130mg, 0.33mmol) was added to tetrahydrofuran (3mL), methanol (1mL) and water (1mL), lithium hydroxide (24mg, 1.0mmol) was added, and stirred at room temperature for 16h. Use 1N hydrochloric acid to adjust pH=4, extract with ethyl acetate (20mL×3), separate the layers, combine the organic phases, dry the organic phases with anhydrous sodium sulfate, filter, and concentrate to obtain the crude compound 1-((1,1' -Diphenyl)-4-ylmethyl)-4-(butane-1-yn-1-yl)-1H-indazole-7-carboxylic acid (120mg, yield 96%).
第四步：6-(1-((1,1’-二苯基)-4-基甲基)-4-(丁烷-1-炔-1-基)-1H-吲唑-7-甲酰胺)螺环[3.3]庚烷-2-甲酸甲酯的制备The fourth step: 6-(1-((1,1'-diphenyl)-4-ylmethyl)-4-(butane-1-yn-1-yl)-1H-indazole-7- Preparation of formamide) spiro[3.3]heptane-2-carboxylic acid methyl ester
室温下将化合物1-((1,1’-二苯基)-4-基甲基)-4-(丁烷-1-炔-1-基)-1H-吲唑-7-甲酸(120mg,0.32mmol)加入到N,N-二甲基甲酰胺(2mL)中，加入2-(6-氨基螺环[3.3]庚烷-2-基)甲酸甲酯盐 酸盐(78mg,0.38mmol)，二异丙基乙胺(123mg,0.95mmol)，HATU(180mg,0.47mmol)，搅拌16h。加入水(20mL)稀释，用乙酸乙酯(20mL×3)萃取，分液，合并有机相，有机相用无水硫酸钠干燥，过滤，浓缩，残留物用薄层硅胶板分离纯化(DCM:甲醇(V/V)＝1:1)得化合物白色固体2-(6-(1-((1,1’-二苯基)-4-基甲基)-4-(丁烷-1-炔-1-基)-1H-吲唑-7-甲酰胺)螺环[3.3]庚烷-2-基)甲酸甲酯(125mg,产率75％)。Compound 1-((1,1'-diphenyl)-4-ylmethyl)-4-(butane-1-yn-1-yl)-1H-indazole-7-carboxylic acid (120mg , 0.32mmol) was added to N,N-dimethylformamide (2mL), and methyl 2-(6-aminospiro[3.3]heptane-2-yl)formate hydrochloride (78mg, 0.38mmol ), diisopropylethylamine (123mg, 0.95mmol), HATU (180mg, 0.47mmol), stirred for 16h. Add water (20mL) to dilute, extract with ethyl acetate (20mL×3), separate the layers, combine the organic phases, dry the organic phases with anhydrous sodium sulfate, filter, concentrate, and the residue is separated and purified with a thin-layer silica gel plate (DCM: Methanol (V/V)=1:1) to obtain compound 2-(6-(1-((1,1'-diphenyl)-4-ylmethyl)-4-(butane-1- Alkyn-1-yl)-1H-indazole-7-carboxamide)spiro[3.3]heptan-2-yl)methyl carboxylate (125 mg, 75% yield).
第五步：6-(1-((1,1’-二苯基)-4-基甲基)-4-(丁烷-1-炔-1-基)-1H-吲唑-7-甲酰胺)螺环[3.3]庚烷-2-甲酸(I-3)的制备The fifth step: 6-(1-((1,1'-diphenyl)-4-ylmethyl)-4-(butane-1-yn-1-yl)-1H-indazole-7- Formamide) spiro[3.3]heptane-2-carboxylic acid (I-3) preparation
Figure PCTCN2022097611-appb-000030
Figure PCTCN2022097611-appb-000030
室温下将化合物2-(6-(1-((1,1’-二苯基)-4-基甲基)-4-(丁烷-1-炔-1-基)-1H-吲唑-7-甲酰胺)螺环[3.3]庚烷-2-基)甲酸甲酯(125mg,0.24mmol)加入到四氢呋喃(3mL)和水(1mL)中，加入氢氧化锂(17.2mg,0.72mmol)，室温搅拌16h。用1N盐酸调节pH＝4，用乙酸乙酯(20mL×3)萃取，分液，合并有机相，有机相用无水硫酸钠干燥，过滤，浓缩，得化合物白色固体6-(1-((1,1’-二苯基)-4-基甲基)-4-(丁烷-1-炔-1-基)-1H-吲唑-7-甲酰胺)螺环[3.3]庚烷-2-甲酸(I-3)(52mg,产率43％)。Compound 2-(6-(1-((1,1'-diphenyl)-4-ylmethyl)-4-(butane-1-yn-1-yl)-1H-indazole -7-carboxamide)spiro[3.3]heptan-2-yl)methyl carboxylate (125mg, 0.24mmol) was added to tetrahydrofuran (3mL) and water (1mL), lithium hydroxide (17.2mg, 0.72mmol ), stirred at room temperature for 16h. Adjust pH=4 with 1N hydrochloric acid, extract with ethyl acetate (20mL×3), separate the layers, combine the organic phases, dry the organic phases with anhydrous sodium sulfate, filter, and concentrate to obtain compound 6-(1-(( 1,1'-diphenyl)-4-ylmethyl)-4-(butane-1-yn-1-yl)-1H-indazole-7-carboxamide)spiro[3.3]heptane- 2-Formic acid (I-3) (52 mg, yield 43%).
1H NMR(400mHz,DMSO-d6)δ12.0(s,1H),8.78(d,1H),8.25(s,1H),7.58(d,2H),7.52(d,2H),7.45(t,2H),7.37(d,2H),7.21(d,1H),7.05(d,2H),5.81(s,2H),4.31-4.25(m,1H),2.94(t,1H),2.58-2.53(m,2H),2.42-2.30(m,1H),2.27-2.19(m,3H),2.11-2.01(m,2H),1.94-1.83(m,2H),1.26(t,3H)。 1 H NMR(400mHz,DMSO-d6)δ12.0(s,1H),8.78(d,1H),8.25(s,1H),7.58(d,2H),7.52(d,2H),7.45(t ,2H),7.37(d,2H),7.21(d,1H),7.05(d,2H),5.81(s,2H),4.31-4.25(m,1H),2.94(t,1H),2.58- 2.53 (m, 2H), 2.42-2.30 (m, 1H), 2.27-2.19 (m, 3H), 2.11-2.01 (m, 2H), 1.94-1.83 (m, 2H), 1.26 (t, 3H).
LC-MS,M/Z(ESI):518.5[M+H] +。 LC-MS, M/Z (ESI): 518.5 [M+H] + .
实施例4化合物I-4的制备The preparation of embodiment 4 compound I-4
合成路线如下所示：The synthetic route is as follows:
Figure PCTCN2022097611-appb-000031
Figure PCTCN2022097611-appb-000031
第一步：1-((1,1’-二苯基)-4-基甲基)-4-溴-1H-吲唑-7-甲酸甲酯的制备Step 1: Preparation of 1-((1,1’-diphenyl)-4-ylmethyl)-4-bromo-1H-indazole-7-carboxylic acid methyl ester
室温下将化合物4-溴-1H-吲唑-7-甲酸甲酯(600mg,2.36mmol)加入到N,N-二甲基甲酰胺(10mL)中，加入4-苯基苄溴(875mg,3.54mmol)，碳酸铯(2.3g,7.09mmol)，室温搅拌16h。加入水(20mL)稀释，用乙酸乙酯(30mL×3)萃取，分液，合并有机相，有机相用无水硫酸钠干燥，过滤，浓缩，残留物用薄层硅胶板分离纯化(石油醚:乙酸乙酯(V/V)＝5:1)得化合物白色固体1-((1,1’-二苯基)-4-基甲基)-4-溴-1H-吲唑-7-甲酸甲酯(420mg,产率42％)。The compound 4-bromo-1H-indazole-7-carboxylic acid methyl ester (600mg, 2.36mmol) was added to N,N-dimethylformamide (10mL) at room temperature, and 4-phenylbenzyl bromide (875mg, 3.54mmol), cesium carbonate (2.3g, 7.09mmol), stirred at room temperature for 16h. Add water (20mL) to dilute, extract with ethyl acetate (30mL×3), separate the layers, combine the organic phases, dry the organic phases with anhydrous sodium sulfate, filter, concentrate, and separate and purify the residue with a thin-layer silica gel plate (petroleum ether : Ethyl acetate (V/V)=5:1) to obtain compound white solid 1-((1,1'-diphenyl)-4-ylmethyl)-4-bromo-1H-indazole-7- Methyl formate (420 mg, 42% yield).
第二步：1-((1,1’-二苯基)-4-基甲基)-4-(环丙烷乙炔基)-1H-吲唑-7-甲酸甲酯的制备The second step: the preparation of 1-((1,1'-diphenyl)-4-ylmethyl)-4-(cyclopropaneethynyl)-1H-indazole-7-carboxylic acid methyl ester
室温下将化合物1-((1,1’-二苯基)-4-基甲基)-4-溴-1H-吲唑-7-甲酸甲酯(550mg,1.31mmol)加入到氮，氮-二甲基乙酰胺(10mL)中，加入醋酸钯(38mg,0.17mmol)，环丙基乙炔(432mg,6.55mmol)，碘化亚铜(25mg,0.13mmol)，碳酸铯(635mg,1.97mmol)，氮气保护下加热至80℃，搅拌16h。冷却至室温，加入水(50mL)稀释，用乙酸乙酯(50mL×3)萃取，分液，合并有机相，有机相用无水硫酸钠干燥，过滤，浓缩，残留物用薄层硅胶板分离纯化(石油醚:乙酸乙酯(V/V)＝10:1)得化合物白色固体1-((1,1’-二苯基)-4-基甲基)-4-(环丙烷乙炔基)-1H-吲唑-7-甲酸甲酯(80mg,产率15％)。Compound 1-((1,1'-diphenyl)-4-ylmethyl)-4-bromo-1H-indazole-7-carboxylic acid methyl ester (550 mg, 1.31 mmol) was added to nitrogen at room temperature, nitrogen - In dimethylacetamide (10mL), add palladium acetate (38mg, 0.17mmol), cyclopropylacetylene (432mg, 6.55mmol), cuprous iodide (25mg, 0.13mmol), cesium carbonate (635mg, 1.97mmol ), heated to 80°C under nitrogen protection, and stirred for 16h. Cool to room temperature, add water (50mL) to dilute, extract with ethyl acetate (50mL×3), separate the liquids, combine the organic phases, dry the organic phases with anhydrous sodium sulfate, filter, concentrate, and separate the residue with a thin-layer silica gel plate Purification (petroleum ether: ethyl acetate (V/V) = 10:1) gave the compound 1-((1,1'-diphenyl)-4-ylmethyl)-4-(cyclopropaneethynyl) as a white solid )-1H-indazole-7-carboxylic acid methyl ester (80 mg, yield 15%).
第三步：1-((1,1’-二苯基)-4-基甲基)-4-(环丙烷乙炔基)-1H-吲唑-7-甲酸的制备Step 3: Preparation of 1-((1,1’-diphenyl)-4-ylmethyl)-4-(cyclopropaneethynyl)-1H-indazole-7-carboxylic acid
室温下将原料1-((1,1’-二苯基)-4-基甲基)-4-(环丙烷乙炔基)-1H-吲唑-7-甲酸甲酯(80mg,0.20mmol)加入到四氢呋喃(5mL)和水(2mL)中，加入氢氧化锂(15mg,0.60mmol)，室温搅拌16h。用1N盐酸调节pH＝4，用乙酸乙酯(20mL×3)萃取，分液，合并有机相，有机相用无水硫酸钠干燥，过滤，浓缩，残留物用薄层硅胶板分离纯化(石油醚:乙酸乙酯(V/V)＝1:1)得化合物白色固体1-((1,1’-二苯基)-4-基甲基)-4-(环丙烷乙炔基)-1H-吲唑-7-甲酸(60mg,产率78％)。Starting material 1-((1,1'-diphenyl)-4-ylmethyl)-4-(cyclopropaneethynyl)-1H-indazole-7-carboxylic acid methyl ester (80mg, 0.20mmol) at room temperature Add to tetrahydrofuran (5mL) and water (2mL), add lithium hydroxide (15mg, 0.60mmol), stir at room temperature for 16h. Use 1N hydrochloric acid to adjust pH=4, extract with ethyl acetate (20mL×3), separate the layers, combine the organic phases, dry the organic phases with anhydrous sodium sulfate, filter, concentrate, and separate and purify the residue with a thin-layer silica gel plate (petroleum Ether: ethyl acetate (V/V) = 1:1) to obtain compound 1-((1,1'-diphenyl)-4-ylmethyl)-4-(cyclopropaneethynyl)-1H as a white solid - Indazole-7-carboxylic acid (60 mg, 78% yield).
第四步：6-(1-((1,1’-二苯基)-4-基甲基)-4-(环丙烷乙炔基)-1H-吲唑-7-甲酰胺)螺环[3.3]庚烷-2-甲酸甲酯的制备The fourth step: 6-(1-((1,1'-diphenyl)-4-ylmethyl)-4-(cyclopropaneethynyl)-1H-indazole-7-carboxamide) spiro[ 3.3] Preparation of methyl heptane-2-carboxylate
室温下将化合物1-((1,1’-二苯基)-4-基甲基)-4-(环丙烷乙炔基)-1H-吲唑-7-甲酸(60mg,0.15mmol)加入到N,N-二甲基甲酰胺(5mL)中，加入2-(6-氨基螺环[3.3]庚烷-2-基)甲酸甲酯盐酸盐 (47mg,0.23mmol)，二异丙基乙胺(59mg,0.46mmol)，HATU(87mg,0.23mmol)，搅拌16h。加入水(20mL)稀释，用乙酸乙酯(20mL×3)萃取，分液，合并有机相，有机相用无水硫酸钠干燥，过滤，浓缩，残留物用薄层硅胶板分离纯化(PE:EA(V/V)＝1:1)得化合物2-(6-(1-((1,1’-二苯基)-4-基甲基)-4-(环丙烷乙炔基)-1H-吲唑-7-甲酰胺)螺环[3.3]庚烷-2-基)甲酸甲酯(40mg,产率48％)。Compound 1-((1,1'-diphenyl)-4-ylmethyl)-4-(cyclopropaneethynyl)-1H-indazole-7-carboxylic acid (60mg, 0.15mmol) was added to In N,N-dimethylformamide (5mL), add 2-(6-aminospiro[3.3]heptane-2-yl)methyl formate hydrochloride (47mg, 0.23mmol), diisopropyl Ethylamine (59mg, 0.46mmol), HATU (87mg, 0.23mmol), stirred for 16h. Add water (20mL) to dilute, extract with ethyl acetate (20mL×3), separate the layers, combine the organic phases, dry the organic phases with anhydrous sodium sulfate, filter, concentrate, and the residue is separated and purified with a thin-layer silica gel plate (PE: EA (V/V)=1:1) to obtain compound 2-(6-(1-((1,1'-diphenyl)-4-ylmethyl)-4-(cyclopropaneethynyl)-1H -indazole-7-carboxamide)spiro[3.3]heptan-2-yl)methyl carboxylate (40 mg, 48% yield).
第五步：6-(1-((1,1’-二苯基)-4-基甲基)-4-(环丙烷乙炔基)-1H-吲唑-7-甲酰胺)螺环[3.3]庚烷-2-甲酸(I-4)的制备The fifth step: 6-(1-((1,1'-diphenyl)-4-ylmethyl)-4-(cyclopropaneethynyl)-1H-indazole-7-carboxamide) spiro[ 3.3] Preparation of heptane-2-carboxylic acid (I-4)
Figure PCTCN2022097611-appb-000032
Figure PCTCN2022097611-appb-000032
室温下将化合物2-(6-(1-((1,1’-二苯基)-4-基甲基)-4-(环丙烷乙炔基)-1H-吲唑-7-甲酰胺)螺环[3.3]庚烷-2-基)甲酸甲酯(40mg,0.073mmol)加入到四氢呋喃(3mL)和水(1mL)中，加入氢氧化锂(5.3mg,0.22mmol)，室温搅拌16h。用1N盐酸调节pH＝4，过滤，干燥得化合物6-(1-((1,1’-二苯基)-4-基甲基)-4-(环丙烷乙炔基)-1H-吲唑-7-甲酰胺)螺环[3.3]庚烷-2-甲酸(I-4)(32mg,产率82％)。Compound 2-(6-(1-((1,1'-diphenyl)-4-ylmethyl)-4-(cyclopropaneethynyl)-1H-indazole-7-carboxamide) Spiro[3.3]heptan-2-yl)methyl carboxylate (40mg, 0.073mmol) was added to tetrahydrofuran (3mL) and water (1mL), lithium hydroxide (5.3mg, 0.22mmol) was added, and stirred at room temperature for 16h. Adjust pH=4 with 1N hydrochloric acid, filter and dry to obtain compound 6-(1-((1,1'-diphenyl)-4-ylmethyl)-4-(cyclopropaneethynyl)-1H-indazole -7-carboxamide) spiro[3.3]heptane-2-carboxylic acid (I-4) (32 mg, 82% yield).
1H NMR(400mHz,DMSO-d6)δ11.9(s,1H),8.76(d,1H),8.24(s,1H),7.58(d,2H),7.52(d,2H),7.43(t,2H),7.33(d,2H),7.18(d,1H),7.03(d,2H),5.80(s,2H),4.30-4.24(m,1H),2.94(t,1H),2.45-2.35(m,1H),2.27-2.18(m,3H),2.11-1.99(m,2H),1.92-1.82(m,2H),1.66(t,1H),0.97-0.88(m,2H),0.88-0.86(m,2H)。 1 H NMR(400mHz,DMSO-d6)δ11.9(s,1H),8.76(d,1H),8.24(s,1H),7.58(d,2H),7.52(d,2H),7.43(t ,2H),7.33(d,2H),7.18(d,1H),7.03(d,2H),5.80(s,2H),4.30-4.24(m,1H),2.94(t,1H),2.45- 2.35(m,1H),2.27-2.18(m,3H),2.11-1.99(m,2H),1.92-1.82(m,2H),1.66(t,1H),0.97-0.88(m,2H), 0.88-0.86(m,2H).
LC-MS,M/Z(ESI):530.5[M+H] +。 LC-MS, M/Z (ESI): 530.5 [M+H] + .
实施例5化合物I-5的制备The preparation of embodiment 5 compound 1-5
合成路线如下所示：The synthetic route is as follows:
Figure PCTCN2022097611-appb-000033
Figure PCTCN2022097611-appb-000033
第一步：1-((1,1’-二苯基)-4-基甲基)-4-溴-1H-吲唑-7-甲酸甲酯的制备Step 1: Preparation of 1-((1,1’-diphenyl)-4-ylmethyl)-4-bromo-1H-indazole-7-carboxylic acid methyl ester
室温下将化合物4-溴-1H-吲唑-7-甲酸甲酯(600mg,2.36mmol)加入到N,N-二甲基甲酰胺(10mL)中，加入4-苯基苄溴(875mg,3.54mmol)，碳酸铯(2.3g,7.09mmol)，室温搅拌16h。加入水(20mL)稀释，用乙酸乙酯(30mL×3)萃取，分液，合并有机相，有机相用无水硫酸钠干燥，过滤，浓缩，残留物用薄层硅胶板分离纯化(石油醚:乙酸乙酯(V/V)＝5:1)得化合物1-((1,1’-二苯基)-4-基甲基)-4-溴-1H-吲唑-7-甲酸甲酯(420mg,产率42％)。The compound 4-bromo-1H-indazole-7-carboxylic acid methyl ester (600mg, 2.36mmol) was added to N,N-dimethylformamide (10mL) at room temperature, and 4-phenylbenzyl bromide (875mg, 3.54mmol), cesium carbonate (2.3g, 7.09mmol), stirred at room temperature for 16h. Add water (20mL) to dilute, extract with ethyl acetate (30mL×3), separate the layers, combine the organic phases, dry the organic phases with anhydrous sodium sulfate, filter, concentrate, and separate and purify the residue with a thin-layer silica gel plate (petroleum ether : Ethyl acetate (V/V)=5:1) to obtain compound 1-((1,1'-diphenyl)-4-ylmethyl)-4-bromo-1H-indazole-7-formic acid Ester (420 mg, 42% yield).
第二步：1-((1,1’-二苯基)-4-基甲基)-4-(3-羟基-3-甲基丁烷-1-炔-1-基)-1H-吲唑-7-甲酸甲酯的制备The second step: 1-((1,1'-diphenyl)-4-ylmethyl)-4-(3-hydroxy-3-methylbutane-1-yn-1-yl)-1H- Preparation of Methyl Indazole-7-carboxylate
室温下将化合物1-((1,1’-二苯基)-4-基甲基)-4-溴-1H-吲唑-7-甲酸甲酯(120mg,0.29mmol)加入到氮，氮-二甲基乙酰胺(5mL)中，加入醋酸钯(9mg,0.04mmol)，2-甲基丁烷-3-炔-2-醇(122mg,1.45mmol)，碘化亚铜(6mg,0.03mmol)，碳酸铯(141mg,0.43mmol)，氮气保护下加热至80℃，搅拌18h。冷却至室温，加入水(50mL)稀释，用乙酸乙酯(50mL×3)萃取，分液，合并有机相，有机相用无水硫酸钠干燥，过滤，浓缩，残留物用薄层硅胶板分离纯化(石油醚:乙酸乙酯(V/V)＝10:1)得化合物1-((1,1’-二苯基)-4-基甲基)-4-(3-羟基-3-甲基丁烷-1-炔-1-基)-1H-吲唑-7-甲酸甲酯(70mg,产率58％)。Compound 1-((1,1'-diphenyl)-4-ylmethyl)-4-bromo-1H-indazole-7-carboxylic acid methyl ester (120 mg, 0.29 mmol) was added to nitrogen at room temperature, nitrogen -Dimethylacetamide (5mL), add palladium acetate (9mg, 0.04mmol), 2-methylbutane-3-yn-2-ol (122mg, 1.45mmol), cuprous iodide (6mg, 0.03 mmol), cesium carbonate (141mg, 0.43mmol), heated to 80°C under nitrogen protection, and stirred for 18h. Cool to room temperature, add water (50mL) to dilute, extract with ethyl acetate (50mL×3), separate the liquids, combine the organic phases, dry the organic phases with anhydrous sodium sulfate, filter, concentrate, and separate the residue with a thin-layer silica gel plate Purification (petroleum ether: ethyl acetate (V/V) = 10:1) yielded compound 1-((1,1'-diphenyl)-4-ylmethyl)-4-(3-hydroxyl-3- Methylbutane-1-yn-1-yl)-1H-indazole-7-carboxylic acid methyl ester (70 mg, 58% yield).
第三步：1-((1,1’-二苯基)-4-基甲基)-4-(3-羟基-3-甲基丁烷-1-炔-1-基)-1H-吲唑-7-甲酸的制备The third step: 1-((1,1'-diphenyl)-4-ylmethyl)-4-(3-hydroxy-3-methylbutane-1-yn-1-yl)-1H- Preparation of indazole-7-carboxylic acid
室温下将原料1-((1,1’-二苯基)-4-基甲基)-4-(3-羟基-3-甲基丁烷-1-炔-1-基)-1H-吲唑-7-甲酸甲酯(70mg,0.16mmol)加入到四氢呋喃(3mL)和水(1mL)中，加入氢氧化锂(12mg,0.50mmol)，室温搅拌16h。用1N盐酸调节pH＝4，过滤，干燥，得化合物粗品1-((1,1’-二苯基)-4-基甲基)-4-(3-羟基-3-甲基丁烷-1-炔-1-基)-1H-吲唑-7-甲酸(40mg,产率59％).The starting material 1-((1,1'-diphenyl)-4-ylmethyl)-4-(3-hydroxy-3-methylbutane-1-yn-1-yl)-1H- Add methyl indazole-7-carboxylate (70mg, 0.16mmol) into tetrahydrofuran (3mL) and water (1mL), add lithium hydroxide (12mg, 0.50mmol), and stir at room temperature for 16h. Use 1N hydrochloric acid to adjust pH=4, filter and dry to obtain the crude product 1-((1,1'-diphenyl)-4-ylmethyl)-4-(3-hydroxy-3-methylbutane- 1-Alkyn-1-yl)-1H-indazole-7-carboxylic acid (40mg, yield 59%).
第四步：6-(1-((1,1’-二苯基)-4-基甲基)-4-(3-羟基-3-甲基丁烷-1-炔-1-基)-1H-吲唑-7-甲酰胺)螺环[3.3]庚烷-2-甲酸甲酯的制备The fourth step: 6-(1-((1,1'-diphenyl)-4-ylmethyl)-4-(3-hydroxy-3-methylbutane-1-yn-1-yl) Preparation of -1H-indazole-7-carboxamide) spiro[3.3]heptane-2-carboxylic acid methyl ester
室温下将化合物1-((1,1’-二苯基)-4-基甲基)-4-(3-羟基-3-甲基丁烷-1-炔-1-基)-1H-吲唑-7-甲酸(40mg,0.10mmol)加入到N,N-二甲基甲酰胺(3mL)中，加入2-(6-氨基螺环[3.3]庚烷-2-基)甲酸甲酯盐酸盐(31mg,0.15mmol)，二异丙基乙胺(39mg,0.30mmol)，HATU(57mg,0.15mmol)，搅拌16h。加入水(20mL)稀释，用乙酸乙酯(20mL×3)萃取，分液，合并有机相，有机相用无水硫酸钠干燥，过滤，浓缩，残留物用薄层硅胶板分离纯化(PE:EA(V/V)＝1:1)得化合物2-(6-(1-((1,1’-二苯基)-4-基甲基)-4-(3-羟基-3-甲基丁烷-1-炔-1-基)-1H-吲唑-7-甲酰胺)螺环[3.3]庚烷-2-基)甲酸甲酯(25mg,产率46％)。Compound 1-((1,1'-diphenyl)-4-ylmethyl)-4-(3-hydroxy-3-methylbutane-1-yn-1-yl)-1H- Indazole-7-carboxylic acid (40 mg, 0.10 mmol) was added to N,N-dimethylformamide (3 mL), and methyl 2-(6-aminospiro[3.3]heptane-2-yl)carboxylate was added Hydrochloride (31mg, 0.15mmol), diisopropylethylamine (39mg, 0.30mmol), HATU (57mg, 0.15mmol), stirred for 16h. Add water (20mL) to dilute, extract with ethyl acetate (20mL×3), separate the layers, combine the organic phases, dry the organic phases with anhydrous sodium sulfate, filter, concentrate, and the residue is separated and purified with a thin-layer silica gel plate (PE: EA (V/V)=1:1) to obtain compound 2-(6-(1-((1,1'-diphenyl)-4-ylmethyl)-4-(3-hydroxyl-3-methyl (25 mg, 46% yield).
第五步：6-(1-((1,1’-二苯基)-4-基甲基)-4-(3-羟基-3-甲基丁烷-1-炔-1-基)-1H-吲唑-7-甲酰胺)螺环[3.3]庚烷-2-甲酸(I-5)的制备The fifth step: 6-(1-((1,1'-diphenyl)-4-ylmethyl)-4-(3-hydroxy-3-methylbutane-1-yn-1-yl) Preparation of -1H-indazole-7-carboxamide) spiro[3.3]heptane-2-carboxylic acid (I-5)
Figure PCTCN2022097611-appb-000034
Figure PCTCN2022097611-appb-000034
室温下将化合物2-(6-(1-((1,1’-二苯基)-4-基甲基)-4-(3-羟基-3-甲基丁烷-1-炔-1-基)-1H-吲唑-7-甲酰胺)螺环[3.3]庚烷-2-基)甲酸甲酯(25mg,0.04mmol)加入到四氢呋喃(3mL)和水(1mL)中，加入氢氧化锂(4mg,0.13mmol)，室温搅拌16h。用1N盐酸调节pH＝4，用乙酸乙酯(20mL×3)萃取，分液，合并有机相，有机相用无水硫酸钠干燥，过滤，浓缩，经酸性制备方法A制备得化合物6-(1-((1,1’-二苯基)-4-基甲基)-4-(3-羟基-3-甲基丁烷-1-炔-1-基)-1H-吲唑-7-甲酰胺)螺环[3.3]庚烷-2-甲酸(I-5)(12mg,产率49％)。Compound 2-(6-(1-((1,1'-diphenyl)-4-ylmethyl)-4-(3-hydroxy-3-methylbutane-1-yne-1 -yl)-1H-indazole-7-carboxamide)spiro[3.3]heptan-2-yl)methyl carboxylate (25 mg, 0.04 mmol) was added to tetrahydrofuran (3 mL) and water (1 mL), and hydrogen Lithium oxide (4mg, 0.13mmol), stirred at room temperature for 16h. Use 1N hydrochloric acid to adjust pH=4, extract with ethyl acetate (20mL×3), separate the layers, combine the organic phases, dry the organic phases with anhydrous sodium sulfate, filter, concentrate, and prepare compound 6-( 1-((1,1'-diphenyl)-4-ylmethyl)-4-(3-hydroxy-3-methylbutane-1-yn-1-yl)-1H-indazole-7 -carboxamide) spiro[3.3]heptane-2-carboxylic acid (1-5) (12 mg, 49% yield).
1H NMR(400mHz,DMSO-d6)δ12.0(s,1H),8.80(d,1H),8.27(s,1H),7.58(d,2H),7.52(d,2H),7.44(t,2H),7.37(d,2H),7.21(d,1H),7.04(d,2H),5.83(s,2H),5.62(s,1H),4.31-4.26(m,1H),2.95(t,1H),2.43-2.31(m,1H),2.27-2.20(m,3H),2.10-2.02(m,2H),1.92-1.84(m,2H),1.55(s,6H)。 1 H NMR (400mHz, DMSO-d6) δ12.0(s,1H),8.80(d,1H),8.27(s,1H),7.58(d,2H),7.52(d,2H),7.44(t ,2H),7.37(d,2H),7.21(d,1H),7.04(d,2H),5.83(s,2H),5.62(s,1H),4.31-4.26(m,1H),2.95( t,1H), 2.43-2.31(m,1H), 2.27-2.20(m,3H), 2.10-2.02(m,2H), 1.92-1.84(m,2H), 1.55(s,6H).
LC-MS,M/Z(ESI):548.5[M+H] +。 LC-MS, M/Z (ESI): 548.5 [M+H] + .
实施例6化合物I-6的制备The preparation of embodiment 6 compound I-6
合成路线如下所示：The synthetic route is as follows:
Figure PCTCN2022097611-appb-000035
Figure PCTCN2022097611-appb-000035
第一步：4-(丙烷-1-炔-1-基)-1H-吲唑-7-乙酸甲酯的制备The first step: the preparation of 4-(propane-1-yn-1-yl)-1H-indazole-7-acetic acid methyl ester
室温下将化合物4-溴-1H-吲唑-7-乙酸甲酯(1.0g,3.94mmol)加入到1,4-二氧六环(10mL)中，加入二三苯基膦二氯化钯(274mg,0.39mmol)，1-丙炔-三正丁基锡(1.52g,4.61mmol)，氮气保护下加热至90℃，搅拌16h。冷却至室温，加入水(50mL)稀释，用乙酸乙酯(50mL×3)萃取，分液，合并有机相，有机相用无水硫酸钠干燥，过滤，浓缩，残留物用薄层硅胶板分离纯化(石油醚:乙酸乙酯(V/V)＝5:1)得化合物4-(丙烷-1-炔-1-基)-1H-吲唑-7-乙酸甲酯(500mg,产率42％)。Add compound 4-bromo-1H-indazole-7-acetic acid methyl ester (1.0g, 3.94mmol) to 1,4-dioxane (10mL) at room temperature, add ditriphenylphosphine palladium dichloride (274mg, 0.39mmol), 1-propyne-tri-n-butyltin (1.52g, 4.61mmol), heated to 90°C under nitrogen protection, and stirred for 16h. Cool to room temperature, add water (50mL) to dilute, extract with ethyl acetate (50mL×3), separate the liquids, combine the organic phases, dry the organic phases with anhydrous sodium sulfate, filter, concentrate, and separate the residue with a thin-layer silica gel plate Purification (petroleum ether:ethyl acetate (V/V)=5:1) gave compound 4-(propane-1-yn-1-yl)-1H-indazole-7-acetic acid methyl ester (500mg, yield 42 %).
第二步：1-((1,1’-二苯基)-4-基甲基)-4-(丙烷-1-炔-1-基)-1H-吲唑-7-乙酸甲酯的制备The second step: 1-((1,1'-diphenyl)-4-ylmethyl)-4-(propane-1-yn-1-yl)-1H-indazole-7-acetic acid methyl ester preparation
室温下将化合物4-(丙烷-1-炔-1-基)-1H-吲唑-7-乙酸甲酯(100mg,0.47mmol)加入到N,N-二甲基甲酰胺(5mL)中，加入4-苯基苄溴(174mg,0.71mmol)，碳酸铯(460mg,1.40mmol)，室温搅拌16h。加入水(20mL)稀释，用乙酸乙酯(20mL×3)萃取，分液，合并有机相，有机相用无水硫酸钠干燥，过滤，浓缩，残留物用薄层硅胶板分离纯化(石油醚:乙酸乙酯(V/V)＝10:1)得化合物1-((1,1’-二苯基)-4-基甲基)-4-(丙烷-1-炔-1-基)-1H-吲唑-7-乙酸甲酯(80mg,产率45％)。Compound 4-(propan-1-yn-1-yl)-1H-indazole-7-acetic acid methyl ester (100 mg, 0.47 mmol) was added into N,N-dimethylformamide (5 mL) at room temperature, Add 4-phenylbenzyl bromide (174mg, 0.71mmol), cesium carbonate (460mg, 1.40mmol), and stir at room temperature for 16h. Add water (20mL) to dilute, extract with ethyl acetate (20mL×3), separate the layers, combine the organic phases, dry the organic phases with anhydrous sodium sulfate, filter, concentrate, and separate and purify the residue with a thin-layer silica gel plate (petroleum ether : Ethyl acetate (V/V)=10:1) to obtain compound 1-((1,1'-diphenyl)-4-ylmethyl)-4-(propane-1-yn-1-yl) - 1H-indazole-7-acetic acid methyl ester (80 mg, yield 45%).
第三步：1-((1,1’-二苯基)-4-基甲基)-4-(丙烷-1-炔-1-基)-1H-吲唑-7-乙酸的制备Step 3: Preparation of 1-((1,1’-diphenyl)-4-ylmethyl)-4-(propane-1-yn-1-yl)-1H-indazole-7-acetic acid
室温下将原料1-((1,1’-二苯基)-4-基甲基)-4-(丙烷-1-炔-1-基)-1H-吲唑-7-乙酸甲酯(80mg,0.21mmol)加入到四氢呋喃(3mL)，甲醇(1mL)和水(1mL)中，加入氢氧化锂(15mg,0.63mmol)，室温搅拌16h。用1N盐酸调节pH＝4，用乙酸乙酯(20mL×3)萃取，分液，合并有机相，有机相用无水硫酸钠干燥，过滤，浓缩，得化合物粗品1-((1,1’-二苯基)-4-基甲基)-4-(丙烷-1-炔-1-基)-1H-吲唑-7-乙酸(50mg,产率65％).The starting material 1-((1,1'-diphenyl)-4-ylmethyl)-4-(propan-1-yn-1-yl)-1H-indazole-7-acetic acid methyl ester ( 80mg, 0.21mmol) was added to tetrahydrofuran (3mL), methanol (1mL) and water (1mL), lithium hydroxide (15mg, 0.63mmol) was added, and stirred at room temperature for 16h. Use 1N hydrochloric acid to adjust pH=4, extract with ethyl acetate (20mL×3), separate the layers, combine the organic phases, dry the organic phases with anhydrous sodium sulfate, filter, and concentrate to obtain the crude compound 1-((1,1' -Diphenyl)-4-ylmethyl)-4-(propane-1-yn-1-yl)-1H-indazole-7-acetic acid (50mg, yield 65%).
第四步：2-(6-(1-((1,1’-二苯基)-4-基甲基)-4-(丙烷-1-炔-1-基)-1H-吲唑-7-甲酰胺)螺环[3.3]庚烷-2-基)乙酸甲酯的制备The fourth step: 2-(6-(1-((1,1'-diphenyl)-4-ylmethyl)-4-(propane-1-yn-1-yl)-1H-indazole- Preparation of 7-carboxamide) spiro[3.3]heptan-2-yl) methyl acetate
室温下将化合物1-((1,1’-二苯基)-4-基甲基)-4-(丙烷-1-炔-1-基)-1H-吲唑-7-乙酸(45mg,0.12mmol)加入到N,N-二甲基甲酰胺(5mL)中，加入2-(6-氨基螺环[3.3]庚烷-2-基)乙酸甲酯盐酸盐(39mg,0.18mmol)，二异丙基乙胺(46mg,0.36mmol)，HATU(68mg,0.18mmol)，搅拌16h。加入水(20mL)稀释，用乙酸乙酯(20mL×3)萃取，分液，合并有机相，有机相用无水硫酸钠干燥，过滤，浓缩，残留物经酸性制备方法A制备得化合物2-(6-(1-((1,1’-二苯基)-4-基甲基)-4-(丙烷-1-炔-1-基)-1H-吲唑-7-甲酰胺)螺环[3.3]庚烷-2-基)乙酸甲酯(40mg,产率63％)。Compound 1-((1,1'-diphenyl)-4-ylmethyl)-4-(propane-1-yn-1-yl)-1H-indazole-7-acetic acid (45 mg, 0.12mmol) was added to N,N-dimethylformamide (5mL), and 2-(6-aminospirocyclo[3.3]heptane-2-yl)methyl acetate hydrochloride (39mg, 0.18mmol) was added , diisopropylethylamine (46mg, 0.36mmol), HATU (68mg, 0.18mmol), stirred for 16h. Add water (20mL) to dilute, extract with ethyl acetate (20mL×3), separate the layers, combine the organic phases, dry the organic phases with anhydrous sodium sulfate, filter, concentrate, and prepare the residue by acidic preparation method A to obtain compound 2- (6-(1-((1,1'-diphenyl)-4-ylmethyl)-4-(propane-1-yn-1-yl)-1H-indazole-7-carboxamide)spiro Cyclo[3.3]heptan-2-yl)methyl acetate (40 mg, 63% yield).
第五步：2-(6-(1-((1,1’-二苯基)-4-基甲基)-4-(丙烷-1-炔-1-基)-1H-吲唑-7-甲酰胺)螺环[3.3]庚烷-2-基)乙酸(I-6)的制备The fifth step: 2-(6-(1-((1,1'-diphenyl)-4-ylmethyl)-4-(propane-1-yn-1-yl)-1H-indazole- Preparation of 7-carboxamide) spiro[3.3]heptan-2-yl) acetic acid (I-6)
Figure PCTCN2022097611-appb-000036
Figure PCTCN2022097611-appb-000036
室温下将化合物2-(6-(1-((1,1’-二苯基)-4-基甲基)-4-(丙烷-1-炔-1-基)-1H-吲唑-7-甲酰胺)螺环[3.3]庚烷-2-基)乙酸甲酯(40mg,0.07mmol)加入到四氢呋喃(3mL)和水(1mL)中，加入氢氧化锂(6.0mg,0.24mmol)，室温搅拌16h。用1N盐酸调节pH＝4，用乙酸乙酯(20mL×3)萃取，分液，合并有机相，有机相用无水硫酸钠干燥，过滤，浓缩，得化合物6-(1-((1,1’-二苯基)-4-基甲基)-4-(丙烷-1-炔-1-基)-1H-吲唑-7-甲酰胺)螺环[3.3]庚烷-2-乙酸(I-6)(35mg,产率92％)。Compound 2-(6-(1-((1,1'-diphenyl)-4-ylmethyl)-4-(propane-1-yn-1-yl)-1H-indazole- 7-Carboxamide)spiro[3.3]heptan-2-yl)methyl acetate (40mg, 0.07mmol) was added to tetrahydrofuran (3mL) and water (1mL), lithium hydroxide (6.0mg, 0.24mmol) was added , stirred at room temperature for 16h. Use 1N hydrochloric acid to adjust pH=4, extract with ethyl acetate (20mL×3), separate the layers, combine the organic phases, dry the organic phases with anhydrous sodium sulfate, filter, and concentrate to obtain compound 6-(1-((1, 1'-diphenyl)-4-ylmethyl)-4-(propane-1-yn-1-yl)-1H-indazole-7-carboxamide)spiro[3.3]heptane-2-acetic acid (I-6) (35 mg, yield 92%).
1H NMR(400mHz,DMSO-d6)δ11.9(s,1H),8.75(d,1H),8.25(s,1H),7.58(d,2H),7.52(d,2H),7.45(t,2H),7.36(d,2H),7.22(d,1H),7.05(d,2H),5.81(s,2H),4.30-4.24(m,1H),2.44-2.35 (m,2H),2.27-2.19(m,4H),2.18(s,3H),1.98-1.81(m,3H),1.76-1.71(m,1H),1.61-1.57(m,1H)。 1 H NMR(400mHz,DMSO-d6)δ11.9(s,1H),8.75(d,1H),8.25(s,1H),7.58(d,2H),7.52(d,2H),7.45(t ,2H),7.36(d,2H),7.22(d,1H),7.05(d,2H),5.81(s,2H),4.30-4.24(m,1H),2.44-2.35(m,2H), 2.27-2.19 (m, 4H), 2.18 (s, 3H), 1.98-1.81 (m, 3H), 1.76-1.71 (m, 1H), 1.61-1.57 (m, 1H).
LC-MS,M/Z(ESI):518.5[M+H] +。 LC-MS, M/Z (ESI): 518.5 [M+H] + .
实施例7化合物I-7的制备The preparation of embodiment 7 compound I-7
合成路线如下所示：The synthetic route is as follows:
Figure PCTCN2022097611-appb-000037
Figure PCTCN2022097611-appb-000037
第一步：5-(丙烷-1-炔-1-基)-1H-吲唑-7-乙酸甲酯的制备The first step: the preparation of 5-(propane-1-yn-1-yl)-1H-indazole-7-acetic acid methyl ester
室温下将化合物5-溴-1H-吲唑-7-乙酸甲酯(1.0g,3.92mmol)加入到1,4-二氧六环(10mL)中，加入二三苯基膦二氯化钯(274mg,0.39mmol)，1-丙炔-三正丁基锡(1.55g,4.70mmol)，氮气保护下加热至90℃，搅拌16h。冷却至室温，加入水(50mL)稀释，用乙酸乙酯(50mL×3)萃取，分液，合并有机相，有机相用无水硫酸钠干燥，过滤，浓缩，残留物用薄层硅胶板分离纯化(石油醚:乙酸乙酯(V/V)＝5:1)得化合物5-(丙烷-1-炔-1-基)-1H-吲唑-7-乙酸甲酯(300mg,产率36％)。Add compound 5-bromo-1H-indazole-7-acetic acid methyl ester (1.0g, 3.92mmol) to 1,4-dioxane (10mL) at room temperature, add ditriphenylphosphine palladium dichloride (274mg, 0.39mmol), 1-propyne-tri-n-butyltin (1.55g, 4.70mmol), heated to 90°C under nitrogen protection, and stirred for 16h. Cool to room temperature, add water (50mL) to dilute, extract with ethyl acetate (50mL×3), separate the liquids, combine the organic phases, dry the organic phases with anhydrous sodium sulfate, filter, concentrate, and separate the residue with a thin-layer silica gel plate Purification (petroleum ether:ethyl acetate (V/V)=5:1) gave compound 5-(propane-1-yn-1-yl)-1H-indazole-7-acetic acid methyl ester (300mg, yield 36 %).
第二步：1-((1,1’-二苯基)-4-基甲基)-5-(丙烷-1-炔-1-基)-1H-吲唑-7-乙酸甲酯的制备The second step: 1-((1,1'-diphenyl)-4-ylmethyl)-5-(propane-1-yn-1-yl)-1H-indazole-7-acetic acid methyl ester preparation
室温下将化合物5-(丙烷-1-炔-1-基)-1H-吲唑-7-乙酸甲酯(100mg,0.47mmol)加入到N,N-二甲基甲酰胺(5mL)中，加入4-苯基苄溴(150mg,0.61mmol)，碳酸铯(460mg,1.40mmol)，室温搅拌16h。加入水(20mL)稀释，用乙酸乙酯(20mL×3)萃取，分液，合并有机相，有机相用无水硫酸钠干燥，过滤，浓缩，残留物用薄层硅胶板分离纯化(石油醚:乙酸乙酯(V/V)＝10:1)得化合物1-((1,1’-二苯基)-4-基甲基)-5-(丙烷-1-炔-1-基)-1H-吲唑-7-乙酸甲酯(50mg,产率28％)。Compound 5-(propan-1-yn-1-yl)-1H-indazole-7-acetic acid methyl ester (100 mg, 0.47 mmol) was added into N,N-dimethylformamide (5 mL) at room temperature, Add 4-phenylbenzyl bromide (150mg, 0.61mmol), cesium carbonate (460mg, 1.40mmol), and stir at room temperature for 16h. Add water (20mL) to dilute, extract with ethyl acetate (20mL×3), separate the layers, combine the organic phases, dry the organic phases with anhydrous sodium sulfate, filter, concentrate, and separate and purify the residue with a thin-layer silica gel plate (petroleum ether : Ethyl acetate (V/V)=10:1) to obtain compound 1-((1,1'-diphenyl)-4-ylmethyl)-5-(propane-1-yn-1-yl) - 1H-indazole-7-acetic acid methyl ester (50 mg, yield 28%).
第三步：1-((1,1’-二苯基)-4-基甲基)-5-(丙烷-1-炔-1-基)-1H-吲唑-7-乙酸的制备Step 3: Preparation of 1-((1,1’-diphenyl)-4-ylmethyl)-5-(propane-1-yn-1-yl)-1H-indazole-7-acetic acid
室温下将1-((1,1’-二苯基)-4-基甲基)-5-(丙烷-1-炔-1-基)-1H-吲唑-7-乙酸甲酯(50mg,0.13mmol)加入到四氢呋喃(3mL)，甲醇(1mL)和水(1mL)中，加入氢氧化锂(10mg,0.40mmol)，室温搅拌4h。用1N盐酸调节pH＝4，用乙酸乙酯(20mL×3)萃取，分液，合并有机相，有机相用无水硫酸钠干燥，过滤，浓缩，得化合物粗品1-((1,1’-二苯基)-4-基甲基)-5-(丙烷-1-炔-1- 基)-1H-吲唑-7-乙酸(40mg,产率83％).1-((1,1'-diphenyl)-4-ylmethyl)-5-(propan-1-yn-1-yl)-1H-indazole-7-acetic acid methyl ester (50mg , 0.13mmol) was added to tetrahydrofuran (3mL), methanol (1mL) and water (1mL), lithium hydroxide (10mg, 0.40mmol) was added, and stirred at room temperature for 4h. Use 1N hydrochloric acid to adjust pH=4, extract with ethyl acetate (20mL×3), separate the layers, combine the organic phases, dry the organic phases with anhydrous sodium sulfate, filter, and concentrate to obtain the crude compound 1-((1,1' -Diphenyl)-4-ylmethyl)-5-(propane-1-yn-1-yl)-1H-indazole-7-acetic acid (40mg, yield 83%).
第四步：2-(6-(1-((1,1’-二苯基)-4-基甲基)-5-(丙烷-1-炔-1-基)-1H-吲唑-7-甲酰胺)螺环[3.3]庚烷-2-基)乙酸甲酯的制备The fourth step: 2-(6-(1-((1,1'-diphenyl)-4-ylmethyl)-5-(propane-1-yn-1-yl)-1H-indazole- Preparation of 7-carboxamide) spiro[3.3]heptan-2-yl) methyl acetate
室温下将化合物1-((1,1’-二苯基)-4-基甲基)-5-(丙烷-1-炔-1-基)-1H-吲唑-7-乙酸(50mg,0.14mmol)加入到N,N-二甲基甲酰胺(5mL)中，加入2-(6-氨基螺环[3.3]庚烷-2-基)乙酸甲酯盐酸盐(46mg,0.21mmol)，二异丙基乙胺(54mg,0.42mmol)，HATU(80mg,0.21mmol)，搅拌16h。加入水(20mL)稀释，用乙酸乙酯(20mL×3)萃取，分液，合并有机相，有机相用无水硫酸钠干燥，过滤，浓缩，残留物用薄层硅胶板分离纯化(石油醚:乙酸乙酯(V/V)＝1:1)得化合物2-(6-(1-((1,1’-二苯基)-4-基甲基)-5-(丙烷-1-炔-1-基)-1H-吲唑-7-甲酰胺)螺环[3.3]庚烷-2-基)乙酸甲酯(50mg,产率69％)。Compound 1-((1,1'-diphenyl)-4-ylmethyl)-5-(propane-1-yn-1-yl)-1H-indazole-7-acetic acid (50mg, 0.14mmol) was added to N,N-dimethylformamide (5mL), and 2-(6-aminospirocyclo[3.3]heptane-2-yl)methyl acetate hydrochloride (46mg, 0.21mmol) was added , diisopropylethylamine (54mg, 0.42mmol), HATU (80mg, 0.21mmol), stirred for 16h. Add water (20mL) to dilute, extract with ethyl acetate (20mL×3), separate the layers, combine the organic phases, dry the organic phases with anhydrous sodium sulfate, filter, concentrate, and separate and purify the residue with a thin-layer silica gel plate (petroleum ether : Ethyl acetate (V/V)=1:1) to obtain compound 2-(6-(1-((1,1'-diphenyl)-4-ylmethyl)-5-(propane-1- Alkyn-1-yl)-1H-indazole-7-carboxamide)spiro[3.3]heptan-2-yl)methyl acetate (50 mg, 69% yield).
第五步：2-(6-(1-((1,1’-二苯基)-4-基甲基)-5-(丙烷-1-炔-1-基)-1H-吲唑-7-甲酰胺)螺环[3.3]庚烷-2-基)乙酸(I-7)的制备The fifth step: 2-(6-(1-((1,1'-diphenyl)-4-ylmethyl)-5-(propane-1-yn-1-yl)-1H-indazole- Preparation of 7-carboxamide) spiro[3.3]heptan-2-yl) acetic acid (I-7)
Figure PCTCN2022097611-appb-000038
Figure PCTCN2022097611-appb-000038
室温下将化合物2-(6-(1-((1,1’-二苯基)-4-基甲基)-5-(丙烷-1-炔-1-基)-1H-吲唑-7-甲酰胺)螺环[3.3]庚烷-2-基)乙酸甲酯(50mg,0.09mmol)加入到四氢呋喃(3mL)和水(1mL)中，加入氢氧化锂(7.0mg,0.29mmol)，室温搅拌16h。用1N盐酸调节pH＝4，用乙酸乙酯(20mL×3)萃取，分液，合并有机相，有机相用无水硫酸钠干燥，过滤，浓缩，残留物经酸性制备方法A制备得化合物2-(6-(1-((1,1’-二苯基)-4-基甲基)-5-(丙烷-1-炔-1-基)-1H-吲唑-7-甲酰胺)螺环[3.3]庚烷-2-基)乙酸(I-7)(45mg,产率92％)。Compound 2-(6-(1-((1,1'-diphenyl)-4-ylmethyl)-5-(propane-1-yn-1-yl)-1H-indazole- 7-Carboxamide)spiro[3.3]heptan-2-yl)methyl acetate (50mg, 0.09mmol) was added to tetrahydrofuran (3mL) and water (1mL), lithium hydroxide (7.0mg, 0.29mmol) was added , stirred at room temperature for 16h. Use 1N hydrochloric acid to adjust pH=4, extract with ethyl acetate (20mL×3), separate the layers, combine the organic phases, dry the organic phases with anhydrous sodium sulfate, filter, concentrate, and prepare the residue by acidic preparation method A to obtain compound 2 -(6-(1-((1,1'-diphenyl)-4-ylmethyl)-5-(propane-1-yn-1-yl)-1H-indazole-7-carboxamide) Spiro[3.3]heptan-2-yl)acetic acid (I-7) (45 mg, 92% yield).
1H NMR(400mHz,DMSO-d6)δ11.9(s,1H),8.75(d,1H),8.25(s,1H),7.58(d,2H),7.52(d,2H),7.45(t,2H),7.35(t,2H),7.21(d,1H),7.05(d,2H),5.81(s,2H),4.30-4.24(m,1H),2.45-2.35(m,2H),2.27-2.19(m,4H),2.17(s,3H),1.98-1.81(m,3H),1.76-1.71(m,1H),1.61-1.57(m,1H)。 1 H NMR(400mHz,DMSO-d6)δ11.9(s,1H),8.75(d,1H),8.25(s,1H),7.58(d,2H),7.52(d,2H),7.45(t ,2H),7.35(t,2H),7.21(d,1H),7.05(d,2H),5.81(s,2H),4.30-4.24(m,1H),2.45-2.35(m,2H), 2.27-2.19 (m, 4H), 2.17 (s, 3H), 1.98-1.81 (m, 3H), 1.76-1.71 (m, 1H), 1.61-1.57 (m, 1H).
LC-MS,M/Z(ESI):518.6[M+H] +。 LC-MS, M/Z (ESI): 518.6 [M+H] + .
实施例8化合物I-8的制备The preparation of embodiment 8 compound I-8
合成路线如下所示：The synthetic route is as follows:
Figure PCTCN2022097611-appb-000039
Figure PCTCN2022097611-appb-000039
第一步：4-(丙烷-1-炔-1-基)-1H-吲唑-7-甲酸甲酯的制备The first step: the preparation of 4-(propane-1-yn-1-yl)-1H-indazole-7-carboxylic acid methyl ester
室温下将化合物4-溴-1H-吲唑-7-甲酸甲酯(1.0g,3.94mmol)加入到1,4-二氧六环(10mL)中，加入二三苯基膦二氯化钯(274mg,0.39mmol)，1-丙炔-三正丁基锡(1.52g,4.61mmol)，氮气保护下加热至90℃，搅拌16h。冷却至室温，加入水(50mL)稀释，用乙酸乙酯(50mL×3)萃取，分液，合并有机相，有机相用无水硫酸钠干燥，过滤，浓缩，残留物用薄层硅胶板分离纯化(石油醚:乙酸乙酯(V/V)＝5:1)得化合物4-(丙烷-1-炔-1-基)-1H-吲唑-7-甲酸甲酯(500mg,产率42％)。Add the compound 4-bromo-1H-indazole-7-carboxylic acid methyl ester (1.0g, 3.94mmol) to 1,4-dioxane (10mL) at room temperature, add ditriphenylphosphine palladium dichloride (274mg, 0.39mmol), 1-propyne-tri-n-butyltin (1.52g, 4.61mmol), heated to 90°C under nitrogen protection, and stirred for 16h. Cool to room temperature, add water (50mL) to dilute, extract with ethyl acetate (50mL×3), separate the liquids, combine the organic phases, dry the organic phases with anhydrous sodium sulfate, filter, concentrate, and separate the residue with a thin-layer silica gel plate Purification (petroleum ether: ethyl acetate (V/V) = 5:1) gave compound 4-(propane-1-yn-1-yl)-1H-indazole-7-carboxylic acid methyl ester (500mg, yield 42 %).
第二步：4-(丙烷-1-炔-1-基)-1-(4-(三氟甲氧基)苄基)-1H-吲唑-7-甲酸甲酯的制备The second step: the preparation of 4-(propane-1-yn-1-yl)-1-(4-(trifluoromethoxy)benzyl)-1H-indazole-7-carboxylic acid methyl ester
室温下将化合物4-(丙烷-1-炔-1-基)-1H-吲唑-7-甲酸甲酯(100mg,0.47mmol)加入到N,N-二甲基甲酰胺(5mL)中，加入4-(三氟甲氧基)苄溴(179mg,0.70mmol)，碳酸铯(456mg,1.40mmol)，室温搅拌16h。加入水(20mL)稀释，用乙酸乙酯(20mL×3)萃取，分液，合并有机相，有机相用无水硫酸钠干燥，过滤，浓缩，残留物用薄层硅胶板分离纯化(石油醚:乙酸乙酯(V/V)＝10:1)得化合物4-(丙烷-1-炔-1-基)-1-(4-(三氟甲氧基)苄基)-1H-吲唑-7-甲酸甲酯(90mg,产率50％)。Compound 4-(propane-1-yn-1-yl)-1H-indazole-7-carboxylic acid methyl ester (100 mg, 0.47 mmol) was added into N,N-dimethylformamide (5 mL) at room temperature, Add 4-(trifluoromethoxy)benzyl bromide (179mg, 0.70mmol), cesium carbonate (456mg, 1.40mmol), and stir at room temperature for 16h. Add water (20mL) to dilute, extract with ethyl acetate (20mL×3), separate the layers, combine the organic phases, dry the organic phases with anhydrous sodium sulfate, filter, concentrate, and separate and purify the residue with a thin-layer silica gel plate (petroleum ether : Ethyl acetate (V/V)=10:1) to obtain compound 4-(propane-1-yn-1-yl)-1-(4-(trifluoromethoxy)benzyl)-1H-indazole - Methyl 7-carboxylate (90 mg, yield 50%).
第三步：4-(丙烷-1-炔-1-基)-1-(4-(三氟甲氧基)苄基)-1H-吲唑-7-甲酸的制备The third step: the preparation of 4-(propane-1-yn-1-yl)-1-(4-(trifluoromethoxy)benzyl)-1H-indazole-7-carboxylic acid
室温下将原料4-(丙烷-1-炔-1-基)-1-(4-(三氟甲氧基)苄基)-1H-吲唑-7-甲酸甲酯(90mg,0.23mmol)加入到四氢呋喃(3mL),水(1mL)中，加入氢氧化锂(17mg,0.70mmol)，室温搅拌16h。用1N盐酸调节pH＝4，用乙酸乙酯(20mL×3)萃取，分液，合并有机相，有机相用无水硫酸钠干燥，过滤，浓缩，得化合物白色固体粗品4-(丙烷-1-炔-1-基)-1-(4-(三氟甲氧基)苄基)-1H-吲唑-7-甲酸(80mg,产率65％).Starting material 4-(propan-1-yn-1-yl)-1-(4-(trifluoromethoxy)benzyl)-1H-indazole-7-carboxylic acid methyl ester (90mg, 0.23mmol) at room temperature Add to tetrahydrofuran (3mL), water (1mL), add lithium hydroxide (17mg, 0.70mmol), stir at room temperature for 16h. Use 1N hydrochloric acid to adjust pH=4, extract with ethyl acetate (20mL×3), separate the layers, combine the organic phases, dry the organic phases with anhydrous sodium sulfate, filter, and concentrate to obtain the crude compound 4-(propane-1 -Alkyn-1-yl)-1-(4-(trifluoromethoxy)benzyl)-1H-indazole-7-carboxylic acid (80mg, yield 65%).
第四步：6-(4-(丙烷-1-炔-1-基)-1-(4-(三氟甲氧基)苄基)-1H-吲唑-7-甲酰胺)螺环[3.3]庚烷-2-甲酸甲酯的制备The fourth step: 6-(4-(propane-1-yn-1-yl)-1-(4-(trifluoromethoxy) benzyl)-1H-indazole-7-carboxamide) spiro[ 3.3] Preparation of methyl heptane-2-carboxylate
室温下将化合物4-(丙烷-1-炔-1-基)-1-(4-(三氟甲氧基)苄基)-1H-吲唑-7-甲酸(80mg,0.21mmol)加入到N,N-二甲基甲酰胺(5mL)中，加入2-(6-氨基螺环[3.3]庚烷-2-基)甲酸甲酯盐酸盐(66mg,0.32mmol)，二异丙基乙胺(83mg,0.64mmol)，HATU(122mg,0.32mmol)，搅拌16h。加入水(20mL)稀释，用乙酸乙酯(20mL×3)萃取，分液，合并有机相，有机相用无水硫酸 钠干燥，过滤，浓缩，残留物用薄层硅胶板分离纯化(石油醚:乙酸乙酯(V/V)＝1:1)得化合物6-(4-(丙烷-1-炔-1-基)-1-(4-(三氟甲氧基)苄基)-1H-吲唑-7-甲酰胺)螺环[3.3]庚烷-2-甲酸甲酯(80mg,产率71％)。Compound 4-(propane-1-yn-1-yl)-1-(4-(trifluoromethoxy)benzyl)-1H-indazole-7-carboxylic acid (80mg, 0.21mmol) was added to To N,N-dimethylformamide (5mL), add 2-(6-aminospiro[3.3]heptane-2-yl)methyl formate hydrochloride (66mg, 0.32mmol), diisopropyl Ethylamine (83mg, 0.64mmol), HATU (122mg, 0.32mmol), stirred for 16h. Add water (20mL) to dilute, extract with ethyl acetate (20mL×3), separate the layers, combine the organic phases, dry the organic phases with anhydrous sodium sulfate, filter, concentrate, and separate and purify the residue with a thin-layer silica gel plate (petroleum ether : ethyl acetate (V/V)=1:1) to obtain compound 6-(4-(propane-1-yn-1-yl)-1-(4-(trifluoromethoxy)benzyl)-1H -indazole-7-carboxamide)spiro[3.3]heptane-2-carboxylic acid methyl ester (80 mg, 71% yield).
第五步：6-(4-(丙烷-1-炔-1-基)-1-(4-(三氟甲氧基)苄基)-1H-吲唑-7-甲酰胺)螺环[3.3]庚烷-2-甲酸(I-8)的制备The fifth step: 6-(4-(propane-1-yn-1-yl)-1-(4-(trifluoromethoxy) benzyl)-1H-indazole-7-carboxamide) spiro[ 3.3] Preparation of heptane-2-carboxylic acid (I-8)
Figure PCTCN2022097611-appb-000040
Figure PCTCN2022097611-appb-000040
室温下将化合物6-(4-(丙烷-1-炔-1-基)-1-(4-(三氟甲氧基)苄基)-1H-吲唑-7-甲酰胺)螺环[3.3]庚烷-2-甲酸甲酯(80mg,0.15mmol)加入到四氢呋喃(3mL)和水(1mL)中，加入氢氧化锂(11mg,0.46mmol)，室温搅拌16h。用1N盐酸调节pH＝4，用乙酸乙酯(20mL×3)萃取，分液，合并有机相，有机相用无水硫酸钠干燥，过滤，浓缩，残留物经酸性制备方法A制备得化合物6-(4-(丙烷-1-炔-1-基)-1-(4-(三氟甲氧基)苄基)-1H-吲唑-7-甲酰胺)螺环[3.3]庚烷-2-甲酸(I-8)(47mg,产率60％)。Compound 6-(4-(propane-1-yn-1-yl)-1-(4-(trifluoromethoxy)benzyl)-1H-indazole-7-carboxamide) spiro[ 3.3] Add methyl heptane-2-carboxylate (80mg, 0.15mmol) to tetrahydrofuran (3mL) and water (1mL), add lithium hydroxide (11mg, 0.46mmol), and stir at room temperature for 16h. Use 1N hydrochloric acid to adjust pH=4, extract with ethyl acetate (20mL×3), separate the layers, combine the organic phases, dry the organic phases with anhydrous sodium sulfate, filter, concentrate, and prepare the residue by acidic preparation method A to obtain compound 6 -(4-(propane-1-yn-1-yl)-1-(4-(trifluoromethoxy)benzyl)-1H-indazole-7-carboxamide)spiro[3.3]heptane- 2-Formic acid (I-8) (47 mg, yield 60%).
1H NMR(400mHz,DMSO-d6)δ12.0(s,1H),8.73(d,1H),8.26(s,1H),7.36(d,1H),7.25(t,3H),7.04(d,2H),5.80(s,2H),4.24-4.18(m,1H),2.94(t,1H),2.40-2.22(m,4H),2.18(s,3H),2.08-2.02(m,2H),1.85-1.79(m,2H)。 1 H NMR(400mHz,DMSO-d6)δ12.0(s,1H),8.73(d,1H),8.26(s,1H),7.36(d,1H),7.25(t,3H),7.04(d ,2H),5.80(s,2H),4.24-4.18(m,1H),2.94(t,1H),2.40-2.22(m,4H),2.18(s,3H),2.08-2.02(m,2H ), 1.85-1.79(m,2H).
LC-MS,M/Z(ESI):512.4[M+H] +。 LC-MS, M/Z (ESI): 512.4 [M+H] + .
实施例9化合物I-9的制备The preparation of embodiment 9 compound I-9
合成路线如下所示：The synthetic route is as follows:
Figure PCTCN2022097611-appb-000041
Figure PCTCN2022097611-appb-000041
第一步：(4-((7-(甲氧羰基)-4-(丙烷-1-炔-1-基)-1H-吲唑-1-基)甲基)苄基)硼酸的制备The first step: the preparation of (4-((7-(methoxycarbonyl)-4-(propane-1-yn-1-yl)-1H-indazol-1-yl)methyl)benzyl)boronic acid
室温下将化合物4-(丙烷-1-炔-1-基)-1H-吲唑-7-甲酸甲酯(190mg,0.89mmol)加入到N,N-二甲基甲酰胺(8mL)中，加入(4-(溴甲基)苄基)硼酸(286mg,1.33mmol)，碳酸铯(867mg,2.66mmol)，室温搅拌16h。加入水(20mL)稀释，用乙酸乙酯(20mL×3)萃取，分液，合并有机 相，有机相用无水硫酸钠干燥，过滤，浓缩，残留物用硅胶柱分离纯化(DCM:甲醇(V/V)＝10:1)得化合物(4-((7-(甲氧羰基)-4-(丙烷-1-炔-1-基)-1H-吲唑-1-基)甲基)苄基)硼酸(210mg,产率68％)。Compound 4-(propane-1-yn-1-yl)-1H-indazole-7-carboxylic acid methyl ester (190 mg, 0.89 mmol) was added to N,N-dimethylformamide (8 mL) at room temperature, Add (4-(bromomethyl)benzyl)boronic acid (286mg, 1.33mmol), cesium carbonate (867mg, 2.66mmol), and stir at room temperature for 16h. Add water (20mL) to dilute, extract with ethyl acetate (20mL×3), separate the layers, combine the organic phase, dry the organic phase with anhydrous sodium sulfate, filter, concentrate, and the residue is separated and purified by silica gel column (DCM: methanol ( V/V)=10:1) to obtain compound (4-((7-(methoxycarbonyl)-4-(propane-1-yn-1-yl)-1H-indazol-1-yl)methyl) Benzyl)boronic acid (210 mg, 68% yield).
第二步：1-(4-(2-甲氧基嘧啶-4-基)苄基)-4-(丙烷-1-炔-1-基)-1H-吲唑-7-甲酸甲酯的制备The second step: 1-(4-(2-methoxypyrimidin-4-yl)benzyl)-4-(propane-1-yn-1-yl)-1H-indazole-7-formic acid methyl ester preparation
室温下将原料(4-((7-(甲氧羰基)-4-(丙烷-1-炔-1-基)-1H-吲唑-1-基)甲基)苄基)硼酸(80mg,0.23mmol)加入到1,4-二氧六环(5mL)和水(1mL)中，加入氯(2-二环己基膦基-2,6-二甲氧基-1,1-联苯基)(2-氨基-1,1-联苯-2-基)钯(II)(17mg,0.02mmol)和磷酸钾(196mg,0.92mmol)，1,1'-双二苯基膦二茂铁二氯化钯(19mg,0.02mmol)，4-溴-2-甲氧基嘧啶(53mg,0.28mmol)，90℃搅拌1h。加入水(20mL)稀释，用乙酸乙酯(20mL×3)萃取，分液，合并有机相，有机相用无水硫酸钠干燥，过滤，浓缩，残留物用薄层硅胶板分离纯化(DCM:甲醇(V/V)＝10:1)得化合物1-(4-(2-甲氧基嘧啶-4-基)苄基)-4-(丙烷-1-炔-1-基)-1H-吲唑-7-甲酸甲酯(70mg,产率74％).The starting material (4-((7-(methoxycarbonyl)-4-(propane-1-yn-1-yl)-1H-indazol-1-yl)methyl)benzyl)boronic acid (80 mg, 0.23mmol) was added to 1,4-dioxane (5mL) and water (1mL), and chloro(2-dicyclohexylphosphino-2,6-dimethoxy-1,1-biphenyl )(2-amino-1,1-biphenyl-2-yl)palladium(II) (17mg, 0.02mmol) and potassium phosphate (196mg, 0.92mmol), 1,1'-bisdiphenylphosphinoferrocene Palladium dichloride (19mg, 0.02mmol), 4-bromo-2-methoxypyrimidine (53mg, 0.28mmol), stirred at 90°C for 1h. Add water (20mL) to dilute, extract with ethyl acetate (20mL×3), separate the layers, combine the organic phases, dry the organic phases with anhydrous sodium sulfate, filter, concentrate, and the residue is separated and purified with a thin-layer silica gel plate (DCM: Methanol (V/V)=10:1) to obtain the compound 1-(4-(2-methoxypyrimidin-4-yl)benzyl)-4-(propane-1-yn-1-yl)-1H- Indazole-7-carboxylic acid methyl ester (70mg, yield 74%).
第三步：1-(4-(2-甲氧基嘧啶-4-基)苄基)-4-(丙烷-1-炔-1-基)-1H-吲唑-7-甲酸的制备Step 3: Preparation of 1-(4-(2-methoxypyrimidin-4-yl)benzyl)-4-(propane-1-yn-1-yl)-1H-indazole-7-carboxylic acid
室温下将原料1-(4-(2-甲氧基嘧啶-4-基)苄基)-4-(丙烷-1-炔-1-基)-1H-吲唑-7-甲酸甲酯(70mg,0.17mmol)加入到四氢呋喃(3mL)，甲醇(1mL)和水(1mL)中，加入氢氧化锂(13mg,0.51mmol)，室温搅拌16h。用1N盐酸调节pH＝4，用乙酸乙酯(10mL×3)萃取，分液，合并有机相，有机相用无水硫酸钠干燥，过滤，浓缩，得化合物粗品1-(4-(2-甲氧基嘧啶-4-基)苄基)-4-(丙烷-1-炔-1-基)-1H-吲唑-7-甲酸(65mg,产率96％).The starting material 1-(4-(2-methoxypyrimidin-4-yl)benzyl)-4-(propane-1-yn-1-yl)-1H-indazole-7-carboxylic acid methyl ester ( 70mg, 0.17mmol) was added to tetrahydrofuran (3mL), methanol (1mL) and water (1mL), lithium hydroxide (13mg, 0.51mmol) was added, and stirred at room temperature for 16h. Use 1N hydrochloric acid to adjust pH=4, extract with ethyl acetate (10mL×3), separate the layers, combine the organic phases, dry the organic phases with anhydrous sodium sulfate, filter, and concentrate to obtain the crude compound 1-(4-(2- Methoxypyrimidin-4-yl)benzyl)-4-(propan-1-yn-1-yl)-1H-indazole-7-carboxylic acid (65 mg, yield 96%).
第四步：6-(1-(4-(2-甲氧基嘧啶-4-基)苄基)-4-(丙烷-1-炔-1-基)-1H-吲唑-7-甲酰胺)螺环[3.3]庚烷-2-甲酸甲酯的制备The fourth step: 6-(1-(4-(2-methoxypyrimidin-4-yl)benzyl)-4-(propane-1-yn-1-yl)-1H-indazole-7-methanol Preparation of amido)spiro[3.3]heptane-2-carboxylic acid methyl ester
室温下将化合物1-(4-(2-甲氧基嘧啶-4-基)苄基)-4-(丙烷-1-炔-1-基)-1H-吲唑-7-甲酸(65mg,0.16mmol)加入到N,N-二甲基甲酰胺(5mL)中，加入2-(6-氨基螺环[3.3]庚烷-2-基)甲酸甲酯盐酸盐(40.3mg,0.19mmol)，二异丙基乙胺(63mg,0.49mmol)，HATU(93mg,0.24mmol)，搅拌16h。加入水(10mL)稀释，用乙酸乙酯(10mL×3)萃取，分液，合并有机相，有机相用无水硫酸钠干燥，过滤，浓缩，残留物用薄层硅胶板分离纯化(DCM:甲醇(V/V)＝10:1)得化合物6-(1-(4-(2-甲氧基嘧啶-4-基)苄基)-4-(丙烷-1-炔-1-基)-1H-吲唑-7-甲酰胺)螺环[3.3]庚烷-2-甲酸甲酯(120mg,产率100％)。Compound 1-(4-(2-methoxypyrimidin-4-yl)benzyl)-4-(propane-1-yn-1-yl)-1H-indazole-7-carboxylic acid (65 mg, 0.16mmol) was added to N,N-dimethylformamide (5mL), and methyl 2-(6-aminospiro[3.3]heptane-2-yl)formate hydrochloride (40.3mg, 0.19mmol ), diisopropylethylamine (63mg, 0.49mmol), HATU (93mg, 0.24mmol), stirred for 16h. Add water (10mL) to dilute, extract with ethyl acetate (10mL×3), separate the layers, combine the organic phase, dry the organic phase with anhydrous sodium sulfate, filter, concentrate, and the residue is separated and purified with a thin-layer silica gel plate (DCM: Methanol (V/V)=10:1) to obtain compound 6-(1-(4-(2-methoxypyrimidin-4-yl)benzyl)-4-(propane-1-yn-1-yl) -1H-indazole-7-carboxamide) spiro[3.3]heptane-2-carboxylic acid methyl ester (120 mg, yield 100%).
第五步：6-(1-(4-(2-甲氧基嘧啶-4-基)苄基)-4-(丙烷-1-炔-1-基)-1H-吲唑-7-甲酰胺)螺环[3.3]庚烷-2-甲酸(I-9)的制备The fifth step: 6-(1-(4-(2-methoxypyrimidin-4-yl)benzyl)-4-(propane-1-yn-1-yl)-1H-indazole-7-methanol Preparation of amide) spiro[3.3]heptane-2-carboxylic acid (I-9)
Figure PCTCN2022097611-appb-000042
Figure PCTCN2022097611-appb-000042
室温下将化合物6-(1-(4-(2-甲氧基嘧啶-4-基)苄基)-4-(丙烷-1-炔-1-基)-1H-吲唑-7-甲酰胺)螺环[3.3]庚烷-2-甲酸甲酯(120mg,0.24mmol)加入到甲醇(1mL)和水(1mL)中，加入氢氧化锂(17mg,0.71mmol)，室温搅拌16h。用1N盐酸调节pH＝4，用乙酸乙酯(20mL×3)萃取，分液，合并有机相，有机相用无水硫酸钠干燥，过滤，浓缩，残留物经酸性制备方法A制备得化合物6-(1-(4-(2-甲氧基嘧啶-4-基)苄基)-4-(丙烷-1-炔-1-基)-1H-吲唑-7-甲酰胺)螺环[3.3]庚烷-2-甲酸(I-9)(50mg,产率43％)。Compound 6-(1-(4-(2-methoxypyrimidin-4-yl)benzyl)-4-(propane-1-yn-1-yl)-1H-indazole-7-methanol Amide) spiro[3.3]heptane-2-carboxylic acid methyl ester (120mg, 0.24mmol) was added to methanol (1mL) and water (1mL), lithium hydroxide (17mg, 0.71mmol) was added, and stirred at room temperature for 16h. Use 1N hydrochloric acid to adjust pH=4, extract with ethyl acetate (20mL×3), separate the layers, combine the organic phases, dry the organic phases with anhydrous sodium sulfate, filter, concentrate, and prepare the residue by acidic preparation method A to obtain compound 6 -(1-(4-(2-methoxypyrimidin-4-yl)benzyl)-4-(propane-1-yn-1-yl)-1H-indazole-7-carboxamide) spiro[ 3.3] Heptane-2-carboxylic acid (I-9) (50 mg, yield 43%).
1H NMR(400mHz,DMSO-d6)δ11.99(s,1H),8.73(d,1H),8.64(d,1H),8.27(s,1H),8.06(d,2H),7.64(d,1H),7.36(d,1H),7.22(d,1H),7.08(d,2H),5.86(s,2H),4.26-4.20(m,1H),3.96(s,3H),2.91-2.87(m,1H),2.38-2.22(m,4H),2.17(s,3H),2.18-1.97(m,2H),1.88-1.79(m,2H). 1 H NMR(400mHz,DMSO-d6)δ11.99(s,1H),8.73(d,1H),8.64(d,1H),8.27(s,1H),8.06(d,2H),7.64(d ,1H),7.36(d,1H),7.22(d,1H),7.08(d,2H),5.86(s,2H),4.26-4.20(m,1H),3.96(s,3H),2.91- 2.87(m,1H),2.38-2.22(m,4H),2.17(s,3H),2.18-1.97(m,2H),1.88-1.79(m,2H).
LC-MS,M/Z(ESI):536.5[M+H] +。 LC-MS, M/Z (ESI): 536.5 [M+H] + .
实施例10化合物I-10的制备The preparation of embodiment 10 compound I-10
合成路线如下所示：The synthetic route is as follows:
Figure PCTCN2022097611-appb-000043
Figure PCTCN2022097611-appb-000043
第一步：(4-((7-(甲氧羰基)-4-(丙烷-1-炔-1-基)-1H-吲唑-1-基)甲基)苄基)硼酸的制备The first step: the preparation of (4-((7-(methoxycarbonyl)-4-(propane-1-yn-1-yl)-1H-indazol-1-yl)methyl)benzyl)boronic acid
室温下将化合物4-(丙烷-1-炔-1-基)-1H-吲唑-7-甲酸甲酯(190mg,0.89mmol)加入到N,N-二甲基甲酰胺(8mL)中，加入(4-(溴甲基)苄基)硼酸(286mg,1.33mmol)，碳酸铯(867mg,2.66mmol)，室温搅拌16h。加入水(20mL)稀释，用乙酸乙酯(20mL×3)萃取，分液，合并有机相，有机相用无水硫酸钠干燥，过滤，浓缩，残留物用硅胶柱分离纯化(DCM:甲醇(V/V)＝10:1)得化合物(4-((7-(甲氧羰基)-4-(丙烷-1-炔-1-基)-1H-吲唑-1-基)甲基)苄基)硼酸(210mg,产率68％)。Compound 4-(propane-1-yn-1-yl)-1H-indazole-7-carboxylic acid methyl ester (190 mg, 0.89 mmol) was added to N,N-dimethylformamide (8 mL) at room temperature, Add (4-(bromomethyl)benzyl)boronic acid (286mg, 1.33mmol), cesium carbonate (867mg, 2.66mmol), and stir at room temperature for 16h. Add water (20mL) to dilute, extract with ethyl acetate (20mL×3), separate the layers, combine the organic phase, dry the organic phase with anhydrous sodium sulfate, filter, concentrate, and the residue is separated and purified by silica gel column (DCM: methanol ( V/V)=10:1) to obtain compound (4-((7-(methoxycarbonyl)-4-(propane-1-yn-1-yl)-1H-indazol-1-yl)methyl) Benzyl)boronic acid (210 mg, 68% yield).
第二步：4-(丙烷-1-炔-1-基)-1-(4-(噻唑-2-基)苄基)-1H-吲唑-7-甲酸甲酯的制备The second step: the preparation of 4-(propane-1-yn-1-yl)-1-(4-(thiazol-2-yl)benzyl)-1H-indazole-7-carboxylic acid methyl ester
室温下将(4-((7-(甲氧羰基)-4-(丙烷-1-炔-1-基)-1H-吲唑-1-基)甲基)苄基)硼酸(80mg,0.23mmol)加入到1,4-二氧六环(10mL)和水(1mL)中，加入氯(2-二环己基膦基-2,6-二甲氧基-1,1-联苯基)(2-氨基-1,1-联苯-2-基)钯(II)(17mg,0.02mmol)和磷酸钾(196mg,0.92mmol)，1,1'-双二苯基膦二茂铁二氯化钯(19mg,0.02mmol)，2-溴噻唑(46mg,0.28mmol)，90℃搅拌1h。加入水(20mL)稀释，用乙酸乙酯(20mL×3)萃取，分液，合并有机相，有机相用无水硫酸钠干 燥，过滤，浓缩，残留物用薄层硅胶板分离纯化(二氯甲烷:甲醇(V/V)＝10:1)得化合物4-(丙烷-1-炔-1-基)-1-(4-(噻唑-2-基)苄基)-1H-吲唑-7-甲酸甲酯(70mg,产率79％).(4-((7-(methoxycarbonyl)-4-(propane-1-yn-1-yl)-1H-indazol-1-yl)methyl)benzyl)boronic acid (80mg, 0.23 mmol) was added to 1,4-dioxane (10 mL) and water (1 mL), and chlorine (2-dicyclohexylphosphino-2,6-dimethoxy-1,1-biphenyl) (2-Amino-1,1-biphenyl-2-yl)palladium(II) (17mg, 0.02mmol) and potassium phosphate (196mg, 0.92mmol), 1,1'-bisdiphenylphosphinoferrocene Palladium chloride (19mg, 0.02mmol), 2-bromothiazole (46mg, 0.28mmol), stirred at 90°C for 1h. Add water (20mL) to dilute, extract with ethyl acetate (20mL×3), separate the layers, combine the organic phases, dry the organic phases with anhydrous sodium sulfate, filter, concentrate, and separate and purify the residue with a thin-layer silica gel plate (dichloro Methane:methanol (V/V)=10:1) to obtain compound 4-(propane-1-yn-1-yl)-1-(4-(thiazol-2-yl)benzyl)-1H-indazole- Methyl 7-formate (70mg, yield 79%).
第三步：4-(丙烷-1-炔-1-基)-1-(4-(噻唑-2-基)苄基)-1H-吲唑-7-甲酸的制备The third step: the preparation of 4-(propane-1-yn-1-yl)-1-(4-(thiazol-2-yl)benzyl)-1H-indazole-7-carboxylic acid
室温下将4-(丙烷-1-炔-1-基)-1-(4-(噻唑-2-基)苄基)-1H-吲唑-7-甲酸甲酯(70mg,0.17mmol)加入到四氢呋喃(3mL)，甲醇(1mL)和水(1mL)中，加入氢氧化锂(13mg,0.51mmol)，室温搅拌16h。用1N盐酸调节pH＝4，用乙酸乙酯(10mL×3)萃取，分液，合并有机相，有机相用无水硫酸钠干燥，过滤，浓缩，得化合物4-(丙烷-1-炔-1-基)-1-(4-(噻唑-2-基)苄基)-1H-吲唑-7-甲酸(65mg,产率94％)。Add 4-(propane-1-yn-1-yl)-1-(4-(thiazol-2-yl)benzyl)-1H-indazole-7-carboxylic acid methyl ester (70mg, 0.17mmol) at room temperature To tetrahydrofuran (3 mL), methanol (1 mL) and water (1 mL), was added lithium hydroxide (13 mg, 0.51 mmol), and stirred at room temperature for 16 h. Use 1N hydrochloric acid to adjust pH=4, extract with ethyl acetate (10mL×3), separate the layers, combine the organic phases, dry the organic phases with anhydrous sodium sulfate, filter, and concentrate to obtain compound 4-(propane-1-yne- 1-yl)-1-(4-(thiazol-2-yl)benzyl)-1H-indazole-7-carboxylic acid (65 mg, 94% yield).
第四步：6-(4-(丙烷-1-炔-1-基)-1-(4-(噻唑-2-基)苄基)-1H-吲唑-7-甲酰胺)螺环[3.3]庚烷-2-甲酸甲酯的制备The fourth step: 6-(4-(propane-1-yn-1-yl)-1-(4-(thiazol-2-yl)benzyl)-1H-indazole-7-carboxamide) spiro[ 3.3] Preparation of methyl heptane-2-carboxylate
室温下将化合物4-(丙烷-1-炔-1-基)-1-(4-(噻唑-2-基)苄基)-1H-吲唑-7-甲酸(65mg,0.17mmol)加入到N,N-二甲基甲酰胺(5mL)中，加入2-(6-氨基螺环[3.3]庚烷-2-基)甲酸甲酯盐酸盐(43mg,0.21mmol)，二异丙基乙胺(67mg,0.52mmol)，HATU(100mg,0.27mmol)，搅拌16h。加入水(10mL)稀释，用乙酸乙酯(10mL×3)萃取，分液，合并有机相，有机相用无水硫酸钠干燥，过滤，浓缩，残留物用薄层硅胶板分离纯化(DCM:甲醇(V/V)＝20:1)得化合物6-(4-(丙烷-1-炔-1-基)-1-(4-(噻唑-2-基)苄基)-1H-吲唑-7-甲酰胺)螺环[3.3]庚烷-2-甲酸甲酯(85mg,产率93％)。Compound 4-(propane-1-yn-1-yl)-1-(4-(thiazol-2-yl)benzyl)-1H-indazole-7-carboxylic acid (65mg, 0.17mmol) was added to To N,N-dimethylformamide (5mL), add 2-(6-aminospiro[3.3]heptane-2-yl)methyl formate hydrochloride (43mg, 0.21mmol), diisopropyl Ethylamine (67mg, 0.52mmol), HATU (100mg, 0.27mmol), stirred for 16h. Add water (10mL) to dilute, extract with ethyl acetate (10mL×3), separate the layers, combine the organic phase, dry the organic phase with anhydrous sodium sulfate, filter, concentrate, and the residue is separated and purified with a thin-layer silica gel plate (DCM: Methanol (V/V)=20:1) to obtain compound 6-(4-(propane-1-yn-1-yl)-1-(4-(thiazol-2-yl)benzyl)-1H-indazole -7-carboxamide) spiro[3.3]heptane-2-carboxylic acid methyl ester (85 mg, 93% yield).
第五步：6-(4-(丙烷-1-炔-1-基)-1-(4-(噻唑-2-基)苄基)-1H-吲唑-7-甲酰胺)螺环[3.3]庚烷-2-甲酸(I-10)的制备The fifth step: 6-(4-(propane-1-yn-1-yl)-1-(4-(thiazol-2-yl)benzyl)-1H-indazole-7-carboxamide) spiro[ 3.3] Preparation of heptane-2-carboxylic acid (I-10)
Figure PCTCN2022097611-appb-000044
Figure PCTCN2022097611-appb-000044
室温下将化合物6-(4-(丙烷-1-炔-1-基)-1-(4-(噻唑-2-基)苄基)-1H-吲唑-7-甲酰胺)螺环[3.3]庚烷-2-甲酸甲酯(85mg,0.16mmol)加入到四氢呋喃(3mL)，甲醇(1mL)和水(1mL)中，加入氢氧化锂(13mg,0.54mmol)，室温搅拌16h。用1N盐酸调节pH＝4，用乙酸乙酯(20mL×3)萃取，分液，合并有机相，有机相用无水硫酸钠干燥，过滤，浓缩，残留物经酸性制备方法A制备得化合物6-(4-(丙烷-1-炔-1-基)-1-(4-(噻唑-2-基)苄基)-1H-吲唑-7-甲酰胺)螺环[3.3]庚烷-2-甲酸(I-10)(55mg,产率66％)。Compound 6-(4-(propane-1-yn-1-yl)-1-(4-(thiazol-2-yl)benzyl)-1H-indazole-7-carboxamide) spiro[ 3.3] Add methyl heptane-2-carboxylate (85mg, 0.16mmol) to tetrahydrofuran (3mL), methanol (1mL) and water (1mL), add lithium hydroxide (13mg, 0.54mmol), and stir at room temperature for 16h. Use 1N hydrochloric acid to adjust pH=4, extract with ethyl acetate (20mL×3), separate the layers, combine the organic phases, dry the organic phases with anhydrous sodium sulfate, filter, concentrate, and prepare the residue by acidic preparation method A to obtain compound 6 -(4-(propane-1-yn-1-yl)-1-(4-(thiazol-2-yl)benzyl)-1H-indazole-7-carboxamide)spiro[3.3]heptane- 2-Formic acid (I-10) (55 mg, yield 66%).
1H NMR(400mHz,DMSO-d6)δ12.0(s,1H),8.72(d,1H),7.89(d,1H),7.82(d,2H),7.76(d,1H),7.36(d,1H),7.22(d,2H),7.05(d,2H),5.82(s,2H),4.26-4.21(m,1H),2.91-2.87(m,1H),2.40-2.22(m,4H),2.18(s,3H),2.18-1.96(m,2H),1.89-1.79(m,2H)。 1 H NMR (400mHz,DMSO-d6)δ12.0(s,1H),8.72(d,1H),7.89(d,1H),7.82(d,2H),7.76(d,1H),7.36(d ,1H),7.22(d,2H),7.05(d,2H),5.82(s,2H),4.26-4.21(m,1H),2.91-2.87(m,1H),2.40-2.22(m,4H ), 2.18(s,3H), 2.18-1.96(m,2H), 1.89-1.79(m,2H).
LC-MS,M/Z(ESI):511.4[M+H] +。 LC-MS, M/Z (ESI): 511.4 [M+H] + .
实施例11化合物I-11的制备The preparation of embodiment 11 compound I-11
合成路线如下所示：The synthetic route is as follows:
Figure PCTCN2022097611-appb-000045
Figure PCTCN2022097611-appb-000045
第一步：1-(4-碘苯基)-4-(丙烷-1-炔-1-基)-1H-吲唑-7-甲酸甲酯的制备Step 1: Preparation of 1-(4-iodophenyl)-4-(propane-1-yn-1-yl)-1H-indazole-7-carboxylic acid methyl ester
室温下将化合物4-(丙烷-1-炔-1-基)-1H-吲唑-7-甲酸甲酯(400mg,1.87mmol)加入到N,N-二甲基甲酰胺(10mL)中，加入1-(溴甲基)-4-碘苯(832mg,2.81mmol)，碳酸铯(1.82g,5.61mmol)，室温搅拌16h。加入水(20mL)稀释，用乙酸乙酯(20mL×3)萃取，分液，合并有机相，有机相用无水硫酸钠干燥，过滤，浓缩，残留物经酸性制备方法A得化合物1-(4-碘苯基)-4-(丙烷-1-炔-1-基)-1H-吲唑-7-甲酸甲酯(300mg,产率37％)。Compound 4-(propane-1-yn-1-yl)-1H-indazole-7-carboxylic acid methyl ester (400 mg, 1.87 mmol) was added into N,N-dimethylformamide (10 mL) at room temperature, Add 1-(bromomethyl)-4-iodobenzene (832mg, 2.81mmol), cesium carbonate (1.82g, 5.61mmol), and stir at room temperature for 16h. Add water (20mL) to dilute, extract with ethyl acetate (20mL×3), separate the layers, combine the organic phases, dry the organic phases with anhydrous sodium sulfate, filter, concentrate, and the residue is subjected to acidic preparation method A to obtain compound 1-( 4-iodophenyl)-4-(propan-1-yn-1-yl)-1H-indazole-7-carboxylic acid methyl ester (300 mg, 37% yield).
第二步：1-(4-(1H-吡唑-1-基)苄基)-4-(丙烷-1-炔-1-基)-1H-吲唑-7-甲酸甲酯的制备The second step: the preparation of 1-(4-(1H-pyrazol-1-yl)benzyl)-4-(propane-1-yn-1-yl)-1H-indazole-7-carboxylic acid methyl ester
室温下将1-(4-碘苯基)-4-(丙烷-1-炔-1-基)-1H-吲唑-7-甲酸甲酯(300mg,0.70mmol)加入到N,N-二甲基甲酰胺(10mL)中，加入1H-吡唑(95mg,1.40mmol)、碘化亚铜(27mg，0.14mmol)和碳酸铯(454mg，1.40mmol)，120℃搅拌72h。加入水(20mL)稀释，用乙酸乙酯(20mL×3)萃取，分液，合并有机相，有机相用无水硫酸钠干燥，过滤，浓缩，残留物用薄层硅胶板分离纯化(石油醚:乙酸乙酯(V/V)＝1:1)得化合物1-(4-(1H-吡唑-1-基)苄基)-4-(丙烷-1-炔-1-基)-1H-吲唑-7-甲酸甲酯(40mg,产率14％).Add 1-(4-iodophenyl)-4-(propane-1-yn-1-yl)-1H-indazole-7-carboxylic acid methyl ester (300mg, 0.70mmol) to N,N-di Add 1H-pyrazole (95mg, 1.40mmol), cuprous iodide (27mg, 0.14mmol) and cesium carbonate (454mg, 1.40mmol) to methylformamide (10mL), and stir at 120°C for 72h. Add water (20mL) to dilute, extract with ethyl acetate (20mL×3), separate the layers, combine the organic phases, dry the organic phases with anhydrous sodium sulfate, filter, concentrate, and separate and purify the residue with a thin-layer silica gel plate (petroleum ether : Ethyl acetate (V/V)=1:1) to obtain compound 1-(4-(1H-pyrazol-1-yl)benzyl)-4-(propane-1-yn-1-yl)-1H - Methyl indazole-7-carboxylate (40mg, yield 14%).
第四步：1-(4-(1H-吡唑-1-基)苄基)-4-(丙烷-1-炔-1-基)-1H-吲唑-7-甲酸的制备Step 4: Preparation of 1-(4-(1H-pyrazol-1-yl)benzyl)-4-(propane-1-yn-1-yl)-1H-indazole-7-carboxylic acid
室温下将1-(4-(1H-吡唑-1-基)苄基)-4-(丙烷-1-炔-1-基)-1H-吲唑-7-甲酸甲酯(40mg,0.11mmol)加入到四氢呋喃(3mL)和水(1mL)中，加入氢氧化锂(8mg,0.33mmol)，室温搅拌16h。用1N盐酸调节pH＝4，用乙酸乙酯(10mL×3)萃取，分液，合并有机相，有机相用无水硫酸钠干燥，过滤，浓缩，得化合物粗品1-(4-(1H-吡唑-1-基)苄基)-4-(丙烷-1-炔-1-基)-1H-吲唑-7-甲酸(35mg,产率90％)。Methyl 1-(4-(1H-pyrazol-1-yl)benzyl)-4-(propane-1-yn-1-yl)-1H-indazole-7-carboxylate (40mg, 0.11 mmol) was added to tetrahydrofuran (3 mL) and water (1 mL), lithium hydroxide (8 mg, 0.33 mmol) was added, and stirred at room temperature for 16 h. Use 1N hydrochloric acid to adjust pH=4, extract with ethyl acetate (10mL×3), separate the layers, combine the organic phases, dry the organic phases with anhydrous sodium sulfate, filter, and concentrate to obtain the crude compound 1-(4-(1H- Pyrazol-1-yl)benzyl)-4-(propan-1-yn-1-yl)-1H-indazole-7-carboxylic acid (35 mg, 90% yield).
第四步：6-(1-(4-(1H-吡唑-1-基)苄基)-4-(丙烷-1-炔-1-基)-1H-吲唑-7-甲酰胺)螺环[3.3]庚烷-2-甲酸甲酯的制备The fourth step: 6-(1-(4-(1H-pyrazol-1-yl)benzyl)-4-(propane-1-yn-1-yl)-1H-indazole-7-carboxamide) Preparation of spiro[3.3]heptane-2-carboxylate methyl ester
室温下将化合物1-(4-(1H-吡唑-1-基)苄基)-4-(丙烷-1-炔-1-基)-1H-吲唑-7-甲酸(35mg,0.10mmol)加入到N,N-二甲基甲酰胺(5mL)中，加入2-(6-氨基螺环[3.3]庚烷-2-基)甲酸甲酯盐酸盐 (31mg,0.15mmol)，二异丙基乙胺(39mg,0.30mmol)，HATU(57mg,0.15mmol)，搅拌16h。加入水(10mL)稀释，用乙酸乙酯(10mL×3)萃取，分液，合并有机相，有机相用无水硫酸钠干燥，过滤，浓缩，残留物用薄层硅胶板分离纯化(石油醚:乙酸乙酯(V/V)＝1:1)得化合物6-(1-(4-(1H-吡唑-1-基)苄基)-4-(丙烷-1-炔-1-基)-1H-吲唑-7-甲酰胺)螺环[3.3]庚烷-2-甲酸甲酯(35mg,产率70％)。Compound 1-(4-(1H-pyrazol-1-yl)benzyl)-4-(propane-1-yn-1-yl)-1H-indazole-7-carboxylic acid (35mg, 0.10mmol ) into N,N-dimethylformamide (5mL), add 2-(6-aminospiro[3.3]heptane-2-yl)methyl formate hydrochloride (31mg, 0.15mmol), di Isopropylethylamine (39mg, 0.30mmol), HATU (57mg, 0.15mmol), stirred for 16h. Add water (10mL) to dilute, extract with ethyl acetate (10mL×3), separate the layers, combine the organic phases, dry the organic phases with anhydrous sodium sulfate, filter, concentrate, and separate and purify the residue with a thin-layer silica gel plate (petroleum ether : Ethyl acetate (V/V)=1:1) to obtain compound 6-(1-(4-(1H-pyrazol-1-yl) benzyl)-4-(propane-1-yn-1-yl )-1H-indazole-7-carboxamide)spiro[3.3]heptane-2-carboxylic acid methyl ester (35 mg, 70% yield).
第五步：6-(1-(4-(1H-吡唑-1-基)苄基)-4-(丙烷-1-炔-1-基)-1H-吲唑-7-甲酰胺)螺环[3.3]庚烷-2-甲酸(I-11)的制备The fifth step: 6-(1-(4-(1H-pyrazol-1-yl)benzyl)-4-(propane-1-yn-1-yl)-1H-indazole-7-carboxamide) Preparation of spiro[3.3]heptane-2-carboxylic acid (I-11)
Figure PCTCN2022097611-appb-000046
Figure PCTCN2022097611-appb-000046
室温下将化合物6-(1-(4-(1H-吡唑-1-基)苄基)-4-(丙烷-1-炔-1-基)-1H-吲唑-7-甲酰胺)螺环[3.3]庚烷-2-甲酸甲酯(35mg,0.07mmol)加入到四氢呋喃(3mL)和水(1mL)中，加入氢氧化锂(5mg,0.21mmol)，室温搅拌16h。用1N盐酸调节pH＝4，用乙酸乙酯(10mL×3)萃取，分液，合并有机相，有机相用无水硫酸钠干燥，过滤，浓缩，残留物经酸性制备方法A制备得化合物6-(1-(4-(1H-吡唑-1-基)苄基)-4-(丙烷-1-炔-1-基)-1H-吲唑-7-甲酰胺)螺环[3.3]庚烷-2-甲酸(I-11)(22mg,产率64％)。Compound 6-(1-(4-(1H-pyrazol-1-yl)benzyl)-4-(propane-1-yn-1-yl)-1H-indazole-7-carboxamide) Methyl spiro[3.3]heptane-2-carboxylate (35mg, 0.07mmol) was added to tetrahydrofuran (3mL) and water (1mL), lithium hydroxide (5mg, 0.21mmol) was added, and stirred at room temperature for 16h. Use 1N hydrochloric acid to adjust pH=4, extract with ethyl acetate (10mL×3), separate the layers, combine the organic phases, dry the organic phases with anhydrous sodium sulfate, filter, concentrate, and prepare the residue by acidic preparation method A to obtain compound 6 -(1-(4-(1H-pyrazol-1-yl)benzyl)-4-(propane-1-yn-1-yl)-1H-indazole-7-carboxamide) spiro[3.3] Heptane-2-carboxylic acid (I-11) (22 mg, yield 64%).
1H NMR(400mHz,DMSO-d6)δ12.00(s,1H),8.76(d,1H),8.41(d,1H),8.25(s,1H),7.70(s，1H),7.68(d,2H),7.40(d,1H),7.07(d,1H),7.04(d,2H),6.51(t,1H),5.79(s,2H),4.30-4.24(m,1H),2.94-2.90(m,1H),2.32-2.30(m,1H),2.27-2.20(m,3H),2.17(s,3H),2.08-2.01(m,2H),1.93-1.83(m,2H)。 1 H NMR (400mHz,DMSO-d6)δ12.00(s,1H),8.76(d,1H),8.41(d,1H),8.25(s,1H),7.70(s,1H),7.68(d ,2H),7.40(d,1H),7.07(d,1H),7.04(d,2H),6.51(t,1H),5.79(s,2H),4.30-4.24(m,1H),2.94- 2.90 (m, 1H), 2.32-2.30 (m, 1H), 2.27-2.20 (m, 3H), 2.17 (s, 3H), 2.08-2.01 (m, 2H), 1.93-1.83 (m, 2H).
LC-MS,M/Z(ESI):494.4[M+H] +。 LC-MS, M/Z (ESI): 494.4 [M+H] + .
实施例12化合物I-12的制备The preparation of embodiment 12 compound I-12
合成路线如下所示：The synthetic route is as follows:
Figure PCTCN2022097611-appb-000047
Figure PCTCN2022097611-appb-000047
第一步：2-氨基-4-溴-3-甲基苯甲酸甲酯的制备The first step: the preparation of 2-amino-4-bromo-3-methylbenzoic acid methyl ester
室温下将化合物2-氨基-4-溴-3-甲基苯甲酸(5.5g,23.9mmol)加入到N,N-二甲基甲酰胺(70mL)中，加入碘甲烷(3.4g,24.0mmol)，碳酸铯(11.7g,35.8mmol)，搅拌20h。加入水(200mL)稀释，用乙酸乙酯(200mL×3)萃取，分液，合并有机相，有机相用无水硫酸钠干燥，过滤，浓缩，残留物用硅胶柱分离纯化(石油醚:乙酸乙酯(V/V)＝4:1)得化合物2-氨基-4-溴-3-甲基苯甲酸甲酯(5.0g,产率85％)。Compound 2-amino-4-bromo-3-methylbenzoic acid (5.5g, 23.9mmol) was added to N,N-dimethylformamide (70mL) at room temperature, and iodomethane (3.4g, 24.0mmol ), cesium carbonate (11.7g, 35.8mmol), stirred for 20h. Add water (200mL) to dilute, extract with ethyl acetate (200mL×3), separate the layers, combine the organic phase, dry the organic phase with anhydrous sodium sulfate, filter, concentrate, and the residue is separated and purified by silica gel column (petroleum ether: acetic acid ethyl ester (V/V)=4:1) to obtain the compound 2-amino-4-bromo-3-methylbenzoic acid methyl ester (5.0 g, yield 85%).
第二步：(3-溴-6-(甲氧羰基)-2-甲基苯基)-1-四氟硼酸重氮盐的制备The second step: the preparation of (3-bromo-6-(methoxycarbonyl)-2-methylphenyl)-1-tetrafluoroborate diazonium salt
室温下将化合物2-氨基-4-溴-3-甲基苯甲酸甲酯(5.0g,20.5mmol)加入到50％HBF 4水溶液(5mL)中，冷却至0℃，滴加亚硝酸钠(1.42g,20.6mmol)的5mL水溶液，0℃下搅拌1h。过滤，干燥，得化合物粗品(3-溴-6-(甲氧羰基)-2-甲基苯基)-1-四氟硼酸重氮盐(6.6g,产率94％)。 The compound 2-amino-4-bromo-3-methylbenzoate (5.0 g, 20.5 mmol) was added to 50% HBF 4 aqueous solution (5 mL) at room temperature, cooled to 0 ° C, and sodium nitrite ( 1.42g, 20.6mmol) in 5mL aqueous solution, stirred at 0°C for 1h. After filtration and drying, the crude compound (3-bromo-6-(methoxycarbonyl)-2-methylphenyl)-1-tetrafluoroborate diazonium salt (6.6 g, yield 94%) was obtained.
第三步：4-溴-1H-吲唑-7-甲酸甲酯的制备The third step: the preparation of 4-bromo-1H-indazole-7-carboxylic acid methyl ester
室温下将化合物(3-溴-6-(甲氧羰基)-2-甲基苯基)-1-四氟硼酸重氮盐(6.6g,19.3mmol)加入到氯仿(70mL)中，加入醋酸钾(3.8g,38.7mmol)，18-冠-6(100mg)，室温搅拌1h。浓缩，残留物用硅胶柱分离纯化(石油醚:乙酸乙酯(V/V)＝4:1)得化合物4-溴-1H-吲唑-7-甲酸甲酯(4.2g,产率94％)。Compound (3-bromo-6-(methoxycarbonyl)-2-methylphenyl)-1-tetrafluoroborate diazonium salt (6.6g, 19.3mmol) was added to chloroform (70mL) at room temperature, and acetic acid was added Potassium (3.8g, 38.7mmol), 18-crown-6 (100mg), stirred at room temperature for 1h. Concentrate, and the residue is separated and purified by silica gel column (petroleum ether:ethyl acetate (V/V)=4:1) to obtain compound 4-bromo-1H-indazole-7-carboxylic acid methyl ester (4.2g, yield 94% ).
第四步：4-(丙烷-1-炔-1-基)-1H-吲唑-7-甲酸甲酯的制备Step 4: Preparation of 4-(propane-1-yn-1-yl)-1H-indazole-7-carboxylic acid methyl ester
室温下将化合物4-溴-1H-吲唑-7-甲酸甲酯(1.0g,3.94mmol)加入到1,4-二氧六环(10mL)中，加入二三苯基膦二氯化钯(274mg,0.39mmol)，1-丙炔-三正丁基锡(1.52g,4.61mmol)，氮气保护下加热至90℃，搅拌16h。冷却至室温，加入水(50mL)稀释，用乙酸乙酯(50mL×3)萃取，分液，合并有机相，有机相用无水硫酸钠干燥，过滤，浓缩，残留物用薄层硅胶板分离纯化(石油醚:乙酸乙酯(V/V)＝5:1)得化合物4-(丙烷-1-炔-1-基)-1H-吲唑-7-甲酸甲酯(500mg,产率42％)。Add the compound 4-bromo-1H-indazole-7-carboxylic acid methyl ester (1.0g, 3.94mmol) to 1,4-dioxane (10mL) at room temperature, add ditriphenylphosphine palladium dichloride (274mg, 0.39mmol), 1-propyne-tri-n-butyltin (1.52g, 4.61mmol), heated to 90°C under nitrogen protection, and stirred for 16h. Cool to room temperature, add water (50mL) to dilute, extract with ethyl acetate (50mL×3), separate the liquids, combine the organic phases, dry the organic phases with anhydrous sodium sulfate, filter, concentrate, and separate the residue with a thin-layer silica gel plate Purification (petroleum ether: ethyl acetate (V/V) = 5:1) gave compound 4-(propane-1-yn-1-yl)-1H-indazole-7-carboxylic acid methyl ester (500mg, yield 42 %).
第五步：1-(4-环丙基苄基)-4-(丙烷-1-炔-1-基)-1H-吲唑-7-甲酸甲酯的制备Step 5: Preparation of 1-(4-cyclopropylbenzyl)-4-(propane-1-yn-1-yl)-1H-indazole-7-carboxylic acid methyl ester
室温下将化合物4-(丙烷-1-炔-1-基)-1H-吲唑-7-甲酸甲酯(150mg,0.70mmol)加入到N,N-二甲基甲酰胺(5mL)中，加入4-环丙基苄溴(200mg,0.95mmol)，碳酸铯(460mg,1.40mmol)，室温搅拌16h。加入水(20mL)稀释，用乙酸乙酯(20mL×3)萃取，分液，合并有机相，有机相用无水硫酸钠干燥，过滤，浓缩，残留物用薄层硅胶板分离纯化(石油醚:乙酸乙酯(V/V)＝10:1)得化合物1-(4-环丙基苄基)-4-(丙烷-1-炔-1-基)-1H-吲唑-7-甲酸甲酯(120mg,产率49％)。Compound 4-(propane-1-yn-1-yl)-1H-indazole-7-carboxylic acid methyl ester (150 mg, 0.70 mmol) was added into N,N-dimethylformamide (5 mL) at room temperature, Add 4-cyclopropylbenzyl bromide (200mg, 0.95mmol), cesium carbonate (460mg, 1.40mmol), and stir at room temperature for 16h. Add water (20mL) to dilute, extract with ethyl acetate (20mL×3), separate the layers, combine the organic phases, dry the organic phases with anhydrous sodium sulfate, filter, concentrate, and separate and purify the residue with a thin-layer silica gel plate (petroleum ether : Ethyl acetate (V/V)=10:1) to obtain compound 1-(4-cyclopropylbenzyl)-4-(propane-1-yn-1-yl)-1H-indazole-7-carboxylic acid Methyl ester (120 mg, 49% yield).
第六步：1-(4-环丙基苄基)-4-(丙烷-1-炔-1-基)-1H-吲唑-7-甲酸的制备Step 6: Preparation of 1-(4-cyclopropylbenzyl)-4-(propane-1-yn-1-yl)-1H-indazole-7-carboxylic acid
室温下将1-(4-环丙基苄基)-4-(丙烷-1-炔-1-基)-1H-吲唑-7-甲酸甲酯(120mg,0.35mmol)加入到四氢呋喃(4mL)和水(1mL)中，加入氢氧化锂(25mg,1.0mmol)，室温搅拌4h。用1N盐酸调节pH＝4，用乙酸乙酯(20mL×3)萃取，分液，合并有机相，有机相用无水硫酸钠干燥，过滤，浓缩，得化合物粗品1-(4-环丙基苄基)-4-(丙烷-1-炔-1-基)-1H-吲唑-7-甲酸(130mg,产率100％)。Add 1-(4-cyclopropylbenzyl)-4-(propane-1-yn-1-yl)-1H-indazole-7-carboxylic acid methyl ester (120mg, 0.35mmol) into tetrahydrofuran (4mL ) and water (1 mL), add lithium hydroxide (25 mg, 1.0 mmol), and stir at room temperature for 4 h. Use 1N hydrochloric acid to adjust pH=4, extract with ethyl acetate (20mL×3), separate the layers, combine the organic phases, dry the organic phases with anhydrous sodium sulfate, filter, and concentrate to obtain the crude compound 1-(4-cyclopropyl Benzyl)-4-(propan-1-yn-1-yl)-1H-indazole-7-carboxylic acid (130 mg, 100% yield).
第七步：6-(1-(4-环丙基苄基)-4-(丙烷-1-炔-1-基)-1H-吲唑-7-甲酰胺)螺环[3.3]庚烷-2-甲酸甲酯的制备The seventh step: 6-(1-(4-cyclopropylbenzyl)-4-(propane-1-yn-1-yl)-1H-indazole-7-carboxamide)spiro[3.3]heptane - Preparation of methyl 2-formate
室温下将化合物1-(4-环丙基苄基)-4-(丙烷-1-炔-1-基)-1H-吲唑-7-甲酸(130mg,0.39mmol)加入到N,N-二甲基甲酰胺(3mL)中，加入2-(6-氨基螺环[3.3]庚烷-2-基)甲酸甲酯盐酸盐(97mg,0.47mmol)，二异丙基乙胺(152mg,1.18mmol)，HATU(225mg,0.59mmol)，搅拌3h。加入水(20mL)稀释，用乙酸乙酯(20mL×3)萃取，分液，合并有机相，有机相用无水硫酸钠干燥，过滤，浓缩，残留物用薄层硅胶板分离纯化(石油醚:乙酸乙酯(V/V)＝10:1)得化合物6-(1-(4-环丙基苄基)-4-(丙烷-1-炔-1-基)-1H-吲唑-7-甲酰胺)螺环[3.3]庚烷-2-甲酸甲酯(156mg,产率82％)。Compound 1-(4-cyclopropylbenzyl)-4-(propane-1-yn-1-yl)-1H-indazole-7-carboxylic acid (130mg, 0.39mmol) was added to N,N- In dimethylformamide (3mL), add 2-(6-aminospirocyclo[3.3]heptane-2-yl)methyl formate hydrochloride (97mg, 0.47mmol), diisopropylethylamine (152mg , 1.18mmol), HATU (225mg, 0.59mmol), stirred for 3h. Add water (20mL) to dilute, extract with ethyl acetate (20mL×3), separate the layers, combine the organic phases, dry the organic phases with anhydrous sodium sulfate, filter, concentrate, and separate and purify the residue with a thin-layer silica gel plate (petroleum ether : ethyl acetate (V/V)=10:1) to obtain compound 6-(1-(4-cyclopropylbenzyl)-4-(propane-1-yn-1-yl)-1H-indazole- 7-Carboxamide) spiro[3.3]heptane-2-carboxylic acid methyl ester (156 mg, 82% yield).
第八步：6-(1-(4-环丙基苄基)-4-(丙烷-1-炔-1-基)-1H-吲唑-7-甲酰胺)螺环[3.3]庚烷-2-甲酸(I-12)的制备The eighth step: 6-(1-(4-cyclopropylbenzyl)-4-(propane-1-yn-1-yl)-1H-indazole-7-carboxamide) spiro[3.3]heptane - Preparation of 2-formic acid (I-12)
Figure PCTCN2022097611-appb-000048
Figure PCTCN2022097611-appb-000048
室温下将化合物6-(1-(4-环丙基苄基)-4-(丙烷-1-炔-1-基)-1H-吲唑-7-甲酰胺)螺环[3.3]庚烷-2-甲酸甲酯(156mg,0.33mmol)加入到四氢呋喃(3mL)，甲醇(1mL)和水(1mL)中，加入氢氧化锂(24mg,1.0mmol)，室温搅拌4h。用1N盐酸调节pH＝4，用乙酸乙酯(20mL×3)萃取，分液，合并有机相，有机相用无水硫酸钠干燥，过滤，浓缩，残留物经酸性制备方法A制备得化合物6-(1-(4-环丙基苄基)-4-(丙烷-1-炔-1-基)-1H-吲唑-7-甲酰胺)螺环[3.3]庚烷-2-甲酸(I-12)(55mg,产率36％)。Compound 6-(1-(4-cyclopropylbenzyl)-4-(propane-1-yn-1-yl)-1H-indazole-7-carboxamide) spiro[3.3]heptane - Methyl 2-carboxylate (156mg, 0.33mmol) was added to tetrahydrofuran (3mL), methanol (1mL) and water (1mL), lithium hydroxide (24mg, 1.0mmol) was added, and stirred at room temperature for 4h. Use 1N hydrochloric acid to adjust pH=4, extract with ethyl acetate (20mL×3), separate the layers, combine the organic phases, dry the organic phases with anhydrous sodium sulfate, filter, concentrate, and prepare the residue by acidic preparation method A to obtain compound 6 -(1-(4-cyclopropylbenzyl)-4-(propane-1-yn-1-yl)-1H-indazole-7-carboxamide) spiro[3.3]heptane-2-carboxylic acid ( 1-12) (55 mg, 36% yield).
1H NMR(400mHz,DMSO-d6)δ11.93(s,1H),8.74(d,1H),8.21(s,1H),7.34(d,1H),7.20(d, 1H),6.92(d,2H),6.85(d,2H),5.70(s,2H),4.29-4.23(m,1H),2.96(t,1H),2.46-2.40(m,1H),2.28-2.22(m,3H),2.17(s,3H),2.15-2.09(m,2H),1.87-1.82(m,3H),0.89-0.87(m,2H)0.57-0.56(m,2H)。 1 H NMR (400mHz,DMSO-d6)δ11.93(s,1H),8.74(d,1H),8.21(s,1H),7.34(d,1H),7.20(d,1H),6.92(d ,2H),6.85(d,2H),5.70(s,2H),4.29-4.23(m,1H),2.96(t,1H),2.46-2.40(m,1H),2.28-2.22(m,3H ), 2.17(s,3H), 2.15-2.09(m,2H), 1.87-1.82(m,3H), 0.89-0.87(m,2H), 0.57-0.56(m,2H).
LC-MS,M/Z(ESI):468.4[M+H] +。 LC-MS, M/Z (ESI): 468.4 [M+H] + .
实施例13化合物I-13的制备The preparation of embodiment 13 compound I-13
合成路线如下所示：The synthetic route is as follows:
Figure PCTCN2022097611-appb-000049
Figure PCTCN2022097611-appb-000049
第一步：1-(4-碘苯基)-4-(丙烷-1-炔-1-基)-1H-吲唑-7-甲酸甲酯的制备Step 1: Preparation of 1-(4-iodophenyl)-4-(propane-1-yn-1-yl)-1H-indazole-7-carboxylic acid methyl ester
室温下将化合物4-(丙烷-1-炔-1-基)-1H-吲唑-7-甲酸甲酯(400mg,1.87mmol)加入到N,N-二甲基甲酰胺(10mL)中，加入1-(溴甲基)-4-碘苯(832mg,2.81mmol)，碳酸铯(1.82g,5.61mmol)，室温搅拌16h。加入水(20mL)稀释，用乙酸乙酯(20mL×3)萃取，分液，合并有机相，有机相用无水硫酸钠干燥，过滤，浓缩，残留物经酸性制备方法A得化合物1-(4-碘苯基)-4-(丙烷-1-炔-1-基)-1H-吲唑-7-甲酸甲酯(300mg,产率37％)。Compound 4-(propane-1-yn-1-yl)-1H-indazole-7-carboxylic acid methyl ester (400 mg, 1.87 mmol) was added into N,N-dimethylformamide (10 mL) at room temperature, Add 1-(bromomethyl)-4-iodobenzene (832mg, 2.81mmol), cesium carbonate (1.82g, 5.61mmol), and stir at room temperature for 16h. Add water (20mL) to dilute, extract with ethyl acetate (20mL×3), separate the layers, combine the organic phases, dry the organic phases with anhydrous sodium sulfate, filter, concentrate, and the residue is subjected to acidic preparation method A to obtain compound 1-( 4-iodophenyl)-4-(propan-1-yn-1-yl)-1H-indazole-7-carboxylic acid methyl ester (300 mg, 37% yield).
第二步：1-(4-(2-甲氧基吡啶-4-基)苯基)-4-(丙烷-1-炔-1-基)-1H-吲唑-7-甲酸甲酯的制备The second step: 1-(4-(2-methoxypyridin-4-yl)phenyl)-4-(propane-1-yn-1-yl)-1H-indazole-7-carboxylic acid methyl ester preparation
室温下将1-(4-碘苯基)-4-(丙烷-1-炔-1-基)-1H-吲唑-7-甲酸甲酯(300mg,0.70mmol)和(2-甲氧基吡啶-4-基)硼酸(161mg，1.05mmol)加入到1,4-二氧六环(10mL)和水(3mL)中，加入氯(2-二环己基膦基2'，6'-二甲氧基-1,1'-联苯)[2-(2'-氨基-1,1'-联苯)]钯(II)(50mg,0.07mmol)和磷酸钾(594mg，2.80mmol)，50℃搅拌16h。加入水(20mL)稀释，用乙酸乙酯(20mL×3)萃取，分液，合并有机相，有机相用无水硫酸钠干燥，过滤，浓缩，残留物用薄层硅胶板分离纯化(石油醚:乙酸乙酯(V/V)＝1:1)得化合物1-(4-(2-甲氧基吡啶-4-基)苯基)-4-(丙烷-1-炔-1-基)-1H-吲唑-7-甲酸甲酯(100mg,产率35％).Methyl 1-(4-iodophenyl)-4-(propane-1-yn-1-yl)-1H-indazole-7-carboxylate (300mg, 0.70mmol) and (2-methoxy Pyridin-4-yl)boronic acid (161 mg, 1.05 mmol) was added to 1,4-dioxane (10 mL) and water (3 mL), and chloro(2-dicyclohexylphosphino 2',6'-di Methoxy-1,1'-biphenyl)[2-(2'-amino-1,1'-biphenyl)]palladium(II) (50 mg, 0.07 mmol) and potassium phosphate (594 mg, 2.80 mmol), Stir at 50°C for 16h. Add water (20mL) to dilute, extract with ethyl acetate (20mL×3), separate the layers, combine the organic phases, dry the organic phases with anhydrous sodium sulfate, filter, concentrate, and separate and purify the residue with a thin-layer silica gel plate (petroleum ether : Ethyl acetate (V/V)=1:1) to obtain compound 1-(4-(2-methoxypyridin-4-yl)phenyl)-4-(propane-1-yn-1-yl) -1H-indazole-7-carboxylic acid methyl ester (100mg, yield 35%).
第三步：1-(4-(2-甲氧基吡啶-4-基)苯基)-4-(丙烷-1-炔-1-基)-1H-吲唑-7-甲酸的制备The third step: the preparation of 1-(4-(2-methoxypyridin-4-yl)phenyl)-4-(propane-1-yn-1-yl)-1H-indazole-7-carboxylic acid
室温下将1-(4-(2-甲氧基吡啶-4-基)苯基)-4-(丙烷-1-炔-1-基)-1H-吲唑-7-甲酸甲酯(100mg,0.24mmol)加入到四氢呋喃(3mL)和水(1mL)中，加入氢氧化锂(18mg,0.73mmol)，室温搅拌 16h。用1N盐酸调节pH＝4，用乙酸乙酯(10mL×3)萃取，分液，合并有机相，有机相用无水硫酸钠干燥，过滤，浓缩，得化合物粗品1-(4-(2-甲氧基吡啶-4-基)苯基)-4-(丙烷-1-炔-1-基)-1H-吲唑-7-甲酸(70mg,产率72％).Methyl 1-(4-(2-methoxypyridin-4-yl)phenyl)-4-(propane-1-yn-1-yl)-1H-indazole-7-carboxylate (100mg , 0.24mmol) was added to tetrahydrofuran (3mL) and water (1mL), lithium hydroxide (18mg, 0.73mmol) was added, and stirred at room temperature for 16h. Use 1N hydrochloric acid to adjust pH=4, extract with ethyl acetate (10mL×3), separate the layers, combine the organic phases, dry the organic phases with anhydrous sodium sulfate, filter, and concentrate to obtain the crude compound 1-(4-(2- Methoxypyridin-4-yl)phenyl)-4-(propane-1-yn-1-yl)-1H-indazole-7-carboxylic acid (70mg, yield 72%).
第四步：6-(1-(4-(2-甲氧基吡啶-4-基)苯基)-4-(丙烷-1-炔-1-基)-1H-吲唑-7-甲酰胺)螺环[3.3]庚烷-2-甲酸甲酯的制备The fourth step: 6-(1-(4-(2-methoxypyridin-4-yl)phenyl)-4-(propane-1-yn-1-yl)-1H-indazole-7-methan Preparation of amido)spiro[3.3]heptane-2-carboxylic acid methyl ester
室温下将化合物1-(4-(2-甲氧基吡啶-4-基)苯基)-4-(丙烷-1-炔-1-基)-1H-吲唑-7-甲酸(70mg,0.18mmol)加入到N,N-二甲基甲酰胺(5mL)中，加入2-(6-氨基螺环[3.3]庚烷-2-基)甲酸甲酯盐酸盐(55mg,0.26mmol)，二异丙基乙胺(67mg,0.52mmol)，HATU(99mg,0.26mmol)，搅拌16h。加入水(10mL)稀释，用乙酸乙酯(10mL×3)萃取，分液，合并有机相，有机相用无水硫酸钠干燥，过滤，浓缩，残留物用薄层硅胶板分离纯化(石油醚:乙酸乙酯(V/V)＝1:1)得化合物6-(1-(4-(2-甲氧基吡啶-4-基)苯基)-4-(丙烷-1-炔-1-基)-1H-吲唑-7-甲酰胺)螺环[3.3]庚烷-2-甲酸甲酯(60mg,产率62％)。Compound 1-(4-(2-methoxypyridin-4-yl)phenyl)-4-(propane-1-yn-1-yl)-1H-indazole-7-carboxylic acid (70mg, 0.18mmol) was added to N,N-dimethylformamide (5mL), and 2-(6-aminospirocyclo[3.3]heptane-2-yl)methyl formate hydrochloride (55mg, 0.26mmol) was added , diisopropylethylamine (67mg, 0.52mmol), HATU (99mg, 0.26mmol), stirred for 16h. Add water (10mL) to dilute, extract with ethyl acetate (10mL×3), separate the layers, combine the organic phases, dry the organic phases with anhydrous sodium sulfate, filter, concentrate, and separate and purify the residue with a thin-layer silica gel plate (petroleum ether : ethyl acetate (V/V)=1:1) to obtain compound 6-(1-(4-(2-methoxypyridin-4-yl)phenyl)-4-(propane-1-yne-1 -yl)-1H-indazole-7-carboxamide)spiro[3.3]heptane-2-carboxylic acid methyl ester (60 mg, yield 62%).
第五步：6-(1-(4-(2-甲氧基吡啶-4-基)苯基)-4-(丙烷-1-炔-1-基)-1H-吲唑-7-甲酰胺)螺环[3.3]庚烷-2-甲酸(I-13)的制备The fifth step: 6-(1-(4-(2-methoxypyridin-4-yl)phenyl)-4-(propane-1-yn-1-yl)-1H-indazole-7-methan Preparation of amide) spiro[3.3]heptane-2-carboxylic acid (I-13)
Figure PCTCN2022097611-appb-000050
Figure PCTCN2022097611-appb-000050
室温下将化合物6-(1-(4-(2-甲氧基吡啶-4-基)苯基)-4-(丙烷-1-炔-1-基)-1H-吲唑-7-甲酰胺)螺环[3.3]庚烷-2-甲酸甲酯(60mg,0.11mmol)加入到四氢呋喃(3mL)和水(1mL)中，加入氢氧化锂(8mg,0.33mmol)，室温搅拌16h。用1N盐酸调节pH＝4，用乙酸乙酯(20mL×3)萃取，分液，合并有机相，有机相用无水硫酸钠干燥，过滤，浓缩，残留物经酸性制备方法A制备得化合物6-(1-(4-(2-甲氧基吡啶-4-基)苯基)-4-(丙烷-1-炔-1-基)-1H-吲唑-7-甲酰胺)螺环[3.3]庚烷-2-甲酸(I-13)(50mg,产率85％)。Compound 6-(1-(4-(2-methoxypyridin-4-yl)phenyl)-4-(propane-1-yn-1-yl)-1H-indazole-7-methan Amide) spiro[3.3]heptane-2-carboxylic acid methyl ester (60mg, 0.11mmol) was added to tetrahydrofuran (3mL) and water (1mL), lithium hydroxide (8mg, 0.33mmol) was added, and stirred at room temperature for 16h. Use 1N hydrochloric acid to adjust pH=4, extract with ethyl acetate (20mL×3), separate the layers, combine the organic phases, dry the organic phases with anhydrous sodium sulfate, filter, concentrate, and prepare the residue by acidic preparation method A to obtain compound 6 -(1-(4-(2-methoxypyridin-4-yl)phenyl)-4-(propane-1-yn-1-yl)-1H-indazole-7-carboxamide) spiro[ 3.3] Heptane-2-carboxylic acid (I-13) (50 mg, 85% yield).
1H NMR(400mHz,DMSO-d6)δ11.97(s,1H),8.75(d,1H),8.26(s,1H),7.64(d,1H),7.36(d，2H),7.34(d,1H),7.22(t,2H),7.06(t,3H),5.82(s,2H),4.27-4.21(m,1H),3.80(s,3H),2.93-2.89(m,1H),2.38-2.22(m,4H),2.13(s,3H),2.18-1.99(m,2H),1.90-1.80(m,2H)。 1 H NMR (400mHz,DMSO-d6)δ11.97(s,1H),8.75(d,1H),8.26(s,1H),7.64(d,1H),7.36(d,2H),7.34(d ,1H),7.22(t,2H),7.06(t,3H),5.82(s,2H),4.27-4.21(m,1H),3.80(s,3H),2.93-2.89(m,1H), 2.38-2.22 (m, 4H), 2.13 (s, 3H), 2.18-1.99 (m, 2H), 1.90-1.80 (m, 2H).
LC-MS,M/Z(ESI):535.4[M+H] +。 LC-MS, M/Z (ESI): 535.4 [M+H] + .
实施例14化合物I-14的制备The preparation of embodiment 14 compound I-14
合成路线如下所示：The synthetic route is as follows:
Figure PCTCN2022097611-appb-000051
Figure PCTCN2022097611-appb-000051
第一步：2-(4-(溴甲基)苄基)丙烷-2-醇的制备The first step: the preparation of 2-(4-(bromomethyl)benzyl)propan-2-ol
室温下将化合物2-(4-甲基苯基)丙烷-2-醇(500mg,3.3mmol)加入到CCl 4(10mL)中，加入氧化二苯甲酰(BPO，16mg,0.06mmol)，N-溴代琥珀酰亚胺(NBS，600mg,3.4mmol)，加热回流搅拌4h。将反应液浓缩，残留物用薄层柱分离纯化(石油醚:乙酸乙酯(V/V)＝4:1)得化合物2-(4-(溴甲基)苄基)丙烷-2-醇(460mg,产率60％)。 Compound 2-(4-methylphenyl)propan-2-ol (500mg, 3.3mmol) was added to CCl 4 (10mL) at room temperature, dibenzoyl oxide (BPO, 16mg, 0.06mmol) was added, N -Bromosuccinimide (NBS, 600mg, 3.4mmol), heated under reflux and stirred for 4h. The reaction solution was concentrated, and the residue was separated and purified by thin-layer column (petroleum ether:ethyl acetate (V/V)=4:1) to obtain compound 2-(4-(bromomethyl)benzyl)propan-2-ol (460 mg, yield 60%).
第二步：1-(4-(2-羟基丙烷-2-基)苄基)-4-(丙烷-1-炔-1-基)-1H-吲唑-7-甲酸甲酯的制备The second step: the preparation of 1-(4-(2-hydroxypropan-2-yl)benzyl)-4-(propane-1-yn-1-yl)-1H-indazole-7-carboxylic acid methyl ester
室温下将化合物4-(丙烷-1-炔-1-基)-1H-吲唑-7-甲酸甲酯(150mg,0.70mmol)加入到N,N-二甲基甲酰胺(5mL)中，加入2-(4-(溴甲基)苄基)丙烷-2-醇(200mg,0.87mmol)，碳酸铯(460mg,1.42mmol)，室温搅拌16h。加入水(20mL)稀释，用乙酸乙酯(20mL×3)萃取，分液，合并有机相，有机相用无水硫酸钠干燥，过滤，浓缩，残留物用薄层柱分离纯化(石油醚:乙酸乙酯(V/V)＝4:1)得化合物1-(4-(2-羟基丙烷-2-基)苄基)-4-(丙烷-1-炔-1-基)-1H-吲唑-7-甲酸甲酯(120mg,产率47％)。Compound 4-(propane-1-yn-1-yl)-1H-indazole-7-carboxylic acid methyl ester (150 mg, 0.70 mmol) was added into N,N-dimethylformamide (5 mL) at room temperature, Add 2-(4-(bromomethyl)benzyl)propan-2-ol (200mg, 0.87mmol), cesium carbonate (460mg, 1.42mmol), and stir at room temperature for 16h. Add water (20mL) to dilute, extract with ethyl acetate (20mL×3), separate the layers, combine the organic phases, dry the organic phases with anhydrous sodium sulfate, filter, concentrate, and the residue is separated and purified by thin-layer column (petroleum ether: Ethyl acetate (V/V)=4:1) to obtain compound 1-(4-(2-hydroxypropan-2-yl)benzyl)-4-(propane-1-yn-1-yl)-1H- Indazole-7-carboxylic acid methyl ester (120mg, yield 47%).
第三步：1-(4-(2-羟基丙烷-2-基)苄基)-4-(丙烷-1-炔-1-基)-1H-吲唑-7-甲酸的制备Step 3: Preparation of 1-(4-(2-hydroxypropan-2-yl)benzyl)-4-(propane-1-yn-1-yl)-1H-indazole-7-carboxylic acid
室温下将1-(4-(2-羟基丙烷-2-基)苄基)-4-(丙烷-1-炔-1-基)-1H-吲唑-7-甲酸甲酯(120mg,0.33mmol)加入到四氢呋喃(4mL)和水(1mL)中，加入氢氧化锂(24mg,1.0mmol)，室温搅拌16h。用1N盐酸调节pH＝4，用乙酸乙酯(10mL×3)萃取，分液，合并有机相，有机相用无水硫酸钠干燥，过滤，浓缩，得化合物粗品1-(4-(2-羟基丙烷-2-基)苄基)-4-(丙烷-1-炔-1-基)-1H-吲唑-7-甲酸(130mg,产率100％).Methyl 1-(4-(2-hydroxypropan-2-yl)benzyl)-4-(propane-1-yn-1-yl)-1H-indazole-7-carboxylate (120mg, 0.33 mmol) was added to tetrahydrofuran (4 mL) and water (1 mL), lithium hydroxide (24 mg, 1.0 mmol) was added, and stirred at room temperature for 16 h. Use 1N hydrochloric acid to adjust pH=4, extract with ethyl acetate (10mL×3), separate the layers, combine the organic phases, dry the organic phases with anhydrous sodium sulfate, filter, and concentrate to obtain the crude compound 1-(4-(2- Hydroxypropan-2-yl)benzyl)-4-(propane-1-yn-1-yl)-1H-indazole-7-carboxylic acid (130mg, yield 100%).
第四步：6-(1-(4-(2-羟基丙烷-2-基)苄基)-4-(丙烷-1-炔-1-基)-1H-吲唑-7-甲酰胺)螺环[3.3]庚烷-2-甲酸甲酯的制备The fourth step: 6-(1-(4-(2-hydroxypropan-2-yl)benzyl)-4-(propane-1-yn-1-yl)-1H-indazole-7-carboxamide) Preparation of spiro[3.3]heptane-2-carboxylate methyl ester
室温下将化合物1-(4-(2-羟基丙烷-2-基)苄基)-4-(丙烷-1-炔-1-基)-1H-吲唑-7-甲酸(110mg,0.32mmol)加入到N,N-二甲基甲酰胺(3mL)中，加入2-(6-氨基螺环[3.3]庚烷-2-基)甲酸甲酯盐酸盐(78mg,0.38mmol)，二异丙基乙胺(122.5mg,0.95mmol)，HATU(180mg,0.47mmol)，搅拌16h。加入水(10mL)稀释，用乙酸乙酯(10mL×3)萃取，分液，合并有机相，有机相用 无水硫酸钠干燥，过滤，浓缩，残留物用薄层硅胶板分离纯化(DCM:甲醇(V/V)＝10:1)得化合物6-(1-(4-(2-羟基丙烷-2-基)苄基)-4-(丙烷-1-炔-1-基)-1H-吲唑-7-甲酰胺)螺环[3.3]庚烷-2-甲酸甲酯(150mg,产率95％)。Compound 1-(4-(2-hydroxypropan-2-yl)benzyl)-4-(propane-1-yn-1-yl)-1H-indazole-7-carboxylic acid (110mg, 0.32mmol ) was added to N,N-dimethylformamide (3mL), 2-(6-aminospirocyclo[3.3]heptane-2-yl)methyl formate hydrochloride (78mg, 0.38mmol) was added, di Isopropylethylamine (122.5mg, 0.95mmol), HATU (180mg, 0.47mmol), stirred for 16h. Add water (10mL) to dilute, extract with ethyl acetate (10mL×3), separate the layers, combine the organic phase, dry the organic phase with anhydrous sodium sulfate, filter, concentrate, and the residue is separated and purified with a thin-layer silica gel plate (DCM: Methanol (V/V)=10:1) to obtain compound 6-(1-(4-(2-hydroxypropan-2-yl)benzyl)-4-(propane-1-yn-1-yl)-1H -indazole-7-carboxamide)spiro[3.3]heptane-2-carboxylic acid methyl ester (150 mg, 95% yield).
第五步：6-(1-(4-(2-羟基丙烷-2-基)苄基)-4-(丙烷-1-炔-1-基)-1H-吲唑-7-甲酰胺)螺环[3.3]庚烷-2-甲酸(I-14)的制备The fifth step: 6-(1-(4-(2-hydroxypropan-2-yl)benzyl)-4-(propane-1-yn-1-yl)-1H-indazole-7-carboxamide) Preparation of spiro[3.3]heptane-2-carboxylic acid (I-14)
Figure PCTCN2022097611-appb-000052
Figure PCTCN2022097611-appb-000052
室温下将化合物6-(1-(4-(2-羟基丙烷-2-基)苄基)-4-(丙烷-1-炔-1-基)-1H-吲唑-7-甲酰胺)螺环[3.3]庚烷-2-甲酸甲酯(150mg,0.30mmol)加入到四氢呋喃(3mL)，甲醇(1mL)和水(1mL)中，加入氢氧化锂(22mg,0.90mmol)，室温搅拌16h。用1N盐酸调节pH＝4，用乙酸乙酯(20mL×3)萃取，分液，合并有机相，有机相用无水硫酸钠干燥，过滤，浓缩，残留物经酸性制备方法A制备得化合物6-(1-(4-(2-羟基丙烷-2-基)苄基)-4-(丙烷-1-炔-1-基)-1H-吲唑-7-甲酰胺)螺环[3.3]庚烷-2-甲酸(I-14)(50mg,产率34％)。Compound 6-(1-(4-(2-hydroxypropan-2-yl)benzyl)-4-(propane-1-yn-1-yl)-1H-indazole-7-carboxamide) Methyl spiro[3.3]heptane-2-carboxylate (150mg, 0.30mmol) was added to tetrahydrofuran (3mL), methanol (1mL) and water (1mL), lithium hydroxide (22mg, 0.90mmol) was added, and stirred at room temperature 16h. Use 1N hydrochloric acid to adjust pH=4, extract with ethyl acetate (20mL×3), separate the layers, combine the organic phases, dry the organic phases with anhydrous sodium sulfate, filter, concentrate, and prepare the residue by acidic preparation method A to obtain compound 6 -(1-(4-(2-hydroxypropan-2-yl)benzyl)-4-(propane-1-yn-1-yl)-1H-indazole-7-carboxamide) spiro[3.3] Heptane-2-carboxylic acid (I-14) (50 mg, yield 34%).
1H NMR(400mHz,DMSO-d6)δ12.0(s,1H),8.78(d,1H),8.21(s,1H),7.35(d，1H),7.31(d,2H),7.21(d,1H),6.92(d,2H),5.72(s,2H),4.92(s,1H),4.30-4.24(m,1H),2.95-2.91(m,1H),2.42-2.17(m,4H),2.17(s,3H),2.10-2.02(m,2H),1.93-1.82(m,2H),1.35(s,6H)。 1 H NMR (400mHz,DMSO-d6)δ12.0(s,1H),8.78(d,1H),8.21(s,1H),7.35(d,1H),7.31(d,2H),7.21(d ,1H),6.92(d,2H),5.72(s,2H),4.92(s,1H),4.30-4.24(m,1H),2.95-2.91(m,1H),2.42-2.17(m,4H ), 2.17(s,3H), 2.10-2.02(m,2H), 1.93-1.82(m,2H), 1.35(s,6H).
LC-MS,M/Z(ESI):486.4[M+H] +。 LC-MS, M/Z (ESI): 486.4 [M+H] + .
实施例15化合物I-15的制备The preparation of embodiment 15 compound I-15
合成路线如下所示：The synthetic route is as follows:
Figure PCTCN2022097611-appb-000053
Figure PCTCN2022097611-appb-000053
第一步：4-(丙烷-1-炔-1-基)-1H-吲唑-7-甲酸甲酯的制备The first step: the preparation of 4-(propane-1-yn-1-yl)-1H-indazole-7-carboxylic acid methyl ester
室温下将化合物4-溴-1H-吲唑-7-甲酸甲酯(1.0g,3.94mmol)加入到1,4-二氧六环(10mL)中，加入二三苯基膦二氯化钯(274mg,0.39mmol)，1-丙炔-三正丁基锡(1.52g,4.61mmol)，氮气保护下加热至90℃，搅拌16h。冷却至室温，加入水(50mL)稀释，用乙酸乙酯(50mL× 3)萃取，分液，合并有机相，有机相用无水硫酸钠干燥，过滤，浓缩，残留物用薄层硅胶板分离纯化(石油醚:乙酸乙酯(V/V)＝5:1)得化合物4-(丙烷-1-炔-1-基)-1H-吲唑-7-甲酸甲酯(500mg,产率42％)。Add the compound 4-bromo-1H-indazole-7-carboxylic acid methyl ester (1.0g, 3.94mmol) to 1,4-dioxane (10mL) at room temperature, add ditriphenylphosphine palladium dichloride (274mg, 0.39mmol), 1-propyne-tri-n-butyltin (1.52g, 4.61mmol), heated to 90°C under nitrogen protection, and stirred for 16h. Cool to room temperature, add water (50mL) to dilute, extract with ethyl acetate (50mL×3), separate the liquids, combine the organic phases, dry the organic phases with anhydrous sodium sulfate, filter, concentrate, and separate the residue with a thin-layer silica gel plate Purification (petroleum ether: ethyl acetate (V/V) = 5:1) gave compound 4-(propane-1-yn-1-yl)-1H-indazole-7-carboxylic acid methyl ester (500mg, yield 42 %).
第二步：4-(丙烷-1-炔-1-基)-1-(4-(三氟甲基)苄基)-1H-吲唑-7-甲酸甲酯的制备The second step: the preparation of 4-(propane-1-yn-1-yl)-1-(4-(trifluoromethyl)benzyl)-1H-indazole-7-carboxylic acid methyl ester
室温下将化合物4-(丙烷-1-炔-1-基)-1H-吲唑-7-甲酸甲酯(100mg,0.47mmol)加入到N,N-二甲基甲酰胺(5mL)中，加入1-(溴甲基)-4-(三氟甲基)苯(168mg,0.70mmol)和碳酸铯(456mg,1.40mmol)，室温搅拌16h。加入水(20mL)稀释，用乙酸乙酯(20mL×3)萃取，分液，合并有机相，有机相用无水硫酸钠干燥，过滤，浓缩，残留物经酸性制备方法A得化合物4-(丙烷-1-炔-1-基)-1-(4-(三氟甲基)苄基)-1H-吲唑-7-甲酸甲酯(120mg,产率69％)。Compound 4-(propane-1-yn-1-yl)-1H-indazole-7-carboxylic acid methyl ester (100 mg, 0.47 mmol) was added into N,N-dimethylformamide (5 mL) at room temperature, Add 1-(bromomethyl)-4-(trifluoromethyl)benzene (168mg, 0.70mmol) and cesium carbonate (456mg, 1.40mmol), and stir at room temperature for 16h. Add water (20mL) to dilute, extract with ethyl acetate (20mL×3), separate the layers, combine the organic phases, dry the organic phases with anhydrous sodium sulfate, filter, concentrate, and the residue is subjected to acidic preparation method A to obtain compound 4-( Propan-1-yn-1-yl)-1-(4-(trifluoromethyl)benzyl)-1H-indazole-7-carboxylic acid methyl ester (120 mg, 69% yield).
第三步：4-(丙烷-1-炔-1-基)-1-(4-(三氟甲基)苄基)-1H-吲唑-7-甲酸的制备The third step: the preparation of 4-(propane-1-yn-1-yl)-1-(4-(trifluoromethyl)benzyl)-1H-indazole-7-carboxylic acid
室温下将原料4-(丙烷-1-炔-1-基)-1-(4-(三氟甲基)苄基)-1H-吲唑-7-甲酸甲酯(120mg,0.32mmol)加入到四氢呋喃(5mL)和水(2mL)中，加入氢氧化锂(23mg,0.97mmol)，室温搅拌16h。用1N盐酸调节pH＝4，用乙酸乙酯(20mL×3)萃取，分液，合并有机相，有机相用无水硫酸钠干燥，过滤，浓缩，得化合物粗品4-(丙烷-1-炔-1-基)-1-(4-(三氟甲氧基)苄基)乙基)-1H-吲唑-7-甲酸(110mg,产率95％)。Starting material 4-(propan-1-yn-1-yl)-1-(4-(trifluoromethyl)benzyl)-1H-indazole-7-carboxylic acid methyl ester (120 mg, 0.32 mmol) was added at room temperature To tetrahydrofuran (5 mL) and water (2 mL), add lithium hydroxide (23 mg, 0.97 mmol), and stir at room temperature for 16 h. Use 1N hydrochloric acid to adjust pH=4, extract with ethyl acetate (20mL×3), separate the layers, combine the organic phases, dry the organic phases with anhydrous sodium sulfate, filter, and concentrate to obtain the crude compound 4-(propane-1-yne -1-yl)-1-(4-(trifluoromethoxy)benzyl)ethyl)-1H-indazole-7-carboxylic acid (110 mg, 95% yield).
第四步：6-(4-(丙烷-1-炔-1-基)-1-(4-(三氟甲基)苄基)-1H-吲唑-7-甲酰胺)螺环[3.3]庚烷-2-甲酸甲酯的制备The fourth step: 6-(4-(propane-1-yn-1-yl)-1-(4-(trifluoromethyl)benzyl)-1H-indazole-7-carboxamide) spirocycle [3.3 ] The preparation of heptane-2-methyl carboxylate
室温下将化合物4-(丙烷-1-炔-1-基)-1-(4-(三氟甲氧基)苄基)乙基)-1H-吲唑-7-甲酸(110mg,0.31mmol)加入到N,N-二甲基甲酰胺(10mL)中，加入2-(6-氨基螺环[3.3]庚烷-2-基)甲酸甲酯盐酸盐(98mg,0.47mmol)，二异丙基乙胺(122mg,0.93mmol)，HATU(180mg,0.47mmol)，搅拌16h。加入水(20mL)稀释，用乙酸乙酯(20mL×3)萃取，分液，合并有机相，有机相用无水硫酸钠干燥，过滤，浓缩，残留物用薄层硅胶板分离纯化(石油醚:乙酸乙酯(V/V)＝1:1)得化合物6-(4-(丙烷-1-炔-1-基)-1-(4-(三氟甲基)苄基)-1H-吲唑-7-甲酰胺)螺环[3.3]庚烷-2-甲酸甲酯(100mg,产率64％)。Compound 4-(propane-1-yn-1-yl)-1-(4-(trifluoromethoxy)benzyl)ethyl)-1H-indazole-7-carboxylic acid (110mg, 0.31mmol ) was added to N,N-dimethylformamide (10mL), and methyl 2-(6-aminospiro[3.3]heptane-2-yl)formate hydrochloride (98mg, 0.47mmol) was added, di Isopropylethylamine (122mg, 0.93mmol), HATU (180mg, 0.47mmol), stirred for 16h. Add water (20mL) to dilute, extract with ethyl acetate (20mL×3), separate the layers, combine the organic phases, dry the organic phases with anhydrous sodium sulfate, filter, concentrate, and separate and purify the residue with a thin-layer silica gel plate (petroleum ether : ethyl acetate (V/V)=1:1) to obtain compound 6-(4-(propane-1-yn-1-yl)-1-(4-(trifluoromethyl)benzyl)-1H- Indazole-7-carboxamide) spiro[3.3]heptane-2-carboxylic acid methyl ester (100 mg, 64% yield).
第五步：6-(4-(丙烷-1-炔-1-基)-1-(4-(三氟甲基)苄基)-1H-吲唑-7-甲酰胺)螺环[3.3]庚烷-2-甲酸(I-15)的制备The fifth step: 6-(4-(propane-1-yn-1-yl)-1-(4-(trifluoromethyl)benzyl)-1H-indazole-7-carboxamide) spirocycle [3.3 ] The preparation of heptane-2-formic acid (I-15)
Figure PCTCN2022097611-appb-000054
Figure PCTCN2022097611-appb-000054
室温下将化合物6-(4-(丙烷-1-炔-1-基)-1-(4-(三氟甲基)苄基)-1H-吲唑-7-甲酰胺)螺环[3.3]庚烷-2-甲酸甲酯(100mg,0.20mmol)加入到四氢呋喃(5mL)和水(2mL)中，加入氢氧化锂(15mg,0.60mmol)，室温搅拌16h。用1N盐酸调节pH＝4，用乙酸乙酯(20mL×3)萃取，分液， 合并有机相，有机相用无水硫酸钠干燥，过滤，浓缩，残留物经酸性制备方法A制备得化合物6-(4-(丙烷-1-炔-1-基)-1-(4-(三氟甲基)苄基)-1H-吲唑-7-甲酰胺)螺环[3.3]庚烷-2-甲酸(I-15)(70mg,产率72％)。Compound 6-(4-(propane-1-yn-1-yl)-1-(4-(trifluoromethyl)benzyl)-1H-indazole-7-carboxamide) spiro[3.3 ]Heptane-2-methyl carboxylate (100mg, 0.20mmol) was added to tetrahydrofuran (5mL) and water (2mL), lithium hydroxide (15mg, 0.60mmol) was added, and stirred at room temperature for 16h. Use 1N hydrochloric acid to adjust pH=4, extract with ethyl acetate (20mL×3), separate the layers, combine the organic phases, dry the organic phases with anhydrous sodium sulfate, filter, concentrate, and prepare the residue by acidic preparation method A to obtain compound 6 -(4-(propane-1-yn-1-yl)-1-(4-(trifluoromethyl)benzyl)-1H-indazole-7-carboxamide)spiro[3.3]heptane-2 - Formic acid (I-15) (70 mg, yield 72%).
1H NMR(400mHz,DMSO-d6)δ12.04(s,1H),8.69(d,1H),8.29(s,1H),7.62(d,2H),7.37(d,1H),7.24(d,1H),7.10(d,2H),5.88(s,2H),4.20-4.15(m,1H),2.94-2.90(d,1H),2.33-2.25(m,4H),2.22(s,3H),2.19-2.00(m,2H)，1.83-1.75(m,2H)。 1 H NMR (400mHz,DMSO-d6)δ12.04(s,1H),8.69(d,1H),8.29(s,1H),7.62(d,2H),7.37(d,1H),7.24(d ,1H),7.10(d,2H),5.88(s,2H),4.20-4.15(m,1H),2.94-2.90(d,1H),2.33-2.25(m,4H),2.22(s,3H ), 2.19-2.00(m,2H), 1.83-1.75(m,2H).
LC-MS,M/Z(ESI):496.4[M+H] +。 LC-MS, M/Z (ESI): 496.4 [M+H] + .
实施例16化合物I-16的制备The preparation of embodiment 16 compound I-16
合成路线如下所示The synthetic route is as follows
Figure PCTCN2022097611-appb-000055
Figure PCTCN2022097611-appb-000055
第一步：1-(溴甲基)-3-环丙基苯的制备The first step: the preparation of 1-(bromomethyl)-3-cyclopropylbenzene
室温下将化合物3-(环丙基苯基)甲醇(300mg,2.0mmol)加入到乙醚(10mL)中，冷却至0℃，加入三溴化膦(550mg,2.2mmol)，室温搅拌2h。加入水(20mL)稀释，用乙醚(20mL×3)萃取，分液，合并有机相，有机相用无水硫酸钠干燥，过滤，浓缩将反应液浓缩，得化合物粗品1-(溴甲基)-3-环丙基苯(320mg,产率75％)。Compound 3-(cyclopropylphenyl)methanol (300mg, 2.0mmol) was added to diethyl ether (10mL) at room temperature, cooled to 0°C, phosphine tribromide (550mg, 2.2mmol) was added, and stirred at room temperature for 2h. Add water (20mL) to dilute, extract with ether (20mL×3), separate the layers, combine the organic phases, dry the organic phases with anhydrous sodium sulfate, filter, concentrate and concentrate the reaction solution to obtain the crude compound 1-(bromomethyl) -3-cyclopropylbenzene (320 mg, yield 75%).
第二步：1-(3-环丙基苄基)-4-(丙烷-1-炔-1-基)-1H-吲唑-7-甲酸甲酯的制备The second step: the preparation of 1-(3-cyclopropylbenzyl)-4-(propane-1-yn-1-yl)-1H-indazole-7-carboxylic acid methyl ester
室温下将化合物4-(丙烷-1-炔-1-基)-1H-吲唑-7-甲酸甲酯(150mg,0.70mmol)加入到N,N-二甲基甲酰胺(5mL)中，加入1-(溴甲基)-3-环丙基苯(300mg,1.42mmol)，碳酸铯(460mg,1.42mmol)，室温搅拌16h。加入水(20mL)稀释，用乙酸乙酯(20mL×3)萃取，分液，合并有机相，有机相用无水硫酸钠干燥，过滤，浓缩，残留物用薄层柱分离纯化(石油醚:乙酸乙酯(V/V)＝4:1)得化合物1-(3-环丙基苄基)-4-(丙烷-1-炔-1-基)-1H-吲唑-7-甲酸甲酯(130mg,产率54％)。Compound 4-(propane-1-yn-1-yl)-1H-indazole-7-carboxylic acid methyl ester (150 mg, 0.70 mmol) was added into N,N-dimethylformamide (5 mL) at room temperature, Add 1-(bromomethyl)-3-cyclopropylbenzene (300mg, 1.42mmol), cesium carbonate (460mg, 1.42mmol), and stir at room temperature for 16h. Add water (20mL) to dilute, extract with ethyl acetate (20mL×3), separate the layers, combine the organic phases, dry the organic phases with anhydrous sodium sulfate, filter, concentrate, and the residue is separated and purified by thin-layer column (petroleum ether: Ethyl acetate (V/V)=4:1) to obtain the compound 1-(3-cyclopropylbenzyl)-4-(propane-1-yn-1-yl)-1H-indazole-7-formic acid Ester (130 mg, 54% yield).
第三步：1-(3-环丙基苄基)-4-(丙烷-1-炔-1-基)-1H-吲唑-7-甲酸的制备Step 3: Preparation of 1-(3-cyclopropylbenzyl)-4-(propane-1-yn-1-yl)-1H-indazole-7-carboxylic acid
室温下将1-(3-环丙基苄基)-4-(丙烷-1-炔-1-基)-1H-吲唑-7-甲酸甲酯(130mg,0.38mmol)加入到四氢呋喃(5mL)和水(2mL)中，加入氢氧化锂(27mg,1.1mmol)，室温搅拌16h。用1N盐酸调节pH＝4，用乙酸乙酯(10mL×3)萃取，分液，合并有机相，有机相用无水硫酸钠干燥， 过滤，浓缩，得化合物粗品1-(3-环丙基苄基)-4-(丙烷-1-炔-1-基)-1H-吲唑-7-甲酸(112mg,产率90％).Add 1-(3-cyclopropylbenzyl)-4-(propane-1-yn-1-yl)-1H-indazole-7-carboxylic acid methyl ester (130 mg, 0.38 mmol) into tetrahydrofuran (5 mL ) and water (2mL), add lithium hydroxide (27mg, 1.1mmol), and stir at room temperature for 16h. Use 1N hydrochloric acid to adjust pH=4, extract with ethyl acetate (10mL×3), separate the layers, combine the organic phases, dry the organic phases with anhydrous sodium sulfate, filter, and concentrate to obtain the crude compound 1-(3-cyclopropyl Benzyl)-4-(propane-1-yn-1-yl)-1H-indazole-7-carboxylic acid (112mg, yield 90%).
第四步：6-(1-(3-环丙基苄基)-4-(丙烷-1-炔-1-基)-1H-吲唑-7-甲酰胺)螺环[3.3]庚烷-2-甲酸甲酯的制备The fourth step: 6-(1-(3-cyclopropylbenzyl)-4-(propane-1-yn-1-yl)-1H-indazole-7-carboxamide) spiro[3.3]heptane - Preparation of methyl 2-formate
室温下将化合物1-(3-环丙基苄基)-4-(丙烷-1-炔-1-基)-1H-吲唑-7-甲酸(112mg,0.34mmol)加入到N,N-二甲基甲酰胺(3mL)中，加入2-(6-氨基螺环[3.3]庚烷-2-基)甲酸甲酯盐酸盐(84mg,0.41mmol)，二异丙基乙胺(135mg,1.05mmol)，HATU(200mg,0.53mmol)，搅拌16h。加入水(10mL)稀释，用乙酸乙酯(10mL×3)萃取，分液，合并有机相，有机相用无水硫酸钠干燥，过滤，浓缩，残留物用薄层硅胶板分离纯化(PE:EA(V/V)＝1:1)得化合物6-(1-(3-环丙基苄基)-4-(丙烷-1-炔-1-基)-1H-吲唑-7-甲酰胺)螺环[3.3]庚烷-2-甲酸甲酯(150mg,产率92％)。Compound 1-(3-cyclopropylbenzyl)-4-(propane-1-yn-1-yl)-1H-indazole-7-carboxylic acid (112mg, 0.34mmol) was added to N,N- In dimethylformamide (3mL), add 2-(6-aminospirocyclo[3.3]heptane-2-yl)methyl formate hydrochloride (84mg, 0.41mmol), diisopropylethylamine (135mg , 1.05mmol), HATU (200mg, 0.53mmol), stirred for 16h. Add water (10mL) to dilute, extract with ethyl acetate (10mL×3), separate the layers, combine the organic phases, dry the organic phases with anhydrous sodium sulfate, filter, concentrate, and the residue is separated and purified with a thin-layer silica gel plate (PE: EA (V/V)=1:1) to obtain compound 6-(1-(3-cyclopropylbenzyl)-4-(propane-1-yn-1-yl)-1H-indazole-7-methan Amide) spiro[3.3]heptane-2-carboxylic acid methyl ester (150 mg, 92% yield).
第五步：6-(1-(3-环丙基苄基)-4-(丙烷-1-炔-1-基)-1H-吲唑-7-甲酰胺)螺环[3.3]庚烷-2-甲酸(I-16)的制备The fifth step: 6-(1-(3-cyclopropylbenzyl)-4-(propane-1-yn-1-yl)-1H-indazole-7-carboxamide)spiro[3.3]heptane - Preparation of 2-formic acid (I-16)
Figure PCTCN2022097611-appb-000056
Figure PCTCN2022097611-appb-000056
室温下将化合物6-(1-(3-环丙基苄基)-4-(丙烷-1-炔-1-基)-1H-吲唑-7-甲酰胺)螺环[3.3]庚烷-2-甲酸甲酯(150mg,0.31mmol)加入到四氢呋喃(5mL)和水(1mL)中，加入氢氧化锂(22mg,0.92mmol)，室温搅拌16h。用1N盐酸调节pH＝4，用乙酸乙酯(20mL×3)萃取，分液，合并有机相，有机相用无水硫酸钠干燥，过滤，浓缩，残留物经酸性制备方法A制备得化合物6-(1-(3-环丙基苄基)-4-(丙烷-1-炔-1-基)-1H-吲唑-7-甲酰胺)螺环[3.3]庚烷-2-甲酸(I-16)(55mg,产率38％)。Compound 6-(1-(3-cyclopropylbenzyl)-4-(propane-1-yn-1-yl)-1H-indazole-7-carboxamide) spiro[3.3]heptane - Methyl 2-carboxylate (150mg, 0.31mmol) was added to tetrahydrofuran (5mL) and water (1mL), lithium hydroxide (22mg, 0.92mmol) was added, and stirred at room temperature for 16h. Use 1N hydrochloric acid to adjust pH=4, extract with ethyl acetate (20mL×3), separate the layers, combine the organic phases, dry the organic phases with anhydrous sodium sulfate, filter, concentrate, and prepare the residue by acidic preparation method A to obtain compound 6 -(1-(3-cyclopropylbenzyl)-4-(propane-1-yn-1-yl)-1H-indazole-7-carboxamide) spiro[3.3]heptane-2-carboxylic acid ( 1-16) (55 mg, 38% yield).
1H NMR(400mHz,DMSO-d6)δ11.97(s,1H),8.77(d,1H),8.23(s,1H),7.36(d,1H),7.21(d,1H),7.07(t,1H),6.86(d,1H),6.77(s,1H),6.64(d,1H),5.72(s,2H),4.25-4.20(m,1H),2.95-2.91(m,1H),2.42-2.17(m,4H),2.17(s,3H),2.10-2.02(m,2H),1.90-1.84(m,3H),0.90(d,2H),0.55(d,2H). 1 H NMR (400mHz,DMSO-d6)δ11.97(s,1H),8.77(d,1H),8.23(s,1H),7.36(d,1H),7.21(d,1H),7.07(t ,1H),6.86(d,1H),6.77(s,1H),6.64(d,1H),5.72(s,2H),4.25-4.20(m,1H),2.95-2.91(m,1H), 2.42-2.17(m,4H),2.17(s,3H),2.10-2.02(m,2H),1.90-1.84(m,3H),0.90(d,2H),0.55(d,2H).
LC-MS,M/Z(ESI):468.5[M+H] +。 LC-MS, M/Z (ESI): 468.5 [M+H] + .
实施例17化合物I-17的制备The preparation of embodiment 17 compound I-17
合成路线如下所示：The synthetic route is as follows:
Figure PCTCN2022097611-appb-000057
Figure PCTCN2022097611-appb-000057
第一步：4-(丙烷-1-炔-1-基)-1H-吲唑-7-甲酸甲酯的制备The first step: the preparation of 4-(propane-1-yn-1-yl)-1H-indazole-7-carboxylic acid methyl ester
室温下将化合物4-溴-1H-吲唑-7-甲酸甲酯(1.0g,3.94mmol)加入到1,4-二氧六环(10mL)中，加入二三苯基膦二氯化钯(274mg,0.39mmol)，1-丙炔-三正丁基锡(1.52g,4.61mmol)，氮气保护下加热至90℃，搅拌16h。冷却至室温，加入水(50mL)稀释，用乙酸乙酯(50mL×3)萃取，分液，合并有机相，有机相用无水硫酸钠干燥，过滤，浓缩，残留物用薄层硅胶板分离纯化(石油醚:乙酸乙酯(V/V)＝5:1)得化合物4-(丙烷-1-炔-1-基)-1H-吲唑-7-甲酸甲酯(500mg,产率42％)。Add the compound 4-bromo-1H-indazole-7-carboxylic acid methyl ester (1.0g, 3.94mmol) to 1,4-dioxane (10mL) at room temperature, add ditriphenylphosphine palladium dichloride (274mg, 0.39mmol), 1-propyne-tri-n-butyltin (1.52g, 4.61mmol), heated to 90°C under nitrogen protection, and stirred for 16h. Cool to room temperature, add water (50mL) to dilute, extract with ethyl acetate (50mL×3), separate the liquids, combine the organic phases, dry the organic phases with anhydrous sodium sulfate, filter, concentrate, and separate the residue with a thin-layer silica gel plate Purification (petroleum ether: ethyl acetate (V/V) = 5:1) gave compound 4-(propane-1-yn-1-yl)-1H-indazole-7-carboxylic acid methyl ester (500mg, yield 42 %).
第二步：4-(丙烷-1-炔-1-基)-1-(3-(三氟甲氧基)苄基)-1H-吲唑-7-甲酸甲酯的制备The second step: the preparation of 4-(propane-1-yn-1-yl)-1-(3-(trifluoromethoxy)benzyl)-1H-indazole-7-carboxylic acid methyl ester
室温下将化合物4-(丙烷-1-炔-1-基)-1H-吲唑-7-甲酸甲酯(150mg,0.70mmol)加入到N,N-二甲基甲酰胺(5mL)中，加入1-(溴甲基)-4-(三氟甲氧基)苯(268mg,1.05mmol)和碳酸铯(683mg,2.10mmol)，室温搅拌16h。加入水(20mL)稀释，用乙酸乙酯(20mL×3)萃取，分液，合并有机相，有机相用无水硫酸钠干燥，过滤，浓缩，残留物经酸性制备方法A得化合物4-(丙烷-1-炔-1-基)-1-(3-(三氟甲氧基)苄基)-1H-吲唑-7-甲酸甲酯(140mg,产率52％)。Compound 4-(propane-1-yn-1-yl)-1H-indazole-7-carboxylic acid methyl ester (150 mg, 0.70 mmol) was added into N,N-dimethylformamide (5 mL) at room temperature, Add 1-(bromomethyl)-4-(trifluoromethoxy)benzene (268mg, 1.05mmol) and cesium carbonate (683mg, 2.10mmol), and stir at room temperature for 16h. Add water (20mL) to dilute, extract with ethyl acetate (20mL×3), separate the layers, combine the organic phases, dry the organic phases with anhydrous sodium sulfate, filter, concentrate, and the residue is subjected to acidic preparation method A to obtain compound 4-( Propan-1-yn-1-yl)-1-(3-(trifluoromethoxy)benzyl)-1H-indazole-7-carboxylic acid methyl ester (140 mg, 52% yield).
第三步：4-(丙烷-1-炔-1-基)-1-(3-(三氟甲氧基)苄基)-1H-吲唑-7-甲酸的制备The third step: the preparation of 4-(propane-1-yn-1-yl)-1-(3-(trifluoromethoxy)benzyl)-1H-indazole-7-carboxylic acid
室温下将4-(丙烷-1-炔-1-基)-1-(3-(三氟甲氧基)苄基)-1H-吲唑-7-甲酸甲酯(140mg,0.36mmol)加入到四氢呋喃(5mL)和水(2mL)中，加入氢氧化锂(26mg,1.08mmol)，室温搅拌16h。用1N盐酸调节pH＝4，用乙酸乙酯(20mL×3)萃取，分液，合并有机相，有机相用无水硫酸钠干燥，过滤，浓缩，得化合物粗品4-(丙烷-1-炔-1-基)-1-(4-(三氟甲氧基)苄基)乙基)-1H-吲唑-7-甲酸(130mg,产率96％)。Add 4-(propane-1-yn-1-yl)-1-(3-(trifluoromethoxy)benzyl)-1H-indazole-7-carboxylic acid methyl ester (140 mg, 0.36 mmol) at room temperature To tetrahydrofuran (5 mL) and water (2 mL), lithium hydroxide (26 mg, 1.08 mmol) was added, and stirred at room temperature for 16 h. Use 1N hydrochloric acid to adjust pH=4, extract with ethyl acetate (20mL×3), separate the layers, combine the organic phases, dry the organic phases with anhydrous sodium sulfate, filter, and concentrate to obtain the crude compound 4-(propane-1-yne -1-yl)-1-(4-(trifluoromethoxy)benzyl)ethyl)-1H-indazole-7-carboxylic acid (130 mg, 96% yield).
第四步：6-(4-(丙烷-1-炔-1-基)-1-(3-(三氟甲氧基)苄基)-1H-吲唑-7-甲酰胺)螺环[3.3]庚烷-2-甲酸甲酯的制备The fourth step: 6-(4-(propane-1-yn-1-yl)-1-(3-(trifluoromethoxy) benzyl)-1H-indazole-7-carboxamide) spiro[ 3.3] Preparation of methyl heptane-2-carboxylate
室温下将化合物4-(丙烷-1-炔-1-基)-1-(4-(三氟甲氧基)苄基)乙基)-1H-吲唑-7-甲酸(130mg,0.35mmol)加入到N,N-二甲基甲酰胺(5mL)中，加入2-(6-氨基螺环[3.3]庚烷-2-基)甲酸甲酯盐酸盐(108mg,0.53mmol)，二异丙基乙胺(136mg,1.05mmol)，HATU(200mg,0.53mmol)，搅拌16h。加入水(20mL)稀释，用乙酸乙酯(20mL×3)萃取，分液，合并有机相，有机相用无水硫酸钠干燥，过滤，浓缩，残留物用薄层硅胶板分离纯化(石油醚:乙酸乙酯(V/V)＝1:1)得化合物6-(4-(丙烷-1-炔-1-基)-1-(3-(三氟甲氧基)苄基)-1H-吲唑-7-甲酰胺)螺环[3.3]庚烷-2-甲酸 甲酯(120mg,产率66％)。Compound 4-(propane-1-yn-1-yl)-1-(4-(trifluoromethoxy)benzyl)ethyl)-1H-indazole-7-carboxylic acid (130mg, 0.35mmol ) was added to N,N-dimethylformamide (5mL), and methyl 2-(6-aminospiro[3.3]heptane-2-yl)formate hydrochloride (108mg, 0.53mmol) was added, di Isopropylethylamine (136mg, 1.05mmol), HATU (200mg, 0.53mmol), stirred for 16h. Add water (20mL) to dilute, extract with ethyl acetate (20mL×3), separate the layers, combine the organic phases, dry the organic phases with anhydrous sodium sulfate, filter, concentrate, and separate and purify the residue with a thin-layer silica gel plate (petroleum ether : ethyl acetate (V/V)=1:1) to obtain compound 6-(4-(propane-1-yn-1-yl)-1-(3-(trifluoromethoxy)benzyl)-1H -indazole-7-carboxamide)spiro[3.3]heptane-2-carboxylic acid methyl ester (120 mg, 66% yield).
第五步：6-(4-(丙烷-1-炔-1-基)-1-(3-(三氟甲氧基)苄基)-1H-吲唑-7-甲酰胺)螺环[3.3]庚烷-2-甲酸(I-17)的制备The fifth step: 6-(4-(propane-1-yn-1-yl)-1-(3-(trifluoromethoxy) benzyl)-1H-indazole-7-carboxamide) spiro[ 3.3] Preparation of heptane-2-carboxylic acid (I-17)
Figure PCTCN2022097611-appb-000058
Figure PCTCN2022097611-appb-000058
室温下将化合物6-(4-(丙烷-1-炔-1-基)-1-(3-(三氟甲氧基)苄基)-1H-吲唑-7-甲酰胺)螺环[3.3]庚烷-2-甲酸甲酯(120mg,0.23mmol)加入到四氢呋喃(5mL)和水(2mL)中，加入氢氧化锂(17mg,0.69mmol)，室温搅拌16h。用1N盐酸调节pH＝4，用乙酸乙酯(20mL×3)萃取，分液，合并有机相，有机相用无水硫酸钠干燥，过滤，浓缩，残留物经酸性制备方法A制备得化合物6-(4-(丙烷-1-炔-1-基)-1-(3-(三氟甲氧基)苄基)-1H-吲唑-7-甲酰胺)螺环[3.3]庚烷-2-甲酸(I-17)(90mg,产率77％)。Compound 6-(4-(propane-1-yn-1-yl)-1-(3-(trifluoromethoxy)benzyl)-1H-indazole-7-carboxamide) spiro[ 3.3] Add methyl heptane-2-carboxylate (120mg, 0.23mmol) to tetrahydrofuran (5mL) and water (2mL), add lithium hydroxide (17mg, 0.69mmol), and stir at room temperature for 16h. Use 1N hydrochloric acid to adjust pH=4, extract with ethyl acetate (20mL×3), separate the layers, combine the organic phases, dry the organic phases with anhydrous sodium sulfate, filter, concentrate, and prepare the residue by acidic preparation method A to obtain compound 6 -(4-(propane-1-yn-1-yl)-1-(3-(trifluoromethoxy)benzyl)-1H-indazole-7-carboxamide)spiro[3.3]heptane- 2-Formic acid (I-17) (90 mg, yield 77%).
1H NMR(400mHz,DMSO-d6)δ11.99(s,1H),8.78(d,1H),8.27(s,1H),7.39(d,2H),7.24(d,2H),6.92(s,2H),5.83(s,2H),4.20-4.19(m,1H),2.94-2.90(m,1H),2.50-2.36(m,4H),2.27(s,3H),2.25-2.03(m,2H)，1.91-1.81(m,2H). 1 H NMR (400mHz,DMSO-d6)δ11.99(s,1H),8.78(d,1H),8.27(s,1H),7.39(d,2H),7.24(d,2H),6.92(s ,2H),5.83(s,2H),4.20-4.19(m,1H),2.94-2.90(m,1H),2.50-2.36(m,4H),2.27(s,3H),2.25-2.03(m ,2H), 1.91-1.81(m,2H).
LC-MS,M/Z(ESI):512.4[M+H] +。 LC-MS, M/Z (ESI): 512.4 [M+H] + .
实施例18化合物I-18的制备The preparation of embodiment 18 compound I-18
合成路线如下所示：The synthetic route is as follows:
Figure PCTCN2022097611-appb-000059
Figure PCTCN2022097611-appb-000059
第一步：4-(丙烷-1-炔-1-基)-1H-吲唑-7-甲酸甲酯的制备The first step: the preparation of 4-(propane-1-yn-1-yl)-1H-indazole-7-carboxylic acid methyl ester
室温下将化合物4-溴-1H-吲唑-7-甲酸甲酯(1.0g,3.94mmol)加入到1,4-二氧六环(10mL)中，加入二三苯基膦二氯化钯(274mg,0.39mmol)，1-丙炔-三正丁基锡(1.52g,4.61mmol)，氮气保护下加热至90℃，搅拌16h。冷却至室温，加入水(50mL)稀释，用乙酸乙酯(50mL×3)萃取，分液，合并有机相，有机相用无水硫酸钠干燥，过滤，浓缩，残留物用薄层硅胶板分离纯化(石油醚:乙酸乙酯(V/V)＝5:1)得化合物4-(丙烷-1-炔-1-基)-1H-吲唑-7-甲酸甲酯(500mg, 产率42％)。Add the compound 4-bromo-1H-indazole-7-carboxylic acid methyl ester (1.0g, 3.94mmol) to 1,4-dioxane (10mL) at room temperature, add ditriphenylphosphine palladium dichloride (274mg, 0.39mmol), 1-propyne-tri-n-butyltin (1.52g, 4.61mmol), heated to 90°C under nitrogen protection, and stirred for 16h. Cool to room temperature, add water (50mL) to dilute, extract with ethyl acetate (50mL×3), separate the liquids, combine the organic phases, dry the organic phases with anhydrous sodium sulfate, filter, concentrate, and separate the residue with a thin-layer silica gel plate Purification (petroleum ether: ethyl acetate (V/V) = 5:1) gave compound 4-(propane-1-yn-1-yl)-1H-indazole-7-carboxylic acid methyl ester (500mg, yield 42 %).
第二步：1-(1-(4-环丙基苯基)乙基)-4-(丙烷-1-炔-1-基)-1H-吲唑-7-甲酸甲酯的制备The second step: the preparation of 1-(1-(4-cyclopropylphenyl)ethyl)-4-(propane-1-yn-1-yl)-1H-indazole-7-carboxylic acid methyl ester
室温下将化合物4-(丙烷-1-炔-1-基)-1H-吲唑-7-甲酸甲酯(500mg,2.34mmol)加入到四氢呋喃(15mL)中，加入1-(1-(4-环丙基苯基)乙基)-1-醇(568mg,3.60mmol)，三苯基膦(1.23g,4.68mmol)，偶氮二甲酸二异丙酯(945mg,4.68mmol)，室温搅拌16h。加入水(20mL)稀释，用乙酸乙酯(20mL×3)萃取，分液，合并有机相，有机相用无水硫酸钠干燥，过滤，浓缩，残留物经酸性制备方法A得化合物1-(1-(4-环丙基苯基)乙基)-4-(丙烷-1-炔-1-基)-1H-吲唑-7-甲酸甲酯(360mg,产率43％)。Compound 4-(propane-1-yn-1-yl)-1H-indazole-7-carboxylic acid methyl ester (500mg, 2.34mmol) was added to tetrahydrofuran (15mL) at room temperature, and 1-(1-(4 -Cyclopropylphenyl) ethyl)-1-ol (568mg, 3.60mmol), triphenylphosphine (1.23g, 4.68mmol), diisopropyl azodicarboxylate (945mg, 4.68mmol), stirring at room temperature 16h. Add water (20mL) to dilute, extract with ethyl acetate (20mL×3), separate the layers, combine the organic phases, dry the organic phases with anhydrous sodium sulfate, filter, concentrate, and the residue is subjected to acidic preparation method A to obtain compound 1-( 1-(4-Cyclopropylphenyl)ethyl)-4-(propan-1-yn-1-yl)-1H-indazole-7-carboxylic acid methyl ester (360 mg, 43% yield).
第三步：1-(1-(4-环丙基苯基)乙基)-4-(丙烷-1-炔-1-基)-1H-吲唑-7-甲酸的制备Step 3: Preparation of 1-(1-(4-cyclopropylphenyl)ethyl)-4-(propane-1-yn-1-yl)-1H-indazole-7-carboxylic acid
室温下将原料1-(1-(4-环丙基苯基)乙基)-4-(丙烷-1-炔-1-基)-1H-吲唑-7-甲酸甲酯(360mg,1.00mmol)加入到四氢呋喃(5mL)和水(2mL)中，加入氢氧化锂(72mg,3.00mmol)，室温搅拌16h。用1N盐酸调节pH＝4，用乙酸乙酯(20mL×3)萃取，分液，合并有机相，有机相用无水硫酸钠干燥，过滤，浓缩，得化合物粗品4-(丙烷-1-炔-1-基)-1-(4-(三氟甲氧基)苯基)乙基)-1H-吲唑-7-甲酸(330mg,产率95％)。The raw material 1-(1-(4-cyclopropylphenyl)ethyl)-4-(propane-1-yn-1-yl)-1H-indazole-7-carboxylic acid methyl ester (360mg, 1.00 mmol) was added to tetrahydrofuran (5 mL) and water (2 mL), lithium hydroxide (72 mg, 3.00 mmol) was added, and stirred at room temperature for 16 h. Use 1N hydrochloric acid to adjust pH=4, extract with ethyl acetate (20mL×3), separate the layers, combine the organic phases, dry the organic phases with anhydrous sodium sulfate, filter, and concentrate to obtain the crude compound 4-(propane-1-yne -1-yl)-1-(4-(trifluoromethoxy)phenyl)ethyl)-1H-indazole-7-carboxylic acid (330 mg, 95% yield).
第四步：6-(1-(1-(4-环丙基苯基)乙基)-4-(丙烷-1-炔-1-基)-1H-吲唑-7-甲酰胺)螺环[3.3]庚烷-2-甲酸甲酯的制备The fourth step: 6-(1-(1-(4-cyclopropylphenyl)ethyl)-4-(propane-1-yn-1-yl)-1H-indazole-7-carboxamide)spiro Preparation of methyl cyclo[3.3]heptane-2-carboxylate
室温下将化合物4-(丙烷-1-炔-1-基)-1-(4-(三氟甲氧基)苯基)乙基)-1H-吲唑-7-甲酸(330mg,0.96mmol)加入到N,N-二甲基甲酰胺(10mL)中，加入2-(6-氨基螺环[3.3]庚烷-2-基)甲酸甲酯盐酸盐(297mg,1.44mmol)，二异丙基乙胺(372mg,2.88mmol)，HATU(547mg,1.44mmol)，搅拌16h。加入水(20mL)稀释，用乙酸乙酯(20mL×3)萃取，分液，合并有机相，有机相用无水硫酸钠干燥，过滤，浓缩，残留物用薄层硅胶板分离纯化(石油醚:乙酸乙酯(V/V)＝1:1)得化合物6-(1-(1-(4-环丙基苯基)乙基)-4-(丙烷-1-炔-1-基)-1H-吲唑-7-甲酰胺)螺环[3.3]庚烷-2-甲酸甲酯(200mg，产率42％)。Compound 4-(propane-1-yn-1-yl)-1-(4-(trifluoromethoxy)phenyl)ethyl)-1H-indazole-7-carboxylic acid (330mg, 0.96mmol ) was added to N,N-dimethylformamide (10mL), 2-(6-aminospirocyclo[3.3]heptane-2-yl)methyl formate hydrochloride (297mg, 1.44mmol) was added, di Isopropylethylamine (372mg, 2.88mmol), HATU (547mg, 1.44mmol), stirred for 16h. Add water (20mL) to dilute, extract with ethyl acetate (20mL×3), separate the layers, combine the organic phases, dry the organic phases with anhydrous sodium sulfate, filter, concentrate, and separate and purify the residue with a thin-layer silica gel plate (petroleum ether : Ethyl acetate (V/V)=1:1) to obtain compound 6-(1-(1-(4-cyclopropylphenyl) ethyl)-4-(propane-1-yn-1-yl) -1H-indazole-7-carboxamide) spiro[3.3]heptane-2-carboxylic acid methyl ester (200 mg, yield 42%).
第五步：6-(1-(1-(4-环丙基苯基)乙基)-4-(丙烷-1-炔-1-基)-1H-吲唑-7-甲酰胺)螺环[3.3]庚烷-2-甲酸(I-18)的制备The fifth step: 6-(1-(1-(4-cyclopropylphenyl)ethyl)-4-(propane-1-yn-1-yl)-1H-indazole-7-carboxamide)spiro Preparation of cyclo[3.3]heptane-2-carboxylic acid (I-18)
Figure PCTCN2022097611-appb-000060
Figure PCTCN2022097611-appb-000060
室温下将化合物6-(1-(1-(4-环丙基苯基)乙基)-4-(丙烷-1-炔-1-基)-1H-吲唑-7-甲酰胺)螺环[3.3]庚烷-2-甲酸甲酯(200mg,0.40mmol)加入到四氢呋喃(5mL)和水(2mL)中，加入氢氧化锂(29mg,1.21mmol)，室温搅拌16h。用1N盐酸调节pH＝4，用乙酸乙酯(20mL×3)萃取，分液，合并有机相，有机相用无水硫酸钠干燥，过滤，浓缩，残留物经酸性制备方法A制备得化合物6-(1-(1-(4-环丙基苯基)乙基)-4-(丙烷-1-炔-1-基)-1H-吲唑-7-甲酰胺)螺环[3.3]庚烷-2-甲 酸(I-18)(178mg，产率92％)。Compound 6-(1-(1-(4-cyclopropylphenyl)ethyl)-4-(propane-1-yn-1-yl)-1H-indazole-7-carboxamide)spiro Methyl cyclo[3.3]heptane-2-carboxylate (200mg, 0.40mmol) was added to tetrahydrofuran (5mL) and water (2mL), lithium hydroxide (29mg, 1.21mmol) was added, and stirred at room temperature for 16h. Use 1N hydrochloric acid to adjust pH=4, extract with ethyl acetate (20mL×3), separate the layers, combine the organic phases, dry the organic phases with anhydrous sodium sulfate, filter, concentrate, and prepare the residue by acidic preparation method A to obtain compound 6 -(1-(1-(4-cyclopropylphenyl)ethyl)-4-(propane-1-yn-1-yl)-1H-indazole-7-carboxamide)spiro[3.3]hept Alkane-2-carboxylic acid (I-18) (178 mg, 92% yield).
1H NMR(400mHz,DMSO-d6)δ12.0(s,1H),8.81(dd,1H),8.22(s,1H),7.28(d,1H),7.18(d,1H),6.90(d,4H),6.22(d,1H),4.36-4.30(m,1H),2.98-2.90(m,1H),2.30-2.29(d,1H),2.27-2.25(m,3H),2.13(s,3H),2.10-1.85(m,4H),1.84(d,3H),1.83-1.82(m,1H),0.88(dd,2H),0.56(dd,2H). 1 H NMR (400mHz,DMSO-d6)δ12.0(s,1H),8.81(dd,1H),8.22(s,1H),7.28(d,1H),7.18(d,1H),6.90(d ,4H),6.22(d,1H),4.36-4.30(m,1H),2.98-2.90(m,1H),2.30-2.29(d,1H),2.27-2.25(m,3H),2.13(s ,3H),2.10-1.85(m,4H),1.84(d,3H),1.83-1.82(m,1H),0.88(dd,2H),0.56(dd,2H).
LC-MS,M/Z(ESI):482.2[M+H] +。 LC-MS, M/Z (ESI): 482.2 [M+H] + .
化合物I-18a的合成Synthesis of compound I-18a
合成路线如下所示：The synthetic route is as follows:
Figure PCTCN2022097611-appb-000061
Figure PCTCN2022097611-appb-000061
第一步：4-(丙烷-1-炔-1-基)-1H-吲唑-7-甲酸甲酯的制备The first step: the preparation of 4-(propane-1-yn-1-yl)-1H-indazole-7-carboxylic acid methyl ester
室温下将化合物4-溴-1H-吲唑-7-乙酸甲酯(1.0g,3.92mmol)加入到1,4-二氧六环(10mL)中，加入二三苯基膦二氯化钯(274mg,0.39mmol)，1-丙炔-三正丁基锡(1.55g,4.70mmol)，氮气保护下加热至90℃，搅拌16h。冷却至室温，加入水(50mL)稀释，用乙酸乙酯(50mL×3)萃取，分液，合并有机相，有机相用无水硫酸钠干燥，过滤，浓缩，残留物用薄层硅胶板分离纯化(石油醚:乙酸乙酯(V/V)＝5:1)得化合物4-(丙烷-1-炔-1-基)-1H-吲唑-7-乙酸甲酯(300mg,产率36％)。Add compound 4-bromo-1H-indazole-7-acetic acid methyl ester (1.0g, 3.92mmol) to 1,4-dioxane (10mL) at room temperature, add ditriphenylphosphine palladium dichloride (274mg, 0.39mmol), 1-propyne-tri-n-butyltin (1.55g, 4.70mmol), heated to 90°C under nitrogen protection, and stirred for 16h. Cool to room temperature, add water (50mL) to dilute, extract with ethyl acetate (50mL×3), separate the liquids, combine the organic phases, dry the organic phases with anhydrous sodium sulfate, filter, concentrate, and separate the residue with a thin-layer silica gel plate Purification (petroleum ether:ethyl acetate (V/V)=5:1) gave compound 4-(propane-1-yn-1-yl)-1H-indazole-7-acetic acid methyl ester (300mg, yield 36 %).
第二步：(R)-1-(1-(4-溴苯基)乙基)-4-(丙烷-1-炔-1-基)-1H-吲唑-7-甲酸甲酯的制备The second step: the preparation of (R)-1-(1-(4-bromophenyl)ethyl)-4-(propane-1-yn-1-yl)-1H-indazole-7-carboxylic acid methyl ester
室温下将4-(丙烷-1-炔-1-基)-1H-吲唑-7-甲酸甲酯(500mg,2.33mmol)和(S)-1-(4-溴苯基)乙-1-醇(845mg,4.2mmol)，三苯基膦(1.22g,4.67mmol)加入到无水四氢呋喃中(10mL)，冷却到0℃,滴加偶氮二甲酸二异丙酯(0.95g,4.67mmol)，室温搅拌16h。加入水(20mL)稀释，用乙酸乙酯(20mL×3)萃取，分液，合并有机相，有机相用无水硫酸钠干燥，过滤，浓缩，残留物薄层硅胶板分离纯化得化合物(R)-1-(1-(4-环丙基苯基)乙基)-(丙烷-1-炔-1-基)-1H-吲唑-7-甲酸甲酯(500mg,产率53.9％).4-(Propan-1-yn-1-yl)-1H-indazole-7-carboxylic acid methyl ester (500mg, 2.33mmol) and (S)-1-(4-bromophenyl)ethane-1 -alcohol (845mg, 4.2mmol), triphenylphosphine (1.22g, 4.67mmol) were added to anhydrous tetrahydrofuran (10mL), cooled to 0°C, diisopropyl azodicarboxylate (0.95g, 4.67 mmol), stirred at room temperature for 16h. Add water (20mL) to dilute, extract with ethyl acetate (20mL×3), separate the layers, combine the organic phases, dry the organic phases with anhydrous sodium sulfate, filter, concentrate, and separate and purify the residue on a thin-layer silica gel plate to obtain the compound (R )-1-(1-(4-cyclopropylphenyl)ethyl)-(propane-1-yn-1-yl)-1H-indazole-7-carboxylic acid methyl ester (500mg, yield 53.9%) .
第三步：(R)-1-(1-(4-环丙基苯基)乙基)-(丙烷-1-炔-1-基)-1H-吲唑-7-甲酸甲酯的制备Step 3: Preparation of (R)-1-(1-(4-cyclopropylphenyl)ethyl)-(propane-1-yn-1-yl)-1H-indazole-7-carboxylic acid methyl ester
室温下将(R)-1-(1-(4-溴苯基)乙基)-4-(丙烷-1-炔-1-基)-1H-吲唑-7-甲酸甲酯(500mg,1.26mmol)和环丙基硼酸(432mg,5.03mmol)加入到1,4-二氧六环(10mL)中，加入磷酸钾溶液(2M,3.2ml)，鼓泡换气后加入1,1'-二(二苯膦基)二茂铁二氯化钯(II)(103mg,0.13mmol)，微波120℃ 反应1h。加入水(20mL)稀释，用乙酸乙酯(20mL×3)萃取，分液，合并有机相，有机相用无水硫酸钠干燥，过滤，浓缩，残留物用薄层硅胶板分离纯化得化合物(R)-1-(1-(4-环丙基苯基)乙基)-4-(丙烷-1-炔-1-基)-1H-吲唑-7-甲酸甲酯(200mg,产率44％).(R)-1-(1-(4-bromophenyl)ethyl)-4-(propane-1-yn-1-yl)-1H-indazole-7-carboxylic acid methyl ester (500mg, 1.26mmol) and cyclopropylboronic acid (432mg, 5.03mmol) were added to 1,4-dioxane (10mL), potassium phosphate solution (2M, 3.2ml) was added, and 1,1' - Bis(diphenylphosphino)ferrocenepalladium(II) dichloride (103mg, 0.13mmol), microwave at 120°C for 1h. Add water (20mL) to dilute, extract with ethyl acetate (20mL×3), separate the layers, combine the organic phases, dry the organic phases with anhydrous sodium sulfate, filter and concentrate, and the residue is separated and purified with a thin-layer silica gel plate to obtain the compound ( R)-1-(1-(4-cyclopropylphenyl)ethyl)-4-(propane-1-yn-1-yl)-1H-indazole-7-carboxylic acid methyl ester (200mg, yield 44%).
第四步：(R)-1-(1-(4-环丙基苯基)乙基)-4-(丙烷-1-炔-1-基)-1H-吲唑-7-甲酸的制备Step 4: Preparation of (R)-1-(1-(4-cyclopropylphenyl)ethyl)-4-(propane-1-yn-1-yl)-1H-indazole-7-carboxylic acid
室温下将化合物(R)-1-(1-(4-环丙基苯基)乙基)-4-(丙烷-1-炔-1-基)-1H-吲唑-7-甲酸甲酯(200mg,0.56mmol)加入到四氢呋喃(5mL)，甲醇(1mL)和水(1mL)中，加入氢氧化锂溶液(2M,0.84ml)，室温搅拌16h。用1N盐酸调节pH＝4，用乙酸乙酯(20mL×3)萃取，分液，合并有机相，有机相用无水硫酸钠干燥，过滤，浓缩，残留物经酸性制备方法A制备得化合物(R)-1-(1-(4-环丙基苯基)乙基)-4-(丙烷-1-炔-1-基)-1H-吲唑-7-甲酸(150mg，产率78％)。Compound (R)-1-(1-(4-cyclopropylphenyl)ethyl)-4-(propane-1-yn-1-yl)-1H-indazole-7-carboxylic acid methyl ester (200mg, 0.56mmol) was added to tetrahydrofuran (5mL), methanol (1mL) and water (1mL), lithium hydroxide solution (2M, 0.84ml) was added, and stirred at room temperature for 16h. Use 1N hydrochloric acid to adjust pH=4, extract with ethyl acetate (20mL×3), separate the layers, combine the organic phases, dry the organic phases with anhydrous sodium sulfate, filter, concentrate, and the residue is prepared by acidic preparation method A to obtain the compound ( R)-1-(1-(4-cyclopropylphenyl)ethyl)-4-(propan-1-yn-1-yl)-1H-indazole-7-carboxylic acid (150 mg, yield 78% ).
第五步：(Sa,R)-6-(1-(1-(4-环丙基苯基)乙基)-4-(丙烷-1-炔-1-基)-1H-吲唑-7-甲酰胺基)螺[3.3]庚烷-2-甲酸甲酯的制备The fifth step: (Sa,R)-6-(1-(1-(4-cyclopropylphenyl)ethyl)-4-(propane-1-yn-1-yl)-1H-indazole- Preparation of methyl 7-formamido)spiro[3.3]heptane-2-carboxylate
将(R)-1-(1-(4-环丙基苯基)乙基)-4-(丙烷-1-炔-1-基)-1H-吲唑-7-甲酸(150mg，0.44mmol)、(Sa)-6-氨基螺[3.3]庚烷-2-甲酸甲酯盐酸盐(110mg，0.52mmol)、二异丙基乙胺(170mg，1.31mmol)和HATU(331mg，0.87mmol)溶于N,N-二甲基甲酰胺(3mL)中，然后室温搅拌16h。向反应液中加入水(15mL)，然后用乙酸乙酯(5mL*3)萃取。有机相合并，用饱和食盐水(5mL*3)洗涤，用无水硫酸钠干燥，浓缩得到粗品。粗品用薄层硅胶板分离纯化(石油醚:乙酸乙酯(V/V)＝1:1)纯化得到产物(Sa,R)-6-(1-(1-(4-环丙基苯基)乙基)-4-(丙烷-1-炔-1-基)-1H-吲唑-7-甲酰胺基)螺[3.3]庚烷-2-甲酸甲酯(150mg，产率69.5％)。LC-MS,M/Z(ESI):496.2[M+H] +。 (R)-1-(1-(4-cyclopropylphenyl)ethyl)-4-(propane-1-yn-1-yl)-1H-indazole-7-carboxylic acid (150mg, 0.44mmol ), (Sa)-6-aminospiro[3.3]heptane-2-carboxylic acid methyl ester hydrochloride (110mg, 0.52mmol), diisopropylethylamine (170mg, 1.31mmol) and HATU (331mg, 0.87mmol ) was dissolved in N,N-dimethylformamide (3 mL), then stirred at room temperature for 16 h. Water (15 mL) was added to the reaction liquid, followed by extraction with ethyl acetate (5 mL*3). The organic phases were combined, washed with saturated brine (5 mL*3), dried over anhydrous sodium sulfate, and concentrated to obtain a crude product. The crude product was separated and purified on a thin-layer silica gel plate (petroleum ether:ethyl acetate (V/V)=1:1) to obtain the product (Sa,R)-6-(1-(1-(4-cyclopropylphenyl )ethyl)-4-(propane-1-yn-1-yl)-1H-indazole-7-carboxamido)spiro[3.3]heptane-2-carboxylic acid methyl ester (150 mg, yield 69.5%) . LC-MS, M/Z (ESI): 496.2 [M+H] + .
第六步：(Sa,R)-6-(1-(1-(4-环丙基苯基)乙基)-4-(丙烷-1-炔-1-基)-1H-吲唑-7-甲酰胺基)螺[3.3]庚烷-2-甲酸(I-18a)The sixth step: (Sa,R)-6-(1-(1-(4-cyclopropylphenyl)ethyl)-4-(propane-1-yn-1-yl)-1H-indazole- 7-formamido)spiro[3.3]heptane-2-carboxylic acid (I-18a)
Figure PCTCN2022097611-appb-000062
Figure PCTCN2022097611-appb-000062
室温下将化合物(Sa,R)-6-(1-(1-(4-环丙基苯基)乙基)-4-(丙烷-1-炔-1-基)-1H-吲唑-7-甲酰胺基)螺[3.3]庚烷-2-甲酸甲酯(150mg,0.30mmol)加入到四氢呋喃(3mL)，甲醇(1mL)和水(1mL)中，加入氢氧化锂溶液(2M,0.45ml)，室温搅拌16h。用1N盐酸调节pH＝4，用乙酸乙酯(20mL×3)萃取，分液，合并有机相，有机相用无水硫酸钠干燥，过滤，浓缩，残留物经酸性制备方法A制备得化合物(Sa,R)-6-(1-(1-(4-环丙基苯基)乙基)-4-(丙烷-1-炔-1-基)-1H-吲唑-7-甲酰胺基)螺[3.3]庚烷-2-甲酸(I-18a，65.1mg,产率44.7％)。Compound (Sa,R)-6-(1-(1-(4-cyclopropylphenyl)ethyl)-4-(propane-1-yn-1-yl)-1H-indazole- 7-Carboxamido)spiro[3.3]heptane-2-carboxylic acid methyl ester (150mg, 0.30mmol) was added to tetrahydrofuran (3mL), methanol (1mL) and water (1mL), and lithium hydroxide solution (2M, 0.45ml), stirred at room temperature for 16h. Use 1N hydrochloric acid to adjust pH=4, extract with ethyl acetate (20mL×3), separate the layers, combine the organic phases, dry the organic phases with anhydrous sodium sulfate, filter, concentrate, and the residue is prepared by acidic preparation method A to obtain the compound ( Sa,R)-6-(1-(1-(4-cyclopropylphenyl)ethyl)-4-(propane-1-yn-1-yl)-1H-indazole-7-carboxamido ) spiro[3.3]heptane-2-carboxylic acid (I-18a, 65.1 mg, 44.7% yield).
1H NMR(400MHz,CDCl 3)δ12.0(s,1H),8.22(s,1H),7.22(d,1H),7.09(d,1H),6.88–6.81(m,3H),6.23(q,1H),5.73(d,1H),4.42(dt,1H),3.09(dd,1H),2.62(dd,1H),2.47–2.39(m,3H),2.32(dd,1H),2.18(d,4H),1.95(dd,4H),1.78(ddd,3H),0.89(ddd,2H),0.58(dt,2H). 1 H NMR (400MHz, CDCl 3 )δ12.0(s,1H),8.22(s,1H),7.22(d,1H),7.09(d,1H),6.88–6.81(m,3H),6.23( q,1H),5.73(d,1H),4.42(dt,1H),3.09(dd,1H),2.62(dd,1H),2.47–2.39(m,3H),2.32(dd,1H),2.18 (d,4H),1.95(dd,4H),1.78(ddd,3H),0.89(ddd,2H),0.58(dt,2H).
LC-MS,M/Z(ESI):482.5[M+H] + LC-MS, M/Z(ESI):482.5[M+H] +
化合物I-18b合成Synthesis of Compound I-18b
合成路线如下所示：The synthetic route is as follows:
Figure PCTCN2022097611-appb-000063
Figure PCTCN2022097611-appb-000063
化合物I-18b的制备方法参考I-18a的制备方法。在第二步将(Sa,S)-1-(4-溴苯基)乙醇换成(R)-1-(4-溴苯基)乙醇,经过同样的六步反应得到化合物(Sa,S)-6-(1-(1-(4-环丙基苯基)乙基)-4-(丙烷-1-炔-1-基)-1H-吲唑-7-甲酰胺基)螺[3.3]庚烷-2-甲酸(I-18b，43.7mg,产率64％)。For the preparation method of compound I-18b, refer to the preparation method of I-18a. In the second step, (Sa, S)-1-(4-bromophenyl)ethanol is replaced with (R)-1-(4-bromophenyl)ethanol, and the compound (Sa,S )-6-(1-(1-(4-cyclopropylphenyl)ethyl)-4-(propane-1-yn-1-yl)-1H-indazole-7-carboxamido)spiro[ 3.3] Heptane-2-carboxylic acid (I-18b, 43.7 mg, 64% yield).
1H NMR(400MHz,CDCl 3)δ12.0(s,1H),8.22(s,1H),7.22(d,1H),7.09(d,1H),6.89–6.82(m,3H),6.24(q,1H),5.77(d,1H),4.43(dq,1H),3.14–3.05(m,1H),2.51(dt,2H),2.42(d,2H),2.34–2.28(m,1H),2.21–2.14(m,4H),1.97(d,3H),1.90–1.70(m,4H),0.93–0.86(m,2H),0.59(dt,2H)。 1 H NMR (400MHz, CDCl 3 )δ12.0(s,1H),8.22(s,1H),7.22(d,1H),7.09(d,1H),6.89–6.82(m,3H),6.24( q,1H),5.77(d,1H),4.43(dq,1H),3.14–3.05(m,1H),2.51(dt,2H),2.42(d,2H),2.34–2.28(m,1H) ,2.21–2.14(m,4H),1.97(d,3H),1.90–1.70(m,4H),0.93–0.86(m,2H),0.59(dt,2H).
LC-MS,M/Z(ESI):482.5[M+H] + LC-MS, M/Z(ESI):482.5[M+H] +
实施例19化合物I-19的制备The preparation of embodiment 19 compound I-19
合成路线如下所示：The synthetic route is as follows:
Figure PCTCN2022097611-appb-000064
Figure PCTCN2022097611-appb-000064
第一步：4-(丙烷-1-炔-1-基)-1H-吲唑-7-甲酸甲酯的制备The first step: the preparation of 4-(propane-1-yn-1-yl)-1H-indazole-7-carboxylic acid methyl ester
室温下将化合物4-溴-1H-吲唑-7-甲酸甲酯(1.0g,3.94mmol)加入到1,4-二氧六环(10mL)中，加入二三苯基膦二氯化钯(274mg,0.39mmol)，1-丙炔-三正丁基锡(1.52g,4.61mmol)，氮气保护下加热至90℃，搅拌16h。冷却至室温，加入水(50mL)稀释，用乙酸乙酯(50mL× 3)萃取，分液，合并有机相，有机相用无水硫酸钠干燥，过滤，浓缩，残留物用薄层硅胶板分离纯化(石油醚:乙酸乙酯(V/V)＝5:1)得化合物4-(丙烷-1-炔-1-基)-1H-吲唑-7-甲酸甲酯(500mg,产率42％)。Add the compound 4-bromo-1H-indazole-7-carboxylic acid methyl ester (1.0g, 3.94mmol) to 1,4-dioxane (10mL) at room temperature, add ditriphenylphosphine palladium dichloride (274mg, 0.39mmol), 1-propyne-tri-n-butyltin (1.52g, 4.61mmol), heated to 90°C under nitrogen protection, and stirred for 16h. Cool to room temperature, add water (50mL) to dilute, extract with ethyl acetate (50mL×3), separate the liquids, combine the organic phases, dry the organic phases with anhydrous sodium sulfate, filter, concentrate, and separate the residue with a thin-layer silica gel plate Purification (petroleum ether: ethyl acetate (V/V) = 5:1) gave compound 4-(propane-1-yn-1-yl)-1H-indazole-7-carboxylic acid methyl ester (500mg, yield 42 %).
第二步：4-(丙烷-1-炔-1-基)-1-(4-(三氟甲氧基)苯基)乙基)-1H-吲唑-7-甲酸甲酯的制备The second step: the preparation of 4-(propane-1-yn-1-yl)-1-(4-(trifluoromethoxy)phenyl)ethyl)-1H-indazole-7-carboxylic acid methyl ester
室温下将化合物4-(丙烷-1-炔-1-基)-1H-吲唑-7-甲酸甲酯(300mg,1.40mmol)加入到四氢呋喃(10mL)中，加入1-(4-(三氟甲氧基)苯基)乙基)-1-醇(433mg,2.10mmol)，三苯基膦(736mg,2.80mmol)，偶氮二甲酸二异丙酯(283mg,2.80mmol)，室温搅拌16h。加入水(20mL)稀释，用乙酸乙酯(20mL×3)萃取，分液，合并有机相，有机相用无水硫酸钠干燥，过滤，浓缩，残留物经酸性制备方法A得化合物4-(丙烷-1-炔-1-基)-1-(4-(三氟甲氧基)苯基)乙基)-1H-吲唑-7-甲酸甲酯(120mg,产率21％)。Compound 4-(propane-1-yn-1-yl)-1H-indazole-7-carboxylic acid methyl ester (300 mg, 1.40 mmol) was added to tetrahydrofuran (10 mL) at room temperature, and 1-(4-(tri Fluoromethoxy)phenyl)ethyl)-1-ol (433mg, 2.10mmol), triphenylphosphine (736mg, 2.80mmol), diisopropyl azodicarboxylate (283mg, 2.80mmol), stirring at room temperature 16h. Add water (20mL) to dilute, extract with ethyl acetate (20mL×3), separate the layers, combine the organic phases, dry the organic phases with anhydrous sodium sulfate, filter, concentrate, and the residue is subjected to acidic preparation method A to obtain compound 4-( Propan-1-yn-1-yl)-1-(4-(trifluoromethoxy)phenyl)ethyl)-1H-indazole-7-carboxylic acid methyl ester (120 mg, 21% yield).
第三步：4-(丙烷-1-炔-1-基)-1-(4-(三氟甲氧基)苯基)乙基)-1H-吲唑-7-甲酸的制备The third step: the preparation of 4-(propane-1-yn-1-yl)-1-(4-(trifluoromethoxy)phenyl)ethyl)-1H-indazole-7-carboxylic acid
室温下将原料4-(丙烷-1-炔-1-基)-1-(4-(三氟甲氧基)苯基)乙基)-1H-吲唑-7-甲酸甲酯(120mg,0.30mmol)加入到四氢呋喃(5mL)和水(2mL)中，加入氢氧化锂(22mg,0.92mmol)，室温搅拌16h。用1N盐酸调节pH＝4，用乙酸乙酯(20mL×3)萃取，分液，合并有机相，有机相用无水硫酸钠干燥，过滤，浓缩，得化合物粗品4-(丙烷-1-炔-1-基)-1-(4-(三氟甲氧基)苯基)乙基)-1H-吲唑-7-甲酸(100mg,产率86％).The starting material 4-(propane-1-yn-1-yl)-1-(4-(trifluoromethoxy)phenyl)ethyl)-1H-indazole-7-carboxylic acid methyl ester (120 mg, 0.30mmol) was added to tetrahydrofuran (5mL) and water (2mL), lithium hydroxide (22mg, 0.92mmol) was added, and stirred at room temperature for 16h. Use 1N hydrochloric acid to adjust pH=4, extract with ethyl acetate (20mL×3), separate the layers, combine the organic phases, dry the organic phases with anhydrous sodium sulfate, filter, and concentrate to obtain the crude compound 4-(propane-1-yne -1-yl)-1-(4-(trifluoromethoxy)phenyl)ethyl)-1H-indazole-7-carboxylic acid (100mg, yield 86%).
LC-MS,M/Z(ESI):389.4[M+H] +。 LC-MS, M/Z (ESI): 389.4 [M+H] + .
第四步：6-(4-(丙烷-1-炔-1-基)-1-(4-(三氟甲氧基)苯基)乙基)-1H-吲唑-7-甲酰胺)螺环[3.3]庚烷-2-甲酸甲酯的制备The fourth step: 6-(4-(propane-1-yn-1-yl)-1-(4-(trifluoromethoxy)phenyl)ethyl)-1H-indazole-7-carboxamide) Preparation of spiro[3.3]heptane-2-carboxylate methyl ester
室温下将化合物4-(丙烷-1-炔-1-基)-1-(4-(三氟甲氧基)苯基)乙基)-1H-吲唑-7-甲酸(100mg,0.26mmol)加入到二异丙基乙胺(5mL)中，加入2-(6-氨基螺环[3.3]庚烷-2-基)甲酸甲酯盐酸盐(80mg,0.39mmol)，二异丙基乙胺(67mg,0.52mmol)，HATU(148mg,0.39mmol)，搅拌16h。加入水(20mL)稀释，用乙酸乙酯(20mL×3)萃取，分液，合并有机相，有机相用无水硫酸钠干燥，过滤，浓缩，残留物用薄层硅胶板分离纯化(石油醚:乙酸乙酯(V/V)＝1:1)得化合物6-(4-(丙烷-1-炔-1-基)-1-(4-(三氟甲氧基)苯基)乙基)-1H-吲唑-7-甲酰胺)螺环[3.3]庚烷-2-甲酸甲酯(80mg,产率58％)。Compound 4-(propane-1-yn-1-yl)-1-(4-(trifluoromethoxy)phenyl)ethyl)-1H-indazole-7-carboxylic acid (100mg, 0.26mmol ) into diisopropylethylamine (5mL), add 2-(6-aminospirocyclo[3.3]heptane-2-yl)methyl formate hydrochloride (80mg, 0.39mmol), diisopropyl Ethylamine (67mg, 0.52mmol), HATU (148mg, 0.39mmol), stirred for 16h. Add water (20mL) to dilute, extract with ethyl acetate (20mL×3), separate the layers, combine the organic phases, dry the organic phases with anhydrous sodium sulfate, filter, concentrate, and separate and purify the residue with a thin-layer silica gel plate (petroleum ether : ethyl acetate (V/V)=1:1) to obtain compound 6-(4-(propane-1-yn-1-yl)-1-(4-(trifluoromethoxy)phenyl)ethyl )-1H-indazole-7-carboxamide)spiro[3.3]heptane-2-carboxylic acid methyl ester (80 mg, 58% yield).
第五步：6-(4-(丙烷-1-炔-1-基)-1-(1-(4-(三氟甲氧基)苯基)乙基)-1H-吲唑-7-甲酰胺)螺环[3.3]庚烷-2-甲酸(I-19)的制备The fifth step: 6-(4-(propane-1-yn-1-yl)-1-(1-(4-(trifluoromethoxy)phenyl)ethyl)-1H-indazole-7- Preparation of formamide) spiro[3.3]heptane-2-carboxylic acid (I-19)
Figure PCTCN2022097611-appb-000065
Figure PCTCN2022097611-appb-000065
室温下将化合物6-(4-(丙烷-1-炔-1-基)-1-(4-(三氟甲氧基)苯基)乙基)-1H-吲唑-7-甲酰胺) 螺环[3.3]庚烷-2-甲酸甲酯(80mg,0.15mmol)加入到四氢呋喃(5mL)和水(2mL)中，加入氢氧化锂(11mg,0.46mmol)，室温搅拌16h。用1N盐酸调节pH＝4，用乙酸乙酯(20mL×3)萃取，分液，合并有机相，有机相用无水硫酸钠干燥，过滤，浓缩，残留物经酸性制备方法A制备得化合物6-(4-(丙烷-1-炔-1-基)-1-(1-(4-(三氟甲氧基)苯基)乙基)-1H-吲唑-7-甲酰胺)螺环[3.3]庚烷-2-甲酸(I-19)(70mg,产率60％)。Compound 6-(4-(propane-1-yn-1-yl)-1-(4-(trifluoromethoxy)phenyl)ethyl)-1H-indazole-7-carboxamide) at room temperature Methyl spiro[3.3]heptane-2-carboxylate (80mg, 0.15mmol) was added to tetrahydrofuran (5mL) and water (2mL), lithium hydroxide (11mg, 0.46mmol) was added, and stirred at room temperature for 16h. Use 1N hydrochloric acid to adjust pH=4, extract with ethyl acetate (20mL×3), separate the layers, combine the organic phases, dry the organic phases with anhydrous sodium sulfate, filter, concentrate, and prepare the residue by acidic preparation method A to obtain compound 6 -(4-(propane-1-yn-1-yl)-1-(1-(4-(trifluoromethoxy)phenyl)ethyl)-1H-indazole-7-carboxamide) spirocycle [3.3] Heptane-2-carboxylic acid (I-19) (70 mg, yield 60%).
1H NMR(400mHz,DMSO-d6)δ12.0(s,1H),8.79(d,1H),8.27(s,1H),7.29(d,1H),7.21(t,3H),7.12(d,2H),6.34(q,1H),4.32-4.26(m,1H),2.96(t,1H),2.33-2.19(m,4H),2.17(s,3H),2.10-1.87(m,4H),1.88(d,3H)。 1 H NMR(400mHz,DMSO-d6)δ12.0(s,1H),8.79(d,1H),8.27(s,1H),7.29(d,1H),7.21(t,3H),7.12(d ,2H),6.34(q,1H),4.32-4.26(m,1H),2.96(t,1H),2.33-2.19(m,4H),2.17(s,3H),2.10-1.87(m,4H ), 1.88(d,3H).
LC-MS,M/Z(ESI):526.6[M+H] +。 LC-MS, M/Z (ESI): 526.6 [M+H] + .
化合物I-19a、I-19b、I-19c、I-19d经化合物I-19手性拆分获得，拆分方法：使用大赛璐AD型号手性拆分柱(规格为：25*250mm,10um)进行手性拆分，流动相A为超临界流体CO 2,流动相B为甲醇(含0.05％二乙醇胺)；梯度条件:流动相B从5％至80％，流动速率为30mL/min；20％的梯度保持10min，40％的梯度保持10min，50％-80％梯度洗脱，保持20min，柱温保持在35℃，柱压保持在100bar。 Compounds I-19a, I-19b, I-19c, and I-19d were obtained by chiral resolution of compound I-19. Resolution method: use Daicel AD type chiral resolution column (specification: 25*250mm, 10um ) for chiral resolution, mobile phase A is supercritical fluid CO 2 , mobile phase B is methanol (containing 0.05% diethanolamine); gradient conditions: mobile phase B is from 5% to 80%, and the flow rate is 30mL/min; The 20% gradient was kept for 10 minutes, the 40% gradient was kept for 10 minutes, the 50%-80% gradient elution was kept for 20 minutes, the column temperature was kept at 35° C., and the column pressure was kept at 100 bar.
化合物I-19a、I-19b、I-19c、I-19d的结构如下所示：The structures of compounds I-19a, I-19b, I-19c, and I-19d are shown below:
Figure PCTCN2022097611-appb-000066
Figure PCTCN2022097611-appb-000066
化合物I-19a保留时间：0.731min，Compound I-19a retention time: 0.731min,
1H NMR(400mHz,DMSO-d6)δ12.0(s,1H),8.80(d,1H),8.27(s,1H),7.32(d,1H),7.25(t,3H),7.12(d,2H),6.35(q,1H),4.33-4.27(m,1H),2.96(t,1H),2.45-2.40(m,1H),2.32-2.19(m,3H),2.17(s,3H),2.14-2.01(m,2H),1.97-1.92(m,1H),1.89(d,3H),1.87-1.83(m,1H). 1 H NMR(400mHz,DMSO-d6)δ12.0(s,1H),8.80(d,1H),8.27(s,1H),7.32(d,1H),7.25(t,3H),7.12(d ,2H),6.35(q,1H),4.33-4.27(m,1H),2.96(t,1H),2.45-2.40(m,1H),2.32-2.19(m,3H),2.17(s,3H ),2.14-2.01(m,2H),1.97-1.92(m,1H),1.89(d,3H),1.87-1.83(m,1H).
LC-MS,M/Z(ESI):526.6[M+H] +。 LC-MS, M/Z (ESI): 526.6 [M+H] + .
化合物I-19b，保留时间：0.968min，Compound I-19b, retention time: 0.968min,
1H NMR(400mHz,DMSO-d6)δ12.05(s,1H),8.80(d,1H),8.27(s,1H),7.32(d,1H),7.24(t,3H),7.12(d,2H),6.34(q,1H),4.33-4.27(m,1H),2.97(t,1H),2.46-2.41(m,1H),2.32-2.19(m,3H),2.17(s,3H),2.14-2.01(m,2H),1.97-1.92(m,1H),1.89(d,3H),1.81-1.76(m,1H). 1 H NMR (400mHz,DMSO-d6)δ12.05(s,1H),8.80(d,1H),8.27(s,1H),7.32(d,1H),7.24(t,3H),7.12(d ,2H),6.34(q,1H),4.33-4.27(m,1H),2.97(t,1H),2.46-2.41(m,1H),2.32-2.19(m,3H),2.17(s,3H ),2.14-2.01(m,2H),1.97-1.92(m,1H),1.89(d,3H),1.81-1.76(m,1H).
LC-MS,M/Z(ESI):526.6[M+H]+。LC-MS, M/Z (ESI): 526.6 [M+H]+.
化合物I-19c，保留时间：1.585min，Compound I-19c, retention time: 1.585min,
1H NMR(400mHz,DMSO-d6)δ12.05(s,1H),8.80(d,1H),8.28(s,1H),7.32(d,1H),7.24(t,3H),7.12(d,2H),6.35(q,1H),4.33-4.27(m,1H),2.98(t,1H),2.49-2.42(m,1H),2.33-2.21(m,3H),2.17(s,3H),2.14-1.99(m,3H),1.89(d,3H),1.82-1.77(m,1H). 1 H NMR (400mHz,DMSO-d6)δ12.05(s,1H),8.80(d,1H),8.28(s,1H),7.32(d,1H),7.24(t,3H),7.12(d ,2H),6.35(q,1H),4.33-4.27(m,1H),2.98(t,1H),2.49-2.42(m,1H),2.33-2.21(m,3H),2.17(s,3H ),2.14-1.99(m,3H),1.89(d,3H),1.82-1.77(m,1H).
LC-MS,M/Z(ESI):526.6[M+H]+。LC-MS, M/Z (ESI): 526.6 [M+H]+.
化合物I-19d，保留时间：3.323min，Compound I-19d, retention time: 3.323min,
1H NMR(400mHz,DMSO-d6)δ11.90(s,1H),8.80(d,1H),8.27(s,1H),7.32(d,1H),7.25(t,3H),7.12(d,2H),6.35(q,1H),4.33-4.27(m,1H),2.95(t,1H),2.45-2.39(m,1H),2.34-2.21(m,3H),2.17(s,3H),2.14-1.92(m,3H),1.89(d,3H),1.87-1.83(m,1H). 1 H NMR(400mHz,DMSO-d6)δ11.90(s,1H),8.80(d,1H),8.27(s,1H),7.32(d,1H),7.25(t,3H),7.12(d ,2H),6.35(q,1H),4.33-4.27(m,1H),2.95(t,1H),2.45-2.39(m,1H),2.34-2.21(m,3H),2.17(s,3H ),2.14-1.92(m,3H),1.89(d,3H),1.87-1.83(m,1H).
LC-MS,M/Z(ESI):526.6[M+H]+。LC-MS, M/Z (ESI): 526.6 [M+H]+.
实施例20化合物I-20a的制备The preparation of embodiment 20 compound I-20a
合成路线如下所示：The synthetic route is as follows:
Figure PCTCN2022097611-appb-000067
Figure PCTCN2022097611-appb-000067
第一步：(R)-1-(4-(4,4,5,5-四甲基-1,3,2-二氧硼烷-2-基)苯基)乙醇的制备Step 1: Preparation of (R)-1-(4-(4,4,5,5-tetramethyl-1,3,2-dioxaborolan-2-yl)phenyl)ethanol
室温下将(R)-1-(4-溴苯基)乙醇(500mg,2.49mmol)和联硼酸频那醇酯(1.26g,4.97mmol)加入到1,4-二氧六环(10mL)中，加入1,1'-二(二苯膦基)二茂铁二氯化钯(II)(180mg,0.25mmol)和醋酸钾(732mg,7.46mmol)，90℃搅拌7h。加入水(30mL)稀释，用乙酸乙酯(20mL×3)萃取，分液，合并有机相，有机相用无水硫酸钠干燥，过滤，浓缩，残留物用薄层硅胶板分离纯化得到(R)-1-(4-(4,4,5,5-四甲基-1,3,2-二氧硼烷-2-基)苯基)乙醇(450mg,产率73％)。(R)-1-(4-Bromophenyl)ethanol (500mg, 2.49mmol) and pinacol diboronate (1.26g, 4.97mmol) were added to 1,4-dioxane (10mL) at room temperature Add 1,1'-bis(diphenylphosphino)ferrocenepalladium(II) chloride (180mg, 0.25mmol) and potassium acetate (732mg, 7.46mmol), and stir at 90°C for 7h. Add water (30mL) to dilute, extract with ethyl acetate (20mL×3), separate the layers, combine the organic phases, dry the organic phases with anhydrous sodium sulfate, filter, concentrate, and the residue is separated and purified on a thin-layer silica gel plate to obtain (R )-1-(4-(4,4,5,5-Tetramethyl-1,3,2-dioxaborolan-2-yl)phenyl)ethanol (450 mg, 73% yield).
第二步：(R)-1-(4-(2-甲氧基嘧啶-4-基)苯基)乙醇的制备The second step: the preparation of (R)-1-(4-(2-methoxypyrimidin-4-yl)phenyl)ethanol
室温下将原料(R)-1-(4-(4,4,5,5-四甲基-1,3,2-二氧硼烷-2-基)苯基)乙醇(400mg,1.6mmol)和4-溴-2-甲氧基嘧啶(366mg,1.94mmol)加入到1,4-二氧六环(10mL)和水(1mL)中，加入1,1'-二(二苯膦基)二茂铁二氯化钯(II)(118mg,0.16mmol)和碳酸钠(1.05g,3.22mmol)，80℃搅拌1h。加入水(20mL)稀释，用乙酸乙酯(20mL×3)萃取，分液，合并有机相，有机相用无水硫酸钠干燥，过滤，浓缩，残留物用薄层硅胶板分离纯化得化合物(R)-1-(4-(2-甲氧基嘧啶-4-基)苯 基)乙醇(260mg,产率70％)。The raw material (R)-1-(4-(4,4,5,5-tetramethyl-1,3,2-dioxaborolan-2-yl)phenyl)ethanol (400mg, 1.6mmol ) and 4-bromo-2-methoxypyrimidine (366mg, 1.94mmol) were added to 1,4-dioxane (10mL) and water (1mL), and 1,1'-bis(diphenylphosphino ) ferrocenepalladium(II) chloride (118mg, 0.16mmol) and sodium carbonate (1.05g, 3.22mmol), stirred at 80°C for 1h. Add water (20mL) to dilute, extract with ethyl acetate (20mL×3), separate the layers, combine the organic phases, dry the organic phases with anhydrous sodium sulfate, filter and concentrate, and the residue is separated and purified with a thin-layer silica gel plate to obtain the compound ( R)-1-(4-(2-methoxypyrimidin-4-yl)phenyl)ethanol (260 mg, 70% yield).
第三步：(S)-1-(1-(4-(2-甲氧基嘧啶-4-基)苯基)乙基)-4-(丙烷-1-炔-1-基)-1H-吲唑-7-甲酸甲酯的制备The third step: (S)-1-(1-(4-(2-methoxypyrimidin-4-yl)phenyl)ethyl)-4-(propane-1-yn-1-yl)-1H - Preparation of indazole-7-methyl carboxylate
室温下将(R)-1-(4-(2-甲氧基嘧啶-4-基)苯基)乙醇(260mg,0.93mmol)和4-(丙烷-1-炔-1-基)-1H-吲唑-7-甲酸甲酯(322mg,1.4mmol)，三苯基膦(860mg,3.27mmol)加入到无水四氢呋喃中(5mL)，冷却到0℃,滴加偶氮二甲酸二异丙酯(660mg,3.27mmol)，室温搅拌16h。加入水(20mL)稀释，用乙酸乙酯(20mL×3)萃取，分液，合并有机相，有机相用无水硫酸钠干燥，过滤，浓缩，残留物薄层硅胶板分离纯化得化合物(S)-1-(1-(4-(2-甲氧基嘧啶-4-基)苯基)乙基)-4-(丙烷-1-炔-1-基)-1H-吲唑-7-甲酸甲酯(120mg,产率30％)。(R)-1-(4-(2-methoxypyrimidin-4-yl)phenyl)ethanol (260mg, 0.93mmol) and 4-(propane-1-yn-1-yl)-1H -Methyl indazole-7-carboxylate (322mg, 1.4mmol), triphenylphosphine (860mg, 3.27mmol) were added to anhydrous tetrahydrofuran (5mL), cooled to 0°C, and diisopropyl azodicarboxylate was added dropwise Ester (660mg, 3.27mmol), stirred at room temperature for 16h. Add water (20mL) to dilute, extract with ethyl acetate (20mL×3), separate the layers, combine the organic phases, dry the organic phases with anhydrous sodium sulfate, filter, concentrate, and the residue is separated and purified on a thin-layer silica gel plate to obtain compound (S )-1-(1-(4-(2-methoxypyrimidin-4-yl)phenyl)ethyl)-4-(propane-1-yn-1-yl)-1H-indazole-7- Methyl formate (120 mg, 30% yield).
第四步：(S)-1-(1-(4-(2-甲氧基嘧啶-4-基)苯基)乙基)-4-(丙烷-1-炔-1-基)-1H-吲唑-7-甲酸的制备The fourth step: (S)-1-(1-(4-(2-methoxypyrimidin-4-yl)phenyl)ethyl)-4-(propane-1-yn-1-yl)-1H - Preparation of indazole-7-carboxylic acid
室温下将化合物(S)-1-(1-(4-(2-甲氧基嘧啶-4-基)苯基)乙基)-4-(丙烷-1-炔-1-基)-1H-吲唑-7-甲酸甲酯(120mg,0.28mmol)加入到四氢呋喃(3mL)，甲醇(1mL)和水(1mL)中，加入氢氧化锂(20mg,0.84mmol)，室温搅拌16h。用1N盐酸调节pH＝4，用乙酸乙酯(20mL×3)萃取，分液，合并有机相，有机相用无水硫酸钠干燥，过滤，浓缩，残留物经酸性制备方法A制备得化合物(S)-1-(1-(4-(2-甲氧基嘧啶-4-基)苯基)乙基)-4-(丙烷-1-炔-1-基)-1H-吲唑-7-甲酸(60mg,产率52％)。Compound (S)-1-(1-(4-(2-methoxypyrimidin-4-yl)phenyl)ethyl)-4-(propane-1-yn-1-yl)-1H -Methyl indazole-7-carboxylate (120mg, 0.28mmol) was added to tetrahydrofuran (3mL), methanol (1mL) and water (1mL), lithium hydroxide (20mg, 0.84mmol) was added, and stirred at room temperature for 16h. Use 1N hydrochloric acid to adjust pH=4, extract with ethyl acetate (20mL×3), separate the layers, combine the organic phases, dry the organic phases with anhydrous sodium sulfate, filter, concentrate, and the residue is prepared by acidic preparation method A to obtain the compound ( S)-1-(1-(4-(2-methoxypyrimidin-4-yl)phenyl)ethyl)-4-(propane-1-yn-1-yl)-1H-indazole-7 - Formic acid (60 mg, 52% yield).
第五步：(Sa,S)-6-(1-(1-(4-(2-甲氧基嘧啶-4-基)苯基)乙基)-4-(丙烷-1-炔-1-基)-1H-吲唑-7-甲酰氨基)螺[3.3]庚烷-2-甲酸甲酯The fifth step: (Sa,S)-6-(1-(1-(4-(2-methoxypyrimidin-4-yl)phenyl)ethyl)-4-(propane-1-yne-1 -yl)-1H-indazole-7-carboxamido)spiro[3.3]heptane-2-carboxylic acid methyl ester
室温下将化合物(S)-1-(1-(4-(2-甲氧基嘧啶-4-基)苯基)乙基)-4-(丙烷-1-炔-1-基)-1H-吲唑-7-甲酸(60mg,0.145mmol)加入到N,N-二甲基甲酰胺(2mL)中，加入(Sa)-2-(6-氨基螺环[3.3]庚烷-2-基)甲酸甲酯盐酸盐(36mg,0.16mmol)，二异丙基乙胺(56mg,0.44mmol)，HATU(83mg,0.22mmol)，搅拌16h。加入水(20mL)稀释，用乙酸乙酯(20mL×3)萃取，分液，合并有机相，有机相用无水硫酸钠干燥，过滤，浓缩，残留物经酸性制备方法A制备得化合物(Sa,S)-6-(1-(1-(4-(2-甲氧基嘧啶-4-基)苯基)乙基)-4-(丙烷-1-炔-1-基)-1H-吲唑-7-甲酰氨基)螺[3.3]庚烷-2-甲酸甲酯(60mg,产率73％)。Compound (S)-1-(1-(4-(2-methoxypyrimidin-4-yl)phenyl)ethyl)-4-(propane-1-yn-1-yl)-1H -Indazole-7-carboxylic acid (60mg, 0.145mmol) was added to N,N-dimethylformamide (2mL), and (Sa)-2-(6-aminospirocyclo[3.3]heptane-2- Base) methyl formate hydrochloride (36mg, 0.16mmol), diisopropylethylamine (56mg, 0.44mmol), HATU (83mg, 0.22mmol), stirred for 16h. Add water (20mL) to dilute, extract with ethyl acetate (20mL×3), separate the layers, combine the organic phases, dry the organic phases with anhydrous sodium sulfate, filter, concentrate, and the residue is prepared by acidic preparation method A to obtain compound (Sa ,S)-6-(1-(1-(4-(2-methoxypyrimidin-4-yl)phenyl)ethyl)-4-(propane-1-yn-1-yl)-1H- Indazole-7-carboxamido)spiro[3.3]heptane-2-carboxylic acid methyl ester (60 mg, yield 73%).
第六步：(Sa,S)-6-(1-(1-(4-(2-甲氧基嘧啶-4-基)苯基)乙基)-4-(丙烷-1-炔-1-基)-1H-吲唑-7-甲酰胺基)螺[3.3]庚烷-2-甲酸(I-20a)The sixth step: (Sa,S)-6-(1-(1-(4-(2-methoxypyrimidin-4-yl)phenyl)ethyl)-4-(propane-1-yne-1 -yl)-1H-indazole-7-carboxamido)spiro[3.3]heptane-2-carboxylic acid (I-20a)
Figure PCTCN2022097611-appb-000068
Figure PCTCN2022097611-appb-000068
室温下将化合物(Sa,S)-甲基6-(1-(1-(4-(2-甲氧基嘧啶-4-基)苯基)乙基)-4-(丙烷-1-炔-1-基)-1H-吲唑-7-甲酰氨基)螺[3.3]庚烷-2-甲酸甲酯(60mg,0.11mmol)加入到四氢呋喃(3mL)，甲醇 (1mL)和水(1mL)中，加入氢氧化锂(8mg,0.32mmol)，室温搅拌16h。用1N盐酸调节pH＝4，用乙酸乙酯(20mL×3)萃取，分液，合并有机相，有机相用无水硫酸钠干燥，过滤，浓缩，残留物经酸性制备方法A制备得化合物(Sa,S)-6-(1-(1-(4-(2-甲氧基嘧啶-4-基)苯基)乙基)-4-(丙烷-1-炔-1-基)-1H-吲唑-7-甲酰胺基)螺[3.3]庚烷-2-甲酸(I-20a)(17.5mg，产率30％)。Compound (Sa,S)-methyl 6-(1-(1-(4-(2-methoxypyrimidin-4-yl)phenyl)ethyl)-4-(propane-1-yne -1-yl)-1H-indazole-7-carboxamido)spiro[3.3]heptane-2-carboxylic acid methyl ester (60mg, 0.11mmol) was added to tetrahydrofuran (3mL), methanol (1mL) and water (1mL ), add lithium hydroxide (8mg, 0.32mmol), and stir at room temperature for 16h. Use 1N hydrochloric acid to adjust pH=4, extract with ethyl acetate (20mL×3), separate the layers, combine the organic phases, dry the organic phases with anhydrous sodium sulfate, filter, concentrate, and the residue is prepared by acidic preparation method A to obtain the compound ( Sa,S)-6-(1-(1-(4-(2-methoxypyrimidin-4-yl)phenyl)ethyl)-4-(propane-1-yn-1-yl)-1H -Indazole-7-carboxamido)spiro[3.3]heptane-2-carboxylic acid (I-20a) (17.5 mg, 30% yield).
1H NMR(400MHz,CDCl 3)δ8.79(d,1H),8.62(d,1H),8.29(d,1H),8.03(d,2H),7.61(d,1H),7.30(d,1H),7.16(dd,3H),6.36(q,1H),4.33(dd,1H),3.95(s,3H),2.93–2.84(m,1H),2.48–2.40(m,1H),2.31–2.20(m,3H),2.18(s,2H),2.09(dd,1H),2.05–1.97(m,2H),1.93(d,3H),1.87–1.77(m,2H)。 1 H NMR (400MHz, CDCl 3 )δ8.79(d,1H),8.62(d,1H),8.29(d,1H),8.03(d,2H),7.61(d,1H),7.30(d, 1H),7.16(dd,3H),6.36(q,1H),4.33(dd,1H),3.95(s,3H),2.93–2.84(m,1H),2.48–2.40(m,1H),2.31 –2.20 (m, 3H), 2.18 (s, 2H), 2.09 (dd, 1H), 2.05 – 1.97 (m, 2H), 1.93 (d, 3H), 1.87 – 1.77 (m, 2H).
LC-MS,M/Z(ESI):550.1[M+H] + LC-MS, M/Z(ESI):550.1[M+H] +
化合物I-20b的制备Preparation of Compound I-20b
合成路线如下所示：The synthetic route is as follows:
Figure PCTCN2022097611-appb-000069
Figure PCTCN2022097611-appb-000069
化合物I-20b的合成方法参考其异构体I-20a的合成方法，将(R)-1-(4-溴苯基)乙醇换成(s)-1-(4-溴苯基)乙醇，经过同样的六步反应得到化合物(Sa,R)-6-(1-(1-(4-(2-甲氧基嘧啶-4-基)苯基)乙基)-4-(丙烷-1-炔-1-基)-1H-吲唑-7-甲酰胺基)螺[3.3]庚烷-2-甲酸(34.5mg,产率59％)。The synthetic method of compound I-20b refers to the synthetic method of its isomer I-20a, changing (R)-1-(4-bromophenyl)ethanol into (s)-1-(4-bromophenyl)ethanol , through the same six-step reaction to obtain the compound (Sa, R)-6-(1-(1-(4-(2-methoxypyrimidin-4-yl)phenyl)ethyl)-4-(propane- 1-yn-1-yl)-1H-indazole-7-carboxamido)spiro[3.3]heptane-2-carboxylic acid (34.5 mg, 59% yield).
1H NMR(400MHz,CDCl 3)δ8.79(d,1H),8.60(d,1H),8.26(d,1H),8.03(d,2H),7.60(d,1H),7.32(d,1H),7.14(dd,3H),6.35(q,1H),4.43(dd,1H),3.95(s,3H),2.92–2.84(m,1H),2.45–2.40(m,1H),2.26–2.20(m,3H),2.18(s,2H),2.09(dd,1H),2.03–1.97(m,2H),1.91(d,3H),1.86–1.77(m,2H). 1 H NMR (400MHz, CDCl 3 )δ8.79(d,1H),8.60(d,1H),8.26(d,1H),8.03(d,2H),7.60(d,1H),7.32(d, 1H),7.14(dd,3H),6.35(q,1H),4.43(dd,1H),3.95(s,3H),2.92–2.84(m,1H),2.45–2.40(m,1H),2.26 –2.20(m,3H),2.18(s,2H),2.09(dd,1H),2.03–1.97(m,2H),1.91(d,3H),1.86–1.77(m,2H).
LC-MS,M/Z(ESI):550.1[M+H] +。 LC-MS, M/Z (ESI): 550.1 [M+H] + .
实施例21化合物I-21的制备The preparation of embodiment 21 compound I-21
合成路线如下所示：The synthetic route is as follows:
Figure PCTCN2022097611-appb-000070
Figure PCTCN2022097611-appb-000070
第一步：4-(丙烷-1-炔-1-基)-1H-吲唑-7-甲酸甲酯的制备The first step: the preparation of 4-(propane-1-yn-1-yl)-1H-indazole-7-carboxylic acid methyl ester
室温下将化合物4-溴-1H-吲唑-7-甲酸甲酯(2.5g,9.8mmol)，四丁基氟化铵(19.6ml,19.6mmol)，碘化亚铜(0.28g,1.47mmol)，二(三苯基膦)二氯化钯(0.688g,0.980mmol)，乙炔基三甲基硅烷(2.2g,19.60mmol)加入到四氢呋喃(20ml)中，氮气保护下室温搅拌12h。待反应完全，加入水(20mL)稀释，用乙酸乙酯(20mL×3)萃取，分液，合并有机相，有机相用无水硫酸钠干燥，过滤，浓缩，残留物用薄层硅胶板分离纯化(石油醚:乙酸乙酯(V/V)＝2:3)得化合物4-(丙-1-烯-1-基)-1H-吲唑-7-甲酸甲酯(1.8g,产率85％)。Compound 4-bromo-1H-indazole-7-carboxylic acid methyl ester (2.5g, 9.8mmol), tetrabutylammonium fluoride (19.6ml, 19.6mmol), cuprous iodide (0.28g, 1.47mmol) at room temperature ), bis(triphenylphosphine)palladium dichloride (0.688g, 0.980mmol), and ethynyltrimethylsilane (2.2g, 19.60mmol) were added to tetrahydrofuran (20ml), and stirred at room temperature for 12h under nitrogen protection. After the reaction is complete, add water (20mL) to dilute, extract with ethyl acetate (20mL×3), separate the layers, combine the organic phase, dry the organic phase with anhydrous sodium sulfate, filter, concentrate, and separate the residue with a thin-layer silica gel plate Purification (petroleum ether: ethyl acetate (V/V) = 2:3) gave compound 4-(prop-1-en-1-yl)-1H-indazole-7-carboxylic acid methyl ester (1.8g, yield 85%).
第二步：1-(3-(4-溴苯基)噁丁环-3-基)-4-(丙烷-1-炔-1-基)-1H-吲唑-7-甲酸甲酯的制备The second step: 1-(3-(4-bromophenyl)oxetan-3-yl)-4-(propane-1-yn-1-yl)-1H-indazole-7-formic acid methyl ester preparation
将化合物4-(丙-1-烯-1-基)-1H-吲唑-7-甲酸甲酯(0.5g,2.33mmol)，3-(4-溴苯基)噁丁环-3-醇(1.6g,7.0mmol)，三苯基膦(1.84g,7.0mmol)加入到四氢呋喃(8ml)中，0℃下滴加偶氮二甲酸二异丙酯(1.4g,7.0mmol)，室温搅拌10h。加入水(20mL)稀释，用乙酸乙酯(10mL×3)萃取，分液，合并有机相，有机相用无水硫酸钠干燥，过滤，浓缩，残留物用薄层硅胶板分离纯化(石油醚:乙酸乙酯(V/V)＝3:1)得化合物1-(3-(4-溴苯基)噁丁环-3-基)-4-(丙烷-1-炔-1-基)-1H-吲唑-7-甲酸甲酯(0.7g,黄色固体，产率70％)。Compound 4-(prop-1-en-1-yl)-1H-indazole-7-carboxylic acid methyl ester (0.5g, 2.33mmol), 3-(4-bromophenyl)oxetane-3-ol (1.6g, 7.0mmol), triphenylphosphine (1.84g, 7.0mmol) were added to tetrahydrofuran (8ml), diisopropyl azodicarboxylate (1.4g, 7.0mmol) was added dropwise at 0°C, and stirred at room temperature 10h. Add water (20mL) to dilute, extract with ethyl acetate (10mL×3), separate the layers, combine the organic phases, dry the organic phases with anhydrous sodium sulfate, filter, concentrate, and separate and purify the residue with a thin-layer silica gel plate (petroleum ether : Ethyl acetate (V/V)=3:1) to obtain compound 1-(3-(4-bromophenyl) oxetan-3-yl)-4-(propane-1-yn-1-yl) -1H-Indazole-7-carboxylic acid methyl ester (0.7 g, yellow solid, yield 70%).
第三步：1-(3-(4-环丙基苯基)噁丁环-3-基)-4-(丙烷-1-炔-1-基)-1H-吲唑-7-甲酸甲酯的制备The third step: 1-(3-(4-cyclopropylphenyl)oxetan-3-yl)-4-(propane-1-yn-1-yl)-1H-indazole-7-formic acid Preparation of esters
将甲基1-(3-(4-溴苯基)噁丁环-3-基)-4-(丙烷-1-炔-1-基)-1H-吲唑-7-甲酸甲酯(300mg,0.71mmol)，环丙基硼酸(300mg,3.53mmol)和四(三苯基膦)钯(80mg，0.071mmol)，氟化钾(123mg,2.12mmol)加入到甲苯(3ml)和水(0.5ml)中,升温至100℃搅拌12h。加入水(20mL)稀释，用乙酸乙酯(10mL×3)萃取，分液，合并有机相，有机相用无水硫酸钠干燥，过滤，浓缩，残留物用薄层硅胶板分离纯化(石油醚:乙酸乙酯(V/V)＝3:1)得化合物固体1-(3-(4-环丙基苯基)噁丁环-3-基)-4-(丙烷-1-炔-1-基)-1H-吲唑-7-甲酸甲酯(180mg,产率66％)。Methyl 1-(3-(4-bromophenyl)oxetan-3-yl)-4-(propane-1-yn-1-yl)-1H-indazole-7-carboxylic acid methyl ester (300mg , 0.71mmol), cyclopropylboronic acid (300mg, 3.53mmol) and tetrakis (triphenylphosphine) palladium (80mg, 0.071mmol), potassium fluoride (123mg, 2.12mmol) was added to toluene (3ml) and water (0.5 ml), heated to 100°C and stirred for 12h. Add water (20mL) to dilute, extract with ethyl acetate (10mL×3), separate the layers, combine the organic phases, dry the organic phases with anhydrous sodium sulfate, filter, concentrate, and separate and purify the residue with a thin-layer silica gel plate (petroleum ether : Ethyl acetate (V/V)=3:1) to obtain compound solid 1-(3-(4-cyclopropylphenyl) oxetan-3-yl)-4-(propane-1-yne-1 -yl)-1H-indazole-7-carboxylic acid methyl ester (180 mg, yield 66%).
第四步：甲基-1-(3-(4-环丙基苯基)噁丁环-3-基)-4-(丙烷-1-炔-1-基)-1H-吲唑-7-甲酸的制备The fourth step: Methyl-1-(3-(4-cyclopropylphenyl)oxetan-3-yl)-4-(propane-1-yn-1-yl)-1H-indazole-7 - Preparation of formic acid
室温下将化合物1-(3-(4-环丙基苯基)噁丁环-3-基)-4-(丙烷-1-炔-1-基)-1H-吲唑-7-甲酸甲酯(180mg,0.47mmol)加入到四氢呋喃(3mL)，甲醇(1mL)和水(1mL)中，加入氢氧化锂(59mg,1.4mmol)，搅拌16h。加入水(10mL)稀释，用1N的盐酸水溶液调节pH为4，用乙酸乙酯(10mL×3)萃取，分液，合并有机相，有机相用无水硫酸钠干燥，过滤，浓缩，残留物用薄层硅胶板分离纯化(石油醚:乙酸乙酯(V/V)＝1:1)得化合物甲基-1-(3-(4-环丙基苯基)噁丁环-3- 基)-4-(丙烷-1-炔-1-基)-1H-吲唑-7-甲酸(120mg,产率70％)。Compound 1-(3-(4-cyclopropylphenyl)oxetan-3-yl)-4-(propane-1-yn-1-yl)-1H-indazole-7-carboxylic acid methyl Ester (180mg, 0.47mmol) was added to tetrahydrofuran (3mL), methanol (1mL) and water (1mL), lithium hydroxide (59mg, 1.4mmol) was added, and stirred for 16h. Add water (10mL) to dilute, adjust the pH to 4 with 1N hydrochloric acid aqueous solution, extract with ethyl acetate (10mL×3), separate the layers, combine the organic phases, dry the organic phases with anhydrous sodium sulfate, filter, concentrate, and the residue Separation and purification with a thin-layer silica gel plate (petroleum ether: ethyl acetate (V/V) = 1:1) to obtain the compound methyl-1-(3-(4-cyclopropylphenyl)oxetan-3-yl )-4-(propan-1-yn-1-yl)-1H-indazole-7-carboxylic acid (120 mg, 70% yield).
第五步：6-(1-(3-(4-环丙基苯基)噁丁环-3-基)-4-(丙烷-1-炔-1-基)-1H-吲唑-7-甲酰胺基)螺[3.3]庚烷-2-甲酸甲酯的制备The fifth step: 6-(1-(3-(4-cyclopropylphenyl)oxetan-3-yl)-4-(propane-1-yn-1-yl)-1H-indazole-7 Preparation of -formamido)spiro[3.3]heptane-2-carboxylic acid methyl ester
室温下将化合物甲基-1-(3-(4-环丙基苯基)噁丁环-3-基)-4-(丙烷-1-炔-1-基)-1H-吲唑-7-甲酸(100mg,0.27mmol)加入到N,N-二甲基甲酰胺(2mL)中，加入2-(6-氨基螺[3.3]庚-2-基)乙酸乙酯盐酸盐(66mg,0.32mmol)，二异丙基乙胺(173mg,1.34mmol)和HATU(155mg,0.40mmol)，室温搅拌16h。加入水(10mL)稀释，用乙酸乙酯(10mL×3)萃取，分液，合并有机相，有机相用无水硫酸钠干燥，过滤，浓缩，残留物用薄层硅胶板分离纯化(石油醚:乙酸乙酯(V/V)＝1:1)得化合物6-(1-(3-(4-环丙基苯基)噁丁环-3-基)-4-(丙烷-1-炔-1-基)-1H-吲唑-7-甲酰胺基)螺[3.3]庚烷-2-甲酸甲酯(100mg,产率70％)。Compound methyl-1-(3-(4-cyclopropylphenyl)oxetan-3-yl)-4-(propane-1-yn-1-yl)-1H-indazole-7 -Formic acid (100mg, 0.27mmol) was added to N,N-dimethylformamide (2mL), and ethyl 2-(6-aminospiro[3.3]hept-2-yl)acetate hydrochloride (66mg, 0.32mmol), diisopropylethylamine (173mg, 1.34mmol) and HATU (155mg, 0.40mmol), stirred at room temperature for 16h. Add water (10mL) to dilute, extract with ethyl acetate (10mL×3), separate the layers, combine the organic phases, dry the organic phases with anhydrous sodium sulfate, filter, concentrate, and separate and purify the residue with a thin-layer silica gel plate (petroleum ether : ethyl acetate (V/V)=1:1) to obtain compound 6-(1-(3-(4-cyclopropylphenyl) oxetan-3-yl)-4-(propane-1-yne -1-yl)-1H-indazole-7-carboxamido)spiro[3.3]heptane-2-carboxylic acid methyl ester (100 mg, 70% yield).
第六步：6-(1-(3-(4-环丙基苯基)噁丁环-3-基)-4-(丙烷-1-炔-1-基)-1H-吲唑-7-甲酰胺基)螺[3.3]庚烷-2-甲酸(I-21)的制备The sixth step: 6-(1-(3-(4-cyclopropylphenyl)oxetan-3-yl)-4-(propane-1-yn-1-yl)-1H-indazole-7 Preparation of -formamido)spiro[3.3]heptane-2-carboxylic acid (I-21)
Figure PCTCN2022097611-appb-000071
Figure PCTCN2022097611-appb-000071
室温下将化合物6-(1-(3-(4-环丙基苯基)噁丁环-3-基)-4-(丙烷-1-炔-1-基)-1H-吲唑-7-甲酰胺基)螺[3.3]庚烷-2-甲酸甲酯(25mg,0.048mmol)加入到四氢呋喃(3mL)，甲醇(1mL)和水(1mL)中，加入氢氧化锂(6mg,0.143mmol)，室温搅拌16h。用1N盐酸调节pH＝4，用乙酸乙酯(10mL×3)萃取，分液，合并有机相，有机相用无水硫酸钠干燥，过滤，浓缩，残留物经酸性制备方法A制备得化合物(Sa)-6-(1-(3-(4-环丙基苯基)噁丁环-3-基)-4-(丙烷-1-炔-1-基)-1H-吲唑-7-甲酰胺基)螺[3.3]庚烷-2-甲酸(I-21)(10mg,产率40％)。Compound 6-(1-(3-(4-cyclopropylphenyl)oxetan-3-yl)-4-(propane-1-yn-1-yl)-1H-indazole-7 -Carboxamido)spiro[3.3]heptane-2-carboxylic acid methyl ester (25mg, 0.048mmol) was added to tetrahydrofuran (3mL), methanol (1mL) and water (1mL), lithium hydroxide (6mg, 0.143mmol ), stirred at room temperature for 16h. Use 1N hydrochloric acid to adjust pH=4, extract with ethyl acetate (10mL×3), separate the layers, combine the organic phases, dry the organic phases with anhydrous sodium sulfate, filter, concentrate, and the residue is prepared by acidic preparation method A to obtain the compound ( Sa)-6-(1-(3-(4-cyclopropylphenyl)oxetan-3-yl)-4-(propane-1-yn-1-yl)-1H-indazole-7- Formamido)spiro[3.3]heptane-2-carboxylic acid (I-21) (10 mg, 40% yield).
1H NMR(400MHz,dmso)δ8.36–8.31(m,2H),7.27–7.20(m,2H),6.94(d,2H),6.72(d,2H),5.30(s,2H),4.83(s,2H),3.91–3.86(m,1H),2.18(s,3H),2.15–2.01(m,5H),1.85–1.66(m,5H),0.91–0.86(m,2H),0.62–0.56(m,2H). 1 H NMR (400MHz,dmso)δ8.36–8.31(m,2H),7.27–7.20(m,2H),6.94(d,2H),6.72(d,2H),5.30(s,2H),4.83 (s,2H),3.91–3.86(m,1H),2.18(s,3H),2.15–2.01(m,5H),1.85–1.66(m,5H),0.91–0.86(m,2H),0.62 –0.56(m,2H).
LC-MS,M/Z(ESI):510[M+H] +。 LC-MS, M/Z (ESI): 510 [M+H] + .
实施例22化合物I-22的制备The preparation of embodiment 22 compound I-22
合成路线如下所示：The synthetic route is as follows:
Figure PCTCN2022097611-appb-000072
Figure PCTCN2022097611-appb-000072
第一步：1-(4-溴苯基)-3,3-二甲氧基环丁烷-1-甲腈The first step: 1-(4-bromophenyl)-3,3-dimethoxycyclobutane-1-carbonitrile
反应瓶中依次加入氢化钠(4.5g,112mmol)和无水N,N-二甲基甲酰胺(100mL)，冰浴下缓慢滴加对溴苯乙腈(10g,51mmol)，搅拌30分钟后往里加入1,3-二溴-2,2-二甲氧基丙烷(13.4g,51mmol)，反应液逐渐升温至60℃搅拌16小时。反应液冷至室温，缓慢加水(150mL)，用乙酸乙酯(50mL×3)萃取。上述有机相合并，用饱和食盐水(50mL×2)洗涤，浓缩后得到粗品1-(4-溴苯基)-3,3-二甲氧基环丁烷-1-甲腈(15g，产率100％)，直接用于下一步反应。Sodium hydride (4.5g, 112mmol) and anhydrous N,N-dimethylformamide (100mL) were successively added into the reaction flask, p-bromophenylacetonitrile (10g, 51mmol) was slowly added dropwise under ice bath, stirred for 30 minutes and then 1,3-dibromo-2,2-dimethoxypropane (13.4 g, 51 mmol) was added, and the reaction solution was gradually heated to 60° C. and stirred for 16 hours. The reaction solution was cooled to room temperature, water (150 mL) was added slowly, and extracted with ethyl acetate (50 mL×3). The above organic phases were combined, washed with saturated brine (50mL×2), and concentrated to obtain the crude product 1-(4-bromophenyl)-3,3-dimethoxycyclobutane-1-carbonitrile (15g, yielding Yield 100%), directly used in next step reaction.
第二步：1-(4-溴苯基)-3-氧杂环丁基-1-甲腈The second step: 1-(4-bromophenyl)-3-oxetanyl-1-carbonitrile
将粗品1-(4-溴苯基)-3,3-二甲氧基环丁烷-1-甲腈(15g,51mmol)溶于丙酮中，加入50％硫酸水溶液(10mL)，回流反应12小时。反应液冷至室温，加入饱和碳酸氢钠溶液(50mL)，旋蒸除去丙酮，水相用乙酸乙酯(50mL×3)萃取。有机相合并后用饱和食盐水(50mL×1)洗涤和无水硫酸钠干燥，浓缩后残留物用柱层析硅胶纯化(石油醚:乙酸乙酯(V/V)＝10:1)得1-(4-溴苯基)-3-氧杂环丁基-1-甲腈(4.6g，浅棕色油状液体，产率47％)。The crude product 1-(4-bromophenyl)-3,3-dimethoxycyclobutane-1-carbonitrile (15g, 51mmol) was dissolved in acetone, 50% sulfuric acid aqueous solution (10mL) was added, and the reflux reaction was carried out for 12 Hour. The reaction solution was cooled to room temperature, saturated sodium bicarbonate solution (50 mL) was added, acetone was removed by rotary evaporation, and the aqueous phase was extracted with ethyl acetate (50 mL×3). The combined organic phases were washed with saturated brine (50mL×1) and dried over anhydrous sodium sulfate. After concentration, the residue was purified by column chromatography on silica gel (petroleum ether:ethyl acetate (V/V)=10:1) to obtain 1 -(4-Bromophenyl)-3-oxetanyl-1-carbonitrile (4.6 g, light brown oily liquid, yield 47%).
第三步：1-(4-溴苯基)-3-甲基烯环丁烷-1-甲腈The third step: 1-(4-bromophenyl)-3-methylenecyclobutane-1-carbonitrile
甲基三苯基溴化膦(9.86g,27.6mmol)溶于四氢呋喃(100mL)中，冰浴下缓慢滴加叔丁醇钾(1mol/L四氢呋喃溶液,27.6mL)，搅拌1小时。将1-(4-溴苯基)-3-氧杂环丁基-1-甲腈(6.9g,27.6mmol)溶于四氢呋喃(10mL)后缓慢加入上述溶液中，逐渐升至室温搅拌过夜16小时。往反应液中加入氯化铵水溶液(20mL)，乙酸乙酯(20mL×3)萃取，有机相合并后用无水硫酸钠干燥，浓缩后的残留物用柱层析硅胶纯化，得到1-(4-溴苯基)-3-甲基烯环丁烷-1-甲腈(4.6g， 产率67％)。Methyltriphenylphosphine bromide (9.86g, 27.6mmol) was dissolved in tetrahydrofuran (100mL), potassium tert-butoxide (1mol/L solution in tetrahydrofuran, 27.6mL) was slowly added dropwise under ice cooling, and stirred for 1 hour. 1-(4-Bromophenyl)-3-oxetanyl-1-carbonitrile (6.9g, 27.6mmol) was dissolved in tetrahydrofuran (10mL) and slowly added to the above solution, gradually raised to room temperature and stirred overnight for 16 Hour. Aqueous ammonium chloride solution (20 mL) was added to the reaction solution, extracted with ethyl acetate (20 mL×3), the organic phases were combined and dried over anhydrous sodium sulfate, and the concentrated residue was purified by column chromatography on silica gel to obtain 1-( 4-Bromophenyl)-3-methylenecyclobutane-1-carbonitrile (4.6 g, 67% yield).
第四步：1-(4-溴苯基)-3-甲基烯环丁烷-1-甲醛The fourth step: 1-(4-bromophenyl)-3-methylenecyclobutane-1-carbaldehyde
1-(4-溴苯基)-3-甲基烯环丁烷-1-甲腈(4.3g,16.2mmol)溶于四氢呋喃(30mL)中，冰浴下缓慢加入二异丙基铝氢(1mol/L四氢呋喃溶液，26mL),慢慢升至室温搅拌过夜。反应液冰浴下缓慢滴加水(5mL)，随后加入稀盐酸(1mol/L水溶液，20mL)，搅拌4小时，用乙酸乙酯(30mL×3)萃取。有机相合并后用无水硫酸钠干燥，浓缩即得粗品1-(4-溴苯基)-3-甲基烯环丁烷-1-甲醛(4.5g，产率95％)。1-(4-bromophenyl)-3-methylalkenecyclobutane-1-carbonitrile (4.3g, 16.2mmol) was dissolved in tetrahydrofuran (30mL), and diisopropylaluminum hydrogen ( 1mol/L tetrahydrofuran solution, 26mL), slowly raised to room temperature and stirred overnight. Water (5 mL) was slowly added dropwise to the reaction solution in an ice bath, followed by dilute hydrochloric acid (1 mol/L aqueous solution, 20 mL), stirred for 4 hours, and extracted with ethyl acetate (30 mL×3). The organic phases were combined, dried over anhydrous sodium sulfate, and concentrated to obtain crude 1-(4-bromophenyl)-3-methylenecyclobutane-1-carbaldehyde (4.5 g, yield 95%).
第五步：1-(4-溴苯基)-3-甲基烯环丁烷-1-甲醇的制备Step 5: Preparation of 1-(4-bromophenyl)-3-methylenecyclobutane-1-methanol
粗品1-(4-溴苯基)-3-甲基烯环丁烷-1-甲醛(4.5g,18mmol)溶于甲醇(30mL)中，冰浴下分批加入硼氢化钠(1.35g,36mmol)，搅拌1小时。往反应液中加入水(30mL)，搅拌10分钟后旋蒸除去甲醇，水相用乙酸乙酯(40mL×3)萃取。有机相合并，无水硫酸钠干燥，浓缩，得到残留物用柱层析纯化，即得1-(4-溴苯基)-3-甲基烯环丁烷-1-甲醇(2.6g，产率57％)。The crude product 1-(4-bromophenyl)-3-methylenecyclobutane-1-carbaldehyde (4.5g, 18mmol) was dissolved in methanol (30mL), and sodium borohydride (1.35g, 36mmol), stirred for 1 hour. Water (30 mL) was added to the reaction solution, and after stirring for 10 minutes, the methanol was removed by rotary evaporation, and the aqueous phase was extracted with ethyl acetate (40 mL×3). The organic phases were combined, dried over anhydrous sodium sulfate, concentrated, and the obtained residue was purified by column chromatography to obtain 1-(4-bromophenyl)-3-methylenecyclobutane-1-methanol (2.6 g, yield rate 57%).
第六步：4-(溴苯基)-1-(碘甲基)-2-氧杂双环[2.2.1]己烷的制备Step 6: Preparation of 4-(bromophenyl)-1-(iodomethyl)-2-oxabicyclo[2.2.1]hexane
1-(4-溴苯基)-3-甲基烯环丁烷-1-甲醇(2.8g,10.8mmol)溶于水(10mL)和甲基叔丁基醚(20mL)中，依次加入碳酸氢钠(1.3g,16.6mol)和碘单质(3.9g,16.6mmol)，室温搅拌过夜。反应液中加入饱和硫代硫酸钠水溶液(10mL)，反应液用二氯甲烷(30mL×3)萃取。有机相合并后依次用饱和食盐水(50mL)洗涤和无水硫酸钠干燥，浓缩后残留物用柱层析纯化，得4-(溴苯基)-1-(碘甲基)-2-氧杂双环[2.2.1]己烷(3.6g，产率86％)。1-(4-Bromophenyl)-3-methylalkenecyclobutane-1-methanol (2.8g, 10.8mmol) was dissolved in water (10mL) and methyl tert-butyl ether (20mL), and carbonic acid was added in sequence Sodium hydrogen (1.3g, 16.6mol) and iodine (3.9g, 16.6mmol) were stirred overnight at room temperature. Saturated aqueous sodium thiosulfate solution (10 mL) was added to the reaction solution, and the reaction solution was extracted with dichloromethane (30 mL×3). The organic phases were combined and washed successively with saturated brine (50 mL) and dried over anhydrous sodium sulfate. After concentration, the residue was purified by column chromatography to obtain 4-(bromophenyl)-1-(iodomethyl)-2-oxo Heterobicyclo[2.2.1]hexane (3.6 g, 86% yield).
第七步：4-(溴苯基)-1-甲基-2-氧杂双环[2.2.1]己烷的制备Step 7: Preparation of 4-(bromophenyl)-1-methyl-2-oxabicyclo[2.2.1]hexane
4-(溴苯基)-1-(碘甲基)-2-氧杂双环[2.2.1]己烷(3.4g,8.97mmol)溶于四氢呋喃(30mL)中，冰浴下缓慢滴加三乙基硼氢化锂(1mol/L四氢呋喃溶液,18mL)，缓慢升至室温搅拌3小时。反应液置于冰浴下依次缓慢滴加水(10mL)和2mol/L氢氧化钠水溶液(10mL)，乙酸乙酯(30mL×3)萃取。有机相合并，无水硫酸钠干燥，浓缩后柱层析纯化得到4-(溴苯基)-1-甲基-2-氧杂双环[2.2.1]己烷(1.9g,84％收率)。4-(bromophenyl)-1-(iodomethyl)-2-oxabicyclo[2.2.1]hexane (3.4g, 8.97mmol) was dissolved in tetrahydrofuran (30mL), and three Lithium ethyl borohydride (1mol/L tetrahydrofuran solution, 18mL), slowly warmed up to room temperature and stirred for 3 hours. The reaction solution was placed in an ice bath, and water (10 mL) and 2 mol/L sodium hydroxide aqueous solution (10 mL) were slowly added dropwise successively, and extracted with ethyl acetate (30 mL×3). The organic phases were combined, dried over anhydrous sodium sulfate, concentrated and purified by column chromatography to obtain 4-(bromophenyl)-1-methyl-2-oxabicyclo[2.2.1]hexane (1.9g, 84% yield ).
LC-MS,M/Z(ESI):252.9[M+H] +。 LC-MS, M/Z (ESI): 252.9 [M+H] + .
第八步：1-(4-(1-甲基-2-氧杂双环[2.2.1]己烷-4-基)苯基)乙烷-1-酮的制备Step 8: Preparation of 1-(4-(1-methyl-2-oxabicyclo[2.2.1]hexane-4-yl)phenyl)ethan-1-one
4-(溴苯基)-1-甲基-2-氧杂双环[2.2.1]己烷(1g,4mmol)，三丁基(1-乙氧基乙烯)锡(1.9g,5.14mmol)和双三苯基磷二氯化钯(0.14g,0.2mmol)于二氧六环(10mL)中，氮气保护下100℃搅拌12小时。反应液冷至室温，加入KF水溶液(20mL)，搅拌2小时，乙酸乙酯(30mL×3)萃取。有机相合并，加入1N盐酸水溶液(20mL)，搅拌1小时，分液，取有机相用无水硫酸钠干燥后浓缩，得到粗品1-(4-(1-甲基-2-氧杂双环[2.2.1]己烷-4-基)苯基)乙烷-1-酮(1.2g，产率100％)，直接用于下一步反应。4-(bromophenyl)-1-methyl-2-oxabicyclo[2.2.1]hexane (1g, 4mmol), tributyl(1-ethoxyethylene)tin (1.9g, 5.14mmol) and bistriphenylphosphinepalladium dichloride (0.14g, 0.2mmol) in dioxane (10mL), stirred at 100°C for 12 hours under nitrogen protection. The reaction solution was cooled to room temperature, KF aqueous solution (20 mL) was added, stirred for 2 hours, and extracted with ethyl acetate (30 mL×3). Combine the organic phases, add 1N hydrochloric acid aqueous solution (20mL), stir for 1 hour, separate the layers, take the organic phase and dry it with anhydrous sodium sulfate and concentrate to obtain the crude product 1-(4-(1-methyl-2-oxabicyclo[ 2.2.1] Hexan-4-yl)phenyl)ethan-1-one (1.2 g, yield 100%) was directly used in the next reaction.
第九步：1-(4-(1-甲基-2-氧杂双环[2.2.1]己烷-4-基)苯基)乙烷-1-醇的制备Step 9: Preparation of 1-(4-(1-methyl-2-oxabicyclo[2.2.1]hexane-4-yl)phenyl)ethan-1-ol
粗品1-(4-(1-甲基-2-氧杂双环[2.2.1]己烷-4-基)苯基)乙烷-1-酮(1.2g,4.62mmol)溶于甲醇(10mL)中，冰浴下分批加入硼氢化钠(264mg,4.62mmol)，搅拌1小时。反应液中加水(10mL)，搅拌10分钟，旋蒸出去甲醇，水相用乙酸乙酯(20mL×3)萃取。有机相合并，无水硫酸钠干燥，浓缩，得到残留物用柱层析纯化，即得1-(4-(1-甲基-2-氧杂双环[2.2.1]己烷-4-基)苯基)乙烷-1-醇(0.67g，产率66％)。The crude product 1-(4-(1-methyl-2-oxabicyclo[2.2.1]hexane-4-yl)phenyl)ethan-1-one (1.2g, 4.62mmol) was dissolved in methanol (10mL ), sodium borohydride (264mg, 4.62mmol) was added in portions under ice-cooling, and stirred for 1 hour. Water (10 mL) was added to the reaction solution, stirred for 10 minutes, methanol was removed by rotary evaporation, and the aqueous phase was extracted with ethyl acetate (20 mL×3). The organic phases were combined, dried over anhydrous sodium sulfate, concentrated, and the obtained residue was purified by column chromatography to obtain 1-(4-(1-methyl-2-oxabicyclo[2.2.1]hexane-4-yl )phenyl)ethan-1-ol (0.67 g, 66% yield).
第十步：1-(4-(1-甲基-2-氧杂双环[2.2.1]己烷-4-基)苯基)乙基)-4-(丙烷-1-炔-1-基)-1H-吲唑-7-甲酸甲酯的制备The tenth step: 1-(4-(1-methyl-2-oxabicyclo[2.2.1]hexane-4-yl)phenyl)ethyl)-4-(propane-1-yne-1- The preparation of methyl)-1H-indazole-7-carboxylate
1-(4-(1-甲基-2-氧杂双环[2.2.1]己烷-4-基)苯基)乙烷-1-醇(336mg,1.54mmol)、甲基4-(丙烷-1-炔-1-基)-1H-吲唑-7-甲酸酯(300mg,1.4mmol)和三苯基膦(478mg,1.82mmol)溶于四氢呋喃(5mL)中，冰浴下缓慢滴加偶氮二异丙酯(368mg，1.82mmol)，随后升至室温搅拌16小时。反应液浓缩，柱层析纯化得1-(4-(1-甲基-2-氧杂双环[2.2.1]己烷-4-基)苯基)乙基)-4-(丙烷-1-炔-1-基)-1H-吲唑-7-甲酸甲酯(500mg，产率86％)。1-(4-(1-methyl-2-oxabicyclo[2.2.1]hexan-4-yl)phenyl)ethan-1-ol (336mg, 1.54mmol), methyl 4-(propane -1-alkyn-1-yl)-1H-indazole-7-carboxylate (300mg, 1.4mmol) and triphenylphosphine (478mg, 1.82mmol) were dissolved in tetrahydrofuran (5mL), slowly dropped under ice bath Azodiisopropyl ester (368 mg, 1.82 mmol) was added, followed by warming to room temperature and stirring for 16 hours. The reaction solution was concentrated and purified by column chromatography to obtain 1-(4-(1-methyl-2-oxabicyclo[2.2.1]hexane-4-yl)phenyl)ethyl)-4-(propane-1 -Alkyn-1-yl)-1H-indazole-7-carboxylic acid methyl ester (500 mg, 86% yield).
LC-MS,M/Z(ESI):415.3[M+H] +。 LC-MS, M/Z (ESI): 415.3 [M+H] + .
第十一步：1-(4-(1-甲基-2-氧杂双环[2.2.1]己烷-4-基)苯基)乙基)-4-(丙烷-1-炔-1-基)-1H-吲唑-7-甲酸锂盐的制备The eleventh step: 1-(4-(1-methyl-2-oxabicyclo[2.2.1]hexane-4-yl)phenyl)ethyl)-4-(propane-1-yne-1 Preparation of -yl)-1H-indazole-7-formic acid lithium salt
甲基1-(4-(1-甲基-2-氧杂双环[2.2.1]己烷-4-基)苯基)乙基)-4-(丙烷-1-炔-1-基)-1H-吲唑-7-甲酸甲酯(500mg，1.21mmol)溶于四氢呋喃/甲醇/水(5mL/1mL/1mL)中，加入一水合氢氧化锂(150mg，3.62mmol)，室温搅拌过夜，反应完浓缩得到1-(4-(1-甲基-2-氧杂双环[2.2.1]己烷-4-基)苯基)乙基)-4-(丙烷-1-炔-1-基)-1H-吲唑-7-甲酸锂盐(610mg，含LiOH，收率100％)。Methyl 1-(4-(1-methyl-2-oxabicyclo[2.2.1]hexan-4-yl)phenyl)ethyl)-4-(propane-1-yn-1-yl) -1H-indazole-7-methyl carboxylate (500mg, 1.21mmol) was dissolved in tetrahydrofuran/methanol/water (5mL/1mL/1mL), lithium hydroxide monohydrate (150mg, 3.62mmol) was added, and stirred overnight at room temperature, The reaction was concentrated to obtain 1-(4-(1-methyl-2-oxabicyclo[2.2.1]hexane-4-yl)phenyl)ethyl)-4-(propane-1-yne-1- base)-1H-indazole-7-carboxylic acid lithium salt (610 mg, containing LiOH, yield 100%).
第十二步：(Sa)-6-(1-(1-(4-(1-1-2-氧杂双环[2.2.1]己烷-4-基)苯基)乙基)-4-(丙烷-1-炔-1-基)-1H-吲唑-7-甲酰胺)螺[3,3]庚烷-2-甲酸甲酯的制备The twelfth step: (Sa)-6-(1-(1-(4-(1-1-2-oxabicyclo[2.2.1]hexane-4-yl)phenyl)ethyl)-4 Preparation of -(propane-1-yn-1-yl)-1H-indazole-7-carboxamide)spiro[3,3]heptane-2-carboxylic acid methyl ester
1-(4-(1-甲基-2-氧杂双环[2.2.1]己烷-4-基)苯基)乙基)-4-(丙烷-1-炔-1-基)-1H-吲唑-7-甲酸锂盐(500mg，1.25mmol)和HATU(760mg，2.00mmol)溶于N,N-二甲基甲酰胺中，冰浴下加入二异丙基乙胺(516mg，4.00mmol)，搅拌15分钟后加入(Sa)-6-氨基螺[3.3]庚烷-2-甲酸甲酯盐酸盐(411mg，2.00mmol)，室温搅拌过夜。反应液中加入水，乙酸乙酯萃取，有机相依次用饱和食盐水洗涤，无水硫酸钠干燥，浓缩后柱层析纯化，得(Sa)-6-(1-(1-(4-(1-1-2-氧杂双环[2.2.1]己烷-4-基)苯基)乙基)-4-(丙烷-1-炔-1-基)-1H-吲唑-7-甲酰胺)螺[3,3]庚烷-2-甲酸甲酯(400mg，产率59％)。1-(4-(1-methyl-2-oxabicyclo[2.2.1]hexan-4-yl)phenyl)ethyl)-4-(propane-1-yn-1-yl)-1H - Lithium salt of indazole-7-carboxylate (500mg, 1.25mmol) and HATU (760mg, 2.00mmol) were dissolved in N,N-dimethylformamide, diisopropylethylamine (516mg, 4.00 mmol), after stirring for 15 minutes, (Sa)-methyl 6-aminospiro[3.3]heptane-2-carboxylate hydrochloride (411 mg, 2.00 mmol) was added, and stirred overnight at room temperature. Water was added to the reaction solution, extracted with ethyl acetate, the organic phase was successively washed with saturated brine, dried over anhydrous sodium sulfate, concentrated and purified by column chromatography to obtain (Sa)-6-(1-(1-(4-( 1-1-2-oxabicyclo[2.2.1]hexan-4-yl)phenyl)ethyl)-4-(propane-1-yn-1-yl)-1H-indazole-7-methan Amide) spiro[3,3]heptane-2-carboxylic acid methyl ester (400 mg, 59% yield).
LC-MS,M/Z(ESI):552.30[M+H] +。 LC-MS, M/Z (ESI): 552.30 [M+H] + .
第十三步：(Sa)-6-(1-(1-(4-(1-1-2-氧杂双环[2.2.1]己烷-4-基)苯基)乙基)-4-(丙烷-1-炔-1-基)-1H-吲唑-7-甲酰胺)螺[3,3]庚烷-2-甲酸(I-22)The thirteenth step: (Sa)-6-(1-(1-(4-(1-1-2-oxabicyclo[2.2.1]hexane-4-yl)phenyl)ethyl)-4 -(propane-1-yn-1-yl)-1H-indazole-7-carboxamide)spiro[3,3]heptane-2-carboxylic acid (I-22)
Figure PCTCN2022097611-appb-000073
Figure PCTCN2022097611-appb-000073
室温下将化合物(Sa)-6-(1-(1-(4-(1-1-2-氧杂双环[2.2.1]己烷-4-基)苯基)乙基)-4-(丙烷-1-炔-1-基)-1H-吲唑-7-甲酰胺)螺[3,3]庚烷-2-甲酸甲酯(50mg,0.09mmol)加入到四氢呋喃(3mL)，甲醇(1mL)和水(1mL)中，加入氢氧化锂(12mg,0.50mmol)，室温搅拌16h。用1N盐酸调节pH＝4，用乙酸乙酯(10mL×3)萃取，分液，合并有机相，有机相用无水硫酸钠干燥，过滤，浓缩，残留物经酸性制备方法A制备得化合物(Sa)-6-(1-(1-(4-(1-1-2-氧杂双环[2.2.1]己烷-4-基)苯基)乙基)-4-(丙烷-1-炔-1-基)-1H-吲唑-7-甲酰胺)螺[3,3]庚烷-2-甲酸(I-22)(35mg，产率72％)。Compound (Sa)-6-(1-(1-(4-(1-1-2-oxabicyclo[2.2.1]hexane-4-yl)phenyl)ethyl)-4- (Propan-1-yn-1-yl)-1H-indazole-7-carboxamide)spiro[3,3]heptane-2-carboxylic acid methyl ester (50 mg, 0.09 mmol) was added to tetrahydrofuran (3 mL), methanol (1 mL) and water (1 mL), add lithium hydroxide (12 mg, 0.50 mmol), and stir at room temperature for 16 h. Use 1N hydrochloric acid to adjust pH=4, extract with ethyl acetate (10mL×3), separate the layers, combine the organic phases, dry the organic phases with anhydrous sodium sulfate, filter, concentrate, and the residue is prepared by acidic preparation method A to obtain the compound ( Sa)-6-(1-(1-(4-(1-1-2-oxabicyclo[2.2.1]hexane-4-yl)phenyl)ethyl)-4-(propane-1- Alkyn-1-yl)-1H-indazole-7-carboxamide)spiro[3,3]heptane-2-carboxylic acid (I-22) (35 mg, 72% yield).
1H NMR(400mHz,DMSO-d6)12.02(s,1H),δ8.81(m,1H),8.24(s,1H),7.28(m,1H),7.19-7.10(m,3H),7.00-6.97(m,2H),6.27(m,1H),4.34(m,1H),3.70(s,2H),2.95(m,1H),2.50(m,1H),2.17-1.87(m,9H),1.86-1.73(m,8H),1.37(s,3H). 1 H NMR(400mHz,DMSO-d6)12.02(s,1H),δ8.81(m,1H),8.24(s,1H),7.28(m,1H),7.19-7.10(m,3H),7.00 -6.97(m,2H),6.27(m,1H),4.34(m,1H),3.70(s,2H),2.95(m,1H),2.50(m,1H),2.17-1.87(m,9H ),1.86-1.73(m,8H),1.37(s,3H).
LC-MS,M/Z(ESI):538.20[M+H] +。 LC-MS, M/Z (ESI): 538.20 [M+H] + .
实施例23化合物I-23的制备The preparation of embodiment 23 compound I-23
合成路线如下所示：The synthetic route is as follows:
Figure PCTCN2022097611-appb-000074
Figure PCTCN2022097611-appb-000074
第一步：(R)-4-(丙烷-1-炔-1-基)-1-(1-(4-(吡咯烷-1-基)苯基)乙基)-1H-吲唑-7-甲酸甲酯的制备The first step: (R)-4-(propane-1-yn-1-yl)-1-(1-(4-(pyrrolidin-1-yl)phenyl)ethyl)-1H-indazole- Preparation of methyl 7-carboxylate
向(R)-1-(1-(4-溴苯基)乙基)-4-(丙烷-1-炔-1-基)-1H-吲唑-7-甲酸甲酯(400mg,1.007mmol)和吡咯烷(358mg,5.03mmol)的1，4-二氧六环(5mL)溶液中加入(1,1’-联苯)-2-基二-叔丁基膦(45.1mg,0.151mmol)、碳酸铯(1968mg,6.04mmol)和醋酸钯(22.61mg，0.101mmol)。反应液置换三次氮气，然后在80℃下搅拌16小时。将反应液倒入水(30mL)中并乙酸乙酯(20mL*3) 萃取。合并有机相，用饱和食盐水(10mL*2)洗涤，并用无水硫酸钠干燥，过滤并浓缩得到粗产物。粗产物通过柱层析(石油醚/乙酸乙酯(V/V)＝1:0-1:3)纯化粗产物以得到(R)-4-(丙烷-1-炔-1-基)-1-(1-(4-(吡咯烷-1-基)苯基)乙基)-1H-吲唑-7-甲酸甲酯(280mg,产率71.8％)。To (R)-1-(1-(4-bromophenyl)ethyl)-4-(propane-1-yn-1-yl)-1H-indazole-7-carboxylic acid methyl ester (400mg, 1.007mmol ) and pyrrolidine (358mg, 5.03mmol) in 1,4-dioxane (5mL) were added (1,1'-biphenyl)-2-yldi-tert-butylphosphine (45.1mg, 0.151mmol ), cesium carbonate (1968mg, 6.04mmol) and palladium acetate (22.61mg, 0.101mmol). The reaction solution was replaced with nitrogen three times, and then stirred at 80° C. for 16 hours. The reaction solution was poured into water (30 mL) and extracted with ethyl acetate (20 mL*3). The organic phases were combined, washed with saturated brine (10 mL*2), dried over anhydrous sodium sulfate, filtered and concentrated to obtain a crude product. The crude product was purified by column chromatography (petroleum ether/ethyl acetate (V/V)=1:0-1:3) to obtain (R)-4-(propane-1-yn-1-yl)- 1-(1-(4-(Pyrrolidin-1-yl)phenyl)ethyl)-1H-indazole-7-carboxylic acid methyl ester (280 mg, 71.8% yield).
LC-MS,M/Z(ESI):388.2[M+H] +。 LC-MS, M/Z (ESI): 388.2 [M+H] + .
第二步：(R)-4-(丙烷-1-炔-1-基)-1-(1-(4-(吡咯烷-1-基)苯基)乙基)-1H-吲唑-7-甲酸的制备The second step: (R)-4-(propane-1-yn-1-yl)-1-(1-(4-(pyrrolidin-1-yl)phenyl)ethyl)-1H-indazole- Preparation of 7-formic acid
向(R)-4-(丙烷-1-炔-1-基)-1-(1-(4-(吡咯烷-1-基)苯基)乙基)-1H-吲唑-7-甲酸甲酯(280mg,0.723mmol)的四氢呋喃(6mL)、甲醇(2mL)和水(2mL)溶液中加入氢氧化锂(87mg,3.61mmol)，然后在25℃搅拌3小时。向反应液中加入水(50mL)并用1N盐酸水溶液调节pH至5～6，然后用乙酸乙酯(20mL*3)萃取。合并有机相，用饱和食盐水(10mL*2)洗涤，并用无水硫酸钠干燥，过滤并浓缩得到产物(R)-4-(丙烷-1-炔-1-基)-1-(1-(4-(吡咯烷-1-基)苯基)乙基)-1H-吲唑-7-甲酸(250mg，产率93％)。To (R)-4-(propane-1-yn-1-yl)-1-(1-(4-(pyrrolidin-1-yl)phenyl)ethyl)-1H-indazole-7-carboxylic acid Lithium hydroxide (87 mg, 3.61 mmol) was added to a solution of methyl ester (280 mg, 0.723 mmol) in tetrahydrofuran (6 mL), methanol (2 mL) and water (2 mL), followed by stirring at 25°C for 3 hours. Water (50 mL) was added to the reaction solution, and the pH was adjusted to 5-6 with 1N aqueous hydrochloric acid solution, followed by extraction with ethyl acetate (20 mL*3). The organic phases were combined, washed with saturated brine (10mL*2), dried over anhydrous sodium sulfate, filtered and concentrated to give the product (R)-4-(propane-1-yn-1-yl)-1-(1- (4-(Pyrrolidin-1-yl)phenyl)ethyl)-1H-indazole-7-carboxylic acid (250 mg, 93% yield).
LC-MS,M/Z(ESI):374.5[M+H] +。 LC-MS, M/Z (ESI): 374.5 [M+H] + .
第三步：(Sa,R)-6-(4-(丙烷-1-炔-1-基)-1-(1-(4-(吡咯烷-1-基)苯基)乙基)-1H-吲唑-7-甲酰胺基)螺[3.3]庚烷-2-甲酸甲酯的制备The third step: (Sa,R)-6-(4-(propane-1-yn-1-yl)-1-(1-(4-(pyrrolidin-1-yl)phenyl)ethyl)- Preparation of 1H-indazole-7-carboxamido)spiro[3.3]heptane-2-carboxylic acid methyl ester
向(R)-4-(丙烷-1-炔-1-基)-1-(1-(4-(吡咯烷-1-基)苯基)乙基)-1H-吲唑-7-甲酸(250mg,0.669mmol)的DMF(2mL)溶液中加入HATU(764mg,2.008mmol)和二异丙基乙胺(260mg,2.008mmol)，并在25℃下搅拌0.5h。然后向反应液中加入(Sa)-6-氨基螺[3.3]庚烷-2-甲酸甲酯(283mg，1.674mmol)，在25℃下继续搅拌2.5小时。反应液浓缩得到粗产物。粗产物通过柱层析(石油醚/乙酸乙酯(V/V)＝1/0-1/3)纯化得到产物(Sa,R)-6-(4-(丙烷-1-炔-1-基)-1-(1-(4-(吡咯烷-1-基)苯基)乙基)-1H-吲唑-7-甲酰胺基)螺[3.3]庚烷-2-甲酸甲酯(200mg，产率56.9％)。To (R)-4-(propane-1-yn-1-yl)-1-(1-(4-(pyrrolidin-1-yl)phenyl)ethyl)-1H-indazole-7-carboxylic acid (250mg, 0.669mmol) in DMF (2mL) was added HATU (764mg, 2.008mmol) and diisopropylethylamine (260mg, 2.008mmol), and stirred at 25°C for 0.5h. Then, methyl (Sa)-6-aminospiro[3.3]heptane-2-carboxylate (283 mg, 1.674 mmol) was added to the reaction solution, and stirring was continued at 25° C. for 2.5 hours. The reaction solution was concentrated to obtain a crude product. The crude product was purified by column chromatography (petroleum ether/ethyl acetate (V/V)=1/0-1/3) to obtain the product (Sa,R)-6-(4-(propane-1-yne-1- Base)-1-(1-(4-(pyrrolidin-1-yl)phenyl)ethyl)-1H-indazole-7-carboxamido)spiro[3.3]heptane-2-carboxylic acid methyl ester ( 200 mg, yield 56.9%).
LC-MS,M/Z(ESI):525.3[M+H] +。 LC-MS, M/Z (ESI): 525.3 [M+H] + .
第四步：(Sa,R)-6-(4-(丙烷-1-炔-1-基)-1-(1-(4-(吡咯烷-1-基)苯基)乙基)-1H-吲唑-7-甲酰胺基)螺[3.3]庚烷-2-甲酸(I-23)The fourth step: (Sa,R)-6-(4-(propane-1-yn-1-yl)-1-(1-(4-(pyrrolidin-1-yl)phenyl)ethyl)- 1H-indazole-7-carboxamido)spiro[3.3]heptane-2-carboxylic acid (I-23)
Figure PCTCN2022097611-appb-000075
Figure PCTCN2022097611-appb-000075
向(Sa,R)-6-(4-(丙烷-1-炔-1-基)-1-(1-(4-(吡咯烷-1-基)苯基)乙基)-1H-吲唑-7-甲酰胺基)螺[3.3]庚烷-2-甲酸甲酯(200mg，0.381mmol)的四氢呋喃(6mL)、甲醇(2mL)和水(2mL)的溶液中加入氢氧化锂(45.6mg，1.906mmol)，然后在25℃搅拌3小时。向反应液中加入水(50mL)并用1N盐酸水溶液调节pH至5～6，然后用乙酸乙酯(20mL*3)萃取。合并有机相，用饱和食盐水(10mL*2)洗涤，用无水硫酸钠干燥，过滤并浓缩得到产物(Sa,R)-6-(4-(丙烷-1-炔-1-基)-1-(1-(4-(吡咯烷-1-基)苯基)乙基)-1H-吲唑-7-甲酰胺基)螺[3.3]庚烷-2-甲酸(I-23)(118.1mg，产率 58.4％)。To (Sa, R)-6-(4-(propane-1-yn-1-yl)-1-(1-(4-(pyrrolidin-1-yl)phenyl)ethyl)-1H-ind Lithium hydroxide (45.6 mg, 1.906 mmol), then stirred at 25°C for 3 hours. Water (50 mL) was added to the reaction solution, and the pH was adjusted to 5-6 with 1N aqueous hydrochloric acid solution, followed by extraction with ethyl acetate (20 mL*3). The organic phases were combined, washed with saturated brine (10mL*2), dried over anhydrous sodium sulfate, filtered and concentrated to give the product (Sa,R)-6-(4-(propane-1-yn-1-yl)- 1-(1-(4-(pyrrolidin-1-yl)phenyl)ethyl)-1H-indazole-7-carboxamido)spiro[3.3]heptane-2-carboxylic acid (I-23)( 118.1 mg, yield 58.4%).
1H NMR(400MHz,dmso)δ12.02(s,1H),8.84(d,1H),8.19(s,1H),7.28(d,1H),7.15(d,1H),6.89(d,1H),6.34(d,1H),6.12(q,1H),4.37(dd,1H),3.21-2.99(m,2H),3.01-2.84(m,1H),2.49-2.40(m,1H),2.37-2.21(m,2H),2.19-2.03(m,3H),1.96(dd,1H),1.92-1.85(m,2H),1.83(d,1H).. 1 H NMR (400MHz,dmso)δ12.02(s,1H),8.84(d,1H),8.19(s,1H),7.28(d,1H),7.15(d,1H),6.89(d,1H ),6.34(d,1H),6.12(q,1H),4.37(dd,1H),3.21-2.99(m,2H),3.01-2.84(m,1H),2.49-2.40(m,1H), 2.37-2.21(m,2H),2.19-2.03(m,3H),1.96(dd,1H),1.92-1.85(m,2H),1.83(d,1H)..
LC-MS,M/Z(ESI):511.60[M+H] +。 LC-MS, M/Z (ESI): 511.60 [M+H] + .
实施例24化合物I-24的制备The preparation of embodiment 24 compound I-24
Figure PCTCN2022097611-appb-000076
Figure PCTCN2022097611-appb-000076
化合物6-(4-(丙烷-1-炔-1-基)-1-(1-(5-(三氟甲氧基)吡啶-2-基)乙基)-1H-吲唑-7-甲酰胺)螺[3,3]庚烷-2-甲酸(I-24)的制备参考化合物I-25制备得到，LC-MS,M/Z(ESI):527.50[M+H] +。 Compound 6-(4-(propane-1-yn-1-yl)-1-(1-(5-(trifluoromethoxy)pyridin-2-yl)ethyl)-1H-indazole-7- Formamide) spiro[3,3]heptane-2-carboxylic acid (I-24) was prepared by referring to compound I-25, LC-MS, M/Z (ESI): 527.50[M+H] + .
实施例25化合物I-25的制备The preparation of embodiment 25 compound I-25
合成路线如下所示：The synthetic route is as follows:
Figure PCTCN2022097611-appb-000077
Figure PCTCN2022097611-appb-000077
第一步：N-甲氧基-N-甲基-6-(三氟甲氧基)吡啶酰胺的制备The first step: the preparation of N-methoxy-N-methyl-6-(trifluoromethoxy)pyridinamide
将6-(三氟甲氧基)烟酸(1.5g，7.24mmol)，1-羟基苯并三氮唑(1.96g，14.5mmol)，1-乙基-3(3-二甲基丙胺)碳二亚胺(2.78g，14.5mmol)溶解在干燥的二氯甲烷中，置换氮气，在0℃下滴加二异丙基乙胺(5.0mL，29.0mmol)，搅拌10min后，将N,O-二甲基羟胺盐酸盐固体(1.41g，14.5mmol)一次性加入到反应液中，搅拌过夜。用二氯甲烷和冰水，食盐水稀释，萃取，干燥有机层，旋干得到粗品。过柱(PE/EA＝5/1)得到N-甲氧基-N-甲基-6-(三氟甲氧基)吡啶 酰胺(1.2g,收率66％)。6-(trifluoromethoxy)nicotinic acid (1.5g, 7.24mmol), 1-hydroxybenzotriazole (1.96g, 14.5mmol), 1-ethyl-3(3-dimethylpropylamine) Carbodiimide (2.78g, 14.5mmol) was dissolved in dry dichloromethane, nitrogen was replaced, diisopropylethylamine (5.0mL, 29.0mmol) was added dropwise at 0°C, and after stirring for 10min, N, O-Dimethylhydroxylamine hydrochloride solid (1.41 g, 14.5 mmol) was added to the reaction solution at one time, and stirred overnight. Dilute with dichloromethane, ice water, and brine, extract, dry the organic layer, and spin dry to obtain the crude product. After passing through the column (PE/EA=5/1), N-methoxy-N-methyl-6-(trifluoromethoxy)pyridine amide (1.2 g, yield 66%) was obtained.
第二步：1-(6-(三氟甲氧基)吡啶-3-基)乙烷-1-酮的制备The second step: the preparation of 1-(6-(trifluoromethoxy)pyridin-3-yl)ethan-1-one
将氮-甲氧基-氮-甲基-6-(三氟甲氧基)吡啶酰胺(1.2g,4.8mmol)溶解在四氢呋喃(30mL)中，然后置换氮气，降温到0℃，甲基酮(2.08mL，6.24mmol)慢慢滴加进去，过程持续10分钟。然后在0℃下继续搅拌3h。饱和氯化铵溶液淬灭反应，乙酸乙酯(30mL*2)萃取，有机相合并，干燥，旋干过柱(PE/EA＝10/1)得到1-(6-(三氟甲氧基)吡啶-3-基)乙烷-1-酮(800mg，收率81％)。Dissolve nitrogen-methoxy-nitrogen-methyl-6-(trifluoromethoxy)pyridine amide (1.2g, 4.8mmol) in tetrahydrofuran (30mL), then replace nitrogen, cool to 0°C, methyl ketone (2.08mL, 6.24mmol) was slowly added dropwise, and the process continued for 10 minutes. Stirring was then continued at 0 °C for 3 h. The reaction was quenched with saturated ammonium chloride solution, extracted with ethyl acetate (30mL*2), the organic phases were combined, dried, and spin-dried through the column (PE/EA=10/1) to obtain 1-(6-(trifluoromethoxy )pyridin-3-yl)ethan-1-one (800 mg, yield 81%).
第三步：1-(6-(三氟甲氧基)吡啶-3-基)乙烷-1-醇的制备The third step: the preparation of 1-(6-(trifluoromethoxy)pyridin-3-yl)ethan-1-ol
1-(6-(三氟甲氧基)吡啶-3-基)乙烷-1-酮(800mg，3.90mmol)溶解在甲醇(20mL)和四氢呋喃(4mL)中，在0℃下分批加入硼氢化钠(295mg，7.8mmol)，0℃下搅拌0.5h，用水淬灭，然后反应液旋干得到粗品，拌样过柱(PE/EA＝1/1)得到1-(6-(三氟甲氧基)吡啶-3-基)乙烷-1-醇(800mg，收率99％)。1-(6-(Trifluoromethoxy)pyridin-3-yl)ethan-1-one (800mg, 3.90mmol) was dissolved in methanol (20mL) and tetrahydrofuran (4mL) and added in portions at 0°C Sodium borohydride (295mg, 7.8mmol), stirred at 0°C for 0.5h, quenched with water, and then the reaction solution was spin-dried to obtain the crude product, which was mixed and passed through the column (PE/EA=1/1) to obtain 1-(6-(tri Fluormethoxy)pyridin-3-yl)ethan-1-ol (800 mg, yield 99%).
第四步：4-(丙烷-1-炔-1-基)-1-(1-(6-(三氟甲氧基)吡啶-3-基)乙烷)-1H-吲唑-7-甲酸甲酯的制备The fourth step: 4-(propane-1-yn-1-yl)-1-(1-(6-(trifluoromethoxy)pyridin-3-yl)ethane)-1H-indazole-7- Preparation of methyl formate
1-(6-(三氟甲氧基)吡啶-3-基)乙烷-1-醇(800mg，3.86mmol)、甲基4-(丙烷-1-炔-1-基)-1H-吲唑-7-甲酸酯(500mg，2.33mmol)和三苯基膦(1.84g，7mmol)溶于四氢呋喃(8mL)中，冰浴下缓慢滴加偶氮二异丙酯(1.42g，7mmol)，随后升至室温搅拌16小时。反应液浓缩，柱层析纯化得4-(丙烷-1-炔-1-基)-1-(1-(6-(三氟甲氧基)吡啶-3-基)乙烷)-1H-吲唑-7-甲酸甲酯(600mg，产率64％)。1-(6-(trifluoromethoxy)pyridin-3-yl)ethan-1-ol (800mg, 3.86mmol), methyl 4-(propane-1-yn-1-yl)-1H-ind Azole-7-carboxylate (500mg, 2.33mmol) and triphenylphosphine (1.84g, 7mmol) were dissolved in tetrahydrofuran (8mL), and azodiisopropyl ester (1.42g, 7mmol) was slowly added dropwise under ice-cooling , followed by warming to room temperature and stirring for 16 hours. The reaction solution was concentrated and purified by column chromatography to obtain 4-(propane-1-yn-1-yl)-1-(1-(6-(trifluoromethoxy)pyridin-3-yl)ethane)-1H- Indazole-7-carboxylic acid methyl ester (600 mg, 64% yield).
第五步：4-(丙烷-1-炔-1-基)-1-(1-(6-(三氟甲氧基)吡啶-3-基)乙烷)-1H-吲唑-7-甲酸的制备The fifth step: 4-(propane-1-yn-1-yl)-1-(1-(6-(trifluoromethoxy)pyridin-3-yl)ethane)-1H-indazole-7- Preparation of formic acid
室温下将化合物4-(丙烷-1-炔-1-基)-1-(1-(6-(三氟甲氧基)吡啶-3-基)乙烷)-1H-吲唑-7-甲酸甲酯(600mg,1.50mmol)加入到四氢呋喃(5mL)，甲醇(1mL)和水(1mL)中，加入氢氧化锂(83mg,3.48mmol)，搅拌16h。加入水(20mL)稀释，用1mol/L的盐酸调节pH为4，用乙酸乙酯(20mL×3)萃取，分液，合并有机相，有机相用无水硫酸钠干燥，过滤，浓缩，残留物用薄层硅胶板分离纯化(石油醚:乙酸乙酯(V/V)＝1:1)得化合物4-(丙烷-1-炔-1-基)-1-(1-(6-(三氟甲氧基)吡啶-3-基)乙烷)-1H-吲唑-7-甲酸(500mg,产率86％)。Compound 4-(propane-1-yn-1-yl)-1-(1-(6-(trifluoromethoxy)pyridin-3-yl)ethane)-1H-indazole-7- Methyl formate (600mg, 1.50mmol) was added to tetrahydrofuran (5mL), methanol (1mL) and water (1mL), lithium hydroxide (83mg, 3.48mmol) was added, and stirred for 16h. Add water (20mL) for dilution, adjust the pH to 4 with 1mol/L hydrochloric acid, extract with ethyl acetate (20mL×3), separate the layers, combine the organic phases, dry the organic phases with anhydrous sodium sulfate, filter, concentrate, and leave The material was separated and purified on a thin-layer silica gel plate (petroleum ether: ethyl acetate (V/V)=1:1) to obtain compound 4-(propane-1-yn-1-yl)-1-(1-(6-( Trifluoromethoxy)pyridin-3-yl)ethane)-1H-indazole-7-carboxylic acid (500 mg, 86% yield).
第六步：(S)-6-(4-(丙烷-1-炔-1-基)-1-(1-(6-(三氟甲氧基)吡啶-3-基)乙基)1H-吲唑-7-甲酰胺)螺[3,3]庚烷-2-甲酸甲酯的制备The sixth step: (S)-6-(4-(propane-1-yn-1-yl)-1-(1-(6-(trifluoromethoxy)pyridin-3-yl)ethyl)1H Preparation of -indazole-7-carboxamide)spiro[3,3]heptane-2-carboxylic acid methyl ester
室温下将化合物4-(丙烷-1-炔-1-基)-1-(1-(6-(三氟甲氧基)吡啶-3-基)乙烷)-1H-吲唑-7-甲酸(100mg,0.26mmol)加入到N,N-二甲基甲酰胺(1mL)中，加入(S)-6-氨基螺环[3.3]庚-2-甲酸甲酯盐酸盐(87mg,0.51mmol)，二异丙基乙胺(133mg,1.03mmol)和HATU(244mg,0.64mmol)，室温搅拌16h。加入水(50mL)稀释，用乙酸乙酯(20mL0用乙萃取，分液，合并有机相，有机相用无水硫酸钠干燥，过滤，浓缩，残留物用薄层硅胶板分离纯化(石油醚:乙酸乙酯(V/V)＝1:1)得化合物(S)-6-(4-(丙烷-1-炔-1-基)-1-(1-(6-(三氟甲氧基)吡啶-3-基)乙基)1H-吲唑-7-甲酰胺)螺[3,3]庚烷-2-甲酸甲酯(100mg,产率72％)。Compound 4-(propane-1-yn-1-yl)-1-(1-(6-(trifluoromethoxy)pyridin-3-yl)ethane)-1H-indazole-7- Formic acid (100mg, 0.26mmol) was added to N,N-dimethylformamide (1mL), and (S)-6-aminospiro[3.3]heptane-2-carboxylic acid methyl ester hydrochloride (87mg, 0.51 mmol), diisopropylethylamine (133mg, 1.03mmol) and HATU (244mg, 0.64mmol), stirred at room temperature for 16h. Add water (50mL) to dilute, extract with ethyl acetate (20mL), separate the layers, combine the organic phases, dry the organic phases with anhydrous sodium sulfate, filter, concentrate, and the residue is separated and purified with a thin-layer silica gel plate (petroleum ether: Ethyl acetate (V/V)=1:1) to obtain compound (S)-6-(4-(propane-1-yn-1-yl)-1-(1-(6-(trifluoromethoxy )pyridin-3-yl)ethyl)lH-indazole-7-carboxamide)spiro[3,3]heptane-2-carboxylic acid methyl ester (100 mg, 72% yield).
第七步：(S)-6-(4-(丙烷-1-炔-1-基)-1-(1-(6-(三氟甲氧基)吡啶-3-基)乙基)1H-吲唑-7-甲酰胺)螺[3,3]庚烷-2-甲酸(I-25)的制备The seventh step: (S)-6-(4-(propane-1-yn-1-yl)-1-(1-(6-(trifluoromethoxy)pyridin-3-yl)ethyl)1H Preparation of -indazole-7-carboxamide) spiro[3,3]heptane-2-carboxylic acid (I-25)
Figure PCTCN2022097611-appb-000078
Figure PCTCN2022097611-appb-000078
室温下将化合物(S)-6-(4-(丙烷-1-炔-1-基)-1-(1-(6-(三氟甲氧基)吡啶-3-基)乙基)1H-吲唑-7-甲酰胺)螺[3,3]庚烷-2-甲酸甲酯(100mg,0.19mmol)加入到四氢呋喃(3mL)，甲醇(1mL)和水(1mL)中，加入氢氧化锂(12mg,0.50mmol)，室温搅拌16h。用1N盐酸调节pH＝4，用乙酸乙酯(10mL×3)萃取，分液，合并有机相，有机相用无水硫酸钠干燥，过滤，浓缩，残留物经酸性制备方法A制备得化合物(S)-6-(4-(丙烷-1-炔-1-基)-1-(1-(6-(三氟甲氧基)吡啶-3-基)乙基)1H-吲唑-7-甲酰胺)螺[3,3]庚烷-2-甲酸(I-25)(90mg,产率92％)。Compound (S)-6-(4-(propane-1-yn-1-yl)-1-(1-(6-(trifluoromethoxy)pyridin-3-yl)ethyl)1H at room temperature -indazole-7-carboxamide)spiro[3,3]heptane-2-carboxylic acid methyl ester (100mg, 0.19mmol) was added to tetrahydrofuran (3mL), methanol (1mL) and water (1mL), and hydrogen Lithium (12mg, 0.50mmol), stirred at room temperature for 16h. Use 1N hydrochloric acid to adjust pH=4, extract with ethyl acetate (10mL×3), separate the layers, combine the organic phases, dry the organic phases with anhydrous sodium sulfate, filter, concentrate, and the residue is prepared by acidic preparation method A to obtain the compound ( S)-6-(4-(propane-1-yn-1-yl)-1-(1-(6-(trifluoromethoxy)pyridin-3-yl)ethyl)1H-indazole-7 -carboxamide) spiro[3,3]heptane-2-carboxylic acid (1-25) (90 mg, 92% yield).
1H NMR(400mHz,DMSO-d6)δ12.00(s,1H),8.83(m,1H),8.27(s,1H),7.99(m,1H),7.51(m,1H),7.33(m,1H),7.21-7.16(m,2H),6.34(m,1H),4.31(m,1H),2.94(m,1H),2.28-1.88(m,12H),1.22(m,1H),0.85(m,1H). 1 H NMR (400mHz,DMSO-d6)δ12.00(s,1H),8.83(m,1H),8.27(s,1H),7.99(m,1H),7.51(m,1H),7.33(m ,1H),7.21-7.16(m,2H),6.34(m,1H),4.31(m,1H),2.94(m,1H),2.28-1.88(m,12H),1.22(m,1H), 0.85(m,1H).
LC-MS,M/Z(ESI):527.50[M+H] +。 LC-MS, M/Z (ESI): 527.50 [M+H] + .
实施例26化合物I-26的制备The preparation of embodiment 26 compound I-26
Figure PCTCN2022097611-appb-000079
Figure PCTCN2022097611-appb-000079
化合物6-(4-(丙烷-1-炔-1-基)-1-(1-(5-(三氟甲氧基)吡嗪-2-基)乙基)1H-吲唑-7-甲酰胺)螺[3,3]庚烷-2-甲酸(I-26)的制备参考化合物I-19的合成方法。LC-MS,M/Z(ESI):528.2[M+H] +。 Compound 6-(4-(propane-1-yn-1-yl)-1-(1-(5-(trifluoromethoxy)pyrazin-2-yl)ethyl)1H-indazole-7- Formamide) spiro[3,3]heptane-2-carboxylic acid (I-26) was prepared by referring to the synthetic method of compound I-19. LC-MS, M/Z (ESI): 528.2 [M+H] + .
实施例27化合物I-27的制备The preparation of embodiment 27 compound I-27
Figure PCTCN2022097611-appb-000080
Figure PCTCN2022097611-appb-000080
化合物6-(4-(丙烷-1-炔-1-基)-1-(1-(2-(三氟甲氧基)嘧啶-5-基)乙基)-1H-吲唑-7-甲酰胺)螺[3,3]庚烷-2-甲酸(I-26)的制备参考化合物I-19的合成方法。LC-MS,M/Z(ESI):528.2[M+H] +。 Compound 6-(4-(propane-1-yn-1-yl)-1-(1-(2-(trifluoromethoxy)pyrimidin-5-yl)ethyl)-1H-indazole-7- Formamide) spiro[3,3]heptane-2-carboxylic acid (I-26) was prepared by referring to the synthetic method of compound I-19. LC-MS, M/Z (ESI): 528.2 [M+H] + .
实施例28化合物I-28的制备The preparation of embodiment 28 compound I-28
合成路线如下所示：The synthetic route is as follows:
Figure PCTCN2022097611-appb-000081
Figure PCTCN2022097611-appb-000081
第一步：1-(1-(5-溴吡啶-2-基)乙基)-4-(丙烷-1-炔-1-基)-1H-吲唑-7-甲酸甲酯的制备Step 1: Preparation of 1-(1-(5-bromopyridin-2-yl)ethyl)-4-(propane-1-yn-1-yl)-1H-indazole-7-carboxylic acid methyl ester
1-(5溴吡啶-2-基)乙醇(943mg，4.67mmol)、4-(丙烷-1-炔-1-基)-1H-吲唑-7-甲酸甲酯(500mg，2.33mmol)和三苯基膦(1.84g，7mmol)溶于四氢呋喃(8mL)中，冰浴下缓慢滴加偶氮二异丙酯(1.42g，7mmol)，随后升至室温搅拌16小时。反应液浓缩，柱层析纯化得1-(1-(5-溴吡啶-2-基)乙基)-4-(丙烷-1-炔-1-基)-1H-吲唑-7-甲酸甲酯(400mg，产率43％)。1-(5-bromopyridin-2-yl)ethanol (943mg, 4.67mmol), 4-(propane-1-yn-1-yl)-1H-indazole-7-carboxylic acid methyl ester (500mg, 2.33mmol) and Triphenylphosphine (1.84g, 7mmol) was dissolved in tetrahydrofuran (8mL), and azobisisopropyl (1.42g, 7mmol) was slowly added dropwise under ice-cooling, followed by warming to room temperature and stirring for 16 hours. The reaction solution was concentrated and purified by column chromatography to obtain 1-(1-(5-bromopyridin-2-yl)ethyl)-4-(propane-1-yn-1-yl)-1H-indazole-7-carboxylic acid Methyl ester (400 mg, 43% yield).
第二步：1-(1-(5-环丙基吡啶-2-基)乙基)-4-(丙烷-1-炔-1-基)-1H-吲唑-7-甲酸甲酯的制备The second step: 1-(1-(5-cyclopropylpyridin-2-yl) ethyl)-4-(propane-1-yn-1-yl)-1H-indazole-7-carboxylic acid methyl ester preparation
室温下将1-(1-(5-溴吡啶-2-基)乙基)-4-(丙烷-1-炔-1-基)-1H-吲唑-7-甲酸甲酯(400mg，1.00mmol)，四三苯基磷钯(116mg，0.1mmol)，环丙基硼酸(431mg，5.02mmol)，氟化钾(321mg，5.52mmol)加入到20mL甲苯和4mL水中，置换氮气，100℃下反应过夜。反应液浓缩，柱层析纯化得到1-(1-(5-环丙基吡啶-2-基)乙基)-4-(丙烷-1-炔-1-基)-1H-吲唑-7-甲酸甲酯(250mg，产率69％)。Methyl 1-(1-(5-bromopyridin-2-yl)ethyl)-4-(propane-1-yn-1-yl)-1H-indazole-7-carboxylate (400 mg, 1.00 mmol), tetrakistriphenylphosphopalladium (116mg, 0.1mmol), cyclopropylboronic acid (431mg, 5.02mmol), potassium fluoride (321mg, 5.52mmol) were added to 20mL of toluene and 4mL of water, replaced with nitrogen, at 100°C React overnight. The reaction solution was concentrated and purified by column chromatography to obtain 1-(1-(5-cyclopropylpyridin-2-yl)ethyl)-4-(propane-1-yn-1-yl)-1H-indazole-7 - Methyl formate (250 mg, 69% yield).
第三步：1-(1-(5-环丙基吡啶-2-基)乙基)-4-(丙烷-1-炔-1-基)-1H-吲唑-7-甲酸的制备Step 3: Preparation of 1-(1-(5-cyclopropylpyridin-2-yl)ethyl)-4-(propane-1-yn-1-yl)-1H-indazole-7-carboxylic acid
室温下将化合物1-(1-(5-环丙基吡啶-2-基)乙基)-4-(丙烷-1-炔-1-基)-1H-吲唑-7-甲酸甲酯(250mg,0.70mmol)加入到四氢呋喃(5mL)，甲醇(1mL)和水(1mL)中，加入氢氧化锂(83mg,3.48mmol)，搅拌16h。加入水(20mL)稀释，用1mol/L的盐酸调节pH为4，用乙酸乙酯(20mL×3)萃取，分液，合并有机相，有机相用无水硫酸钠干燥，过滤，浓缩，残留物用薄层硅胶板分离纯化(石油醚:乙酸乙酯(V/V)＝1:1)得到1-(1-(5-环丙基吡啶-2-基)乙基)-4-(丙烷-1-炔-1-基)-1H-吲唑-7-甲酸(240mg,产率100％)。Compound 1-(1-(5-cyclopropylpyridin-2-yl)ethyl)-4-(propane-1-yn-1-yl)-1H-indazole-7-carboxylic acid methyl ester ( 250mg, 0.70mmol) was added to tetrahydrofuran (5mL), methanol (1mL) and water (1mL), lithium hydroxide (83mg, 3.48mmol) was added, and stirred for 16h. Add water (20mL) for dilution, adjust the pH to 4 with 1mol/L hydrochloric acid, extract with ethyl acetate (20mL×3), separate the layers, combine the organic phases, dry the organic phases with anhydrous sodium sulfate, filter, concentrate, and leave The material was separated and purified on a thin-layer silica gel plate (petroleum ether:ethyl acetate (V/V)=1:1) to obtain 1-(1-(5-cyclopropylpyridin-2-yl)ethyl)-4-( Propan-1-yn-1-yl)-1H-indazole-7-carboxylic acid (240 mg, 100% yield).
第四步：(Sa)-6-(1-(1-(5-环丙基吡啶-2-基)乙基)-4-(丙基-1-炔-1-基)-1H-吲唑-7-甲酰胺)螺[3.3]庚烷-2-甲酸甲酯的制备The fourth step: (Sa)-6-(1-(1-(5-cyclopropylpyridin-2-yl)ethyl)-4-(propyl-1-yn-1-yl)-1H-ind Preparation of oxazole-7-carboxamide)spiro[3.3]heptane-2-carboxylate methyl ester
室温下将化合物1-(1-(5-环丙基吡啶-2-基)乙基)-4-(丙烷-1-炔-1-基)-1H-吲唑-7-甲酸(50mg,0.15mmol)加入到N,N-二甲基甲酰胺(1mL)中，加入(Sa)-6-氨基螺环[3.3]庚-2-甲酸甲酯盐酸盐(37mg,0.22mmol)，二异丙基乙胺(75mg,0.58mmol)和HATU(110mg,0.29mmol)，室温搅拌16h。加入水(50mL)稀释，用乙酸乙酯(20mL×3)萃取，分液，合并有机相，有机相用 无水硫酸钠干燥，过滤，浓缩，残留物用薄层硅胶板分离纯化(石油醚:乙酸乙酯(V/V)＝1:1)得化合物(Sa)-6-(1-(1-(5-环丙基吡啶-2-基)乙基)-4-(丙基-1-炔-1-基)-1H-吲唑-7-甲酰胺)螺[3.3]庚烷-2-甲酸甲酯(50mg,产率70％)。Compound 1-(1-(5-cyclopropylpyridin-2-yl)ethyl)-4-(propane-1-yn-1-yl)-1H-indazole-7-carboxylic acid (50mg, 0.15mmol) was added to N,N-dimethylformamide (1mL), and (Sa)-6-aminospiro[3.3]heptane-2-carboxylic acid methyl ester hydrochloride (37mg, 0.22mmol) was added, di Isopropylethylamine (75mg, 0.58mmol) and HATU (110mg, 0.29mmol), stirred at room temperature for 16h. Add water (50mL) to dilute, extract with ethyl acetate (20mL×3), separate the layers, combine the organic phases, dry the organic phases with anhydrous sodium sulfate, filter, concentrate, and separate and purify the residue with a thin-layer silica gel plate (petroleum ether : ethyl acetate (V/V)=1:1) to obtain compound (Sa)-6-(1-(1-(5-cyclopropylpyridin-2-yl) ethyl)-4-(propyl- 1-Alkyn-1-yl)-1H-indazole-7-carboxamide)spiro[3.3]heptane-2-carboxylic acid methyl ester (50 mg, 70% yield).
第五步：(Sa)-6-(1-(1-(5-环丙基吡啶-2-基)乙基)-4-(丙基-1-炔-1-基)-1H-吲唑-7-甲酰胺)螺[3.3]庚烷-2-甲酸(I-28)The fifth step: (Sa)-6-(1-(1-(5-cyclopropylpyridin-2-yl)ethyl)-4-(propyl-1-yn-1-yl)-1H-ind Azole-7-carboxamide) spiro[3.3]heptane-2-carboxylic acid (I-28)
Figure PCTCN2022097611-appb-000082
Figure PCTCN2022097611-appb-000082
室温下将化合物(Sa)-6-(1-(1-(5-环丙基吡啶-2-基)乙基)-4-(丙基-1-炔-1-基)-1H-吲唑-7-甲酰胺)螺[3.3]庚烷-2-甲酸甲酯(50mg,0.10mmol)加入到四氢呋喃(3mL)，甲醇(1mL)和水(1mL)中，加入氢氧化锂(12mg,0.50mmol)，室温搅拌16h。用1N盐酸调节pH＝4，用乙酸乙酯(10mL×3)萃取，分液，合并有机相，有机相用无水硫酸钠干燥，过滤，浓缩，残留物经酸性制备方法A制备得化合物(Sa)-6-(1-(1-(5-环丙基吡啶-2-基)乙基)-4-(丙基-1-炔-1-基)-1H-吲唑-7-甲酰胺)螺[3.3]庚烷-2-甲酸(I-28)(45mg,产率93％)。Compound (Sa)-6-(1-(1-(5-cyclopropylpyridin-2-yl)ethyl)-4-(propyl-1-yn-1-yl)-1H-ind Azole-7-carboxamide) spiro[3.3]heptane-2-carboxylate (50mg, 0.10mmol) was added to tetrahydrofuran (3mL), methanol (1mL) and water (1mL), and lithium hydroxide (12mg, 0.50mmol), stirred at room temperature for 16h. Use 1N hydrochloric acid to adjust pH=4, extract with ethyl acetate (10mL×3), separate the layers, combine the organic phases, dry the organic phases with anhydrous sodium sulfate, filter, concentrate, and the residue is prepared by acidic preparation method A to obtain the compound ( Sa)-6-(1-(1-(5-cyclopropylpyridin-2-yl)ethyl)-4-(propyl-1-yn-1-yl)-1H-indazole-7-methyl Amide) spiro[3.3]heptane-2-carboxylic acid (I-28) (45 mg, 93% yield).
1H NMR(400mHz,DMSO-d6)δ8.71(m,1H),8.32(m,1H),8.23(s,1H),7.35(m,1H),7.30(m,1H),7.20(m,1H),6.61(m,1H),6.29(m,1H),4.18(m,1H),2.95(m,1H),2.15-1.87(m,14H),0.97(m,1H),0.96-0.94(m,2H),0.68-0.66(m,2H). 1 H NMR (400mHz, DMSO-d6) δ8.71(m,1H),8.32(m,1H),8.23(s,1H),7.35(m,1H),7.30(m,1H),7.20(m ,1H),6.61(m,1H),6.29(m,1H),4.18(m,1H),2.95(m,1H),2.15-1.87(m,14H),0.97(m,1H),0.96- 0.94(m,2H),0.68-0.66(m,2H).
LC-MS,M/Z(ESI):483.54[M+H] +。 LC-MS, M/Z (ESI): 483.54 [M+H] + .
实施例29化合物I-29a的制备The preparation of embodiment 29 compound I-29a
合成路线如下所示：The synthetic route is as follows:
Figure PCTCN2022097611-appb-000083
Figure PCTCN2022097611-appb-000083
第一步：1-(5-氯吡嗪-2-基)-乙烷-1-醇的制备The first step: the preparation of 1-(5-chloropyrazin-2-yl)-ethan-1-ol
0℃下把1-(5-氯吡嗪-2-基)乙烷-1-酮(3g，19.2mmol)溶解到乙醇(30mL)中，然后慢慢分批加入硼氢化钠(1.45g,38.3mmol)，恢复室温搅拌1h。滴加冰水淬灭反应，反应液旋干得到粗品，柱层析(PE/EA＝2/1)得到1-(5-氯吡嗪-2-基)-乙烷-1-醇(2.2g,产率72％)。Dissolve 1-(5-chloropyrazin-2-yl)ethan-1-one (3g, 19.2mmol) in ethanol (30mL) at 0°C, then slowly add sodium borohydride (1.45g, 38.3mmol), returned to room temperature and stirred for 1h. The reaction was quenched by adding ice water dropwise, and the reaction solution was spin-dried to obtain the crude product. Column chromatography (PE/EA=2/1) gave 1-(5-chloropyrazin-2-yl)-ethane-1-ol (2.2 g, yield 72%).
第二步：1-(5-环丙基吡嗪-2-基)-乙烷-1-醇的制备The second step: the preparation of 1-(5-cyclopropylpyrazin-2-yl)-ethan-1-ol
室温下将1-(2-氯吡嗪-5-基)-乙烷-1-醇(1.2g，7.57mmol)加入到1,4-二氧六环(30mL)和水(6mL)中，加入环丙基硼酸(3.25g,37.8mmol)，碳酸钠(2.41g，22.7mmol)，1,1'-双二苯基膦二茂铁二氯化钯(1.11g,1.51mmol)，氮气保护下加热至100℃，搅拌14h。反应液浓缩，柱层析纯化得到1-(5-环丙基吡嗪-2-基)-乙烷-1-醇(1.3g，产率84％)。1-(2-Chloropyrazin-5-yl)-ethan-1-ol (1.2 g, 7.57 mmol) was added to 1,4-dioxane (30 mL) and water (6 mL) at room temperature, Add cyclopropylboronic acid (3.25g, 37.8mmol), sodium carbonate (2.41g, 22.7mmol), 1,1'-bisdiphenylphosphinoferrocenepalladium dichloride (1.11g, 1.51mmol), nitrogen protection Heated to 100°C and stirred for 14h. The reaction solution was concentrated and purified by column chromatography to obtain 1-(5-cyclopropylpyrazin-2-yl)-ethan-1-ol (1.3 g, yield 84%).
第三步：1-(1-(5-环丙基吡嗪-2-基)乙基)-4-(丙烷-1-炔-1-基)-1H-吲唑-7-甲酸甲酯的制备The third step: 1-(1-(5-cyclopropylpyrazin-2-yl)ethyl)-4-(propane-1-yn-1-yl)-1H-indazole-7-carboxylic acid methyl ester preparation of
1-(5-环丙基吡嗪-2-基)-乙烷-1-醇(613mg，3.73mmol)、甲基4-(丙烷-1-炔-1-基)-1H-吲唑-7-甲酸酯(400mg，1.87mmol)和三苯基膦(1.47g，5.60mmol)溶于四氢呋喃(10mL)中，冰浴下缓慢滴加偶氮二异丙酯(1.13g，5.60mmol)，随后升至室温搅拌16小时。反应液浓缩，柱层析纯化得1-(1-(5-环丙基吡嗪-2-基)乙基)-4-(丙烷-1-炔-1-基)-1H-吲唑-7-甲酸甲酯(400mg，产率30％)。1-(5-cyclopropylpyrazin-2-yl)-ethan-1-ol (613 mg, 3.73 mmol), methyl 4-(propan-1-yn-1-yl)-1H-indazole- 7-Carboxylate (400mg, 1.87mmol) and triphenylphosphine (1.47g, 5.60mmol) were dissolved in tetrahydrofuran (10mL), and azobisisopropyl ester (1.13g, 5.60mmol) was slowly added dropwise under ice-cooling , followed by warming to room temperature and stirring for 16 hours. The reaction solution was concentrated and purified by column chromatography to obtain 1-(1-(5-cyclopropylpyrazin-2-yl)ethyl)-4-(propane-1-yn-1-yl)-1H-indazole- 7-Methyl carboxylate (400 mg, 30% yield).
第四步：1-(1-(5-环丙基吡嗪-2-基)乙基)-4-(丙烷-1-炔-1-基)-1H-吲唑-7-甲酸的制备Step 4: Preparation of 1-(1-(5-cyclopropylpyrazin-2-yl)ethyl)-4-(propane-1-yn-1-yl)-1H-indazole-7-carboxylic acid
室温下将化合物1-(1-(5-环丙基吡嗪-2-基)乙基)-4-(丙烷-1-炔-1-基)-1H-吲唑-7-甲酸甲酯(200mg,0.56mmol)加入到四氢呋喃(5mL)，甲醇(1mL)和水(1mL)中，加入氢氧化锂(66mg,2.80mmol)，搅拌16h。加入水(20mL)稀释，用1mol/L的盐酸调节pH为4，用乙酸乙酯(20mL×3)萃取，分液，合并有机相，有机相用无水硫酸钠干燥，过滤，浓缩，残留物用薄层硅胶板分离纯化(石油醚:乙酸乙酯(V/V)＝1:1)得化合物1-(1-(5-环丙基吡嗪-2-基)乙基)-4-(丙烷-1-炔 -1-基)-1H-吲唑-7-甲酸(190mg,产率99％)。Compound 1-(1-(5-cyclopropylpyrazin-2-yl)ethyl)-4-(propane-1-yn-1-yl)-1H-indazole-7-carboxylic acid methyl ester (200mg, 0.56mmol) was added into tetrahydrofuran (5mL), methanol (1mL) and water (1mL), lithium hydroxide (66mg, 2.80mmol) was added, and stirred for 16h. Add water (20mL) for dilution, adjust the pH to 4 with 1mol/L hydrochloric acid, extract with ethyl acetate (20mL×3), separate the layers, combine the organic phases, dry the organic phases with anhydrous sodium sulfate, filter, concentrate, and leave The material was separated and purified with a thin-layer silica gel plate (petroleum ether:ethyl acetate (V/V)=1:1) to obtain compound 1-(1-(5-cyclopropylpyrazin-2-yl)ethyl)-4 -(propan-1-yn-1-yl)-1H-indazole-7-carboxylic acid (190 mg, 99% yield).
第五步：(Sa)-6-(1-(1-(5-环丙基吡嗪-2-基)乙基)-4-(丙烷-1-炔-1-基)-1H-吲唑-7-甲酰胺)螺环[3.3]庚烷-2-基甲酸甲酯的制备The fifth step: (Sa)-6-(1-(1-(5-cyclopropylpyrazin-2-yl)ethyl)-4-(propane-1-yn-1-yl)-1H-ind Preparation of azole-7-carboxamide)spiro[3.3]heptane-2-ylcarboxylic acid methyl ester
室温下将化合物1-(1-(5-环丙基吡嗪-2-基)乙基)-4-(丙烷-1-炔-1-基)-1H-吲唑-7-甲酸(190mg,0.55mmol)加入到N,N-二甲基甲酰胺(5mL)中，加入(Sa)-6-氨基螺环[3.3]庚-2-基-甲酸甲酯盐酸盐(280mg,1.65mmol)，二异丙基乙胺(354mg,2.74mmol)和HATU(417mg,1.10mmol)，室温搅拌16h。加入水(50mL)稀释，用乙酸乙酯(20mL×3)萃取，分液，合并有机相，有机相用无水硫酸钠干燥，过滤，浓缩，残留物用薄层硅胶板分离纯化(石油醚:乙酸乙酯(V/V)＝1:1)得化合物(Sa)-6-(1-(1-(5-环丙基吡嗪-2-基)乙基)-4-(丙烷-1-炔-1-基)-1H-吲唑-7-甲酰胺)螺环[3.3]庚烷-2-基甲酸甲酯(150mg,产率55％)。Compound 1-(1-(5-cyclopropylpyrazin-2-yl)ethyl)-4-(propane-1-yn-1-yl)-1H-indazole-7-carboxylic acid (190mg , 0.55mmol) was added to N,N-dimethylformamide (5mL), and (Sa)-6-aminospiro[3.3]hept-2-yl-formic acid methyl ester hydrochloride (280mg, 1.65mmol ), diisopropylethylamine (354mg, 2.74mmol) and HATU (417mg, 1.10mmol), stirred at room temperature for 16h. Add water (50mL) to dilute, extract with ethyl acetate (20mL×3), separate the layers, combine the organic phases, dry the organic phases with anhydrous sodium sulfate, filter, concentrate, and separate and purify the residue with a thin-layer silica gel plate (petroleum ether : ethyl acetate (V/V)=1:1) to obtain compound (Sa)-6-(1-(1-(5-cyclopropylpyrazin-2-yl) ethyl)-4-(propane- 1-Alkyn-1-yl)-1H-indazole-7-carboxamide)spiro[3.3]heptan-2-ylcarboxylic acid methyl ester (150 mg, 55% yield).
第六步：(Sa,R)-6-(1-(1-(5-环丙基吡嗪-2-基)乙基)-4-(丙烷-1-炔-1-基)-1H-吲唑-7-甲酰胺)螺环[3.3]庚烷-2-基甲酸甲酯的制备The sixth step: (Sa,R)-6-(1-(1-(5-cyclopropylpyrazin-2-yl)ethyl)-4-(propane-1-yn-1-yl)-1H Preparation of -indazole-7-carboxamide) spiro[3.3]heptane-2-ylcarboxylic acid methyl ester
将化合物(Sa)-6-(1-(1-(5-环丙基吡嗪-2-基)乙基)-4-(丙烷-1-炔-1-基)-1H-吲唑-7-甲酰胺)螺环[3.3]庚烷-2(S)-基甲酸甲酯(150mg,0.30mmol)进行手性拆分方法(IC-3-IPA(DEA)-40-3ML-35T，r.t.＝0.949min)制备得化合物(Sa,R)-6-(1-(1-(5-环丙基吡嗪-2-基)乙基)-4-(丙烷-1-炔-1-基)-1H-吲唑-7-甲酰胺)螺环[3.3]庚烷-2-基甲酸甲酯(50mg,产率33％)。Compound (Sa)-6-(1-(1-(5-cyclopropylpyrazin-2-yl)ethyl)-4-(propane-1-yn-1-yl)-1H-indazole- 7-carboxamide) spiro[3.3]heptane-2(S)-methyl carboxylate (150mg, 0.30mmol) for chiral resolution (IC-3-IPA(DEA)-40-3ML-35T, r.t.=0.949min) prepared compound (Sa,R)-6-(1-(1-(5-cyclopropylpyrazin-2-yl)ethyl)-4-(propane-1-yne-1- yl)-1H-indazole-7-carboxamide)spiro[3.3]heptan-2-ylcarboxylate (50 mg, yield 33%).
第七步：(Sa,R)-6-(1-(1-(5-环丙基吡嗪-2-基)乙基)-4-(丙烷-1-炔-1-基)-1H-吲唑-7-甲酰胺)螺环[3.3]庚烷-2-基甲酸(I-29a)The seventh step: (Sa,R)-6-(1-(1-(5-cyclopropylpyrazin-2-yl)ethyl)-4-(propane-1-yn-1-yl)-1H -indazole-7-carboxamide) spiro[3.3]heptane-2-ylcarboxylic acid (I-29a)
Figure PCTCN2022097611-appb-000084
Figure PCTCN2022097611-appb-000084
室温下将化合物(Sa,R)-6-(1-(1-(5-环丙基吡嗪-2-基)乙基)-4-(丙烷-1-炔-1-基)-1H-吲唑-7-甲酰胺)螺环[3.3]庚烷-2-基甲酸甲酯(50mg,0.10mmol)加入到四氢呋喃(3mL)，甲醇(1mL)和水(1mL)中，加入氢氧化锂(12mg,0.50mmol)，室温搅拌16h。用1N盐酸调节pH＝4，用乙酸乙酯(10mL×3)萃取，分液，合并有机相，有机相用无水硫酸钠干燥，过滤，浓缩，残留物经酸性制备方法A制备得化合物(Sa,R)-6-(1-(1-(5-环丙基吡嗪-2-基)乙基)-4-(丙烷-1-炔-1-基)-1H-吲唑-7-甲酰胺)螺环[3.3]庚烷-2-基甲酸(I-29a)(45mg，产率93％)。Compound (Sa,R)-6-(1-(1-(5-cyclopropylpyrazin-2-yl)ethyl)-4-(propane-1-yn-1-yl)-1H at room temperature -indazole-7-carboxamide) spiro[3.3]heptane-2-ylcarboxylate (50mg, 0.10mmol) was added to tetrahydrofuran (3mL), methanol (1mL) and water (1mL), and hydrogen Lithium (12mg, 0.50mmol), stirred at room temperature for 16h. Use 1N hydrochloric acid to adjust pH=4, extract with ethyl acetate (10mL×3), separate the layers, combine the organic phases, dry the organic phases with anhydrous sodium sulfate, filter, concentrate, and the residue is prepared by acidic preparation method A to obtain the compound ( Sa,R)-6-(1-(1-(5-cyclopropylpyrazin-2-yl)ethyl)-4-(propane-1-yn-1-yl)-1H-indazole-7 -carboxamide) spiro[3.3]heptan-2-ylcarboxylic acid (I-29a) (45 mg, 93% yield).
1H NMR(400mHz,DMSO-d6)δ8.75(d,1H),8.43(s,1H),8.24(s,1H),7.88(s,1H),7.32(m,1H),7.22(m,1H),6.33(m,1H),4.20(m,1H),2.95(m,1H),2.19(m,1H),2.17-1.90(m,13H),1.75(m,1H),1.01-0.96(m,2H),0.88-0.84(m,2H). 1 H NMR (400mHz,DMSO-d6)δ8.75(d,1H),8.43(s,1H),8.24(s,1H),7.88(s,1H),7.32(m,1H),7.22(m ,1H),6.33(m,1H),4.20(m,1H),2.95(m,1H),2.19(m,1H),2.17-1.90(m,13H),1.75(m,1H),1.01- 0.96(m,2H),0.88-0.84(m,2H).
LC-MS,M/Z(ESI):484.30[M+H] +。 LC-MS, M/Z (ESI): 484.30 [M+H] + .
化合物I-29b的制备Preparation of Compound I-29b
合成路线如下所示：The synthetic route is as follows:
Figure PCTCN2022097611-appb-000085
Figure PCTCN2022097611-appb-000085
第一步：(Sa,S)-6-(1-(1-(5-环丙基吡嗪-2-基)乙基)-4-(丙烷-1-炔-1-基)-1H-吲唑-7-甲酰胺)螺环[3.3]庚烷-2-甲酸甲酯的制备The first step: (Sa,S)-6-(1-(1-(5-cyclopropylpyrazin-2-yl)ethyl)-4-(propane-1-yn-1-yl)-1H Preparation of -indazole-7-carboxamide) spiro[3.3]heptane-2-carboxylic acid methyl ester
将化合物(Sa)-6-(1-(1-(5-环丙基吡嗪-2-基)乙基)-4-(丙烷-1-炔-1-基)-1H-吲唑-7-甲酰胺)螺环[3.3]庚烷-2-甲酸甲酯(150mg,0.30mmol)进行手性拆分方法(IC-3-IPA(DEA)-40-3ML-35T，r.t.＝1.676min)制备得化合物(Sa,S)-6-(1-(1-(5-环丙基吡嗪-2-基)乙基)-4-(丙烷-1-炔-1-基)-1H-吲唑-7-甲酰胺)螺环[3.3]庚烷-2-甲酸甲酯(50mg，产率33％)。Compound (Sa)-6-(1-(1-(5-cyclopropylpyrazin-2-yl)ethyl)-4-(propane-1-yn-1-yl)-1H-indazole- 7-carboxamide) spiro[3.3]heptane-2-carboxylate (150mg, 0.30mmol) chiral resolution method (IC-3-IPA(DEA)-40-3ML-35T, r.t.=1.676min ) prepared compound (Sa, S)-6-(1-(1-(5-cyclopropylpyrazin-2-yl) ethyl)-4-(propane-1-yn-1-yl)-1H -indazole-7-carboxamide) spiro[3.3]heptane-2-carboxylic acid methyl ester (50 mg, 33% yield).
第二步：(Sa,S)-6-(1-(1-(5-环丙基吡嗪-2-基)乙基)-4-(丙烷-1-炔-1-基)-1H-吲唑-7-甲酰胺)螺环[3.3]庚烷-2-甲酸(I-29b)的制备The second step: (Sa,S)-6-(1-(1-(5-cyclopropylpyrazin-2-yl)ethyl)-4-(propane-1-yn-1-yl)-1H Preparation of -indazole-7-carboxamide) spiro[3.3]heptane-2-carboxylic acid (I-29b)
Figure PCTCN2022097611-appb-000086
Figure PCTCN2022097611-appb-000086
室温下将化合物(Sa,S)-6-(1-(1-(5-环丙基吡嗪-2-基)乙基)-4-(丙烷-1-炔-1-基)-1H-吲唑-7-甲酰胺)螺环[3.3]庚烷-2-甲酸甲酯(50mg,0.10mmol)加入到四氢呋喃(3mL)，甲醇(1mL)和水(1mL)中，加入氢氧化锂(12mg,0.50mmol)，室温搅拌16h。用1N盐酸调节pH＝4，用乙酸乙酯(10mL×3)萃取，分液，合并有机相，有机相用无水硫酸钠干燥，过滤，浓缩，残留物经酸性制备方法A制备得化合物(Sa,S)-6-(1-(1-(5-环丙基吡嗪-2-基)乙基)-4-(丙烷-1-炔-1-基)-1H-吲唑-7-甲酰胺)螺环[3.3]庚烷-2-甲酸(I-29b)(45mg,产率93％)。Compound (Sa,S)-6-(1-(1-(5-cyclopropylpyrazin-2-yl)ethyl)-4-(propane-1-yn-1-yl)-1H at room temperature -indazole-7-carboxamide) spiro[3.3]heptane-2-carboxylate (50mg, 0.10mmol) was added to tetrahydrofuran (3mL), methanol (1mL) and water (1mL) and lithium hydroxide was added (12mg, 0.50mmol), stirred at room temperature for 16h. Use 1N hydrochloric acid to adjust pH=4, extract with ethyl acetate (10mL×3), separate the layers, combine the organic phases, dry the organic phases with anhydrous sodium sulfate, filter, concentrate, and the residue is prepared by acidic preparation method A to obtain the compound ( Sa,S)-6-(1-(1-(5-cyclopropylpyrazin-2-yl)ethyl)-4-(propane-1-yn-1-yl)-1H-indazole-7 -carboxamide) spiro[3.3]heptane-2-carboxylic acid (I-29b) (45 mg, 93% yield).
1H NMR(400mHz,DMSO-d6)δ12.03(s,1H),8.76(d,1H),8.45(s,1H),8.24(s,1H),7.88(s,1H),7.33(m,1H),7.23(m,1H),6.33(m,1H),4.20(m,1H),2.95(m,1H),2.51-1.90(m,15H),1.01-0.97(m,2H),0.88-0.86(m,2H). 1 H NMR(400mHz,DMSO-d6)δ12.03(s,1H),8.76(d,1H),8.45(s,1H),8.24(s,1H),7.88(s,1H),7.33(m ,1H),7.23(m,1H),6.33(m,1H),4.20(m,1H),2.95(m,1H),2.51-1.90(m,15H),1.01-0.97(m,2H), 0.88-0.86(m,2H).
LC-MS,M/Z(ESI):484.30[M+H] +。 LC-MS, M/Z (ESI): 484.30 [M+H] + .
实施例30化合物I-30a、I-30b的制备The preparation of embodiment 30 compound I-30a, I-30b
合成路线如下所示：The synthetic route is as follows:
Figure PCTCN2022097611-appb-000087
Figure PCTCN2022097611-appb-000087
第一步：1-(1-(6-溴吡啶-3-基)乙基)-4-(丙烷-1-炔-1-基)-1H-吲唑-7-甲酸甲酯的制备Step 1: Preparation of 1-(1-(6-bromopyridin-3-yl)ethyl)-4-(propane-1-yn-1-yl)-1H-indazole-7-carboxylic acid methyl ester
将4-丙炔基-1H-7-吲唑甲酯(500mg，2.334mmol)，1-(6-溴吡啶-3-基)乙醇(943mg，4.67mmol)和三苯基膦(1837mg，7.00mmol)溶于四氢呋喃(10mL)中，在0℃下缓慢滴入偶氮二甲酸二异丙酯(1416mg，7.00mmol)，然后在25℃下反应16h。将反应液浓缩得到粗品。粗品经过柱层析(石油醚/乙酸乙酯(V/V)＝10:1-1:1)得到产物1-(1-(6-溴吡啶-3-基)乙基)-4-(丙烷-1-炔-1-基)-1H-吲唑-7-甲酸甲酯(450mg，产率48.4％)。4-propynyl-1H-7-indazole methyl ester (500mg, 2.334mmol), 1-(6-bromopyridin-3-yl)ethanol (943mg, 4.67mmol) and triphenylphosphine (1837mg, 7.00 mmol) was dissolved in tetrahydrofuran (10 mL), and diisopropyl azodicarboxylate (1416 mg, 7.00 mmol) was slowly added dropwise at 0 °C, and then reacted at 25 °C for 16 h. The reaction solution was concentrated to obtain a crude product. The crude product was subjected to column chromatography (petroleum ether/ethyl acetate (V/V)=10:1-1:1) to obtain the product 1-(1-(6-bromopyridin-3-yl)ethyl)-4-( Propan-1-yn-1-yl)-1H-indazole-7-carboxylic acid methyl ester (450 mg, 48.4% yield).
LC-MS,M/Z(ESI):399.3[M+H] +。 LC-MS, M/Z (ESI): 399.3 [M+H] + .
第二步：1-(1-(6-环丙基吡啶-3-基)乙基)-4-(丙烷-1-炔-1-基)-1H-吲唑-7-甲酸甲酯的制备The second step: 1-(1-(6-cyclopropylpyridin-3-yl) ethyl)-4-(propane-1-yn-1-yl)-1H-indazole-7-carboxylic acid methyl ester preparation
将1-(1-(6-溴吡啶-3-基)乙基)-4-(丙烷-1-炔-1-基)-1H-吲唑-7-甲酸甲酯(450mg，1.130mmol)、环丙基硼酸(485mg，5.65mmol)、碳酸钾(625mg，4.52mmol)和[1,1'-双(二苯基膦基)二茂铁]二氯化钯(83mg，0.113mmol)溶于1,4-二氧六环(6mL)和水(2mL)中，然后在氮气保护下升至100℃反应16h。将反应液倒入水(50mL)中，用乙酸乙酯(20mL*3)萃取。有机相合并，用饱和食盐水(20mL*2)洗涤，用无水硫酸钠干燥，过滤浓缩得到棕色粗品。粗品经过柱层析(乙酸乙酯：石油醚(V/V)＝10:1-1:1)得到产物1-(1-(6-环丙基吡啶-3-基)乙基)-4-(丙烷-1-炔-1-基)-1H-吲唑-7-甲酸甲酯(180mg，产率44.3％)。1-(1-(6-bromopyridin-3-yl)ethyl)-4-(propane-1-yn-1-yl)-1H-indazole-7-carboxylic acid methyl ester (450mg, 1.130mmol) , cyclopropylboronic acid (485mg, 5.65mmol), potassium carbonate (625mg, 4.52mmol) and [1,1'-bis(diphenylphosphino)ferrocene]palladium dichloride (83mg, 0.113mmol) In 1,4-dioxane (6mL) and water (2mL), then rise to 100°C under nitrogen protection for 16h. The reaction solution was poured into water (50 mL), extracted with ethyl acetate (20 mL*3). The organic phases were combined, washed with saturated brine (20 mL*2), dried over anhydrous sodium sulfate, filtered and concentrated to obtain a brown crude product. The crude product was subjected to column chromatography (ethyl acetate:petroleum ether (V/V)=10:1-1:1) to obtain the product 1-(1-(6-cyclopropylpyridin-3-yl)ethyl)-4 -(Propan-1-yn-1-yl)-1H-indazole-7-carboxylic acid methyl ester (180 mg, 44.3% yield).
LC-MS,M/Z(ESI):360.4[M+H] +。 LC-MS, M/Z (ESI): 360.4 [M+H] + .
第三步：1-(1-(6-环丙基吡啶-3-基)乙基)-4-(丙烷-1-炔-1-基)-1H-吲唑-7-甲酸的制备Step 3: Preparation of 1-(1-(6-cyclopropylpyridin-3-yl)ethyl)-4-(propane-1-yn-1-yl)-1H-indazole-7-carboxylic acid
将1-(1-(6-环丙基吡啶-3-基)乙基)-4-(丙烷-1-炔-1-基)-1H-吲唑-7-甲酸甲酯(160mg，0.445mmol)溶于四氢呋喃(3mL)、甲醇(1mL)和水(1mL)中，然后加入氢氧化锂(53.3mg，2.226mmol)，室温反应16h。向反应液中加入水(50mL),用1N的盐酸水溶液将pH调至5～6，并用乙酸乙酯(20mL*3)萃取。有机相合并后用饱和食盐水(10mL*2)洗涤，无水硫酸钠干燥，过滤，浓缩得到产物1-(1-(6-环丙基吡啶-3-基)乙基)-4-(丙烷-1-炔-1-基)-1H-吲唑-7-甲酸(150mg，产率98％)。Methyl 1-(1-(6-cyclopropylpyridin-3-yl)ethyl)-4-(propane-1-yn-1-yl)-1H-indazole-7-carboxylate (160 mg, 0.445 mmol) was dissolved in tetrahydrofuran (3 mL), methanol (1 mL) and water (1 mL), then lithium hydroxide (53.3 mg, 2.226 mmol) was added and reacted at room temperature for 16 h. Water (50 mL) was added to the reaction solution, the pH was adjusted to 5-6 with 1N aqueous hydrochloric acid, and extracted with ethyl acetate (20 mL*3). The combined organic phases were washed with saturated brine (10mL*2), dried over anhydrous sodium sulfate, filtered, and concentrated to obtain the product 1-(1-(6-cyclopropylpyridin-3-yl)ethyl)-4-( Propan-1-yn-1-yl)-1H-indazole-7-carboxylic acid (150 mg, 98% yield).
LC-MS,M/Z(ESI):346.3[M+H] +。 LC-MS, M/Z (ESI): 346.3 [M+H] + .
第四步：6-(1-(1-(6-环丙基吡啶-3-基)乙基)-4-(丙烷-1-炔-1-基)-1H-吲唑-7-甲酰胺基)螺[3.3]庚烷-2-甲酸甲酯的制备The fourth step: 6-(1-(1-(6-cyclopropylpyridin-3-yl)ethyl)-4-(propane-1-yn-1-yl)-1H-indazole-7-methan Preparation of amido)spiro[3.3]heptane-2-carboxylic acid methyl ester
将1-(1-(6-环丙基吡啶-3-基)乙基)-4-(丙烷-1-炔-1-基)-1H-吲唑-7-甲酸(150mg，0.434mmol)、6-氨基螺[3.3]庚烷-2-甲酸甲酯(184mg，1.086mmol)、二异丙基乙胺(168mg，1.303mmol)和HATU(495mg，1.303mmol)溶于N,N-二甲基甲酰胺(3mL)中，然后室温搅拌3h。将反应液加入水(50mL)中，用乙酸乙酯(10mL*3)萃取。有机相合并后用饱和食盐水(10mL*3)洗涤，无水硫酸钠干燥，过滤，浓缩得到粗品。粗品经过薄层硅胶板分离纯化(石油醚:乙酸乙酯(V/V)＝1:1)得到6-(1-(1-(6-环丙基吡啶-3-基)乙基)-4-(丙烷-1-炔-1-基)-1H-吲唑-7-甲酰胺基)螺[3.3]庚烷-2-甲酸甲酯(120mg，产率55.6％)。1-(1-(6-cyclopropylpyridin-3-yl)ethyl)-4-(propane-1-yn-1-yl)-1H-indazole-7-carboxylic acid (150mg, 0.434mmol) , methyl 6-aminospiro[3.3]heptane-2-carboxylate (184mg, 1.086mmol), diisopropylethylamine (168mg, 1.303mmol) and HATU (495mg, 1.303mmol) were dissolved in N,N-di Methylformamide (3 mL), then stirred at room temperature for 3 h. The reaction solution was added to water (50 mL), and extracted with ethyl acetate (10 mL*3). The combined organic phases were washed with saturated brine (10 mL*3), dried over anhydrous sodium sulfate, filtered, and concentrated to obtain a crude product. The crude product was separated and purified on a thin-layer silica gel plate (petroleum ether:ethyl acetate (V/V)=1:1) to obtain 6-(1-(1-(6-cyclopropylpyridin-3-yl)ethyl)- 4-(Propan-1-yn-1-yl)-1H-indazole-7-carboxamido)spiro[3.3]heptane-2-carboxylic acid methyl ester (120 mg, 55.6% yield).
LC-MS,M/Z(ESI):497.4[M+H] +。 LC-MS, M/Z (ESI): 497.4 [M+H] + .
第五步：(R)-6-(1-(1-(6-环丙基吡啶-3-基)乙基)-4-(丙烷-1-炔-1-基)-1H-吲唑-7-甲酰胺基)螺[3.3]庚烷-2-甲酸甲酯和(S)-6-(1-(1-(6-环丙基吡啶-3-基)乙基)-4-(丙烷-1-炔-1-基)-1H-吲唑-7-甲酰胺基)螺[3.3]庚烷-2-甲酸甲酯的制备The fifth step: (R)-6-(1-(1-(6-cyclopropylpyridin-3-yl)ethyl)-4-(propane-1-yn-1-yl)-1H-indazole -7-Carboxamido)spiro[3.3]heptane-2-carboxylic acid methyl ester and (S)-6-(1-(1-(6-cyclopropylpyridin-3-yl)ethyl)-4- Preparation of (propane-1-yn-1-yl)-1H-indazole-7-carboxamido)spiro[3.3]heptane-2-carboxylic acid methyl ester
将6-(1-(1-(6-环丙基吡啶-3-基)乙基)-4-(丙烷-1-炔-1-基)-1H-吲唑-7-甲酰胺基)螺[3.3]庚烷-2-甲酸甲酯(120mg，0.242mmol)经过拆分得到(Sa,R)-6-(1-(1-(6-环丙基吡啶-3-基)乙基)-4-(丙烷-1-炔-1-基)-1H-吲唑-7-甲酰胺基)螺[3.3]庚烷-2-甲酸甲酯(50mg，0.101mmol)和(Sa,S)-6-(1-(1-(6-环丙基吡啶-3-基)乙基)-4-(丙烷-1-炔-1-基)-1H-吲唑-7-甲酰胺基)螺[3.3]庚烷-2-甲酸甲酯(50mg，0.101mmol)。6-(1-(1-(6-cyclopropylpyridin-3-yl)ethyl)-4-(propane-1-yn-1-yl)-1H-indazole-7-carboxamido) Spiro[3.3]heptane-2-carboxylic acid methyl ester (120mg, 0.242mmol) was resolved to obtain (Sa,R)-6-(1-(1-(6-cyclopropylpyridin-3-yl)ethyl )-4-(propane-1-yn-1-yl)-1H-indazole-7-carboxamido)spiro[3.3]heptane-2-carboxylic acid methyl ester (50mg, 0.101mmol) and (Sa,S )-6-(1-(1-(6-cyclopropylpyridin-3-yl)ethyl)-4-(propane-1-yn-1-yl)-1H-indazole-7-carboxamide ) spiro[3.3]heptane-2-carboxylic acid methyl ester (50 mg, 0.101 mmol).
拆分方法:使用Column:Chiralpak AD-3型号手性拆分柱(规格为：50×4.6mm,3um)进行手性拆分，流动相A为超临界流体CO 2,流动相B为异丙醇(含0.05％二乙醇胺)；梯度条件:流动相B为40％，流动速率为3mL/min；柱温保持在35℃，柱压保持在100bar。其中，(R)-6-(1-(1-(6-环丙基吡啶-3-基)乙基)-4-(丙烷-1-炔-1-基)-1H-吲唑-7-甲酰胺基)螺[3.3]庚烷-2-甲酸甲酯保留时间为0.754min。(S)-6-(1-(1-(6-环丙基吡啶-3-基)乙基)-4-(丙烷-1-炔-1-基)-1H-吲唑-7-甲酰胺基)螺[3.3]庚烷-2-甲酸甲酯保留时间为1.448min。 Resolution method: Use Column: Chiralpak AD-3 type chiral resolution column (specification: 50×4.6mm, 3um) for chiral resolution, mobile phase A is supercritical fluid CO 2 , mobile phase B is isopropyl Alcohol (containing 0.05% diethanolamine); gradient conditions: mobile phase B is 40%, flow rate is 3mL/min; column temperature is maintained at 35°C, and column pressure is maintained at 100bar. Among them, (R)-6-(1-(1-(6-cyclopropylpyridin-3-yl)ethyl)-4-(propane-1-yn-1-yl)-1H-indazole-7 -Carboxamido)spiro[3.3]heptane-2-carboxylic acid methyl ester retention time is 0.754min. (S)-6-(1-(1-(6-cyclopropylpyridin-3-yl)ethyl)-4-(propane-1-yn-1-yl)-1H-indazole-7-methanol The retention time of amido)spiro[3.3]heptane-2-carboxylic acid methyl ester was 1.448min.
第六步：(Sa,R)-6-(1-(1-(6-环丙基吡啶-3-基)乙基)-4-(丙烷-1-炔-1-基)-1H-吲唑-7-甲酰胺基)螺[3.3]庚烷-2-甲酸(I-30a)的制备The sixth step: (Sa,R)-6-(1-(1-(6-cyclopropylpyridin-3-yl)ethyl)-4-(propane-1-yn-1-yl)-1H- Preparation of indazole-7-carboxamido)spiro[3.3]heptane-2-carboxylic acid (I-30a)
Figure PCTCN2022097611-appb-000088
Figure PCTCN2022097611-appb-000088
将(Sa,R)-6-(1-(1-(6-环丙基吡啶-3-基)乙基)-4-(丙烷-1-炔-1-基)-1H-吲唑-7-甲酰胺基)螺[3.3]庚烷-2-甲酸甲酯(50mg，0.101mmol)溶于四氢呋喃(3mL)、甲醇(1mL)和水(1mL)中，加入氢氧化锂(12.06mg，0.503mmol)，反应液在25℃搅拌16h。向反应液中加入水(50mL),然后用1N盐酸水溶液调pH至5～6，乙酸乙酯(20mL*3)萃取。有机相合并，用饱和食盐水(20mL*2)洗涤，用无水硫酸钠干燥，过滤，浓缩得到产物(Sa,R)-6-(1-(1-(6-环丙基吡啶-3-基)乙基)-4-(丙烷-1-炔-1-基)-1H-吲唑-7-甲酰胺基)螺[3.3]庚烷-2-甲酸(I-30a)(46.5mg，产率96％)。(Sa,R)-6-(1-(1-(6-cyclopropylpyridin-3-yl)ethyl)-4-(propane-1-yn-1-yl)-1H-indazole- 7-Carboxamido)spiro[3.3]heptane-2-carboxylic acid methyl ester (50 mg, 0.101 mmol) was dissolved in tetrahydrofuran (3 mL), methanol (1 mL) and water (1 mL), and lithium hydroxide (12.06 mg, 0.503mmol), the reaction solution was stirred at 25°C for 16h. Water (50 mL) was added to the reaction liquid, then the pH was adjusted to 5-6 with 1N aqueous hydrochloric acid solution, and extracted with ethyl acetate (20 mL*3). The organic phases were combined, washed with saturated brine (20mL*2), dried over anhydrous sodium sulfate, filtered, and concentrated to obtain the product (Sa,R)-6-(1-(1-(6-cyclopropylpyridine-3 -yl)ethyl)-4-(propane-1-yn-1-yl)-1H-indazole-7-carboxamido)spiro[3.3]heptane-2-carboxylic acid (I-30a) (46.5mg , yield 96%).
1H NMR(400MHz,dmso)δ11.98(s,1H),8.83(d,1H),8.25(s,1H),8.05(d,1H),7.31(d,1H),7.26–7.21(m,1H),7.20(d,1H),7.13(d,1H),6.23(q,1H),4.30(dq,1H),3.00–2.89(m,1H),2.47–2.40(m,1H),2.34–2.20(m,3H),2.18–2.12(m,3H),2.11–1.94(m,4H),1.90–1.79(m,4H),0.87(dt,2H),0.83–0.79(m,2H).1H NMR(400MHz,dmso)δ11.98(s,1H),8.83(d,1H),8.25(s,1H),8.05(d,1H),7.31(d,1H),7.26–7.21(m, 1H),7.20(d,1H),7.13(d,1H),6.23(q,1H),4.30(dq,1H),3.00–2.89(m,1H),2.47–2.40(m,1H),2.34 –2.20(m,3H),2.18–2.12(m,3H),2.11–1.94(m,4H),1.90–1.79(m,4H),0.87(dt,2H),0.83–0.79(m,2H) .
LC-MS,M/Z(ESI):483.3[M+H] +。 LC-MS, M/Z (ESI): 483.3 [M+H] + .
第七步：(Sa,S)-6-(1-(1-(6-环丙基吡啶-3-基)乙基)-4-(丙烷-1-炔-1-基)-1H-吲唑-7-甲酰胺基)螺[3.3]庚烷-2-甲酸(I-30b)的制备Step 7: (Sa,S)-6-(1-(1-(6-cyclopropylpyridin-3-yl)ethyl)-4-(propane-1-yn-1-yl)-1H- Preparation of indazole-7-carboxamido)spiro[3.3]heptane-2-carboxylic acid (I-30b)
Figure PCTCN2022097611-appb-000089
Figure PCTCN2022097611-appb-000089
将(Sa,S)-6-(1-(1-(6-环丙基吡啶-3-基)乙基)-4-(丙烷-1-炔-1-基)-1H-吲唑-7-甲酰胺基)螺[3.3]庚烷-2-甲酸甲酯(50mg，0.101mmol)溶于四氢呋喃(3mL)、甲醇(1mL)和水(1mL)中，加入氢氧化锂(12.06mg，0.503mmol)，反应液在25℃搅拌16h。向反应液中加入水(50mL),然后用1N盐酸水溶液调pH至5～6，乙酸乙酯(20mL*3)萃取。有机相合并，用饱和食盐水(20mL*2)洗涤，用无水硫酸钠干燥，过滤，浓缩得到产物(Sa,S)-6-(1-(1-(6-环丙基吡啶-3-基)乙基)-4-(丙烷-1-炔-1-基)-1H-吲唑-7-甲酰胺基)螺[3.3]庚烷-2-甲酸(I-30b)(47.3mg，产率97％)。(Sa,S)-6-(1-(1-(6-cyclopropylpyridin-3-yl)ethyl)-4-(propane-1-yn-1-yl)-1H-indazole- 7-Carboxamido)spiro[3.3]heptane-2-carboxylic acid methyl ester (50 mg, 0.101 mmol) was dissolved in tetrahydrofuran (3 mL), methanol (1 mL) and water (1 mL), and lithium hydroxide (12.06 mg, 0.503mmol), the reaction solution was stirred at 25°C for 16h. Water (50 mL) was added to the reaction liquid, then the pH was adjusted to 5-6 with 1N aqueous hydrochloric acid solution, and extracted with ethyl acetate (20 mL*3). The organic phases were combined, washed with saturated brine (20mL*2), dried over anhydrous sodium sulfate, filtered, and concentrated to obtain the product (Sa,S)-6-(1-(1-(6-cyclopropylpyridine-3 -yl)ethyl)-4-(propane-1-yn-1-yl)-1H-indazole-7-carboxamido)spiro[3.3]heptane-2-carboxylic acid (I-30b) (47.3mg , yield 97%).
1H NMR(400MHz,dmso)δ11.99(s,1H),8.82(d,1H),8.23(s,1H),8.03(d,1H),7.28(d,1H),7.23(dd,1H),7.17(d,1H),7.12(d,1H),6.22(q,1H),4.27(dq,1H),2.98–2.87(m,1H),2.46–2.36(m,1H),2.25(dd,3H),2.18–2.11(m,3H),2.09–1.82(m,8H),0.86(dt,2H),0.80(dt,2H). 1 H NMR (400MHz,dmso)δ11.99(s,1H),8.82(d,1H),8.23(s,1H),8.03(d,1H),7.28(d,1H),7.23(dd,1H ),7.17(d,1H),7.12(d,1H),6.22(q,1H),4.27(dq,1H),2.98–2.87(m,1H),2.46–2.36(m,1H),2.25( dd,3H),2.18–2.11(m,3H),2.09–1.82(m,8H),0.86(dt,2H),0.80(dt,2H).
LC-MS,M/Z(ESI):483.3[M+H] +。 LC-MS, M/Z (ESI): 483.3 [M+H] + .
实施例31化合物I-31a的制备Preparation of Example 31 Compound I-31a
合成路线如下所示：The synthetic route is as follows:
Figure PCTCN2022097611-appb-000090
Figure PCTCN2022097611-appb-000090
第一步：1-(2-环丙基嘧啶-5-基)-乙烷-1-醇的制备Step 1: Preparation of 1-(2-cyclopropylpyrimidin-5-yl)-ethan-1-ol
室温下将1-(2-氯嘧啶-5-基)-乙烷-1-醇(2g，12.6mmol)，四三苯基磷钯(1.46g，1.26mmol)，环丙基硼酸(10.8g，126mmol)，氟化钾(7.3g，126mmol)加入到20mL甲苯和4mL水中，置换氮气，100℃下反应过夜。反应液浓缩，柱层析纯化得到1-(2-环丙基嘧啶-5-基)-乙烷-1-醇(800mg，产率39％)。1-(2-chloropyrimidin-5-yl)-ethan-1-ol (2g, 12.6mmol), tetrakistriphenylphosphopalladium (1.46g, 1.26mmol), cyclopropylboronic acid (10.8g , 126mmol), potassium fluoride (7.3g, 126mmol) were added to 20mL of toluene and 4mL of water, replaced with nitrogen, and reacted overnight at 100°C. The reaction solution was concentrated and purified by column chromatography to obtain 1-(2-cyclopropylpyrimidin-5-yl)-ethan-1-ol (800 mg, yield 39%).
第二步：1-(1-(2-环丙基嘧啶-5-基)乙基)-4-(丙烷-1-炔-1-基)-1H-吲唑-7-甲酸甲酯的制备The second step: 1-(1-(2-cyclopropylpyrimidin-5-yl) ethyl)-4-(propane-1-yn-1-yl)-1H-indazole-7-carboxylic acid methyl ester preparation
1-(2-环丙基嘧啶-5-基)-乙烷-1-醇(537mg，3.27mmol)、甲基4-(丙烷-1-炔-1-基)-1H-吲唑-7-甲酸酯(280mg，1.31mmol)和三苯基膦(1.37g，5.23mmol)溶于四氢呋喃(10mL)中，冰浴下缓慢滴加偶氮二异丙酯(1.06g，5.23mmol)，随后升至室温搅拌16小时。反应液浓缩，柱层析纯化得1-(1-(2-环丙基嘧啶-5-基)乙基)-4-(丙烷-1-炔-1-基)-1H-吲唑-7-甲酸甲酯(200mg，产率43％)。1-(2-cyclopropylpyrimidin-5-yl)-ethan-1-ol (537mg, 3.27mmol), methyl 4-(propane-1-yn-1-yl)-1H-indazole-7 - formate (280mg, 1.31mmol) and triphenylphosphine (1.37g, 5.23mmol) were dissolved in tetrahydrofuran (10mL), and azobisisopropyl (1.06g, 5.23mmol) was slowly added dropwise under ice-cooling, Then warm to room temperature and stir for 16 hours. The reaction solution was concentrated and purified by column chromatography to obtain 1-(1-(2-cyclopropylpyrimidin-5-yl)ethyl)-4-(propane-1-yn-1-yl)-1H-indazole-7 - Methyl formate (200 mg, 43% yield).
第三步：1-(1-(2-环丙基嘧啶-5-基)乙基)-4-(丙烷-1-炔-1-基)-1H-吲唑-7-甲酸的制备Step 3: Preparation of 1-(1-(2-cyclopropylpyrimidin-5-yl)ethyl)-4-(propane-1-yn-1-yl)-1H-indazole-7-carboxylic acid
室温下将化合物1-(1-(2-环丙基嘧啶-5-基)乙基)-4-(丙烷-1-炔-1-基)-1H-吲唑-7-甲酸甲酯(200mg,0.56mmol)加入到四氢呋喃(5mL)，甲醇(1mL)和水(1mL)中，加入氢氧化锂(66mg,2.80mmol)，搅拌16h。加入水(20mL)稀释，用1mol/L的盐酸调节pH为4，用乙酸乙酯(20mL×3)萃取，分液，合并有机相，有机相用无水硫酸钠干燥，过滤，浓缩，残留物用薄层硅胶板分离纯化(石油醚:乙酸乙酯(V/V)＝1:1)得化合物1-(1-(2-环丙基嘧啶-5-基)乙基)-4-(丙烷-1-炔-1-基)-1H-吲唑-7-甲酸(190mg,产率99％)。Compound 1-(1-(2-cyclopropylpyrimidin-5-yl)ethyl)-4-(propane-1-yn-1-yl)-1H-indazole-7-carboxylic acid methyl ester ( 200mg, 0.56mmol) was added to tetrahydrofuran (5mL), methanol (1mL) and water (1mL), lithium hydroxide (66mg, 2.80mmol) was added, and stirred for 16h. Add water (20mL) for dilution, adjust the pH to 4 with 1mol/L hydrochloric acid, extract with ethyl acetate (20mL×3), separate the layers, combine the organic phases, dry the organic phases with anhydrous sodium sulfate, filter, concentrate, and leave The material was separated and purified on a thin-layer silica gel plate (petroleum ether:ethyl acetate (V/V)=1:1) to obtain compound 1-(1-(2-cyclopropylpyrimidin-5-yl)ethyl)-4- (Propan-1-yn-1-yl)-1H-indazole-7-carboxylic acid (190 mg, 99% yield).
第四步：(Sa)-6-(1-(1-(2-环丙基嘧啶-5-基)乙基)-4-(丙烷-1-炔-1-基)-1H-吲唑-7-甲酰胺)螺环[3.3]庚烷-2-甲酸甲酯的制备The fourth step: (Sa)-6-(1-(1-(2-cyclopropylpyrimidin-5-yl)ethyl)-4-(propane-1-yn-1-yl)-1H-indazole Preparation of -7-carboxamide) spiro[3.3]heptane-2-carboxylic acid methyl ester
室温下将化合物1-(1-(2-环丙基嘧啶-5-基)乙基)-4-(丙烷-1-炔-1-基)-1H-吲唑-7-甲酸(190mg,0.55mmol)加入到N,N-二甲基甲酰胺(5mL)中，加入(Sa)-6-氨基螺环[3.3]庚-2-基-甲酸甲酯盐酸盐(280mg,1.65mmol)，二异丙基乙胺(354mg,2.74mmol)和HATU(417mg,1.10mmol)，室温搅拌16h。加入水(50mL)稀释，用乙酸乙酯(20mL×3)萃取，分液，合并有机相，有机相用无水硫酸钠干燥，过滤，浓缩，残留物用薄层硅胶板分离纯化(石油醚:乙酸乙酯(V/V)＝1:1) 得化合物(Sa)-6-(1-(1-(2-环丙基嘧啶-5-基)乙基)-4-(丙烷-1-炔-1-基)-1H-吲唑-7-甲酰胺)螺环[3.3]庚烷-2-甲酸甲酯(150mg,产率55％)。Compound 1-(1-(2-cyclopropylpyrimidin-5-yl)ethyl)-4-(propane-1-yn-1-yl)-1H-indazole-7-carboxylic acid (190mg, 0.55mmol) was added to N,N-dimethylformamide (5mL), and (Sa)-6-aminospiro[3.3]hept-2-yl-formic acid methyl ester hydrochloride (280mg, 1.65mmol) was added , diisopropylethylamine (354mg, 2.74mmol) and HATU (417mg, 1.10mmol), stirred at room temperature for 16h. Add water (50mL) to dilute, extract with ethyl acetate (20mL×3), separate the layers, combine the organic phases, dry the organic phases with anhydrous sodium sulfate, filter, concentrate, and separate and purify the residue with a thin-layer silica gel plate (petroleum ether : ethyl acetate (V/V)=1:1) to obtain compound (Sa)-6-(1-(1-(2-cyclopropylpyrimidin-5-yl) ethyl)-4-(propane-1 -Alkyn-1-yl)-1H-indazole-7-carboxamide)spiro[3.3]heptane-2-carboxylic acid methyl ester (150 mg, 55% yield).
第五步：(Sa,R)-6-(1-(1-(2-环丙基嘧啶-5-基)乙基)-4-(丙烷-1-炔-1-基)-1H-吲唑-7-甲酰胺)螺环[3.3]庚烷-2-甲酸甲酯的制备The fifth step: (Sa,R)-6-(1-(1-(2-cyclopropylpyrimidin-5-yl)ethyl)-4-(propane-1-yn-1-yl)-1H- Preparation of indazole-7-carboxamide) spiro[3.3]heptane-2-carboxylate methyl ester
将化合物(Sa)-6-(1-(1-(2-环丙基嘧啶-5-基)乙基)-4-(丙烷-1-炔-1-基)-1H-吲唑-7-甲酰胺)螺环[3.3]庚烷-2-甲酸甲酯(150mg,0.30mmol)进行手性拆分方法(IC-3-IPA(DEA)-5-40-3ML-35T，r.t.＝1.969min)制备得化合物(Sa,R)-6-(1-(1-(2-环丙基嘧啶-5-基)乙基)-4-(丙烷-1-炔-1-基)-1H-吲唑-7-甲酰胺)螺环[3.3]庚烷-2-甲酸甲酯(50mg,产率33％)。Compound (Sa)-6-(1-(1-(2-cyclopropylpyrimidin-5-yl)ethyl)-4-(propane-1-yn-1-yl)-1H-indazole-7 -Carboxamide) spiro[3.3]heptane-2-methyl carboxylate (150mg, 0.30mmol) for chiral resolution (IC-3-IPA(DEA)-5-40-3ML-35T, r.t.=1.969 min) Compound (Sa,R)-6-(1-(1-(2-cyclopropylpyrimidin-5-yl)ethyl)-4-(propane-1-yn-1-yl)-1H -indazole-7-carboxamide)spiro[3.3]heptane-2-carboxylic acid methyl ester (50 mg, 33% yield).
第六步：(Sa,R)-6-(1-(1-(2-环丙基嘧啶-5-基)乙基)-4-(丙烷-1-炔-1-基)-1H-吲唑-7-甲酰胺)螺环[3.3]庚烷-2-甲酸(I-31a)The sixth step: (Sa,R)-6-(1-(1-(2-cyclopropylpyrimidin-5-yl)ethyl)-4-(propane-1-yn-1-yl)-1H- Indazole-7-carboxamide) spiro[3.3]heptane-2-carboxylic acid (I-31a)
Figure PCTCN2022097611-appb-000091
Figure PCTCN2022097611-appb-000091
室温下将化合物(Sa,R)-6-(1-(1-(2-环丙基嘧啶-5-基)乙基)-4-(丙烷-1-炔-1-基)-1H-吲唑-7-甲酰胺)螺环[3.3]庚烷-2-甲酸甲酯(50mg,0.10mmol)加入到四氢呋喃(3mL)，甲醇(1mL)和水(1mL)中，加入氢氧化锂(12mg,0.50mmol)，室温搅拌16h。用1N盐酸调节pH＝4，用乙酸乙酯(10mL×3)萃取，分液，合并有机相，有机相用无水硫酸钠干燥，过滤，浓缩，残留物经酸性制备方法A制备得化合物(Sa,R)-6-(1-(1-(2-环丙基嘧啶-5-基)乙基)-4-(丙烷-1-炔-1-基)-1H-吲唑-7-甲酰胺)螺环[3.3]庚烷-2-甲酸(I-31a)(45mg,产率93％)。Compound (Sa,R)-6-(1-(1-(2-cyclopropylpyrimidin-5-yl)ethyl)-4-(propane-1-yn-1-yl)-1H- Indazole-7-carboxamide) spiro[3.3]heptane-2-carboxylate (50 mg, 0.10 mmol) was added to tetrahydrofuran (3 mL), methanol (1 mL) and water (1 mL), and lithium hydroxide ( 12mg, 0.50mmol), stirred at room temperature for 16h. Use 1N hydrochloric acid to adjust pH=4, extract with ethyl acetate (10mL×3), separate the layers, combine the organic phases, dry the organic phases with anhydrous sodium sulfate, filter, concentrate, and the residue is prepared by acidic preparation method A to obtain the compound ( Sa,R)-6-(1-(1-(2-cyclopropylpyrimidin-5-yl)ethyl)-4-(propane-1-yn-1-yl)-1H-indazol-7- Formamide) spiro[3.3]heptane-2-carboxylic acid (I-31a) (45 mg, 93% yield).
1H NMR(400mHz,DMSO-d6)δ12.01(s,1H),8.65(d,1H),8.26-8.24(m,3H),7.33(m,1H),7.21(m,1H),6.21(m,1H),4.29(m,1H),2.94(m,1H),2.49(m,1H),2.48-2.18(m,14H),0.97-0.95(m,2H),0.91-0.88(m,2H)。 1 H NMR(400mHz,DMSO-d6)δ12.01(s,1H),8.65(d,1H),8.26-8.24(m,3H),7.33(m,1H),7.21(m,1H),6.21 (m,1H),4.29(m,1H),2.94(m,1H),2.49(m,1H),2.48-2.18(m,14H),0.97-0.95(m,2H),0.91-0.88(m ,2H).
LC-MS,M/Z(ESI):484.30[M+H] +。 LC-MS, M/Z (ESI): 484.30 [M+H] + .
化合物I-31b的制备Preparation of compound I-31b
合成路线如下所示The synthetic route is as follows
Figure PCTCN2022097611-appb-000092
Figure PCTCN2022097611-appb-000092
第一步：(Sa)-6-(1-(1-(2-环丙基嘧啶-5-基)乙基)-4-(丙烷-1-炔-1-基)-1H-吲唑-7-甲酰胺)螺环[3.3]庚烷-2-甲酸甲酯的制备The first step: (Sa)-6-(1-(1-(2-cyclopropylpyrimidin-5-yl)ethyl)-4-(propane-1-yn-1-yl)-1H-indazole Preparation of -7-carboxamide) spiro[3.3]heptane-2-carboxylic acid methyl ester
将化合物(Sa)-6-(1-(1-(2-环丙基嘧啶-5-基)乙基)-4-(丙烷-1-炔-1-基)-1H-吲唑-7-甲酰胺)螺环[3.3]庚烷-2-甲酸甲酯(150mg,0.30mmol)进行手性拆分方法(IC-3-IPA(DEA)-5-40-3ML-35T，r.t.＝2.552min)制备得化合物(Sa,S)-6-(1-(1-(2-环丙基嘧啶-5-基)乙基)-4-(丙烷-1-炔-1-基)-1H-吲唑-7-甲酰胺)螺环[3.3]庚烷-2-甲酸甲酯(50mg，产率33％)。Compound (Sa)-6-(1-(1-(2-cyclopropylpyrimidin-5-yl)ethyl)-4-(propane-1-yn-1-yl)-1H-indazole-7 -formamide) spiro[3.3]heptane-2-carboxylic acid methyl ester (150mg, 0.30mmol) was carried out chiral resolution method (IC-3-IPA(DEA)-5-40-3ML-35T, r.t.=2.552 min) prepared compound (Sa, S)-6-(1-(1-(2-cyclopropylpyrimidin-5-yl)ethyl)-4-(propane-1-yn-1-yl)-1H -indazole-7-carboxamide) spiro[3.3]heptane-2-carboxylic acid methyl ester (50 mg, 33% yield).
第二步：(Sa,S)-6-(1-(1-(2-环丙基嘧啶-5-基)乙基)-4-(丙烷-1-炔-1-基)-1H-吲唑-7-甲酰胺)螺环[3.3]庚烷-2-甲酸(I-31b)The second step: (Sa,S)-6-(1-(1-(2-cyclopropylpyrimidin-5-yl)ethyl)-4-(propane-1-yn-1-yl)-1H- Indazole-7-carboxamide) spiro[3.3]heptane-2-carboxylic acid (I-31b)
Figure PCTCN2022097611-appb-000093
Figure PCTCN2022097611-appb-000093
室温下将化合物(Sa,S)-6-(1-(1-(2-环丙基嘧啶-5-基)乙基)-4-(丙烷-1-炔-1-基)-1H-吲唑-7-甲酰胺)螺环[3.3]庚烷-2-甲酸甲酯(50mg,0.10mmol)加入到四氢呋喃(3mL)，甲醇(1mL)和水(1mL)中，加入氢氧化锂(12mg,0.50mmol)，室温搅拌16h。用1N盐酸调节pH＝4，用乙酸乙酯(10mL×3)萃取，分液，合并有机相，有机相用无水硫酸钠干燥，过滤，浓缩，残留物经酸性制备方法A制备得化合物(Sa,S)-6-(1-(1-(2-环丙基嘧啶-5-基)乙基)-4-(丙烷-1-炔-1-基)-1H-吲唑-7-甲酰胺)螺环[3.3]庚烷-2-甲酸(I-31b)(45mg，产率93％)。Compound (Sa,S)-6-(1-(1-(2-cyclopropylpyrimidin-5-yl)ethyl)-4-(propane-1-yn-1-yl)-1H- Indazole-7-carboxamide) spiro[3.3]heptane-2-carboxylate (50 mg, 0.10 mmol) was added to tetrahydrofuran (3 mL), methanol (1 mL) and water (1 mL), and lithium hydroxide ( 12mg, 0.50mmol), stirred at room temperature for 16h. Use 1N hydrochloric acid to adjust pH=4, extract with ethyl acetate (10mL×3), separate the layers, combine the organic phases, dry the organic phases with anhydrous sodium sulfate, filter, concentrate, and the residue is prepared by acidic preparation method A to obtain the compound ( Sa,S)-6-(1-(1-(2-cyclopropylpyrimidin-5-yl)ethyl)-4-(propane-1-yn-1-yl)-1H-indazol-7- Formamide) spiro[3.3]heptane-2-carboxylic acid (I-31b) (45 mg, 93% yield).
1H NMR(400mHz,DMSO-d6)δ12.03(s,1H),8.88(d,1H),8.28-8.27(m,3H),7.34(m,1H),7.23(m,1H),6.25(m,1H),4.31(m,1H),2.99(m,1H),2.28-1.88(m,15H),1.01-0.96(m,2H),0.93-0.90(m,2H)。 1 H NMR(400mHz,DMSO-d6)δ12.03(s,1H),8.88(d,1H),8.28-8.27(m,3H),7.34(m,1H),7.23(m,1H),6.25 (m,1H), 4.31(m,1H), 2.99(m,1H), 2.28-1.88(m,15H), 1.01-0.96(m,2H), 0.93-0.90(m,2H).
LC-MS,M/Z(ESI):484.30[M+H] +。 LC-MS, M/Z (ESI): 484.30 [M+H] + .
实施例32化合物I-32a、I-32b的制备The preparation of embodiment 32 compound I-32a, I-32b
合成路线如下所示：The synthetic route is as follows:
Figure PCTCN2022097611-appb-000094
Figure PCTCN2022097611-appb-000094
Figure PCTCN2022097611-appb-000095
Figure PCTCN2022097611-appb-000095
第一步：1-(1-(5-溴嘧啶-2-基)乙基)-4-(丙烷-1-炔-1-基)-1H-7-吲唑-7-甲酸甲酯的制备The first step: 1-(1-(5-bromopyrimidin-2-yl)ethyl)-4-(propane-1-yn-1-yl)-1H-7-indazole-7-carboxylic acid methyl ester preparation
将4-丙炔基-1H-7-吲唑甲酯(563mg，2.63mmol)，1-(5-溴嘧啶-2-基)乙醇(800mg，3.94mmol)和三苯基膦(2067mg，7.88mmol)溶于四氢呋喃(15mL)中，在0℃下缓慢滴入偶氮二甲酸二异丙酯(1593mg，7.88mmol)，然后在25℃下反应16h。将反应液浓缩得到粗品。粗品经过柱层析(石油醚/乙酸乙酯(V/V)＝10:1-1:1)得到产物1-(1-(5-溴嘧啶-2-基)乙基)-4-(丙烷-1-炔-1-基)-1H-7-吲唑-7-甲酸甲酯(600mg，产率57.2％)。4-propynyl-1H-7-indazole methyl ester (563mg, 2.63mmol), 1-(5-bromopyrimidin-2-yl)ethanol (800mg, 3.94mmol) and triphenylphosphine (2067mg, 7.88 mmol) was dissolved in tetrahydrofuran (15 mL), and diisopropyl azodicarboxylate (1593 mg, 7.88 mmol) was slowly added dropwise at 0 °C, and then reacted at 25 °C for 16 h. The reaction solution was concentrated to obtain a crude product. The crude product was subjected to column chromatography (petroleum ether/ethyl acetate (V/V)=10:1-1:1) to obtain the product 1-(1-(5-bromopyrimidin-2-yl)ethyl)-4-( Propan-1-yn-1-yl)-1H-7-indazole-7-carboxylic acid methyl ester (600 mg, 57.2% yield).
LC-MS,M/Z(ESI):400.3[M+H] +。 LC-MS, M/Z (ESI): 400.3 [M+H] + .
第二步：1-(1-(5-环丙基嘧啶-2-基)乙基)-4-(丙烷-1-炔-1-基)-1H-吲唑-7-甲酸甲酯的制备The second step: 1-(1-(5-cyclopropylpyrimidin-2-yl) ethyl)-4-(propane-1-yn-1-yl)-1H-indazole-7-carboxylic acid methyl ester preparation
将1-(1-(5-溴嘧啶-2-基)乙基)-4-(丙烷-1-炔-1-基)-1H-7-吲唑-7-甲酸甲酯(600mg，1.503mmol)、环丙基硼酸(645mg，7.51mmol)、氟化钾(262mg，4.51mmol)和四(三苯基膦)钯(174mg，0.150mmol)溶于甲苯(10mL)和水(2mL)中，然后在氮气保护下升至100℃反应16h。将反应液倒入水(50mL)中，用乙酸乙酯(20mL*3)萃取。有机相合并，用饱和食盐水(20mL*2)洗涤，用无水硫酸钠干燥，过滤浓缩得到粗品。粗品经过柱层析(乙酸乙酯：石油醚(V/V)＝10:1-1:1)得到产物1-(1-(5-环丙基嘧啶-2-基)乙基)-4-(丙烷-1-炔-1-基)-1H-吲唑-7-甲酸甲酯(400mg，产率73.8％)。1-(1-(5-bromopyrimidin-2-yl)ethyl)-4-(propane-1-yn-1-yl)-1H-7-indazole-7-carboxylic acid methyl ester (600mg, 1.503 mmol), cyclopropylboronic acid (645 mg, 7.51 mmol), potassium fluoride (262 mg, 4.51 mmol), and tetrakis(triphenylphosphine)palladium (174 mg, 0.150 mmol) were dissolved in toluene (10 mL) and water (2 mL) , and then raised to 100°C for 16h under the protection of nitrogen. The reaction solution was poured into water (50 mL), extracted with ethyl acetate (20 mL*3). The organic phases were combined, washed with saturated brine (20 mL*2), dried over anhydrous sodium sulfate, filtered and concentrated to obtain a crude product. The crude product was subjected to column chromatography (ethyl acetate:petroleum ether (V/V)=10:1-1:1) to obtain the product 1-(1-(5-cyclopropylpyrimidin-2-yl)ethyl)-4 -(Propan-1-yn-1-yl)-1H-indazole-7-carboxylic acid methyl ester (400 mg, 73.8% yield).
LC-MS,M/Z(ESI):360.2[M+H] +。 LC-MS, M/Z (ESI): 360.2 [M+H] + .
第三步：1-(1-(5-环丙基嘧啶-2-基)乙基)-4-(丙烷-1-炔-1-基)-1H-吲唑-7-甲酸的制备Step 3: Preparation of 1-(1-(5-cyclopropylpyrimidin-2-yl)ethyl)-4-(propane-1-yn-1-yl)-1H-indazole-7-carboxylic acid
将1-(1-(5-环丙基嘧啶-2-基)乙基)-4-(丙烷-1-炔-1-基)-1H-吲唑-7-甲酸甲酯(400mg，1.11mmol)溶于四氢呋喃(6mL)、甲醇(2mL)和水(2mL)中，然后加入氢氧化锂(133mg，5.55mmol)，室温反应16h。向反应液中加入水(50mL),用1N的盐酸水溶液将pH调至5～6并用乙酸乙酯(20mL*3)萃取。有机相合并后用饱和食盐水(10mL*2)洗涤，无水硫酸钠干燥，过滤，浓缩得到产物1-(1-(5-环丙基嘧啶-2-基)乙基)-4-(丙烷-1-炔-1-基)-1H-吲唑-7-甲酸(320mg，产率83％)。1-(1-(5-cyclopropylpyrimidin-2-yl)ethyl)-4-(propane-1-yn-1-yl)-1H-indazole-7-carboxylic acid methyl ester (400mg, 1.11 mmol) was dissolved in tetrahydrofuran (6 mL), methanol (2 mL) and water (2 mL), then lithium hydroxide (133 mg, 5.55 mmol) was added and reacted at room temperature for 16 h. Water (50 mL) was added to the reaction solution, the pH was adjusted to 5-6 with 1N aqueous hydrochloric acid and extracted with ethyl acetate (20 mL*3). The combined organic phases were washed with saturated brine (10mL*2), dried over anhydrous sodium sulfate, filtered, and concentrated to obtain the product 1-(1-(5-cyclopropylpyrimidin-2-yl)ethyl)-4-( Propan-1-yn-1-yl)-1H-indazole-7-carboxylic acid (320 mg, 83% yield).
LC-MS,M/Z(ESI):347.3[M+H] +。 LC-MS, M/Z (ESI): 347.3 [M+H] + .
第四步：(Sa)-6-(1-(1-(5-环丙基嘧啶-2-基)乙基)-4-(丙烷-1-炔-1-基)-1H-吲唑-7-甲酰胺基)螺[3.3]庚烷-2-甲酸甲酯的制备The fourth step: (Sa)-6-(1-(1-(5-cyclopropylpyrimidin-2-yl)ethyl)-4-(propane-1-yn-1-yl)-1H-indazole Preparation of -7-formamido)spiro[3.3]heptane-2-carboxylic acid methyl ester
将1-(1-(5-环丙基嘧啶-2-基)乙基)-4-(丙烷-1-炔-1-基)-1H-吲唑-7-甲酸(320mg，0.924mmol)、6-氨基螺[3.3]庚烷-2-甲酸甲酯(391mg，2.310mmol)、二异丙基乙胺(358mg，2.77mmol)和HATU(1054mg，2.77mmol)溶于N,N-二甲基甲酰胺(5mL)中，然后室温搅拌3h。将反应液加入水(50mL)中，用乙酸乙酯(10mL*3)萃取。有机相合并后用饱和食盐水(10mL*3)洗涤，无水硫酸钠干燥，过滤，浓缩得到粗品。粗品经过薄层硅胶板分离纯化(石油醚:乙酸乙酯(V/V)＝1:1)得到(Sa)-6-(1-(1-(5-环丙基嘧啶-2-基)乙基)-4-(丙烷-1-炔-1-基)-1H-吲唑-7甲酰胺基)螺[3.3]庚烷-2-甲酸甲酯(350mg，产率76％)。1-(1-(5-cyclopropylpyrimidin-2-yl)ethyl)-4-(propane-1-yn-1-yl)-1H-indazole-7-carboxylic acid (320mg, 0.924mmol) , methyl 6-aminospiro[3.3]heptane-2-carboxylate (391mg, 2.310mmol), diisopropylethylamine (358mg, 2.77mmol) and HATU (1054mg, 2.77mmol) were dissolved in N,N-di Methylformamide (5 mL), then stirred at room temperature for 3 h. The reaction solution was added to water (50 mL), and extracted with ethyl acetate (10 mL*3). The combined organic phases were washed with saturated brine (10 mL*3), dried over anhydrous sodium sulfate, filtered, and concentrated to obtain a crude product. The crude product was separated and purified on a thin-layer silica gel plate (petroleum ether:ethyl acetate (V/V)=1:1) to obtain (Sa)-6-(1-(1-(5-cyclopropylpyrimidin-2-yl) Ethyl)-4-(propan-1-yn-1-yl)-1H-indazole-7carboxamido)spiro[3.3]heptane-2-carboxylic acid methyl ester (350 mg, 76% yield).
LC-MS,M/Z(ESI):498.4[M+H] +。 LC-MS, M/Z (ESI): 498.4 [M+H] + .
第五步：(Sa,R)-6-(1-(1-(5-环丙基嘧啶-2-基)乙基)-4-(丙烷-1-炔-1-基)-1H-吲唑-7-甲酰胺基)螺[3.3]庚烷-2-甲酸甲酯和(Sa,S)-6-(1-(1-(5-环丙基嘧啶-2-基)乙基)-4-(丙烷-1-炔-1-基)-1H-吲唑-7-甲酰胺基)螺[3.3]庚烷-2-甲酸甲酯的制备The fifth step: (Sa,R)-6-(1-(1-(5-cyclopropylpyrimidin-2-yl)ethyl)-4-(propane-1-yn-1-yl)-1H- Indazole-7-carboxamido)spiro[3.3]heptane-2-carboxylate methyl ester and (Sa,S)-6-(1-(1-(5-cyclopropylpyrimidin-2-yl)ethyl Preparation of )-4-(propane-1-yn-1-yl)-1H-indazole-7-carboxamido)spiro[3.3]heptane-2-carboxylic acid methyl ester
Figure PCTCN2022097611-appb-000096
Figure PCTCN2022097611-appb-000096
将6-(1-(1-(5-环丙基嘧啶-2-基)乙基)-4-(丙烷-1-炔-1-基)-1H-吲唑-7-甲酰胺基)螺[3.3]庚烷-2-甲酸甲酯(350mg，0.703mmol)经过SFC拆分得到(R)-6-(1-(1-(5-环丙基嘧啶-2-基)乙基)-4-(丙烷-1-炔-1-基)-1H-吲唑-7-甲酰胺基)螺[3.3]庚烷-2-甲酸甲酯(80mg，0.161mmol)和(S)-6-(1-(1-(5-环丙基嘧啶-2-基)乙基)-4-(丙烷-1-炔-1-基)-1H-吲唑-7-甲酰胺基)螺[3.3]庚烷-2-甲酸甲酯(80mg，0.161mmol)。SFC拆分方法：使用Kromasil(S,S)Whelk-O1型号手性拆分柱(规格为：50×4.6mm,3.5um)进行手性拆分，流动相A为超临界流体CO 2,流动相B为乙醇(含0.05％二乙醇胺)；梯度条件:流动相B从5％to 80％，流动速率为10mL/min；40％的梯度保持10min，50％的梯度保持10min，柱温保持在35℃，柱压保持在100bar。 6-(1-(1-(5-cyclopropylpyrimidin-2-yl)ethyl)-4-(propane-1-yn-1-yl)-1H-indazole-7-carboxamido) Methyl spiro[3.3]heptane-2-carboxylate (350mg, 0.703mmol) was resolved by SFC to obtain (R)-6-(1-(1-(5-cyclopropylpyrimidin-2-yl)ethyl) -4-(propane-1-yn-1-yl)-1H-indazole-7-carboxamido)spiro[3.3]heptane-2-carboxylic acid methyl ester (80mg, 0.161mmol) and (S)-6 -(1-(1-(5-cyclopropylpyrimidin-2-yl)ethyl)-4-(propane-1-yn-1-yl)-1H-indazole-7-carboxamido)spiro[ 3.3] Heptane-2-carboxylic acid methyl ester (80 mg, 0.161 mmol). SFC resolution method: use Kromasil (S, S) Whelk-O1 type chiral resolution column (specification: 50×4.6mm, 3.5um) for chiral resolution, mobile phase A is supercritical fluid CO 2 , flow Phase B is ethanol (containing 0.05% diethanolamine); Gradient conditions: mobile phase B is from 5% to 80%, flow rate is 10mL/min; 40% gradient keeps 10min, 50% gradient keeps 10min, column temperature keeps at At 35°C, the column pressure was maintained at 100 bar.
(Sa,R)-6-(1-(1-(5-环丙基嘧啶-2-基)乙基)-4-(丙烷-1-炔-1-基)-1H-吲唑-7-甲酰胺基)螺[3.3]庚烷-2-甲酸甲酯的保留时间：1.045min。(Sa,R)-6-(1-(1-(5-cyclopropylpyrimidin-2-yl)ethyl)-4-(propane-1-yn-1-yl)-1H-indazole-7 Retention time of methyl -formamido)spiro[3.3]heptane-2-carboxylate: 1.045 min.
(Sa,S)-6-(1-(1-(5-环丙基嘧啶-2-基)乙基)-4-(丙烷-1-炔-1-基)-1H-吲唑-7-甲酰胺基)螺[3.3]庚烷-2-甲酸甲酯的保留时间：1.796min。(Sa,S)-6-(1-(1-(5-cyclopropylpyrimidin-2-yl)ethyl)-4-(propane-1-yn-1-yl)-1H-indazole-7 Retention time of methyl -formamido)spiro[3.3]heptane-2-carboxylate: 1.796 min.
第六步：(Sa,R)-6-(1-(1-(5-环丙基嘧啶-2-基)乙基)-4-(丙烷-1-炔-1-基)-1H-吲唑-7-甲酰胺基)螺[3.3]庚烷-2-甲酸(I-32a)The sixth step: (Sa,R)-6-(1-(1-(5-cyclopropylpyrimidin-2-yl)ethyl)-4-(propane-1-yn-1-yl)-1H- Indazole-7-carboxamido)spiro[3.3]heptane-2-carboxylic acid (I-32a)
Figure PCTCN2022097611-appb-000097
Figure PCTCN2022097611-appb-000097
将(Sa,R)-6-(1-(1-(5-环丙基嘧啶-2-基)乙基)-4-(丙烷-1-炔-1-基)-1H-吲唑-7-甲酰胺基)螺[3.3]庚烷-2-甲酸甲酯(80mg，0.161mmol)溶于四氢呋喃(3mL)、甲醇(1mL)和水(1mL)中，加入氢氧化锂(19.25mg，0.804mmol)，反应液在25℃搅拌16h。向反应液中加入水(50mL),然后用1N盐酸水溶液调pH至5～6，乙酸乙酯(20mL*3)萃取。有机相合并，用饱和食盐水(20mL*2)洗涤，用无水硫酸钠干燥，过滤，浓缩得到产物(Sa,R)-6-(1-(1-(5-环丙基嘧啶-2-基)乙基)-4-(丙烷-1-炔-1-基)-1H-吲唑-7-甲酰胺基)螺[3.3]庚烷-2-甲酸(I-32a)(33.9mg，产率43％)。(Sa,R)-6-(1-(1-(5-cyclopropylpyrimidin-2-yl)ethyl)-4-(propane-1-yn-1-yl)-1H-indazole- Methyl 7-carboxamido)spiro[3.3]heptane-2-carboxylate (80 mg, 0.161 mmol) was dissolved in tetrahydrofuran (3 mL), methanol (1 mL) and water (1 mL), and lithium hydroxide (19.25 mg, 0.804mmol), the reaction solution was stirred at 25°C for 16h. Water (50 mL) was added to the reaction liquid, then the pH was adjusted to 5-6 with 1N aqueous hydrochloric acid solution, and extracted with ethyl acetate (20 mL*3). The organic phases were combined, washed with saturated brine (20mL*2), dried over anhydrous sodium sulfate, filtered, and concentrated to obtain the product (Sa,R)-6-(1-(1-(5-cyclopropylpyrimidine-2 -yl)ethyl)-4-(propane-1-yn-1-yl)-1H-indazole-7-carboxamido)spiro[3.3]heptane-2-carboxylic acid (I-32a) (33.9mg , yield 43%).
1H NMR(400MHz,dmso)δ12.00(s,1H),8.85(d,1H),8.25(d,3H),7.32(d,1H),7.20(d,1H),6.20(dd,1H),4.26(dd,1H),2.94-2.90(m,1H),2.49-1.86(m,15H),1.03-0.90(m,2H),0.82-0.66(m,2H). 1 H NMR (400MHz,dmso)δ12.00(s,1H),8.85(d,1H),8.25(d,3H),7.32(d,1H),7.20(d,1H),6.20(dd,1H ),4.26(dd,1H),2.94-2.90(m,1H),2.49-1.86(m,15H),1.03-0.90(m,2H),0.82-0.66(m,2H).
LC-MS,M/Z(ESI):484.5[M+H] +。 LC-MS, M/Z (ESI): 484.5 [M+H] + .
第七步：(Sa,S)-6-(1-(1-(5-环丙基嘧啶-2-基)乙基)-4-(丙烷-1-炔-1-基)-1H-吲唑-7-甲酰胺基)螺[3.3]庚烷-2-甲酸(I-32b)的制备The seventh step: (Sa,S)-6-(1-(1-(5-cyclopropylpyrimidin-2-yl)ethyl)-4-(propane-1-yn-1-yl)-1H- Preparation of indazole-7-carboxamido)spiro[3.3]heptane-2-carboxylic acid (I-32b)
Figure PCTCN2022097611-appb-000098
Figure PCTCN2022097611-appb-000098
将(Sa,S)-6-(1-(1-(5-环丙基嘧啶-2-基)乙基)-4-(丙烷-1-炔-1-基)-1H-吲唑-7-甲酰胺基)螺[3.3]庚烷-2-甲酸甲酯(80mg，0.161mmol)溶于四氢呋喃(3mL)、甲醇(1mL)和水(1mL)中，加入氢氧化锂(19.25mg，0.804mmol)，反应液在25℃搅拌16h。向反应液中加入水(50mL),然后用1N盐酸水溶液调pH至5～6，乙酸乙酯(20mL*3)萃取。有机相合并，用饱和食盐水(20mL*2)洗涤，用无水硫酸钠干燥，过滤，浓缩得到产物(Sa,S)-6-(1-(1-(5-环丙基嘧啶-2-基)乙基)-4-(丙烷-1-炔-1-基)-1H-吲唑-7-甲酰胺基)螺[3.3]庚烷-2-甲酸(I-32b)(35.9mg，产率45.6％)。(Sa,S)-6-(1-(1-(5-cyclopropylpyrimidin-2-yl)ethyl)-4-(propane-1-yn-1-yl)-1H-indazole- Methyl 7-carboxamido)spiro[3.3]heptane-2-carboxylate (80 mg, 0.161 mmol) was dissolved in tetrahydrofuran (3 mL), methanol (1 mL) and water (1 mL), and lithium hydroxide (19.25 mg, 0.804mmol), the reaction solution was stirred at 25°C for 16h. Water (50 mL) was added to the reaction liquid, then the pH was adjusted to 5-6 with 1N aqueous hydrochloric acid solution, and extracted with ethyl acetate (20 mL*3). The organic phases were combined, washed with saturated brine (20mL*2), dried over anhydrous sodium sulfate, filtered, and concentrated to obtain the product (Sa,S)-6-(1-(1-(5-cyclopropylpyrimidine-2 -yl)ethyl)-4-(propane-1-yn-1-yl)-1H-indazole-7-carboxamido)spiro[3.3]heptane-2-carboxylic acid (I-32b) (35.9mg , yield 45.6%).
1H NMR(400MHz,dmso)δ12.00(s,1H),8.84(d,1H),8.25(d,3H),7.33(d,1H),7.20(d,1H),6.22(dd,1H),4.27(dd,1H),2.93-2.90(m,1H),2.51-1.86(m,15H),1.02-0.90(m,2H),0.82-0.65(m,2H). 1 H NMR (400MHz,dmso)δ12.00(s,1H),8.84(d,1H),8.25(d,3H),7.33(d,1H),7.20(d,1H),6.22(dd,1H ),4.27(dd,1H),2.93-2.90(m,1H),2.51-1.86(m,15H),1.02-0.90(m,2H),0.82-0.65(m,2H).
LC-MS,M/Z(ESI):484.5[M+H] +。 LC-MS, M/Z (ESI): 484.5 [M+H] + .
实施例33化合物I-33的制备The preparation of embodiment 33 compound I-33
Figure PCTCN2022097611-appb-000099
Figure PCTCN2022097611-appb-000099
参考化合物I-19的合成方法。LC-MS,M/Z(ESI):528.2[M+H] +。 Refer to the synthetic method of compound I-19. LC-MS, M/Z (ESI): 528.2 [M+H] + .
实施例34化合物I-34a的制备The preparation of embodiment 34 compound I-34a
合成路线如下所示：The synthetic route is as follows:
Figure PCTCN2022097611-appb-000100
Figure PCTCN2022097611-appb-000100
第一步：1-(4-(二氟甲基)苯基)乙烷-1-酮的制备The first step: the preparation of 1-(4-(difluoromethyl)phenyl)ethan-1-one
1-溴-4-(二氟甲基)苯(1g,4.83mmol)，三丁基(1-乙氧基乙烯)锡(2.27g,6.28mmol)和双(三苯基磷)二氯化钯(0.17g,0.242mmol)于二氧六环(20mL)中，氮气保护下100℃搅拌12小时。反应液冷至室温，加入KF水溶液(20mL)，搅拌2小时，乙酸乙酯(30mL×3)萃取。有机相合并，加入1N盐酸水溶液(20mL)，搅拌1小时，分液，取有机相用无水硫酸钠干燥后浓缩，得到粗品1-(4-(二氟甲基)苯基)乙烷-1-酮(1g，产率122％)，直接用于下一步反应。1-Bromo-4-(difluoromethyl)benzene (1g, 4.83mmol), tributyl(1-ethoxyethylene)tin (2.27g, 6.28mmol) and bis(triphenylphosphine) dichloride Palladium (0.17 g, 0.242 mmol) was dissolved in dioxane (20 mL) and stirred at 100° C. for 12 hours under nitrogen protection. The reaction solution was cooled to room temperature, KF aqueous solution (20 mL) was added, stirred for 2 hours, and extracted with ethyl acetate (30 mL×3). Combine the organic phases, add 1N hydrochloric acid aqueous solution (20 mL), stir for 1 hour, separate the layers, take the organic phase and dry it with anhydrous sodium sulfate and concentrate to obtain the crude product 1-(4-(difluoromethyl)phenyl)ethane- 1-Kone (1 g, yield 122%) was directly used in the next reaction.
第二步：1-(4-(二氟甲基)苯基)乙烷-1-醇的制备The second step: the preparation of 1-(4-(difluoromethyl)phenyl)ethan-1-ol
粗品1-(4-(二氟甲基)苯基)乙烷-1-酮(1g,5.88mmol)溶于甲醇(10mL)中，冰浴下分批加入硼氢化钠(445mg,11.75mmol)，搅拌1小时。反应液中加水(10mL)，搅拌10分钟，旋蒸出去甲醇，水相用乙酸乙酯(20mL×3)萃取。有机相合并，无水硫酸钠干燥，浓缩，得到残留物用柱层析纯化，即得1-(4-(二氟甲基)苯基)乙烷-1-醇(0.65g，产率64％)。Crude 1-(4-(difluoromethyl)phenyl)ethan-1-one (1g, 5.88mmol) was dissolved in methanol (10mL), and sodium borohydride (445mg, 11.75mmol) was added in portions under ice-cooling , and stirred for 1 hour. Water (10 mL) was added to the reaction solution, stirred for 10 minutes, methanol was removed by rotary evaporation, and the aqueous phase was extracted with ethyl acetate (20 mL×3). The organic phases were combined, dried over anhydrous sodium sulfate, concentrated, and the obtained residue was purified by column chromatography to obtain 1-(4-(difluoromethyl)phenyl)ethan-1-ol (0.65g, yield 64 %).
第三步：1-(1-(4-(二氟甲基)苯基)乙基)-4-(丙烷-1-炔-1-基)-1H-吲唑-7-甲酸甲酯的制备The third step: 1-(1-(4-(difluoromethyl)phenyl)ethyl)-4-(propane-1-yn-1-yl)-1H-indazole-7-formic acid methyl ester preparation
1-(4-(二氟甲基)苯基)乙烷-1-醇(600mg,3.48mmol)、4-(丙烷-1-炔-1-基)-1H-吲唑-7-甲酸甲酯(439mg,2.05mmol)和三苯基膦(1.6g,6.15mmol)溶于四氢呋喃(5mL)中，冰浴下缓慢滴加偶氮二甲酸二异丙酯(1.2g,6.15mmol)，随后升至室温搅拌16小时。反应液浓缩，柱层析纯化得1-(1-(4-(二氟甲基)苯基)乙基)-4-(丙烷-1-炔-1-基)-1H-吲唑-7-甲酸甲酯(450mg，产率60％)。1-(4-(Difluoromethyl)phenyl)ethan-1-ol (600mg, 3.48mmol), 4-(propane-1-yn-1-yl)-1H-indazole-7-carboxylic acid Ester (439mg, 2.05mmol) and triphenylphosphine (1.6g, 6.15mmol) were dissolved in tetrahydrofuran (5mL), and diisopropyl azodicarboxylate (1.2g, 6.15mmol) was slowly added dropwise under ice-cooling, followed by Warm to room temperature and stir for 16 hours. The reaction solution was concentrated and purified by column chromatography to obtain 1-(1-(4-(difluoromethyl)phenyl)ethyl)-4-(propane-1-yn-1-yl)-1H-indazole-7 - Methyl formate (450 mg, 60% yield).
LC-MS,M/Z(ESI):368.2[M+H] +。 LC-MS, M/Z (ESI): 368.2 [M+H] + .
第四步：1-(1-(4-(二氟甲基)苯基)乙基)-4-(丙烷-1-炔-1-基)-1H-吲唑-7-甲酸的制备Step 4: Preparation of 1-(1-(4-(difluoromethyl)phenyl)ethyl)-4-(propane-1-yn-1-yl)-1H-indazole-7-carboxylic acid
1-(1-(4-(二氟甲基)苯基)乙基)-4-(丙烷-1-炔-1-基)-1H-吲唑-7-甲酸甲酯(450mg,1.22mmol) 溶于四氢呋喃/甲醇/水(5mL/1mL/1mL)中，加入一水合氢氧化锂(154mg,3.66mmol)，室温搅拌16小时。反应液浓缩，加入水(5mL)，再加稀盐酸(1mol/L水溶液)调节溶液pH值约2，有大量白色固体产生，过滤收集白色固体，干燥后得到1-(1-(4-(二氟甲基)苯基)乙基)-4-(丙烷-1-炔-1-基)-1H-吲唑-7-甲酸(433mg，收率100％)。1-(1-(4-(Difluoromethyl)phenyl)ethyl)-4-(propane-1-yn-1-yl)-1H-indazole-7-carboxylic acid methyl ester (450mg, 1.22mmol ) was dissolved in tetrahydrofuran/methanol/water (5mL/1mL/1mL), added lithium hydroxide monohydrate (154mg, 3.66mmol), and stirred at room temperature for 16 hours. The reaction solution was concentrated, water (5mL) was added, and then diluted hydrochloric acid (1mol/L aqueous solution) was added to adjust the pH value of the solution to about 2. A large amount of white solid was produced. The white solid was collected by filtration and dried to obtain 1-(1-(4-( Difluoromethyl)phenyl)ethyl)-4-(propan-1-yn-1-yl)-1H-indazole-7-carboxylic acid (433 mg, yield 100%).
第五步：(Sa)-6-(1-(1-(4-(二氟甲基)苯基)乙基)-4-(丙烷-1-炔-1-基)-1H-吲唑-7-甲酰胺)螺环[3.3]庚烷-2-甲酸甲酯的制备The fifth step: (Sa)-6-(1-(1-(4-(difluoromethyl)phenyl)ethyl)-4-(propane-1-yn-1-yl)-1H-indazole Preparation of -7-carboxamide) spiro[3.3]heptane-2-carboxylic acid methyl ester
1-(1-(4-(二氟甲基)苯基)乙基)-4-(丙烷-1-炔-1-基)-1H-吲唑-7-甲酸(420mg,1.185mmol)和HATU(901mg,2.37mmol)溶于N,N-二甲基甲酰胺(10mL)中，冰浴下加入N,N-二异丙基乙胺(766mg,5.93mmol)，搅拌15分钟后加入6-氨基螺[3.3]庚烷-2-甲酸甲酯盐酸盐(731mg,3.56mmol)，室温搅拌16小时。反应液中加入水(30mL)，乙酸乙酯(30mL×3)萃取，有机相依次用饱和食盐水(50mL)洗涤，无水硫酸钠干燥，浓缩后柱层析纯化，得(Sa)-6-(1-(1-(4-(二氟甲基)苯基)乙基)-4-(丙烷-1-炔-1-基)-1H-吲唑-7-甲酰胺)螺环[3.3]庚烷-2-甲酸甲酯(480mg，产率80％)。1-(1-(4-(difluoromethyl)phenyl)ethyl)-4-(propan-1-yn-1-yl)-1H-indazole-7-carboxylic acid (420mg, 1.185mmol) and HATU (901mg, 2.37mmol) was dissolved in N,N-dimethylformamide (10mL), N,N-diisopropylethylamine (766mg, 5.93mmol) was added under ice-cooling, and after stirring for 15 minutes, 6 - Aminospiro[3.3]heptane-2-carboxylic acid methyl ester hydrochloride (731 mg, 3.56 mmol), stirred at room temperature for 16 hours. Add water (30mL) to the reaction solution, extract with ethyl acetate (30mL×3), wash the organic phase successively with saturated brine (50mL), dry over anhydrous sodium sulfate, concentrate and purify by column chromatography to obtain (Sa)-6 -(1-(1-(4-(difluoromethyl)phenyl)ethyl)-4-(propane-1-yn-1-yl)-1H-indazole-7-carboxamide) spiro[ 3.3] Heptane-2-carboxylic acid methyl ester (480 mg, 80% yield).
LC-MS,M/Z(ESI):506.2[M+H] +。 LC-MS, M/Z (ESI): 506.2 [M+H] + .
第六步：(Sa,R)-6-(1-(1-(4-(二氟甲基)苯基)乙基)-4-(丙烷-1-炔-1-基)-1H-吲唑-7-甲酰胺)螺环[3.3]庚烷-2-甲酸甲酯的制备The sixth step: (Sa,R)-6-(1-(1-(4-(difluoromethyl)phenyl)ethyl)-4-(propane-1-yn-1-yl)-1H- Preparation of indazole-7-carboxamide) spiro[3.3]heptane-2-carboxylate methyl ester
(Sa)-6-(1-(1-(4-(二氟甲基)苯基)乙基)-4-(丙烷-1-炔-1-基)-1H-吲唑-7-甲酰胺)螺环[3.3]庚烷-2-甲酸甲酯(130F)(180mg,0.356mmol)用SFC拆分，得到(Sa,R)-6-(1-(1-(4-(二氟甲基)苯基)乙基)-4-(丙烷-1-炔-1-基)-1H-吲唑-7-甲酰胺)螺环[3.3]庚烷-2-甲酸甲酯(80mg,产率44％)。SFC拆分方法：使用Chiralpak AD-3型号手性拆分柱(规格为：50×4.6mm,3um)进行手性拆分，流动相A为超临界流体CO 2,流动相B为异丙醇(含0.05％二乙醇胺)；梯度条件:流动相B为40％的梯度保持10min，柱温保持在35℃，柱压保持在100bar，出峰时间：0.581min。 (Sa)-6-(1-(1-(4-(difluoromethyl)phenyl)ethyl)-4-(propane-1-yn-1-yl)-1H-indazole-7-methan Amide)spiro[3.3]heptane-2-carboxylic acid methyl ester (130F) (180mg, 0.356mmol) was resolved by SFC to give (Sa,R)-6-(1-(1-(4-(difluoro Methyl)phenyl)ethyl)-4-(propane-1-yn-1-yl)-1H-indazole-7-carboxamide)spiro[3.3]heptane-2-carboxylic acid methyl ester (80mg, Yield 44%). SFC resolution method: use Chiralpak AD-3 chiral resolution column (specification: 50×4.6mm, 3um) for chiral resolution, mobile phase A is supercritical fluid CO 2 , mobile phase B is isopropanol (containing 0.05% diethanolamine); Gradient conditions: the mobile phase B is 40% of the gradient to maintain 10min, the column temperature is maintained at 35 ° C, the column pressure is maintained at 100bar, the peak time: 0.581min.
第七步：(Sa,R)-6-(1-(1-(4-(二氟甲基)苯基)乙基)-4-(丙烷-1-炔-1-基)-1H-吲唑-7-甲酰胺)螺环[3.3]庚烷-2-甲酸(I-34a)The seventh step: (Sa,R)-6-(1-(1-(4-(difluoromethyl)phenyl)ethyl)-4-(propane-1-yn-1-yl)-1H- Indazole-7-carboxamide) spiro[3.3]heptane-2-carboxylic acid (I-34a)
Figure PCTCN2022097611-appb-000101
Figure PCTCN2022097611-appb-000101
(Sa,R)-6-(1-(1-(4-(二氟甲基)苯基)乙基)-4-(丙烷-1-炔-1-基)-1H-吲唑-7-甲酰胺)螺环[3.3]庚烷-2-甲酸甲酯(80mg,0.158mmol)溶于四氢呋喃/甲醇/水(4mL/0.8mL/0.8mL)中，加入一水合氢氧化锂(33mg,0.79mmol)，室温搅拌3小时。反应液浓缩，加入水(3mL)，再加稀盐酸(1mol/L水溶液)调节溶液pH值约2，有大量白色固体产生，过滤收集白色固体，干燥后得到(Sa,R)-6-(1-(1-(4-(二氟甲基)苯基)乙基)-4-(丙烷-1-炔-1-基)-1H-吲唑-7-甲酰胺)螺环[3.3]庚烷-2-甲酸(I-34a)(73.3mg，收率94％)。(Sa,R)-6-(1-(1-(4-(Difluoromethyl)phenyl)ethyl)-4-(propane-1-yn-1-yl)-1H-indazole-7 -formamide) spiro[3.3]heptane-2-carboxylic acid methyl ester (80mg, 0.158mmol) was dissolved in tetrahydrofuran/methanol/water (4mL/0.8mL/0.8mL), added lithium hydroxide monohydrate (33mg, 0.79 mmol), stirred at room temperature for 3 hours. The reaction solution was concentrated, water (3mL) was added, and then diluted hydrochloric acid (1mol/L aqueous solution) was added to adjust the pH value of the solution to about 2. A large amount of white solid was produced, and the white solid was collected by filtration and dried to obtain (Sa,R)-6-( 1-(1-(4-(Difluoromethyl)phenyl)ethyl)-4-(propane-1-yn-1-yl)-1H-indazole-7-carboxamide)spiro[3.3] Heptane-2-carboxylic acid (I-34a) (73.3 mg, yield 94%).
LC-MS,M/Z(ESI):492.4[M+H] +。 LC-MS, M/Z (ESI): 492.4 [M+H] + .
1H NMR(400MHz,CDCl 3)δ8.25(s,1H),7.31(d,2H),7.24(d,1H),7.12(d,1H),7.04(d,2H),6.71–6.34(m,2H),5.75(d,1H),4.48–4.34(m,1H),3.15–3.02(m,1H),2.66–2.57(m,1H),2.47–2.36(m,3H),2.30(dd,1H),2.24–2.13(m,4H),2.01(d,3H),1.95–1.89(m,1H),1.65(dd,1H). 1 H NMR (400MHz, CDCl 3 )δ8.25(s,1H),7.31(d,2H),7.24(d,1H),7.12(d,1H),7.04(d,2H),6.71–6.34( m,2H),5.75(d,1H),4.48–4.34(m,1H),3.15–3.02(m,1H),2.66–2.57(m,1H),2.47–2.36(m,3H),2.30( dd,1H),2.24–2.13(m,4H),2.01(d,3H),1.95–1.89(m,1H),1.65(dd,1H).
化合物I-34b的制备Preparation of Compound I-34b
合成路线如下所示：The synthetic route is as follows:
Figure PCTCN2022097611-appb-000102
Figure PCTCN2022097611-appb-000102
第一步：(Sa,S)-6-(1-(1-(4-(二氟甲基)苯基)乙基)-4-(丙烷-1-炔-1-基)-1H-吲唑-7-甲酰胺)螺环[3.3]庚烷-2-甲酸甲酯的制备The first step: (Sa,S)-6-(1-(1-(4-(difluoromethyl)phenyl)ethyl)-4-(propane-1-yn-1-yl)-1H- Preparation of indazole-7-carboxamide) spiro[3.3]heptane-2-carboxylate methyl ester
(Sa)-6-(1-(1-(4-(二氟甲基)苯基)乙基)-4-(丙烷-1-炔-1-基)-1H-吲唑-7-甲酰胺)螺环[3.3]庚烷-2-甲酸甲酯(180mg,0.356mmol)用SFC拆分，得到(Sa,S)-6-(1-(1-(4-(二氟甲基)苯基)乙基)-4-(丙烷-1-炔-1-基)-1H-吲唑-7-甲酰胺)螺环[3.3]庚烷-2-甲酸甲酯(80mg,产率44％)。SFC拆分方法：使用Chiralpak AD-3型号手性拆分柱(规格为：50×4.6mm,3um)进行手性拆分，流动相A为超临界流体CO 2,流动相B为异丙醇(含0.05％二乙醇胺)；梯度条件:流动相B为40％的梯度保持10min，柱温保持在35℃，柱压保持在100bar，出峰时间：2.245min。 (Sa)-6-(1-(1-(4-(difluoromethyl)phenyl)ethyl)-4-(propane-1-yn-1-yl)-1H-indazole-7-methan Amide)spiro[3.3]heptane-2-carboxylic acid methyl ester (180mg, 0.356mmol) was resolved by SFC to give (Sa,S)-6-(1-(1-(4-(difluoromethyl) Phenyl)ethyl)-4-(propane-1-yn-1-yl)-1H-indazole-7-carboxamide)spiro[3.3]heptane-2-carboxylic acid methyl ester (80mg, yield 44 %). SFC resolution method: use Chiralpak AD-3 chiral resolution column (specification: 50×4.6mm, 3um) for chiral resolution, mobile phase A is supercritical fluid CO 2 , mobile phase B is isopropanol (Containing 0.05% diethanolamine); Gradient conditions: mobile phase B is 40% gradient and kept for 10min, column temperature was kept at 35°C, column pressure was kept at 100bar, peak eluting time: 2.245min.
LC-MS,M/Z(ESI):506.3[M+H] +。 LC-MS, M/Z (ESI): 506.3 [M+H] + .
第七步：(Sa,S)-6-(1-(1-(4-(二氟甲基)苯基)乙基)-4-(丙烷-1-炔-1-基)-1H-吲唑-7-甲酰胺)螺环[3.3]庚烷-2-甲酸(I-34b)The seventh step: (Sa,S)-6-(1-(1-(4-(difluoromethyl)phenyl)ethyl)-4-(propane-1-yn-1-yl)-1H- Indazole-7-carboxamide) spiro[3.3]heptane-2-carboxylic acid (I-34b)
(Sa,S)-6-(1-(1-(4-(二氟甲基)苯基)乙基)-4-(丙烷-1-炔-1-基)-1H-吲唑-7-甲酰胺)螺环[3.3]庚烷-2-甲酸甲酯(80mg,0.158mmol)溶于四氢呋喃/甲醇/水(4mL/0.8mL/0.8mL)中，加入一水合氢氧化锂(33mg,0.791mmol)，室温搅拌3小时。反应液浓缩，加入水(3mL)，再加稀盐酸(1mol/L水溶液)调节溶液pH值约2，干燥后得到(Sa,S)-6-(1-(1-(4-(二氟甲基)苯基)乙基)-4-(丙烷-1-炔-1-基)-1H-吲唑-7-甲酰胺)螺环[3.3]庚烷-2-甲酸(I-34b)(62.5mg，收率80％)。(Sa,S)-6-(1-(1-(4-(Difluoromethyl)phenyl)ethyl)-4-(propane-1-yn-1-yl)-1H-indazole-7 -formamide) spiro[3.3]heptane-2-carboxylic acid methyl ester (80mg, 0.158mmol) was dissolved in tetrahydrofuran/methanol/water (4mL/0.8mL/0.8mL), added lithium hydroxide monohydrate (33mg, 0.791 mmol), stirred at room temperature for 3 hours. The reaction solution was concentrated, water (3mL) was added, and then dilute hydrochloric acid (1mol/L aqueous solution) was added to adjust the pH value of the solution to about 2. After drying, (Sa,S)-6-(1-(1-(4-(difluoro Methyl)phenyl)ethyl)-4-(propane-1-yn-1-yl)-1H-indazole-7-carboxamide)spiro[3.3]heptane-2-carboxylic acid (I-34b) (62.5 mg, yield 80%).
LC-MS,M/Z(ESI):492.5[M+H] +。 LC-MS, M/Z (ESI): 492.5 [M+H] + .
1H NMR(400MHz,CDCl 3)δ8.25(s,1H),7.32(d,2H),7.24(d,1H),7.12(d,1H),7.04(d,J＝8.0Hz,2H),6.73–6.35(m,2H),5.75(d,1H),4.47–4.33(m,1H),3.09(p,1H),2.55–2.45(m,2H),2.41(d,2H),2.30(dd,1H),2.21–2.11(m,4H),2.01(d,3H),1.91–1.83(m,1H),1.73–1.64(dd,1H). 1 H NMR (400MHz, CDCl 3 )δ8.25(s,1H),7.32(d,2H),7.24(d,1H),7.12(d,1H),7.04(d,J=8.0Hz,2H) ,6.73–6.35(m,2H),5.75(d,1H),4.47–4.33(m,1H),3.09(p,1H),2.55–2.45(m,2H),2.41(d,2H),2.30 (dd,1H),2.21–2.11(m,4H),2.01(d,3H),1.91–1.83(m,1H),1.73–1.64(dd,1H).
实施例35化合物I-35a的制备Preparation of Example 35 Compound I-35a
合成路线如下所示：The synthetic route is as follows:
Figure PCTCN2022097611-appb-000103
Figure PCTCN2022097611-appb-000103
第一步：1-((1R)-1-(4-(3-氮杂双环[3.1.0]己烷-3-基)苯基)乙基)-4-(丙烷-1-炔-1-基)-1H-吲唑-7-甲酸甲酯的制备The first step: 1-((1R)-1-(4-(3-azabicyclo[3.1.0]hexan-3-yl)phenyl)ethyl)-4-(propane-1-yne- Preparation of 1-yl)-1H-indazole-7-carboxylic acid methyl ester
(R)-1-(1-(4-溴苯基)乙基)-4-(丙烷-1-炔-1-基)-1H-吲唑-7-甲酸甲酯(220mg，0.554mmol),3-氮杂双环[3.1.0]己烷盐酸盐(265mg,2.22mmol)，醋酸钯(12.5mg,0.055mmol)，碳酸铯(1.1g,3.32mmol)，2-(二叔丁基膦)联苯(19.8mg,0.066mmol)于二氧六环(3mL)中，氮气保护下80℃搅拌16小时。反应液浓缩，残留物用柱层析纯化，得1-((1R)-1-(4-(3-氮杂双环[3.1.0]己烷-3-基)苯基)乙基)-4-(丙烷-1-炔-1-基)-1H-吲唑-7-甲酸甲酯(175mg，产率79％)。(R)-1-(1-(4-bromophenyl)ethyl)-4-(propane-1-yn-1-yl)-1H-indazole-7-carboxylic acid methyl ester (220mg, 0.554mmol) , 3-Azabicyclo[3.1.0]hexane hydrochloride (265mg, 2.22mmol), palladium acetate (12.5mg, 0.055mmol), cesium carbonate (1.1g, 3.32mmol), 2-(di-tert-butyl Phosphine)biphenyl (19.8mg, 0.066mmol) in dioxane (3mL) was stirred at 80°C for 16 hours under nitrogen protection. The reaction solution was concentrated, and the residue was purified by column chromatography to obtain 1-((1R)-1-(4-(3-azabicyclo[3.1.0]hexane-3-yl)phenyl)ethyl)- 4-(Propan-1-yn-1-yl)-1H-indazole-7-carboxylic acid methyl ester (175 mg, 79% yield).
LC-MS,M/Z(ESI):400.4[M+H] +。 LC-MS, M/Z (ESI): 400.4 [M+H] + .
第二步：1-((1R)-1-(4-(3-氮杂双环[3.1.0]己烷-3-基)苯基)乙基)-4-(丙烷-1-炔-1-基)-1H-吲唑-7-甲酸锂盐的制备The second step: 1-((1R)-1-(4-(3-azabicyclo[3.1.0]hexan-3-yl)phenyl)ethyl)-4-(propane-1-yne- Preparation of 1-yl)-1H-indazole-7-carboxylate lithium salt
1-((1R)-1-(4-(3-氮杂双环[3.1.0]己烷-3-基)苯基)乙基)-4-(丙烷-1-炔-1-基)-1H-吲唑-7-甲酸甲酯(170mg,0.426mmol)溶于四氢呋喃/甲醇/水(3mL/0.6mL/0.6mL)中，加入一水合氢氧化锂(54mg,1.28mmol)，室温搅拌4小时。反应液浓缩干燥得到1-((1R)-1-(4-(3-氮杂双环[3.1.0]己烷-3-基)苯基)乙基)-4-(丙烷-1-炔-1-基)-1H-吲唑-7-甲酸锂盐(220mg，含LiOH，收率134％)。1-((1R)-1-(4-(3-Azabicyclo[3.1.0]hexan-3-yl)phenyl)ethyl)-4-(propane-1-yn-1-yl) -1H-indazole-7-methyl carboxylate (170mg, 0.426mmol) was dissolved in tetrahydrofuran/methanol/water (3mL/0.6mL/0.6mL), added lithium hydroxide monohydrate (54mg, 1.28mmol), stirred at room temperature 4 hours. The reaction solution was concentrated and dried to obtain 1-((1R)-1-(4-(3-azabicyclo[3.1.0]hexane-3-yl)phenyl)ethyl)-4-(propane-1-yne -1-yl)-1H-indazole-7-carboxylic acid lithium salt (220 mg, containing LiOH, yield 134%).
LC-MS,M/Z(ESI):400.4[M+H] +。 LC-MS, M/Z (ESI): 400.4 [M+H] + .
第三步：(Sa)-6-(1-((1R)-1-(4-(3-氮杂双环[3.1.0]己烷-3-基)苯基)乙基)-4-(丙烷-1-炔-1-基)-1H-吲唑-7-甲酰胺)螺[3.3]庚烷-2-甲酸甲酯的制备The third step: (Sa)-6-(1-((1R)-1-(4-(3-azabicyclo[3.1.0]hexane-3-yl)phenyl)ethyl)-4- Preparation of (propane-1-yn-1-yl)-1H-indazole-7-carboxamide)spiro[3.3]heptane-2-carboxylic acid methyl ester
(Sa)-1-((1R)-1-(4-(3-氮杂双环[3.1.0]己烷-3-基)苯基)乙基)-4-(丙烷-1-炔-1-基)-1H-吲唑-7-甲酸锂盐(200mg,0.52mmol)和HATU(395mg,1.04mmol)溶于N,N-二甲基甲酰胺(5mL)中，冰浴下加入二异丙基乙胺(268mg,2.08mmol)，搅拌15分钟后加入甲基6-氨基螺[3.3]庚烷-2-甲酸酯盐酸盐(213mg,1.04mmol)，室温搅拌12小时。反应液中加入水(20mL)，乙酸乙酯(10mL×3)萃取，有机相依次用饱和食盐水(10mL×2)洗涤，无水硫酸钠干燥，浓缩后柱层析纯化，得(Sa)-6-(1-((1R)-1-(4-(3-氮杂双环[3.1.0]己烷-3-基)苯基)乙基)-4-(丙烷-1-炔-1-基)-1H-吲唑-7-甲酰胺)螺[3.3]庚烷-2-甲酸甲酯(150mg，产率54％)。(Sa)-1-((1R)-1-(4-(3-Azabicyclo[3.1.0]hexane-3-yl)phenyl)ethyl)-4-(propane-1-yne- 1-yl)-1H-indazole-7-carboxylic acid lithium salt (200mg, 0.52mmol) and HATU (395mg, 1.04mmol) were dissolved in N,N-dimethylformamide (5mL), and di Isopropylethylamine (268mg, 2.08mmol) was stirred for 15 minutes, then methyl 6-aminospiro[3.3]heptane-2-carboxylate hydrochloride (213mg, 1.04mmol) was added, and stirred at room temperature for 12 hours. Add water (20mL) to the reaction solution, extract with ethyl acetate (10mL×3), wash the organic phase successively with saturated brine (10mL×2), dry over anhydrous sodium sulfate, concentrate and purify by column chromatography to obtain (Sa) -6-(1-((1R)-1-(4-(3-Azabicyclo[3.1.0]hexane-3-yl)phenyl)ethyl)-4-(propane-1-yne- 1-yl)-1H-indazole-7-carboxamide)spiro[3.3]heptane-2-carboxylic acid methyl ester (150 mg, 54% yield).
LC-MS,M/Z(ESI):537.7[M+H] +。 LC-MS, M/Z (ESI): 537.7 [M+H] + .
第四步：(Sa)-6-(1-((1R)-1-(4-(3-氮杂双环[3.1.0]己烷-3-基)苯基)乙基)-4-(丙烷-1-炔-1-基)-1H-吲唑-7-甲酰胺)螺[3.3]庚烷-2-甲酸的制备The fourth step: (Sa)-6-(1-((1R)-1-(4-(3-azabicyclo[3.1.0]hexan-3-yl)phenyl)ethyl)-4- Preparation of (propane-1-yn-1-yl)-1H-indazole-7-carboxamide)spiro[3.3]heptane-2-carboxylic acid
(Sa)-6-(1-((1R)-1-(4-(3-氮杂双环[3.1.0]己烷-3-基)苯基)乙基)-4-(丙烷-1-炔-1-基)-1H-吲唑-7-甲酰胺)螺[3.3]庚烷-2-甲酸甲酯(80mg,0.15mmol)溶于四氢呋喃/甲醇/水(3mL/0.6mL/0.6mL)中，加入一水合氢氧化锂(31mg,0.75mmol)，室温搅拌2小时。反应液浓缩，加入水(3mL)，再加稀盐酸(1mol/L水溶液)调节溶液pH值约5，有大量白色固体产生，过滤收集白色固体，干燥后得到(Sa)-6-(1-((1R)-1-(4-(3-氮杂双环[3.1.0]己烷-3-基)苯基)乙基)-4-(丙烷-1-炔-1-基)-1H-吲唑-7-甲酰胺)螺[3.3]庚烷-2-甲酸(I-35a)(63.2mg，收率81％)。(Sa)-6-(1-((1R)-1-(4-(3-azabicyclo[3.1.0]hexane-3-yl)phenyl)ethyl)-4-(propane-1 -alkyn-1-yl)-1H-indazole-7-carboxamide)spiro[3.3]heptane-2-carboxylic acid methyl ester (80mg, 0.15mmol) dissolved in tetrahydrofuran/methanol/water (3mL/0.6mL/0.6 mL), add lithium hydroxide monohydrate (31 mg, 0.75 mmol), and stir at room temperature for 2 hours. The reaction solution was concentrated, water (3mL) was added, and then diluted hydrochloric acid (1mol/L aqueous solution) was added to adjust the pH value of the solution to about 5. A large amount of white solid was produced, and the white solid was collected by filtration and dried to obtain (Sa)-6-(1- ((1R)-1-(4-(3-Azabicyclo[3.1.0]hexane-3-yl)phenyl)ethyl)-4-(propane-1-yn-1-yl)-1H -indazole-7-carboxamide) spiro[3.3]heptane-2-carboxylic acid (I-35a) (63.2 mg, yield 81%).
LC-MS,M/Z(ESI):523.6[M+H] +。 LC-MS, M/Z (ESI): 523.6 [M+H] + .
1H NMR(400MHz,dmso)δ12.02(s,1H),8.83(d,1H),8.18(s,1H),7.27(d,1H),7.15(d,1H),6.86(d,2H),6.34(d,2H),6.11(q,1H),4.35(q,1H),3.40–3.36(m,2H),3.09-3.01(m,2H),2.99-2.85(m,1H),2.48-2.42(m,1H),2.36–2.21(m,3H),2.19–2.01(m,6H),1.94(dd,1H),1.81(d,3H),1.68–1.58(m,2H),0.71–0.61(m,1H),0.15(q,1H). 1 H NMR (400MHz,dmso)δ12.02(s,1H),8.83(d,1H),8.18(s,1H),7.27(d,1H),7.15(d,1H),6.86(d,2H ),6.34(d,2H),6.11(q,1H),4.35(q,1H),3.40–3.36(m,2H),3.09-3.01(m,2H),2.99-2.85(m,1H), 2.48-2.42(m,1H),2.36–2.21(m,3H),2.19–2.01(m,6H),1.94(dd,1H),1.81(d,3H),1.68–1.58(m,2H), 0.71–0.61(m,1H),0.15(q,1H).
化合物I-35b的制备Preparation of Compound I-35b
合成路线如下所示：The synthetic route is as follows:
Figure PCTCN2022097611-appb-000104
Figure PCTCN2022097611-appb-000104
第一步：1-((1S)-1-(4-(3-氮杂双环[3.1.0]己烷-3-基)苯基)乙基)-4-(丙烷-1-炔-1-基)-1H-吲唑-7-甲酸甲酯的制备The first step: 1-((1S)-1-(4-(3-azabicyclo[3.1.0]hexan-3-yl)phenyl)ethyl)-4-(propane-1-yne- Preparation of 1-yl)-1H-indazole-7-carboxylic acid methyl ester
1-((1S)-1-(4-(3-溴-3-基)苯基)乙基)-4-(丙烷-1-炔-1-基)-1H-吲唑-7-甲酸甲酯(250mg,0.63mmol),3-氮杂双环[3.1.0]己烷盐酸酸(301mg,2.52mmol)，醋酸钯(14mg,0.063mmol)，碳酸铯(1.23g,3.38mmol)，2-(二叔丁基膦)联苯(22.5mg,0.076mmol)于二氧六环(3mL)中，氮气保护下80℃搅拌16小时。反应液浓缩，残留物用柱层析纯化，得1-((1S)-1-(4-(3-氮杂双环[3.1.0]己烷-3-基)苯基)乙基)-4-(丙烷-1-炔-1-基)-1H-吲唑-7-甲酸甲酯(180mg，产率72％)。1-((1S)-1-(4-(3-bromo-3-yl)phenyl)ethyl)-4-(propane-1-yn-1-yl)-1H-indazole-7-carboxylic acid Methyl ester (250mg, 0.63mmol), 3-azabicyclo[3.1.0]hexane hydrochloric acid (301mg, 2.52mmol), palladium acetate (14mg, 0.063mmol), cesium carbonate (1.23g, 3.38mmol), 2 -(Di-tert-butylphosphine)biphenyl (22.5mg, 0.076mmol) in dioxane (3mL) was stirred at 80°C for 16 hours under nitrogen protection. The reaction solution was concentrated, and the residue was purified by column chromatography to obtain 1-((1S)-1-(4-(3-azabicyclo[3.1.0]hexane-3-yl)phenyl)ethyl)- 4-(Propan-1-yn-1-yl)-1H-indazole-7-carboxylic acid methyl ester (180 mg, 72% yield).
第二步：1-((1S)-1-(4-(3-氮杂双环[3.1.0]己烷-3-基)苯基)乙基)-4-(丙烷-1-炔-1-基)-1H-吲唑-7-甲酸锂盐的制备The second step: 1-((1S)-1-(4-(3-azabicyclo[3.1.0]hexan-3-yl)phenyl)ethyl)-4-(propane-1-yne- Preparation of 1-yl)-1H-indazole-7-carboxylate lithium salt
1-((1S)-1-(4-(3-氮杂双环[3.1.0]己烷-3-基)苯基)乙基)-4-(丙烷-1-炔-1-基)-1H-吲唑-7-甲酸甲酯(175mg,0.44mmol)溶于四氢呋喃/甲醇/水(3mL/0.6mL/0.6mL)中，加入一水合氢氧化锂(55.1mg,1.31mmol)，室温搅拌16小时。反应液浓缩干燥得到1-((1S)-1-(4-(3-氮杂双环[3.1.0]己烷-3-基)苯基)乙基)-4-(丙烷-1-炔-1-基)-1H-吲唑-7-甲酸锂盐(210mg，含LiOH，产率124％)。1-((1S)-1-(4-(3-Azabicyclo[3.1.0]hexan-3-yl)phenyl)ethyl)-4-(propane-1-yn-1-yl) -1H-Indazole-7-methyl carboxylate (175mg, 0.44mmol) was dissolved in tetrahydrofuran/methanol/water (3mL/0.6mL/0.6mL), added lithium hydroxide monohydrate (55.1mg, 1.31mmol), room temperature Stir for 16 hours. The reaction solution was concentrated and dried to obtain 1-((1S)-1-(4-(3-azabicyclo[3.1.0]hexane-3-yl)phenyl)ethyl)-4-(propane-1-yne -1-yl)-1H-indazole-7-carboxylic acid lithium salt (210 mg with LiOH, 124% yield).
第三步：6-(1-((1S)-1-(4-(3-氮杂双环[3.1.0]己烷-3-基)苯基)乙基)-4-(丙烷-1-炔-1-基)-1H-吲唑-7-甲酰胺)螺[3.3]庚烷-2-甲酸甲酯的制备The third step: 6-(1-((1S)-1-(4-(3-azabicyclo[3.1.0]hexane-3-yl)phenyl)ethyl)-4-(propane-1 Preparation of -alkyn-1-yl)-1H-indazole-7-carboxamide)spiro[3.3]heptane-2-carboxylic acid methyl ester
1-((1S)-1-(4-(3-氮杂双环[3.1.0]己烷-3-基)苯基)乙基)-4-(丙烷-1-炔-1-基)-1H-吲唑-7-甲酸锂盐(230mg,0.6mmol)和HATU(454mg,1.19mmol)溶于N,N-二甲基甲酰胺(5mL)中，冰浴下加入N,N-二异丙基乙胺(308mg,2.39mmol)，搅拌15分钟后加入(Sa)-6-氨基螺[3.3]庚烷-2-甲酸甲酯盐酸盐(245mg,1.19mmol)，室温搅拌3小时。反应液中加入水(20mL)，乙酸乙酯(10mL×3)萃取，有机相依次用饱和食盐水(10mL×2)洗涤，无水硫酸钠干燥，浓缩后柱层析纯化，得(Sa)-6-(1-((1S)-1-(4-(3-氮杂双环[3.1.0]己烷-3-基)苯基)乙基)-4-(丙烷-1-炔-1-基)-1H-吲唑-7-甲酰胺)螺[3.3]庚烷-2-甲酸甲酯(150mg，产率47％)。1-((1S)-1-(4-(3-Azabicyclo[3.1.0]hexan-3-yl)phenyl)ethyl)-4-(propane-1-yn-1-yl) -1H-indazole-7-carboxylic acid lithium salt (230mg, 0.6mmol) and HATU (454mg, 1.19mmol) were dissolved in N,N-dimethylformamide (5mL), and N,N-di Isopropylethylamine (308mg, 2.39mmol), stirred for 15 minutes, then added (Sa)-6-aminospiro[3.3]heptane-2-carboxylic acid methyl ester hydrochloride (245mg, 1.19mmol), stirred at room temperature for 3 hours . Add water (20mL) to the reaction solution, extract with ethyl acetate (10mL×3), wash the organic phase successively with saturated brine (10mL×2), dry over anhydrous sodium sulfate, concentrate and purify by column chromatography to obtain (Sa) -6-(1-((1S)-1-(4-(3-Azabicyclo[3.1.0]hexane-3-yl)phenyl)ethyl)-4-(propane-1-yne- 1-yl)-1H-indazole-7-carboxamide)spiro[3.3]heptane-2-carboxylic acid methyl ester (150 mg, 47% yield).
LC-MS,M/Z(ESI):537.7[M+H] +。 LC-MS, M/Z (ESI): 537.7 [M+H] + .
第四步：(Sa)-6-(1-((1S)-1-(4-(3-氮杂双环[3.1.0]己烷-3-基)苯基)乙基)-4-(丙烷-1-炔-1-基)-1H-吲唑-7-甲酰胺)螺[3.3]庚烷-2-甲酸(I-35b)The fourth step: (Sa)-6-(1-((1S)-1-(4-(3-azabicyclo[3.1.0]hexan-3-yl)phenyl)ethyl)-4- (Propan-1-yn-1-yl)-1H-indazole-7-carboxamide)spiro[3.3]heptane-2-carboxylic acid (I-35b)
Figure PCTCN2022097611-appb-000105
Figure PCTCN2022097611-appb-000105
(Sa)-6-(1-((1S)-1-(4-(3-氮杂双环[3.1.0]己烷-3-基)苯基)乙基)-4-(丙烷-1-炔-1-基)-1H-吲唑-7-甲酰胺)螺[3.3]庚烷-2-甲酸甲酯(75mg,0.14mmol)溶于四氢呋喃/甲醇/水(3mL/0.6mL/0.6mL)中，加入一水合氢氧化锂(29.3mg,0.7mmol)，室温搅拌16小时。反应液浓缩，加入水(3mL)，再加稀盐酸(1mol/L水溶液)调节溶液pH值约5，有大量白色固体产生，过滤收集白色固体，干燥后得到(Sa)-6-(1-((1S)-1-(4-(3-氮杂双环[3.1.0]己烷-3-基)苯基)乙基)-4-(丙烷-1-炔-1-基)-1H-吲唑-7-甲酰胺)螺[3.3]庚烷-2-甲酸(I-35b)(66mg，收率90％)。(Sa)-6-(1-((1S)-1-(4-(3-azabicyclo[3.1.0]hexane-3-yl)phenyl)ethyl)-4-(propane-1 -alkyn-1-yl)-1H-indazole-7-carboxamide)spiro[3.3]heptane-2-carboxylic acid methyl ester (75mg, 0.14mmol) dissolved in tetrahydrofuran/methanol/water (3mL/0.6mL/0.6 mL), add lithium hydroxide monohydrate (29.3 mg, 0.7 mmol), and stir at room temperature for 16 hours. The reaction solution was concentrated, water (3mL) was added, and then diluted hydrochloric acid (1mol/L aqueous solution) was added to adjust the pH value of the solution to about 5. A large amount of white solid was produced, and the white solid was collected by filtration and dried to obtain (Sa)-6-(1- ((1S)-1-(4-(3-Azabicyclo[3.1.0]hexane-3-yl)phenyl)ethyl)-4-(propane-1-yn-1-yl)-1H -indazole-7-carboxamide) spiro[3.3]heptane-2-carboxylic acid (I-35b) (66 mg, yield 90%).
LC-MS,M/Z(ESI):523.7[M+H] +。 LC-MS, M/Z (ESI): 523.7 [M+H] + .
1H NMR(400MHz,DMSO-d6)δ12.00(s,1H),8.84(d,1H),8.18(s,1H),7.27(d,1H),7.15(d,1H),6.86(d,2H),6.35(d,2H),6.11(q,1H),4.35(q,J＝8.3Hz,1H),3.40–3.37(m,2H),3.08–3.02(m,2H),3.01–2.91(m,1H),2.49–2.44(m,1H),2.39–2.20(m,3H),2.16(s,3H),2.15–1.95(m,4H),1.81(d,3H),1.67–1.59(m,2H),0.71–0.62(m,1H),0.15(q,1H). 1 H NMR(400MHz,DMSO-d6)δ12.00(s,1H),8.84(d,1H),8.18(s,1H),7.27(d,1H),7.15(d,1H),6.86(d ,2H),6.35(d,2H),6.11(q,1H),4.35(q,J＝8.3Hz,1H),3.40–3.37(m,2H),3.08–3.02(m,2H),3.01– 2.91(m,1H),2.49–2.44(m,1H),2.39–2.20(m,3H),2.16(s,3H),2.15–1.95(m,4H),1.81(d,3H),1.67– 1.59(m,2H),0.71–0.62(m,1H),0.15(q,1H).
实施例36化合物I-36的制备The preparation of embodiment 36 compound I-36
合成路线如下所示：The synthetic route is as follows:
Figure PCTCN2022097611-appb-000106
Figure PCTCN2022097611-appb-000106
第一步：(R)-1-(1-(4-(6-(二氟甲基)吡啶-2-基)苯基)乙基)-4-(丙烷-1-炔-1-基)-1H-吲唑-7-甲酸甲酯的制备The first step: (R)-1-(1-(4-(6-(difluoromethyl)pyridin-2-yl)phenyl)ethyl)-4-(propane-1-yn-1-yl ) Preparation of -1H-indazole-7-methyl carboxylate
向(R)-4-(丙烷-1-炔-1-基)-1-(1-4-(4,4,5,5-四甲基-1,3,2-二氧杂硼烷-2-基)苯基)乙基)-1H-吲唑-7-甲酸甲酯(200mg，0.450mmol)的甲苯(2mL)和水(0.4mL)溶液中加入2-溴-6-(二氟甲基)吡啶(62.4mg,0.300mmol)、碳酸钾(124mg,0.900mmol)和[1,1'-双(二苯基膦基)二茂铁]二氯化钯(21.96mg,0.030mmol)。反应液置换3次氮气，并在80℃下搅拌18小时。将反应液倒入水(20mL)中并用乙酸乙酯(10mL*3)萃取。合并有机层，用无水硫酸钠干燥，过滤并浓缩得到粗产物。粗产物通过柱层析(石油醚/乙酸乙酯(V/V)＝30/1-1/1)纯化得到(R)-1-(1-(4-(6-(二氟甲基)吡啶-2-基)苯基)乙基)-4-(丙烷-1-炔-1-基)-1H-吲唑-7-甲酸甲酯(120mg，产率90％)。To (R)-4-(propane-1-yn-1-yl)-1-(1-4-(4,4,5,5-tetramethyl-1,3,2-dioxaborane -2-yl)phenyl)ethyl)-1H-indazole-7-carboxylic acid methyl ester (200 mg, 0.450 mmol) in toluene (2 mL) and water (0.4 mL) was added 2-bromo-6-(di Fluoromethyl)pyridine (62.4mg, 0.300mmol), potassium carbonate (124mg, 0.900mmol) and [1,1'-bis(diphenylphosphino)ferrocene]palladium dichloride (21.96mg, 0.030mmol ). The reaction solution was replaced with nitrogen three times, and stirred at 80° C. for 18 hours. The reaction solution was poured into water (20 mL) and extracted with ethyl acetate (10 mL*3). The organic layers were combined, dried over anhydrous sodium sulfate, filtered and concentrated to give crude product. The crude product was purified by column chromatography (petroleum ether/ethyl acetate (V/V)=30/1-1/1) to obtain (R)-1-(1-(4-(6-(difluoromethyl) Pyridin-2-yl)phenyl)ethyl)-4-(propan-1-yn-1-yl)-1H-indazole-7-carboxylic acid methyl ester (120 mg, 90% yield).
LC-MS,M/Z(ESI)：446.1[M+H] +。 LC-MS, M/Z (ESI): 446.1 [M+H] + .
第二步：(R)-1-(1-(4-(6-(二氟甲基)吡啶-2-基)苯基)乙基)-4-(丙烷-1-炔-1-基)-1H-吲唑-7-甲酸的制备The second step: (R)-1-(1-(4-(6-(difluoromethyl)pyridin-2-yl)phenyl)ethyl)-4-(propane-1-yn-1-yl )-1H-indazole-7-carboxylic acid preparation
向(R)-1-(1-(4-(6-(二氟甲基)吡啶-2-基)苯基)乙基)-4-(丙烷-1-炔-1-基)-1H-吲唑-7-甲酸甲酯(120mg,0.269mmol)的四氢呋喃(6mL)、甲醇(2mL)和水(2mL)溶液中加入氢氧化锂(32.3mg,1.347mmol)，然后在室温下搅拌18h。向反应液中加入水(50mL)，并通过1N盐酸水溶液调节pH至5～6，然后用乙酸乙酯(20mL*2)萃取。合并有机层，用饱和食盐水(10mL*2)洗涤，用无水硫酸钠干燥，过滤并浓缩得到(R)-1-(1-(4-(6-(二氟甲基)吡啶-2-基)苯基)乙基)-4-(丙烷-1-炔-1-基)-1H-吲唑-7-甲酸(100mg，产率86％)。To (R)-1-(1-(4-(6-(difluoromethyl)pyridin-2-yl)phenyl)ethyl)-4-(propane-1-yn-1-yl)-1H Add lithium hydroxide (32.3mg, 1.347mmol) to a solution of methyl indazole-7-carboxylate (120mg, 0.269mmol) in tetrahydrofuran (6mL), methanol (2mL) and water (2mL), then stir at room temperature for 18h . Water (50 mL) was added to the reaction solution, and the pH was adjusted to 5-6 by 1N aqueous hydrochloric acid solution, and then extracted with ethyl acetate (20 mL*2). The organic layers were combined, washed with saturated brine (10mL*2), dried over anhydrous sodium sulfate, filtered and concentrated to obtain (R)-1-(1-(4-(6-(difluoromethyl)pyridine-2 -yl)phenyl)ethyl)-4-(propan-1-yn-1-yl)-1H-indazole-7-carboxylic acid (100 mg, 86% yield).
LC-MS,M/Z(ESI)：432.1[M+H] +。 LC-MS, M/Z (ESI): 432.1 [M+H] + .
第三步：(Sa,R)-6-(1-(1-(4-(6-(二氟甲基)吡啶-2-基)苯基)乙基)-4-(丙烷-1-炔-1-基)-1H-吲唑 -7-羧酰胺)螺[3.3]庚烷-2-甲酸甲酯的制备The third step: (Sa,R)-6-(1-(1-(4-(6-(difluoromethyl)pyridin-2-yl)phenyl)ethyl)-4-(propane-1- Preparation of alkyn-1-yl)-1H-indazole-7-carboxamide)spiro[3.3]heptane-2-carboxylic acid methyl ester
向(R)-1-(1-(4-(6-(二氟甲基)吡啶-2-基)苯基)乙基)-4-(丙烷-1-炔-1-基)-1H-吲唑-7-甲酸(100mg，0.232mmol)的N,N-二甲基甲酰胺(2mL)溶液中加入HATU(264mg，0.695mmol)和二异丙基乙胺(90mg，0.695mmol)并在室温下搅拌1小时。然后向反应液中加入(Sa)-6-氨基螺[3.3]庚烷-2-甲酸甲酯(58.8mg，0.348mmol)并在室温下搅拌2小时。将反应液倒水(50mL)中并用乙酸乙酯(10mL*3)萃取。合并有机相，用饱和食盐水(10mL*2)洗涤，用无水硫酸钠干燥，过滤并浓缩得到粗产物。粗产物通过薄层硅胶板分离纯化(石油醚/乙酸乙酯(V/V)＝3/1)纯化得到(Sa,R)-6-(1-(1-(4-(2-(二氟甲基))吡啶-4-基)苯基)乙基)-4-(丙烷-1-炔-1-基)-1H-吲唑-7-甲酰胺基)螺[3.3]庚烷-2-甲酸甲酯(100mg，产率74％)。To (R)-1-(1-(4-(6-(difluoromethyl)pyridin-2-yl)phenyl)ethyl)-4-(propane-1-yn-1-yl)-1H -Indazole-7-carboxylic acid (100mg, 0.232mmol) in N,N-dimethylformamide (2mL) was added HATU (264mg, 0.695mmol) and diisopropylethylamine (90mg, 0.695mmol) and Stir at room temperature for 1 hour. Then, (Sa)-methyl 6-aminospiro[3.3]heptane-2-carboxylate (58.8 mg, 0.348 mmol) was added to the reaction liquid and stirred at room temperature for 2 hours. The reaction solution was poured into water (50 mL) and extracted with ethyl acetate (10 mL*3). The organic phases were combined, washed with saturated brine (10 mL*2), dried over anhydrous sodium sulfate, filtered and concentrated to obtain a crude product. The crude product was separated and purified by thin-layer silica gel plate (petroleum ether/ethyl acetate (V/V)=3/1) to obtain (Sa,R)-6-(1-(1-(4-(2-(di Fluoromethyl))pyridin-4-yl)phenyl)ethyl)-4-(propane-1-yn-1-yl)-1H-indazole-7-carboxamido)spiro[3.3]heptane- Methyl 2-carboxylate (100 mg, 74% yield).
LC-MS,M/Z(ESI)：583.3[M+H] +。 LC-MS, M/Z (ESI): 583.3 [M+H] + .
第四步：(Sa,R)-6-(1-(1-(4-(6-(二氟甲基)吡啶-2-基)苯基)乙基)-4-(丙烷-1-炔-1-基)-1H-吲唑-7-甲酰氨基)螺[3.3]庚烷-2-甲酸(I-36)的制备The fourth step: (Sa,R)-6-(1-(1-(4-(6-(difluoromethyl)pyridin-2-yl)phenyl)ethyl)-4-(propane-1- Preparation of alkyn-1-yl)-1H-indazole-7-carboxamido)spiro[3.3]heptane-2-carboxylic acid (I-36)
Figure PCTCN2022097611-appb-000107
Figure PCTCN2022097611-appb-000107
向(R)-6-(1-(1-(4-(2-(二氟甲基))吡啶-4-基)苯基)乙基)-4-(丙烷-1-炔-1-基)-1H-吲唑-7-甲酰胺基)螺[3.3]庚烷-2-甲酸甲酯(100mg，0.172mmol)的四氢呋喃(3mL)、甲醇(1mL)和水(1mL)的溶液中加入氢氧化锂(20.55mg，0.858mmol)，然后在25℃下搅拌18小时。向反应液中加入水(50mL)并通过1N盐酸水溶液调节pH至5～6，然后用EA(10mL*3)萃取。合并有机相，用饱和食盐水(10mL*2)洗涤，用无水硫酸钠干燥，过滤并浓缩得到(Sa,R)-6-(1-(1-(4-(2-(二氟甲基))吡啶-4-基)苯基)乙基)-4-(丙烷-1-炔-1-基)-1H-吲唑-7-甲酰胺基)螺[3.3]庚烷-2-甲酸(I-36)(95.9mg，产率95％)。To (R)-6-(1-(1-(4-(2-(difluoromethyl))pyridin-4-yl)phenyl)ethyl)-4-(propane-1-yne-1- In a solution of methyl)-1H-indazole-7-carboxamido)spiro[3.3]heptane-2-carboxylate (100mg, 0.172mmol) in tetrahydrofuran (3mL), methanol (1mL) and water (1mL) Lithium hydroxide (20.55 mg, 0.858 mmol) was added, followed by stirring at 25°C for 18 hours. Water (50 mL) was added to the reaction solution and the pH was adjusted to 5-6 by 1N aqueous hydrochloric acid solution, and then extracted with EA (10 mL*3). The organic phases were combined, washed with saturated brine (10mL*2), dried over anhydrous sodium sulfate, filtered and concentrated to obtain (Sa,R)-6-(1-(1-(4-(2-(difluoromethane Base))pyridin-4-yl)phenyl)ethyl)-4-(propane-1-yn-1-yl)-1H-indazole-7-carboxamido)spiro[3.3]heptane-2- Formic acid (I-36) (95.9 mg, 95% yield).
1H NMR(400MHz,dmso)δ12.03(s,1H),8.81(d,1H),8.28(s,1H),8.09-8.02(m,2H),7.94(d,2H),7.66-7.59(m,1H),7.31(d,1H),7.19(d,1H),7.15(d,1H),6.96(t,1H),6.35(q,1H),4.34(dq,1H),2.94(p,1H),2.48-2.42(m,1H),2.37-2.20(m,3H),2.20-1.98(m,6H),1.92(t,3H),1.86(dd,1H)。 1 H NMR (400MHz,dmso)δ12.03(s,1H),8.81(d,1H),8.28(s,1H),8.09-8.02(m,2H),7.94(d,2H),7.66-7.59 (m,1H),7.31(d,1H),7.19(d,1H),7.15(d,1H),6.96(t,1H),6.35(q,1H),4.34(dq,1H),2.94( p,1H), 2.48-2.42(m,1H), 2.37-2.20(m,3H), 2.20-1.98(m,6H), 1.92(t,3H), 1.86(dd,1H).
LC-MS,M/Z(ESI):569.3[M+H] +。 LC-MS, M/Z (ESI): 569.3 [M+H] + .
实施例37化合物I-37的制备The preparation of embodiment 37 compound I-37
合成路线如下所示：The synthetic route is as follows:
Figure PCTCN2022097611-appb-000108
Figure PCTCN2022097611-appb-000108
第一步：(R)-(1-(1-(4-(3,3-二氟吡咯烷-1-基)苯基)乙基)-4-(丙烷-1-炔-1-基)-1H-吲唑-7-甲酸甲酯The first step: (R)-(1-(1-(4-(3,3-difluoropyrrolidin-1-yl)phenyl)ethyl)-4-(propane-1-yn-1-yl )-1H-indazole-7-carboxylic acid methyl ester
向(R)-1-(1-(4-溴苯基)乙基)-4-(丙烷-1-炔-1-基)-1H-吲唑-7-甲酸甲酯(500mg,1.26mmol)和3,3-二氟吡咯烷(900mg,6.29mmol)的1，4-二氧六环(10mL)溶液中加入(1,1’-联苯)-2-基二-叔丁基膦(150mg,0.50mmol)、碳酸铯(2.46g,7.55mmol)和醋酸钯(57mg，0.25mmol)。反应液置换三次氮气，然后在80℃下搅拌16小时。将反应液倒入水(50mL)中并乙酸乙酯(30mL*3)萃取。合并有机相，用饱和食盐水(20mL*2)洗涤，并用无水硫酸钠干燥，过滤并浓缩得到粗产物。粗产物通过柱层析(石油醚/乙酸乙酯(V/V)＝1:0-1:3)纯化得到(R)-(1-(1-(4-(3,3-二氟吡咯烷-1-基)苯基)乙基)-4-(丙烷-1-炔-1-基)-1H-吲唑-7-甲酸甲酯(150mg,产率22％)。To (R)-1-(1-(4-bromophenyl)ethyl)-4-(propane-1-yn-1-yl)-1H-indazole-7-carboxylic acid methyl ester (500mg, 1.26mmol ) and 3,3-difluoropyrrolidine (900mg, 6.29mmol) in 1,4-dioxane (10mL) solution was added (1,1'-biphenyl)-2-yldi-tert-butylphosphine (150mg, 0.50mmol), cesium carbonate (2.46g, 7.55mmol) and palladium acetate (57mg, 0.25mmol). The reaction solution was replaced with nitrogen three times, and then stirred at 80° C. for 16 hours. The reaction solution was poured into water (50 mL) and extracted with ethyl acetate (30 mL*3). The organic phases were combined, washed with saturated brine (20 mL*2), dried over anhydrous sodium sulfate, filtered and concentrated to obtain a crude product. The crude product was purified by column chromatography (petroleum ether/ethyl acetate (V/V)=1:0-1:3) to obtain (R)-(1-(1-(4-(3,3-difluoropyrrole Alk-1-yl)phenyl)ethyl)-4-(propan-1-yn-1-yl)-1H-indazole-7-carboxylic acid methyl ester (150 mg, 22% yield).
第二步：(R)-(1-(1-(4-(3,3-二氟吡咯烷-1-基)苯基)乙基)-4-(丙烷-1-炔-1-基)-1H-吲唑-7-甲酸The second step: (R)-(1-(1-(4-(3,3-difluoropyrrolidin-1-yl)phenyl)ethyl)-4-(propane-1-yn-1-yl )-1H-indazole-7-carboxylic acid
向(R)-(1-(1-(4-(3,3-二氟吡咯烷-1-基)苯基)乙基)-4-(丙烷-1-炔-1-基)-1H-吲唑-7-甲酸甲酯(150mg,0.28mmol)的四氢呋喃(6mL)、甲醇(2mL)和水(2mL)溶液中加入氢氧化锂(34mg,1.42mmol)，然后在25℃搅拌3小时。向反应液中加入水(50mL)并用1N盐酸水溶液调节pH至5～6，然后用乙酸乙酯(20mL*3)萃取。合并有机相，用饱和食盐水(10mL*2)洗涤，并用无水硫酸钠干燥，过滤并浓缩得到(R)-(1-(1-(4-(3,3-二氟吡咯烷-1-基)苯基)乙基)-4-(丙烷-1-炔-1-基)-1H-吲唑-7-甲酸(110mg，产率95％)。To (R)-(1-(1-(4-(3,3-difluoropyrrolidin-1-yl)phenyl)ethyl)-4-(propane-1-yn-1-yl)-1H Add lithium hydroxide (34mg, 1.42mmol) to a solution of methyl indazole-7-carboxylate (150mg, 0.28mmol) in tetrahydrofuran (6mL), methanol (2mL) and water (2mL), then stir at 25°C for 3 hours Add water (50mL) to the reaction solution and adjust the pH to 5-6 with 1N hydrochloric acid aqueous solution, then extract with ethyl acetate (20mL*3). Combine the organic phases, wash with saturated brine (10mL*2), and wash with over sodium sulfate, filtered and concentrated to give (R)-(1-(1-(4-(3,3-difluoropyrrolidin-1-yl)phenyl)ethyl)-4-(propane-1- Alkyn-1-yl)-1H-indazole-7-carboxylic acid (110 mg, 95% yield).
第三步：(Sa,R)-6-(1-(1-(4-(3,3-二氟吡咯烷-1-基)苯基)乙基)-4-(丙烷-1-炔-1-基)-1H-吲唑-7-甲酰胺基)螺[3.3]庚烷-2(S)-甲酸甲酯The third step: (Sa,R)-6-(1-(1-(4-(3,3-difluoropyrrolidin-1-yl)phenyl)ethyl)-4-(propane-1-yne -1-yl)-1H-indazole-7-carboxamido)spiro[3.3]heptane-2(S)-methyl carboxylate
向(R)-(1-(1-(4-(3,3-二氟吡咯烷-1-基)苯基)乙基)-4-(丙烷-1-炔-1-基)-1H-吲唑-7-甲酸(100mg,0.24mmol)的N,N-二甲基甲酰胺(2mL)溶液中加入HATU(139mg,0.37mmol)和二异丙基乙胺(158mg,1.22mmol)，并在25℃下搅拌0.5h。然后向反应液中加入(Sa)-6-氨基螺[3.3]庚烷-2-甲酸甲酯(100mg，0.49mmol)，在25℃下继续搅拌2.5小时。反应液浓缩得到粗产物。粗产物通过柱层析(石油醚/乙酸乙酯(V/V)＝1/0-1/3)纯化得到(Sa,R)-6-(1-(1-(4-(3,3-二氟吡咯烷-1-基)苯基)乙基)-4-(丙烷-1-炔-1-基)-1H-吲唑-7-甲酰胺基)螺[3.3]庚烷-2(S)-甲酸甲酯 (100mg，产率73％)。To (R)-(1-(1-(4-(3,3-difluoropyrrolidin-1-yl)phenyl)ethyl)-4-(propane-1-yn-1-yl)-1H -Indazole-7-carboxylic acid (100mg, 0.24mmol) in N,N-dimethylformamide (2mL) solution was added HATU (139mg, 0.37mmol) and diisopropylethylamine (158mg, 1.22mmol), And stirred at 25° C. for 0.5 h. Then, (Sa)-methyl 6-aminospiro[3.3]heptane-2-carboxylate (100 mg, 0.49 mmol) was added to the reaction solution, and stirring was continued at 25° C. for 2.5 hours. The reaction solution was concentrated to obtain a crude product.The crude product was purified by column chromatography (petroleum ether/ethyl acetate (V/V)=1/0-1/3) to obtain (Sa,R)-6-(1-(1- (4-(3,3-difluoropyrrolidin-1-yl)phenyl)ethyl)-4-(propane-1-yn-1-yl)-1H-indazole-7-carboxamido)spiro [3.3] Heptane-2(S)-methyl carboxylate (100 mg, yield 73%).
第四步：(Sa,R)-6-(1-(1-(4-(3,3-二氟吡咯烷-1-基)苯基)乙基)-4-(丙烷-1-炔-1-基)-1H-吲唑-7-甲酰胺基)螺[3.3]庚烷-2(S)-甲酸(I-37)The fourth step: (Sa,R)-6-(1-(1-(4-(3,3-difluoropyrrolidin-1-yl)phenyl)ethyl)-4-(propane-1-yne -1-yl)-1H-indazole-7-carboxamido)spiro[3.3]heptane-2(S)-carboxylic acid (I-37)
Figure PCTCN2022097611-appb-000109
Figure PCTCN2022097611-appb-000109
向(Sa,R)-6-(1-(1-(4-(3,3-二氟吡咯烷-1-基)苯基)乙基)-4-(丙烷-1-炔-1-基)-1H-吲唑-7-甲酰胺基)螺[3.3]庚烷-2(S)-甲酸甲酯(100mg，0.18mmol)的四氢呋喃(6mL)、甲醇(2mL)和水(2mL)的溶液中加入氢氧化锂(45.6mg，1.906mmol)，然后在25℃搅拌3小时。向反应液中加入水(50mL)并用1N盐酸水溶液调节pH至5～6，然后用乙酸乙酯(20mL*3)萃取。合并有机相，用饱和食盐水(10mL*2)洗涤，用无水硫酸钠干燥，过滤并浓缩得到(Sa,R)-6-(1-(1-(4-(3,3-二氟吡咯烷-1-基)苯基)乙基)-4-(丙烷-1-炔-1-基)-1H-吲唑-7-甲酰胺基)螺[3.3]庚烷-2(S)-甲酸(I-37)(20mg，产率21％)。To (Sa,R)-6-(1-(1-(4-(3,3-difluoropyrrolidin-1-yl)phenyl)ethyl)-4-(propane-1-yne-1- yl)-1H-indazole-7-carboxamido)spiro[3.3]heptane-2(S)-methyl carboxylate (100 mg, 0.18 mmol) in tetrahydrofuran (6 mL), methanol (2 mL) and water (2 mL) Lithium hydroxide (45.6mg, 1.906mmol) was added to the solution, followed by stirring at 25°C for 3 hours. Water (50 mL) was added to the reaction solution, and the pH was adjusted to 5-6 with 1N aqueous hydrochloric acid solution, followed by extraction with ethyl acetate (20 mL*3). The organic phases were combined, washed with saturated brine (10mL*2), dried over anhydrous sodium sulfate, filtered and concentrated to obtain (Sa,R)-6-(1-(1-(4-(3,3-difluoro Pyrrolidin-1-yl)phenyl)ethyl)-4-(propane-1-yn-1-yl)-1H-indazole-7-carboxamido)spiro[3.3]heptane-2(S) - Formic acid (I-37) (20 mg, 21% yield).
1H NMR(400MHz,dmso)δ8.82(d,1H),8.18(s,1H),7.26(d,1H),7.14(d,1H),6.91(d,1H),6.42(d,1H),6.13(q,1H),4.34(dd,1H),3.56(t,4H),2.93(t,1H),2.30–1.81(m,16H). 1 H NMR (400MHz,dmso)δ8.82(d,1H),8.18(s,1H),7.26(d,1H),7.14(d,1H),6.91(d,1H),6.42(d,1H ), 6.13(q,1H), 4.34(dd,1H), 3.56(t,4H), 2.93(t,1H), 2.30–1.81(m,16H).
LC-MS,M/Z(ESI):511.60[M+H] +。 LC-MS, M/Z (ESI): 511.60 [M+H] + .
实施例38化合物I-38的制备Preparation of Example 38 Compound I-38
合成路线如下所示：The synthetic route is as follows:
Figure PCTCN2022097611-appb-000110
Figure PCTCN2022097611-appb-000110
第一步：(1-(4-溴苯基)乙氧基)(叔丁基)二甲基硅烷的制备Step 1: Preparation of (1-(4-bromophenyl)ethoxy)(tert-butyl)dimethylsilane
将1-(4-溴苯基)-1-乙醇(3g,14.92mmol)和咪唑(2g,29.8mmol)溶于N,N-二甲基甲酰胺(30mL)中，冰浴条件下加入叔丁基二甲基氯硅烷(3.37g,22.38mmol)，反应液继续搅拌2小时。往反应液中加入水(200mL),用乙酸乙酯(50mL×3)萃取。有机相合并后用饱和食盐水(50mL×2)洗涤，浓缩后用柱层析纯化，得到(1-(4-溴苯基)乙氧基)(叔丁基)二甲基硅烷(4.3g，产率91％)。Dissolve 1-(4-bromophenyl)-1-ethanol (3g, 14.92mmol) and imidazole (2g, 29.8mmol) in N,N-dimethylformamide (30mL), add t- Butyldimethylsilyl chloride (3.37g, 22.38mmol), the reaction solution was stirred for 2 hours. Water (200 mL) was added to the reaction liquid, and extracted with ethyl acetate (50 mL×3). The combined organic phases were washed with saturated brine (50mL×2), concentrated and purified by column chromatography to obtain (1-(4-bromophenyl)ethoxy)(tert-butyl)dimethylsilane (4.3g , yield 91%).
第二步：叔丁基-(1-(4-环丁基苯基)乙氧基)二甲基硅烷的制备The second step: the preparation of tert-butyl-(1-(4-cyclobutylphenyl)ethoxy)dimethylsilane
(1-(4-溴苯基)乙氧基)(叔丁基)二甲基硅烷(1g,3.17mmol)、环丁基氟硼酸钾(617mg,3.81mmol)、醋酸钯(36mg,0.16mmol)、正丁基二(1-金刚烷基)膦(91mg,0.254mmol)和碳酸铯(3.1g,9.51mmol)于甲苯(10mL)和水(1mL)中，氮气保护条件下100℃搅拌16小时。往反应液中加入水(10mL)，用乙酸乙酯(10mL×2)萃取。上述有机相合并，用无水硫酸钠干燥，浓缩后得到粗品叔丁基-(1-(4-环丁基苯基)乙氧基)二甲基硅烷(0.67g，产率73％)。(1-(4-bromophenyl)ethoxy)(tert-butyl)dimethylsilane (1g, 3.17mmol), potassium cyclobutylfluoroborate (617mg, 3.81mmol), palladium acetate (36mg, 0.16mmol ), n-butylbis(1-adamantyl)phosphine (91mg, 0.254mmol) and cesium carbonate (3.1g, 9.51mmol) in toluene (10mL) and water (1mL), stirred at 100°C for 16 Hour. Water (10 mL) was added to the reaction liquid, and extracted with ethyl acetate (10 mL×2). The above organic phases were combined, dried over anhydrous sodium sulfate, and concentrated to obtain crude tert-butyl-(1-(4-cyclobutylphenyl)ethoxy)dimethylsilane (0.67 g, yield 73%).
第三步：1-(4-环丁基苯基)乙氧基-1-醇的制备The third step: the preparation of 1-(4-cyclobutylphenyl)ethoxy-1-ol
粗品叔丁基-(1-(4-环丁基苯基)乙氧基)二甲基硅烷(0.87g,3mmol)溶于四氢呋喃中，加入四丁基氟化铵(9mL,1M/L四氢呋喃溶液)，室温搅拌12小时。反应液浓缩，柱层析纯化得到1-(4-环丁基苯基)乙氧基-1-醇(0.3g，产率57％)。Crude tert-butyl-(1-(4-cyclobutylphenyl)ethoxy)dimethylsilane (0.87g, 3mmol) was dissolved in tetrahydrofuran, and tetrabutylammonium fluoride (9mL, 1M/L tetrahydrofuran solution), stirred at room temperature for 12 hours. The reaction solution was concentrated and purified by column chromatography to obtain 1-(4-cyclobutylphenyl)ethoxy-1-ol (0.3 g, yield 57%).
第四步：(1-(1-(4-环丁基苯基)乙基)-4-(丙烷-1-炔-1-基)-1H-吲唑-7-甲酸甲酯的制备Step 4: Preparation of (1-(1-(4-cyclobutylphenyl)ethyl)-4-(propane-1-yn-1-yl)-1H-indazole-7-carboxylic acid methyl ester
1-(4-环丁基苯基)乙氧基-1-醇(240mg,1.362mmol)、甲基4-(丙烷-1-炔-1-基)-1H-吲唑-7-甲酸酯(146mg,0.681mmol)和三苯基膦(357mg,1.362mmol)溶于四氢呋喃(20mL)中，冰浴下缓慢滴加DIAD(275mg,1.362mmol)，随后升至室温搅拌16小时。反应液浓缩，柱层析纯化得(1-(1-(4-环丁基苯基)乙基)-4-(丙烷-1-炔-1-基)-1H-吲唑-7-甲酸甲酯(100mg，产率39％)。1-(4-cyclobutylphenyl)ethoxy-1-ol (240mg, 1.362mmol), methyl 4-(propane-1-yn-1-yl)-1H-indazole-7-carboxylic acid Ester (146mg, 0.681mmol) and triphenylphosphine (357mg, 1.362mmol) were dissolved in tetrahydrofuran (20mL), and DIAD (275mg, 1.362mmol) was slowly added dropwise under ice-cooling, followed by warming to room temperature and stirring for 16 hours. The reaction solution was concentrated and purified by column chromatography to obtain (1-(1-(4-cyclobutylphenyl)ethyl)-4-(propane-1-yn-1-yl)-1H-indazole-7-carboxylic acid Methyl ester (100 mg, 39% yield).
第五步：(1-(1-(4-环丁基苯基)乙基)-4-(丙烷-1-炔-1-基)-1H-吲唑-7-甲酸的制备Step 5: Preparation of (1-(1-(4-cyclobutylphenyl)ethyl)-4-(propane-1-yn-1-yl)-1H-indazole-7-carboxylic acid
(1-(1-(4-环丁基苯基)乙基)-4-(丙烷-1-炔-1-基)-1H-吲唑-7-甲酸甲酯(100mg,0.268mmol)溶于四氢呋喃/甲醇/水(2mL/0.4mL/0.4mL)中，加入一水合氢氧化锂(56mg,1.342mmol)，室温搅拌16小时。反应液浓缩，加入水(5mL)，再加稀盐酸(1mol/L水溶液)调节溶液pH值约2，有白色固体产生，过滤收集白色固体，干燥后得到(1-(1-(4-环丁基苯基)乙基)-4-(丙烷-1-炔-1-基)-1H-吲唑-7-甲酸(80mg，收率83％)。(1-(1-(4-cyclobutylphenyl)ethyl)-4-(propane-1-yn-1-yl)-1H-indazole-7-carboxylic acid methyl ester (100mg, 0.268mmol) In tetrahydrofuran/methanol/water (2mL/0.4mL/0.4mL), add lithium hydroxide monohydrate (56mg, 1.342mmol), and stir at room temperature for 16 hours. The reaction solution is concentrated, water (5mL) is added, and then dilute hydrochloric acid ( 1mol/L aqueous solution) to adjust the pH value of the solution to about 2, a white solid is produced, the white solid is collected by filtration, and dried to obtain (1-(1-(4-cyclobutylphenyl)ethyl)-4-(propane-1 -Alkyn-1-yl)-1H-indazole-7-carboxylic acid (80 mg, yield 83%).
第六步：(Sa)-6-(1-(1-(4-环丁基苯基)乙基)-4-(丙烷-1-炔-1-基)-1H-吲唑-7-甲酰胺)螺[3.3]庚烷-2-甲酸甲酯的制备The sixth step: (Sa)-6-(1-(1-(4-cyclobutylphenyl)ethyl)-4-(propane-1-yn-1-yl)-1H-indazole-7- Formamide) the preparation of spiro[3.3]heptane-2-carboxylic acid methyl ester
(1-(1-(4-环丁基苯基)乙基)-4-(丙烷-1-炔-1-基)-1H-吲唑-7-甲酸(80mg,0.223mmol)和N,N,N′,N′-四甲基-O-(7-氮杂苯并三唑-1-基)六氟磷酸脲(255mg,0.67mmol)溶于N,N-二甲基甲酰胺(3mL)中，冰浴下加入N,N-二异丙基乙胺(144mg,1.116mmol)，搅拌15分钟后加入(Sa)-6-氨基螺[3.3]庚烷-2-甲酸甲酯盐酸盐(138mg,0.67mmol)，室温搅拌16小时。反应液中加入水(10mL)，乙酸乙酯(10mL×3)萃取，有机相依次用饱和食盐水(10mL)洗涤，无水硫 酸钠干燥，浓缩后柱层析纯化，得(Sa)-6-(1-(1-(4-环丁基苯基)乙基)-4-(丙烷-1-炔-1-基)-1H-吲唑-7-甲酰胺)螺[3.3]庚烷-2-甲酸甲酯(70mg，产率62％)。(1-(1-(4-cyclobutylphenyl)ethyl)-4-(propane-1-yn-1-yl)-1H-indazole-7-carboxylic acid (80mg, 0.223mmol) and N, N,N',N'-tetramethyl-O-(7-azabenzotriazol-1-yl)urea hexafluorophosphate (255mg, 0.67mmol) was dissolved in N,N-dimethylformamide ( 3mL), add N,N-diisopropylethylamine (144mg, 1.116mmol) under ice-cooling, and add (Sa)-6-aminospiro[3.3]heptane-2-carboxylic acid methyl ester salt after stirring for 15 minutes Acetate (138mg, 0.67mmol), stirred at room temperature for 16 hours. Added water (10mL) to the reaction solution, extracted with ethyl acetate (10mL×3), the organic phase was washed with saturated brine (10mL) successively, and dried over anhydrous sodium sulfate , concentrated and purified by column chromatography to obtain (Sa)-6-(1-(1-(4-cyclobutylphenyl)ethyl)-4-(propane-1-yn-1-yl)-1H- Indazole-7-carboxamide) spiro[3.3]heptane-2-carboxylic acid methyl ester (70 mg, 62% yield).
LC-MS,M/Z(ESI):510.5[M+H] +。 LC-MS, M/Z (ESI): 510.5 [M+H] + .
第七步：(Sa)-6-(1-(1-(4-环丁基苯基)乙基)-4-(丙烷-1-炔-1-基)-1H-吲唑-7-甲酰胺)螺[3.3]庚烷-2-甲酸(I-38)The seventh step: (Sa)-6-(1-(1-(4-cyclobutylphenyl)ethyl)-4-(propane-1-yn-1-yl)-1H-indazole-7- Formamide) spiro[3.3]heptane-2-carboxylic acid (I-38)
(Sa)-6-(1-(1-(4-环丁基苯基)乙基)-4-(丙烷-1-炔-1-基)-1H-吲唑-7-甲酰胺)螺[3.3]庚烷-2-甲酸甲酯(70mg,0.137mmol)溶于四氢呋喃/甲醇/水(3mL/0.6mL/0.6mL)中，加入一水合氢氧化锂(28.8mg,0.687mmol)，室温搅拌16小时。反应液浓缩，反相制备液相色谱纯化，冻干后得到(Sa)-6-(1-(1-(4-环丁基苯基)乙基)-4-(丙烷-1-炔-1-基)-1H-吲唑-7-甲酰胺)螺[3.3]庚烷-2-甲酸(I-38)(22mg，收率32％)。(Sa)-6-(1-(1-(4-cyclobutylphenyl)ethyl)-4-(propane-1-yn-1-yl)-1H-indazole-7-carboxamide)spiro [3.3] Dissolve methyl heptane-2-carboxylate (70mg, 0.137mmol) in tetrahydrofuran/methanol/water (3mL/0.6mL/0.6mL), add lithium hydroxide monohydrate (28.8mg, 0.687mmol), room temperature Stir for 16 hours. The reaction solution was concentrated, purified by reverse-phase preparative liquid chromatography, and (Sa)-6-(1-(1-(4-cyclobutylphenyl)ethyl)-4-(propane-1-yne- 1-yl)-1H-indazole-7-carboxamide)spiro[3.3]heptane-2-carboxylic acid (I-38) (22 mg, yield 32%).
LC-MS,M/Z(ESI):338.3[M+H] +。 LC-MS, M/Z (ESI): 338.3 [M+H] + .
1H NMR(400MHz,DMSO-d6)δ8.79(dd,1H),8.23(s,1H),7.28(d,1H),7.17(d,1H),7.07(dd,2H),6.95(dd,2H),6.24(q,1H),4.40–4.25(m,1H),3.00–2.89(m,1H),2.48–2.41(m,1H),2.37–1.82(m,19H),1.79–1.70(m,1H). 1 H NMR(400MHz,DMSO-d6)δ8.79(dd,1H),8.23(s,1H),7.28(d,1H),7.17(d,1H),7.07(dd,2H),6.95(dd ,2H),6.24(q,1H),4.40–4.25(m,1H),3.00–2.89(m,1H),2.48–2.41(m,1H),2.37–1.82(m,19H),1.79–1.70 (m,1H).
实施例39化合物I-39a的制备Preparation of Example 39 Compound I-39a
合成路线如下所示：The synthetic route is as follows:
Figure PCTCN2022097611-appb-000111
Figure PCTCN2022097611-appb-000111
第一步：1-(4-(二氟甲氧基)苯基)乙醇的制备The first step: the preparation of 1-(4-(difluoromethoxy)phenyl)ethanol
在0°下向1-(4-(二氟甲氧基)苯基)乙酮(500mg,2.69mmol)的甲醇(5mL)溶液中加入硼氢化钠(203mg,5.37mmol)，然后在0℃下搅拌1h。将反应液缓慢倒入饱和氯化铵水溶液(50mL)中并用二氯甲烷(20mL*2)萃取。合并有机相，用饱和食盐水(10mL*2)洗涤，用无水硫酸钠干燥，过滤并浓缩得到1-(4-(二氟甲氧基)苯基)乙醇(400mg，产率79％)。To a solution of 1-(4-(difluoromethoxy)phenyl)ethanone (500mg, 2.69mmol) in methanol (5mL) was added sodium borohydride (203mg, 5.37mmol) at 0°C, and then Stir for 1h. The reaction solution was slowly poured into saturated aqueous ammonium chloride (50 mL) and extracted with dichloromethane (20 mL*2). The organic phases were combined, washed with saturated brine (10mL*2), dried over anhydrous sodium sulfate, filtered and concentrated to give 1-(4-(difluoromethoxy)phenyl)ethanol (400mg, yield 79%) .
第二步：1-(1-(4-(二氟甲氧基)苯基)乙基)-4-(丙烷-1-炔-1-基)-1H-吲唑-7-甲酸甲酯的制备The second step: 1-(1-(4-(difluoromethoxy)phenyl)ethyl)-4-(propane-1-yn-1-yl)-1H-indazole-7-carboxylic acid methyl ester preparation of
向1-(4-(二氟甲氧基)苯基)乙醇(400mg,2.126mmol)的四氢呋喃(10mL)溶液中加入4-(丙烷-1-炔-1-基)-1H-吲唑-7-甲酸甲酯(228mg,1.063mmol)，三苯基膦(836mg，3.19mmol)。然后在0℃下将偶氮二甲酸二异丙酯(645mg，3.19mmol)逐滴加入反应液中，并在25℃下搅拌16小时。浓缩反应液得到粗产物。粗产物通过柱层析(石油醚/乙酸乙酯(V/V)＝1/0-1/1)纯化得到1-(1-(4-(二氟甲氧基)苯基)乙基)-4-(丙烷-1-炔-1-基)-1H-吲唑-7-甲酸甲酯(200mg，产率49.0％)。To a solution of 1-(4-(difluoromethoxy)phenyl)ethanol (400 mg, 2.126 mmol) in tetrahydrofuran (10 mL) was added 4-(propane-1-yn-1-yl)-1H-indazole- Methyl 7-carboxylate (228 mg, 1.063 mmol), triphenylphosphine (836 mg, 3.19 mmol). Then diisopropyl azodicarboxylate (645 mg, 3.19 mmol) was added dropwise to the reaction solution at 0°C, and stirred at 25°C for 16 hours. The reaction solution was concentrated to obtain a crude product. The crude product was purified by column chromatography (petroleum ether/ethyl acetate (V/V)=1/0-1/1) to give 1-(1-(4-(difluoromethoxy)phenyl)ethyl) -4-(Propan-1-yn-1-yl)-1H-indazole-7-carboxylic acid methyl ester (200 mg, yield 49.0%).
LC-MS,M/Z(ESI):385.4[M+H] +。 LC-MS, M/Z (ESI): 385.4 [M+H] + .
第三步：1-(1-(4-(二氟甲氧基)苯基)乙基)-4-(丙烷-1-炔-1-基)-1H-吲唑-7-甲酸的制备Step 3: Preparation of 1-(1-(4-(difluoromethoxy)phenyl)ethyl)-4-(propane-1-yn-1-yl)-1H-indazole-7-carboxylic acid
向1-(1-(4-(二氟甲氧基)苯基)乙基)-4-(丙烷-1-炔-1-基)-1H-吲唑-7-甲酸甲酯(200mg,0.520mmol)的四氢呋喃(6mL)、甲醇(2mL)和水(2mL)溶液中加入氢氧化锂(62.3mg，2.60mmol)，然后在25℃下搅拌16小时。向反应液中加入水(50mL)并用1N盐酸水溶液调节pH至5～6，并用EA(20mL*3)萃取。合并有机相，用无水硫酸钠干燥，过滤，浓缩得到1-(1-(4-(二氟甲氧基)苯基)乙基)-4-(丙烷-1-炔-1-基)-1H-吲唑-7-甲酸(180mg，产率93％)。To 1-(1-(4-(difluoromethoxy)phenyl)ethyl)-4-(propane-1-yn-1-yl)-1H-indazole-7-carboxylic acid methyl ester (200mg, Lithium hydroxide (62.3 mg, 2.60 mmol) was added to a solution of 0.520 mmol) in tetrahydrofuran (6 mL), methanol (2 mL) and water (2 mL), followed by stirring at 25° C. for 16 hours. Water (50 mL) was added to the reaction solution and the pH was adjusted to 5-6 with 1N aqueous hydrochloric acid solution, and extracted with EA (20 mL*3). The organic phases were combined, dried over anhydrous sodium sulfate, filtered and concentrated to give 1-(1-(4-(difluoromethoxy)phenyl)ethyl)-4-(propan-1-yn-1-yl) -1H-indazole-7-carboxylic acid (180 mg, yield 93%).
LC-MS,M/Z(ESI):371.3[M+H] +。 LC-MS, M/Z (ESI): 371.3 [M+H] + .
第四步：(Sa)-6-(1-(1-(4-(二氟甲氧基)苯基)乙基)-4-(丙烷-1-炔-1-基)-1H-吲唑-7-甲酰胺基)螺[3.3]庚烷-2-甲酸甲酯的制备The fourth step: (Sa)-6-(1-(1-(4-(difluoromethoxy)phenyl)ethyl)-4-(propane-1-yn-1-yl)-1H-ind Preparation of azole-7-carboxamido)spiro[3.3]heptane-2-carboxylic acid methyl ester
将1-(1-(4-(二氟甲氧基)苯基)乙基)-4-(丙烷-1-炔-1-基)-1H-吲唑-7-甲酸(180mg，0.486mmol)、(Sa)-6-氨基螺[3.3]庚烷-2-甲酸甲酯(206mg，1.215mmol)、二异丙基乙胺(188mg，1.458mmol)和HATU(554mg，1.458mmol)溶于N,N-二甲基甲酰胺(3mL)中，然后室温搅拌3h。将反应液加入水(50mL)中，用乙酸乙酯(10mL*3)萃取。有机相合并后用饱和食盐水(10mL*3)洗涤，无水硫酸钠干燥，过滤，浓缩得到粗品。粗品经过薄层硅胶板分离纯化(石油醚:乙酸乙酯(V/V)＝1:1)得到(Sa)-6-(1-(1-(4-(二氟甲氧基)苯基)乙基)-4-(丙烷-1-炔-1-基)-1H-吲唑-7-甲酰胺基)螺[3.3]庚烷-2-甲酸甲酯(170mg，产率67.1％)。1-(1-(4-(Difluoromethoxy)phenyl)ethyl)-4-(propane-1-yn-1-yl)-1H-indazole-7-carboxylic acid (180mg, 0.486mmol ), (Sa)-methyl 6-aminospiro[3.3]heptane-2-carboxylate (206mg, 1.215mmol), diisopropylethylamine (188mg, 1.458mmol) and HATU (554mg, 1.458mmol) were dissolved in N,N-dimethylformamide (3 mL), then stirred at room temperature for 3 h. The reaction solution was added to water (50 mL), and extracted with ethyl acetate (10 mL*3). The combined organic phases were washed with saturated brine (10 mL*3), dried over anhydrous sodium sulfate, filtered, and concentrated to obtain a crude product. The crude product was separated and purified on a thin-layer silica gel plate (petroleum ether:ethyl acetate (V/V)=1:1) to obtain (Sa)-6-(1-(1-(4-(difluoromethoxy)phenyl )ethyl)-4-(propane-1-yn-1-yl)-1H-indazole-7-carboxamido)spiro[3.3]heptane-2-carboxylic acid methyl ester (170 mg, yield 67.1%) .
LC-MS,M/Z(ESI):522.4[M+H] +。 LC-MS, M/Z (ESI): 522.4 [M+H] + .
第五步：(Sa)-6-(1-(1-(4-(二氟甲氧基)苯基)乙基)-4-(丙烷-1-炔-1-基)-1H-吲唑-7-甲酰胺基)螺[3.3]庚烷-2-甲酸的制备The fifth step: (Sa)-6-(1-(1-(4-(difluoromethoxy)phenyl)ethyl)-4-(propane-1-yn-1-yl)-1H-ind Preparation of azole-7-carboxamido)spiro[3.3]heptane-2-carboxylic acid
将(Sa)-6-(1-(1-(4-(二氟甲氧基)苯基)乙基)-4-(丙烷-1-炔-1-基)-1H-吲唑-7-甲酰胺基)螺[3.3]庚烷-2-甲酸甲酯(170mg，0.326mmol)溶于四氢呋喃(6mL)、甲醇(2mL)和水(2mL)中，加入氢氧化锂(39mg，1.630mmol)，反应液在25℃搅拌16h。向反应液中加入水(50mL),然后用1N盐酸水溶液调pH至5～6，乙酸乙酯(20mL*3)萃取。有机相合并，用饱和食盐水(20mL*2)洗涤，用无水硫酸钠干燥，过滤，浓缩得到产物(Sa)-6-(1-(1-(4-(二氟甲氧基)苯基)乙基)-4-(丙烷-1-炔-1-基)-1H-吲唑-7-甲酰胺基)螺[3.3]庚烷-2-甲酸(121.4mg，产率98％)。(Sa)-6-(1-(1-(4-(difluoromethoxy)phenyl)ethyl)-4-(propane-1-yn-1-yl)-1H-indazole-7 -Carboxamido)spiro[3.3]heptane-2-carboxylic acid methyl ester (170 mg, 0.326 mmol) was dissolved in tetrahydrofuran (6 mL), methanol (2 mL) and water (2 mL), lithium hydroxide (39 mg, 1.630 mmol) was added ), and the reaction solution was stirred at 25°C for 16h. Water (50 mL) was added to the reaction liquid, then the pH was adjusted to 5-6 with 1N aqueous hydrochloric acid solution, and extracted with ethyl acetate (20 mL*3). The organic phases were combined, washed with saturated brine (20mL*2), dried over anhydrous sodium sulfate, filtered, and concentrated to obtain the product (Sa)-6-(1-(1-(4-(difluoromethoxy)benzene yl)ethyl)-4-(propane-1-yn-1-yl)-1H-indazole-7-carboxamido)spiro[3.3]heptane-2-carboxylic acid (121.4 mg, yield 98%) .
LC-MS,M/Z(ESI):508.4[M+H] + LC-MS, M/Z(ESI):508.4[M+H] +
第六步：化合物I-39a和化合物I-39b的制备Step 6: Preparation of Compound I-39a and Compound I-39b
Figure PCTCN2022097611-appb-000112
Figure PCTCN2022097611-appb-000112
(Sa)-6-(1-(1-(4-(二氟甲氧基)苯基)乙基)-4-(丙烷-1-炔-1-基)-1H-吲唑-7-甲酰胺基)螺[3.3]庚烷-2-甲酸(121.4mg,0.234mmol)经过SFC拆分得到(Sa,R)-6-(1-(1-(4-(二氟甲氧基)苯基)乙基)-4-(丙烷-1-炔-1-基)-1H-吲唑-7-甲酰胺基)螺[3.3]庚烷-2-甲酸(I-39a)(25.9mg，白色固体产物)和(Sa,S)-6-(1-(1-(4-(二氟甲氧基)苯基)乙基)-4-(丙烷-1-炔-1-基)-1H-吲唑-7-甲酰胺基)螺[3.3]庚烷-2-甲酸(I-39b)(26.1mg，白色固体产物)。SFC拆分方法：使用(S,S)Whelk-O1型号手性拆分柱(规格为：50×4.6mm,3.5um)进行手性拆分，流动相A为超临界流体CO 2,流动相B为甲醇(含0.05％二乙醇胺)；梯度条件:流动相B为5％-40％的梯度保持10min，柱温保持在35℃，柱压保持在100bar。I-39a保留时间：1.903min；I-39b保留时间：2.231min。 (Sa)-6-(1-(1-(4-(Difluoromethoxy)phenyl)ethyl)-4-(propane-1-yn-1-yl)-1H-indazole-7- Formamido)spiro[3.3]heptane-2-carboxylic acid (121.4mg, 0.234mmol) was resolved by SFC to obtain (Sa,R)-6-(1-(1-(4-(difluoromethoxy) Phenyl)ethyl)-4-(propane-1-yn-1-yl)-1H-indazole-7-carboxamido)spiro[3.3]heptane-2-carboxylic acid (I-39a) (25.9mg , white solid product) and (Sa,S)-6-(1-(1-(4-(difluoromethoxy)phenyl)ethyl)-4-(propan-1-yn-1-yl) - 1H-indazole-7-carboxamido)spiro[3.3]heptane-2-carboxylic acid (I-39b) (26.1 mg, white solid product). SFC resolution method: use (S,S)Whelk-O1 chiral resolution column (specification: 50×4.6mm, 3.5um) for chiral resolution, mobile phase A is supercritical fluid CO 2 , mobile phase B is methanol (containing 0.05% diethanolamine); Gradient conditions: mobile phase B is 5%-40% gradient for 10min, column temperature is maintained at 35°C, and column pressure is maintained at 100bar. I-39a retention time: 1.903 min; I-39b retention time: 2.231 min.
I-39a： 1H NMR(400MHz,dmso)δ11.99(s,1H),8.81(d,1H),8.25(s,1H),7.36–7.25(m,1H),7.21–7.12(m,1H),7.14–6.90(m,4H),6.27(q,1H),4.32(dq,1H),3.01–2.90(m,1H),2.48–2.39(m,1H),2.37–1.84(m,14H). I-39a: 1 H NMR (400MHz, dmso) δ11.99 (s, 1H), 8.81 (d, 1H), 8.25 (s, 1H), 7.36–7.25 (m, 1H), 7.21–7.12 (m, 1H),7.14–6.90(m,4H),6.27(q,1H),4.32(dq,1H),3.01–2.90(m,1H),2.48–2.39(m,1H),2.37–1.84(m, 14H).
LC-MS,M/Z(ESI):508.2[M+H] +。 LC-MS, M/Z (ESI): 508.2 [M+H] + .
I-39b： 1H NMR(400MHz,dmso)δ12.04–11.67(m,1H),8.81(d,1H),8.25(s,1H),7.35–7.26(m,1H),7.19(d,1H),7.10–6.84(m,4H),6.26(q,1H),4.32(dq,1H),3.05–2.83(m,1H),2.49–2.39(m,1H),2.34–1.76(m,14H). I-39b: 1 H NMR (400MHz, dmso) δ12.04–11.67 (m, 1H), 8.81 (d, 1H), 8.25 (s, 1H), 7.35–7.26 (m, 1H), 7.19 (d, 1H),7.10–6.84(m,4H),6.26(q,1H),4.32(dq,1H),3.05–2.83(m,1H),2.49–2.39(m,1H),2.34–1.76(m, 14H).
LC-MS,M/Z(ESI):508.2[M+H] +。 LC-MS, M/Z (ESI): 508.2 [M+H] + .
实施例40化合物I-40的制备The preparation of embodiment 40 compound I-40
合成路线如下所示：The synthetic route is as follows:
Figure PCTCN2022097611-appb-000113
Figure PCTCN2022097611-appb-000113
第一步：(R)-4-(丙烷-1-炔-1-基)-1-(1-4-(4,4,5,5-四甲基-1,3,2-二氧杂硼烷-2-基)苯基)乙基)- 1H-吲唑-7-甲酸甲酯的制备The first step: (R)-4-(propane-1-yn-1-yl)-1-(1-4-(4,4,5,5-tetramethyl-1,3,2-dioxo Preparation of Borolin-2-yl)phenyl)ethyl)-1H-indazole-7-carboxylic acid methyl ester
向(R)-1-(1-(4-溴苯基)乙基)-4-(丙烷-1-炔-1-基)-1H-吲唑-7-甲酸甲酯(400mg,1.007mmol)的二氧六环(1mL)溶液中加入频哪醇硼酸酯(384mg,1.510mmol)、乙酸钾(296mg,3.02mmol)和[1,1'-双(二苯基膦基)二茂铁]二氯化钯(73.7mg,0.101mmol)，然后在80℃下搅拌16小时。将反应液倒入水(50mL)中并用乙酸乙酯(10mL*2)萃取。合并有机层，用无水硫酸钠干燥，过滤并浓缩得到粗产物。粗产物通过柱层析(石油醚/乙酸乙酯(V/V)＝30/1-1/1)纯化得到产物(R)-4-(丙烷-1-炔-1-基)-1-(1-4-(4,4,5,5-四甲基-1,3,2-二氧杂硼烷-2-基)苯基)乙基)-1H-吲唑-7-甲酸甲酯(350mg,产率78％)。To (R)-1-(1-(4-bromophenyl)ethyl)-4-(propane-1-yn-1-yl)-1H-indazole-7-carboxylic acid methyl ester (400mg, 1.007mmol ) in dioxane (1mL) were added pinacol borate (384mg, 1.510mmol), potassium acetate (296mg, 3.02mmol) and [1,1'-bis(diphenylphosphino)dicene Iron]palladium dichloride (73.7mg, 0.101mmol), then stirred at 80°C for 16 hours. The reaction solution was poured into water (50 mL) and extracted with ethyl acetate (10 mL*2). The organic layers were combined, dried over anhydrous sodium sulfate, filtered and concentrated to give crude product. The crude product was purified by column chromatography (petroleum ether/ethyl acetate (V/V)=30/1-1/1) to obtain the product (R)-4-(propane-1-yn-1-yl)-1- (1-4-(4,4,5,5-tetramethyl-1,3,2-dioxaborolan-2-yl)phenyl)ethyl)-1H-indazole-7-carboxylic acid Ester (350 mg, 78% yield).
LC-MS,M/Z(ESI)：445.3[M+H] +。 LC-MS, M/Z (ESI): 445.3 [M+H] + .
第二步：(R)-1-(1-(4-(2-(二氟甲基)吡啶-4-基)苯基)乙基)-4-(丙烷-1-炔-1-基)-1H-吲唑-7-甲酸甲酯的制备The second step: (R)-1-(1-(4-(2-(difluoromethyl)pyridin-4-yl)phenyl)ethyl)-4-(propane-1-yn-1-yl ) Preparation of -1H-indazole-7-methyl carboxylate
向(R)-4-(丙烷-1-炔-1-基)-1-(1-4-(4,4,5,5-四甲基-1,3,2-二氧杂硼烷-2-基)苯基)乙基)-1H-吲唑-7-甲酸甲酯(150mg，0.338mmol)的甲苯(2mL)和水(0.4mL)溶液中加入4-溴-2-(二氟甲基)吡啶(58.5mg,0.281mmol)、碳酸钾(117mg,0.844mmol)和四(三苯基膦)钯(32.5mg,0.028mmol)。反应液置换3次氮气，并在80℃下搅拌18小时。将反应液倒入水(10mL)中并用乙酸乙酯(5mL*2)萃取。合并有机层，用无水硫酸钠干燥，过滤并浓缩得到粗产物。粗产物通过柱层析(石油醚/乙酸乙酯(V/V)＝30/1-1/1)纯化得到(R)-1-(1-(4-(2-(二氟甲基)吡啶-4-基)苯基)乙基)-4-(丙烷-1-炔-1-基)-1H-吲唑-7-甲酸甲酯(100mg，80％产率)。To (R)-4-(propane-1-yn-1-yl)-1-(1-4-(4,4,5,5-tetramethyl-1,3,2-dioxaborane -2-yl)phenyl)ethyl)-1H-indazole-7-carboxylic acid methyl ester (150 mg, 0.338 mmol) in toluene (2 mL) and water (0.4 mL) was added 4-bromo-2-(di Fluoromethyl)pyridine (58.5 mg, 0.281 mmol), potassium carbonate (117 mg, 0.844 mmol) and tetrakis(triphenylphosphine)palladium (32.5 mg, 0.028 mmol). The reaction solution was replaced with nitrogen three times, and stirred at 80° C. for 18 hours. The reaction solution was poured into water (10 mL) and extracted with ethyl acetate (5 mL*2). The organic layers were combined, dried over anhydrous sodium sulfate, filtered and concentrated to give crude product. The crude product was purified by column chromatography (petroleum ether/ethyl acetate (V/V)=30/1-1/1) to obtain (R)-1-(1-(4-(2-(difluoromethyl) Pyridin-4-yl)phenyl)ethyl)-4-(propan-1-yn-1-yl)-1H-indazole-7-carboxylic acid methyl ester (100 mg, 80% yield).
LC-MS,M/Z(ESI)：446.4[M+H] +。 LC-MS, M/Z (ESI): 446.4 [M+H] + .
第三步：(R)-1-(1-(4-(2-(二氟甲基)吡啶-4-基)苯基)乙基)-4-(丙烷-1-炔-1-基)-1H-吲唑-7-甲酸的制备The third step: (R)-1-(1-(4-(2-(difluoromethyl)pyridin-4-yl)phenyl)ethyl)-4-(propane-1-yn-1-yl )-1H-indazole-7-carboxylic acid preparation
向(R)-1-(1-(4-(2-(二氟甲基)吡啶-4-基)苯基)乙基)-4-(丙烷-1-炔-1-基)-1H-吲唑-7-甲酸甲酯(100mg,0.224mmol)的四氢呋喃(6mL)、甲醇(2mL)和水(2mL)溶液中加入氢氧化锂(26.9mg,1.122mmol)，然后在室温下搅拌3h。向反应液中加入水(50mL)，并通过1N盐酸水溶液调节pH至5～6，然后用乙酸乙酯(20mL*2)萃取。合并有机层，用饱和食盐水(10mL*2)洗涤，用无水硫酸钠干燥，过滤并浓缩得到(R)-1-(1-(4-(2-(二氟甲基)吡啶-4-基)苯基)乙基)-4-(丙烷-1-炔-1-基)-1H-吲唑-7-甲酸(90mg，产率93％)。To (R)-1-(1-(4-(2-(difluoromethyl)pyridin-4-yl)phenyl)ethyl)-4-(propane-1-yn-1-yl)-1H Add lithium hydroxide (26.9mg, 1.122mmol) to a solution of methyl indazole-7-carboxylate (100mg, 0.224mmol) in tetrahydrofuran (6mL), methanol (2mL) and water (2mL), then stir at room temperature for 3h . Water (50 mL) was added to the reaction solution, and the pH was adjusted to 5-6 by 1N aqueous hydrochloric acid solution, and then extracted with ethyl acetate (20 mL*2). The organic layers were combined, washed with saturated brine (10mL*2), dried over anhydrous sodium sulfate, filtered and concentrated to obtain (R)-1-(1-(4-(2-(difluoromethyl)pyridine-4 -yl)phenyl)ethyl)-4-(propan-1-yn-1-yl)-1H-indazole-7-carboxylic acid (90 mg, 93% yield).
LC-MS,M/Z(ESI)：432.5[M+H] +。 LC-MS, M/Z (ESI): 432.5 [M+H] + .
第四步：(Sa,R)-6-(1-(1-(4-(2-(二氟甲基)吡啶-4-基)苯基)乙基)-4-(丙烷-1-炔-1-基)-1H-吲唑-7-羧酰胺)螺[3.3]庚烷-2-甲酸甲酯的制备The fourth step: (Sa,R)-6-(1-(1-(4-(2-(difluoromethyl)pyridin-4-yl)phenyl)ethyl)-4-(propane-1- Preparation of alkyn-1-yl)-1H-indazole-7-carboxamide)spiro[3.3]heptane-2-carboxylic acid methyl ester
向(R)-1-(1-(4-(2-(二氟甲基)吡啶-4-基)苯基)乙基)-4-(丙烷-1-炔-1-基)-1H-吲唑-7-甲酸(90mg，0.209mmol)的N,N-二甲基甲酰胺(2mL)溶液中加入HATU(238mg，0.626mmol)和二异丙基乙胺(81mg，0.626mmol)并在室温下搅拌1小时。然后向反应液中加入(Sa)-6-氨基螺[3.3]庚烷-2-甲酸甲酯(53mg，0.313mmol)并在室温下搅拌17小时。将反应液倒水(20mL)中并用乙酸乙酯(10mL*3)萃取。合并有机相，用饱和食盐水(10mL*2)洗涤，用无水硫酸钠干燥，过滤并浓缩得到粗产物。粗产物通过薄层硅胶板分离纯化(石油醚/乙酸乙酯(V/V)＝3/1)纯化得 到(Sa,R)-6-(1-(1-(4-(2-(二氟甲基))吡啶-4-基)苯基)乙基)-4-(丙烷-1-炔-1-基)-1H-吲唑-7-甲酰胺基)螺[3.3]庚烷-2-甲酸甲酯(20mg，产率16.46％)。To (R)-1-(1-(4-(2-(difluoromethyl)pyridin-4-yl)phenyl)ethyl)-4-(propane-1-yn-1-yl)-1H -Indazole-7-carboxylic acid (90mg, 0.209mmol) in N,N-dimethylformamide (2mL) was added HATU (238mg, 0.626mmol) and diisopropylethylamine (81mg, 0.626mmol) and Stir at room temperature for 1 hour. Then, (Sa)-methyl 6-aminospiro[3.3]heptane-2-carboxylate (53 mg, 0.313 mmol) was added to the reaction solution and stirred at room temperature for 17 hours. The reaction solution was poured into water (20 mL) and extracted with ethyl acetate (10 mL*3). The organic phases were combined, washed with saturated brine (10 mL*2), dried over anhydrous sodium sulfate, filtered and concentrated to obtain a crude product. The crude product was separated and purified by thin-layer silica gel plate (petroleum ether/ethyl acetate (V/V)=3/1) to obtain (Sa,R)-6-(1-(1-(4-(2-(di Fluoromethyl))pyridin-4-yl)phenyl)ethyl)-4-(propane-1-yn-1-yl)-1H-indazole-7-carboxamido)spiro[3.3]heptane- Methyl 2-carboxylate (20 mg, 16.46% yield).
LC-MS,M/Z(ESI)：583.3[M+H] +。 LC-MS, M/Z (ESI): 583.3 [M+H] + .
第五步：(Sa,R)-6-(1-(1-(4-(2-(二氟甲基)吡啶-4-基)苯基)乙基)-4-(丙烷-1-炔-1-基)-1H-吲唑-7-甲酰氨基)螺[3.3]庚烷-2-甲酸(I-40)The fifth step: (Sa,R)-6-(1-(1-(4-(2-(difluoromethyl)pyridin-4-yl)phenyl)ethyl)-4-(propane-1- Alkyn-1-yl)-1H-indazole-7-carboxamido)spiro[3.3]heptane-2-carboxylic acid (I-40)
Figure PCTCN2022097611-appb-000114
Figure PCTCN2022097611-appb-000114
向(Sa,R)-6-(1-(1-(4-(2-(二氟甲基))吡啶-4-基)苯基)乙基)-4-(丙烷-1-炔-1-基)-1H-吲唑-7-甲酰胺基)螺[3.3]庚烷-2-甲酸甲酯(20mg，0.034mmol)的四氢呋喃(3mL)、甲醇(1mL)和水(1mL)的溶液中加入氢氧化锂(4.11mg，0.172mmol)，然后在25℃下搅拌18小时。向反应液中加入水(50mL)并通过1N盐酸水溶液调节pH至5～6，然后用EA(10mL*3)萃取。合并有机相，用饱和食盐水(10mL*2)洗涤，用无水硫酸钠干燥，过滤并浓缩得到产物(Sa,R)-6-(1-(1-(4-(2-(二氟甲基))吡啶-4-基)苯基)乙基)-4-(丙烷-1-炔-1-基)-1H-吲唑-7-甲酰胺基)螺[3.3]庚烷-2-甲酸(I-40)(15mg，产率72.9％)。To (Sa,R)-6-(1-(1-(4-(2-(difluoromethyl))pyridin-4-yl)phenyl)ethyl)-4-(propane-1-yne- 1-yl)-1H-indazole-7-carboxamido)spiro[3.3]heptane-2-carboxylic acid methyl ester (20 mg, 0.034 mmol) in tetrahydrofuran (3 mL), methanol (1 mL) and water (1 mL) Lithium hydroxide (4.11 mg, 0.172 mmol) was added to the solution, followed by stirring at 25°C for 18 hours. Water (50 mL) was added to the reaction solution and the pH was adjusted to 5-6 by 1N aqueous hydrochloric acid solution, and then extracted with EA (10 mL*3). The organic phases were combined, washed with saturated brine (10mL*2), dried over anhydrous sodium sulfate, filtered and concentrated to obtain the product (Sa,R)-6-(1-(1-(4-(2-(difluoro Methyl))pyridin-4-yl)phenyl)ethyl)-4-(propane-1-yn-1-yl)-1H-indazole-7-carboxamido)spiro[3.3]heptane-2 - Formic acid (I-40) (15 mg, 72.9% yield).
1H NMR(400MHz,dmso)δ12.02(s,1H),8.80(d,1H),8.71(d,1H),8.29(s,1H),7.88(s,1H),7.81(d,1H),7.72(d,2H),7.31(d,1H),7.24-7.07(m,3H),6.35(q,1H),4.32(dt,1H),3.01-2.84(m,1H),2.45(dd,1H),2.29(ddd,3H),2.16-1.99(m,5H),1.92(d,2H),1.84(dd,1H),1.36-1.07(m,3H). 1 H NMR (400MHz,dmso)δ12.02(s,1H),8.80(d,1H),8.71(d,1H),8.29(s,1H),7.88(s,1H),7.81(d,1H ),7.72(d,2H),7.31(d,1H),7.24-7.07(m,3H),6.35(q,1H),4.32(dt,1H),3.01-2.84(m,1H),2.45( dd,1H),2.29(ddd,3H),2.16-1.99(m,5H),1.92(d,2H),1.84(dd,1H),1.36-1.07(m,3H).
LC-MS，M/Z(ESI):569.30[M+H] +。 LC-MS, M/Z (ESI): 569.30 [M+H] + .
下列化合物的制备参考化合物I-18或I-18a的制备方法得到。The preparation of the following compounds can be obtained by referring to the preparation method of compound I-18 or I-18a.
Figure PCTCN2022097611-appb-000115
Figure PCTCN2022097611-appb-000115
Figure PCTCN2022097611-appb-000116
Figure PCTCN2022097611-appb-000116
测试例1：EP2拮抗作用测定实验Test Example 1: EP2 Antagonism Determination Experiment
化合物对EP2拮抗作用在高表达人源EP2受体的CHO稳转细胞株上进行。细胞经胰蛋白酶消化后，重悬于缓冲液中(1×HBSS,0.1％BSA,20mM HEPES和500μM IBMX)，每孔接种8000个细胞于384孔板，接种体积为15μL。使用DMSO试剂将待测化合物、EP2完全拮抗剂TG4-155和PGE2分别配制成10mM母液，用实验缓冲液制备8X浓度的待测化合物工作液、EP2完全拮抗剂TG4-155工作液和PGE2工作液，随后分别加入2.5μL 8X化合物工作液、8X的EP2完全拮抗剂TG4-155和DMSO(终浓度0.2％)至上述384孔板中，于37℃孵育10min。每孔加入2.5μL的8X浓度的激动剂PGE 2工作液至上述384孔板中(PGE 2终浓度为0.3nM)，于37℃孵育30min。反应完成后，使用试剂盒(Perkin Elmer,Cat#TRF0263)中的cAMP检测缓冲液1：50稀释Eu-cAMP，每孔加入10μL的Eu-cAMP，使用cAMP检测缓冲液1：150稀释ULight-anti-cAMP，每孔加入10μL的ULight-anti-cAMP，室温孵育1h。在Envision 2105读板器上读取665nm和615nm波长的数据。计算测试化合物的拮抗作用(IC 50值)。 The antagonistic effect of the compound on EP2 was performed on a CHO stably transfected cell line highly expressing the human EP2 receptor. After trypsinization, the cells were resuspended in buffer (1×HBSS, 0.1% BSA, 20 mM HEPES and 500 μM IBMX), and 8000 cells were inoculated in a 384-well plate in a volume of 15 μL per well. Use DMSO reagent to prepare the test compound, EP2 complete antagonist TG4-155 and PGE2 respectively into 10mM stock solution, and use the experimental buffer to prepare 8X concentration of the test compound working solution, EP2 complete antagonist TG4-155 working solution and PGE2 working solution , followed by adding 2.5 μL of 8X compound working solution, 8X complete EP2 antagonist TG4-155 and DMSO (final concentration 0.2%) to the above-mentioned 384-well plate, and incubated at 37° C. for 10 min. Add 2.5 μL of agonist PGE 2 working solution at 8X concentration to each well of the above-mentioned 384-well plate (final concentration of PGE 2 is 0.3 nM), and incubate at 37° C. for 30 min. After the reaction is complete, dilute Eu-cAMP with cAMP detection buffer 1:50 in the kit (Perkin Elmer, Cat#TRF0263), add 10 μL of Eu-cAMP to each well, and dilute ULight-anti with cAMP detection buffer 1:150 -cAMP, add 10 μL of ULight-anti-cAMP to each well, and incubate at room temperature for 1 hour. Data were read at 665nm and 615nm wavelengths on an Envision 2105 plate reader. Antagonism ( IC50 values) of test compounds were calculated.
表1测试化合物对EP2的拮抗作用Table 1 Antagonism of test compounds to EP2
测试化合物test compound IC 50(nM) IC 50 (nM)
I-1I-1 148148
I-3I-3 166166
I-4I-4 247247
I-5I-5 340340
I-6I-6 482482
I-7I-7 3131
I-8I-8 592592
I-9I-9 353353
I-10I-10 466466
I-11I-11 12571257
I-12I-12 474474
I-13I-13 4949
I-14I-14 86788678
I-15I-15 19651965
I-16I-16 10001000
I-17I-17 10001000
I-18I-18 3737
I-19I-19 3030
I-19aI-19a 9898
I-19bI-19b 4.604.60
I-19cI-19c >1000>1000
I-19dI-19d >1000>1000
I-28I-28 319319
I-32bI-32b >1000>1000
I-29bI-29b >1000>1000
I-31aI-31a 395395
I-31bI-31b >1000>1000
I-20aI-20a >1000>1000
I-20bI-20b 1414
I-21I-21 486486
I-22I-22 9494
I-25I-25 245245
I-18aI-18a 1717
I-18bI-18b >1000>1000
I-34aI-34a 4848
I-34bI-34b >1000>1000
I-35aI-35a 88
I-35bI-35b >1000>1000
I-23I-23 >16>16
I-37I-37 1818
I-39aI-39a >1000>1000
I-39bI-39b 3838
I-40I-40 23twenty three
I-36I-36 1616
结果表明：本发明化合物对EP2具有很好的拮抗作用。The results show that the compound of the present invention has very good antagonistic effect on EP2.
测试例2：对EP4受体钙流抑制效果测定Test Example 2: Determination of Inhibitory Effect on EP4 Receptor Calcium Flow
化合物对EP4钙流抑制效果在过表达人EP4受体的293细胞上进行。将生长良好的细胞重悬于细胞培养基中，调整细胞密度为1×10 6细胞每毫升。将细胞悬浮液以20μL/孔接种于2块多聚赖氨酸包被的384孔板(20,000个细胞/孔)，置于37℃，5％CO 2培养箱中过夜。准备2X Fluo-4 Direct TM(Invitrogen,Cat#F10471)上样缓冲液：向1mL FLIPR缓冲液中加入77mg丙磺舒，浓度为250mM。每管Fluo-4 Direct TM crystals(F10471)加入10mL FLIPR缓冲液，和0.2mL的250mM的丙磺舒，涡旋并避光静止5min。 The inhibitory effect of compounds on EP4 calcium flow was carried out on 293 cells overexpressing human EP4 receptor. The well-grown cells were resuspended in cell culture medium, and the cell density was adjusted to 1×10 6 cells per milliliter. The cell suspension was inoculated on two polylysine-coated 384-well plates (20,000 cells/well) at 20 μL/well, and placed in a 5% CO 2 incubator at 37° C. overnight. Prepare 2X Fluo-4 Direct ™ (Invitrogen, Cat#F10471) loading buffer: add 77 mg probenecid to 1 mL FLIPR buffer at a concentration of 250 mM. Add 10 mL of FLIPR buffer and 0.2 mL of 250 mM probenecid to each tube of Fluo-4 Direct TM crystals (F10471), vortex and keep still for 5 min in the dark.
从培养箱中取出一块细胞板并去除培养基，加入20μL分析缓冲液和2X Fluo-4 Direct TM免洗上样缓冲液至384孔细胞培养板，最终体积为40μL。在37℃，5％CO 2培养箱中孵育50分钟，室温孵育10分钟，放入FLIPR。将10μL缓冲液转移至细胞板，读取荧光信号。将激动剂PGE 2在DMSO溶剂中配制成10mM的储备液，使用缓冲液梯度稀释10个浓度点的6X工作液。将10μL激动剂PGE 2转移至细胞板，读取荧光信号，计算EC 80值。 Remove one cell plate from the incubator and remove the medium, add 20 μL of Assay Buffer and 2X Fluo-4 Direct ™ No-Wash Loading Buffer to a 384-well cell culture plate for a final volume of 40 μL. Incubate for 50 min in a 37 °C, 5% CO2 incubator, 10 min at room temperature, and place in the FLIPR. Transfer 10 μL of the buffer to the cell plate and read the fluorescent signal. The agonist PGE 2 was prepared as a 10 mM stock solution in DMSO solvent, and the 6X working solution at 10 concentration points was serially diluted with buffer. Transfer 10 μL of the agonist PGE 2 to the cell plate, read the fluorescence signal, and calculate the EC 80 value.
准备6X EC 80浓度的激动剂PGE 2，并将待测化合物在DMSO溶剂中配制成10mM的储备液，使用缓冲液梯度稀释10个浓度点6X化合物工作液。 Prepare the agonist PGE 2 at the concentration of 6X EC 80 , prepare the compound to be tested into a 10 mM stock solution in DMSO solvent, and use the buffer solution to serially dilute the 6X compound working solution at 10 concentration points.
另取一块细胞板去除培养基，加入20μL分析缓冲液和2X Fluo-4 Direct TM免洗上样缓冲液。在37℃，5％CO 2培养箱中孵育50分钟，室温孵育10分钟，放入FLIPR。将10μL化合物工作液、DMSO、EP4完全拮抗剂转移至细胞板，读取荧光信号。将10μL 6X EC 80浓度的激动剂PGE 2转移至细胞板，读取荧光信号，计算化合物对EP4钙流抑制的IC 50值。 Take another cell plate to remove the culture medium, add 20 μL assay buffer and 2X Fluo-4 Direct TM no-wash loading buffer. Incubate for 50 min in a 37 °C, 5% CO2 incubator, 10 min at room temperature, and place in the FLIPR. Transfer 10 μL compound working solution, DMSO, and EP4 complete antagonist to the cell plate, and read the fluorescence signal. Transfer 10 μL of the 6X EC 80 concentration of the agonist PGE 2 to the cell plate, read the fluorescence signal, and calculate the IC 50 value of the compound on the inhibition of EP4 calcium flow.
表2测试化合物对EP4钙流抑制作用Table 2 The inhibitory effect of test compounds on EP4 calcium flux
测试化合物test compound IC 50(nM) IC 50 (nM)
I-1I-1 2525
I-2I-2 1515
I-3I-3 5959
I-4I-4 355355
I-5I-5 500500
I-6I-6 8585
I-7I-7 24twenty four
I-8I-8 1414
I-9I-9 9292
I-10I-10 111111
I-11I-11 5050
I-12I-12 77
I-13I-13 5555
I-14I-14 137137
I-15I-15 3434
I-16I-16 7474
I-17I-17 8080
I-18I-18 3636
I-19I-19 2828
I-19aI-19a 422422
I-19bI-19b 1010
I-19cI-19c >1000>1000
I-19dI-19d 256256
I-28I-28 182182
I-31aI-31a 144144
I-31bI-31b 727727
I-32bI-32b >1000>1000
I-29aI-29a 3838
I-29bI-29b >1000>1000
I-20aI-20a 178178
I-20bI-20b 44
I-22I-22 88
I-25I-25 4444
I-18aI-18a 22
I-18bI-18b >1000>1000
I-34aI-34a 33
I-34bI-34b >1000>1000
I-35aI-35a 66
I-35bI-35b 340340
I-23I-23 44
I-37I-37 77
I-39aI-39a >1000>1000
I-39bI-39b 44
I-40I-40 1212
I-36I-36 1717
实验结果表明，本发明化合物对EP4钙流显示较好抑制作用。The experimental results show that the compound of the present invention has a better inhibitory effect on EP4 calcium flow.
测试例3：放射性配体EP2受体结合测定Test Example 3: Radioligand EP2 Receptor Binding Assay
使用重组人EP2受体膜蛋白(Perkin Elmer#ES-562-M400UA，从过表达人EP2受体的293细胞制备)进行放射性配体EP2结合测定。使用DMSO溶剂将待测化合物配制成10μM 的储备液，用结合测定缓冲液(50mM Tris-HCl,pH 7.4,10mM MgCl2,0.5mM EDTA)将待测化合物和放射性配体[ 3H]-PGE 2(Perkin Elmer#NET428025UC)分别配制成10×工作液。将10μL化合物工作液、1％的DMSO、PGE 2工作液(终浓度10μM)分别加到测定板中，加入80μL的EP2受体膜蛋白(10μg/孔)和10μL的放射性配体[ 3H]-PGE 2(Perkin Elmer#NET428025UC)(终浓度7.5nM)，25℃孵育1.5小时。使用Cell Harvester通过0.3％PEI包被的GF/C板过滤反应混合物，使用冰的洗涤缓冲液(50mM Tris-HCl,pH 7.4)清洗滤板3次，在37℃下将滤板干燥2小时，干燥后，加入50μL的scintillation cocktail，密封滤板顶部。使用Topcount读取过滤器上捕获的 3H计数。 Radioligand EP2 binding assays were performed using recombinant human EP2 receptor membrane protein (Perkin Elmer #ES-562-M400UA, prepared from 293 cells overexpressing human EP2 receptor). The compound to be tested was prepared as a 10 μM stock solution using DMSO solvent, and the compound to be tested and the radioligand [ 3 H]-PGE 2 (Perkin Elmer#NET428025UC) were prepared into 10 × working solution. Add 10 μL compound working solution, 1% DMSO, PGE 2 working solution (final concentration 10 μM) to the assay plate, add 80 μL EP2 receptor membrane protein (10 μg/well) and 10 μL radioligand [ 3 H] - PGE2 (Perkin Elmer # NET428025UC ) (final concentration 7.5nM), incubate at 25°C for 1.5 hours. The reaction mixture was filtered through a 0.3% PEI-coated GF/C plate using a Cell Harvester, the filter plate was washed 3 times with ice washing buffer (50 mM Tris-HCl, pH 7.4), and the filter plate was dried at 37° C. for 2 hours. After drying, add 50 μL of scintillation cocktail to seal the top of the filter plate. The 3 H count captured on the filter was read using Topcount.
使用GraphPad Prism 5分析数据，按以下公式计算抑制率：The data were analyzed using GraphPad Prism 5, and the inhibition rate was calculated according to the following formula:
抑制率(％)＝100-(测试组-PGE 2组)/(DMSO组-PGE 2组)*100 Inhibition rate (%)=100-(test group-PGE 2 group)/(DMSO group-PGE 2 group)*100
根据化合物不同浓度的抑制率，计算化合物通过放射性配体EP2结合测定的IC 50和Ki值。 According to the inhibition rate of different concentrations of the compound, the IC 50 and Ki values of the compound determined by radioligand EP2 binding were calculated.
实验数据表明：本发明化合物与EP2受体具有很好的亲和力。The experimental data shows that the compound of the present invention has a good affinity with the EP2 receptor.
测试例4：热力学溶解度实验Test Example 4: Thermodynamic Solubility Experiment
配制pH 1.6的禁食状态模拟胃液FaSSGF(1L溶液含80μM的牛黄胆酸钠,20μM的卵磷脂,0.1g的胃蛋白酶,34.2mM的氯化钠)、pH 6.5的禁食状态模拟肠液FaSSIF(1L溶液含3mM的牛黄胆酸钠,0.2mM的卵磷脂,38.4mM的氢氧化钠,68.62mM的氯化钠,19.12mM的马来酸)和pH 7.4的磷酸缓冲盐溶液PBS(1L溶液含100mM的磷酸盐缓冲液,11g碳酸氢二钠,3.5g的二水合磷酸二氢钠)作为测定缓冲液。Prepare fasting state simulated gastric juice FaSSGF at pH 1.6 (1L solution contains 80 μM sodium taurocholate, 20 μM lecithin, 0.1 g pepsin, 34.2 mM sodium chloride), fasting state simulated intestinal fluid FaSSIF at pH 6.5 ( 1L solution contains 3mM sodium taurocholate, 0.2mM lecithin, 38.4mM sodium hydroxide, 68.62mM sodium chloride, 19.12mM maleic acid) and pH 7.4 phosphate buffered saline PBS (1L solution contains 100mM phosphate buffer, 11g disodium bicarbonate, 3.5g sodium dihydrogen phosphate dihydrate) as the assay buffer.
准确称取化合物，使用不同pH的测定缓冲液将化合物配置成4mg/mL的工作液，1000rpm振荡1小时，室温平衡过夜。样品置于离心机12000rmp离心10min，除去未溶解颗粒。将上清转移至新的离心管，通过LCMSMS检测上清中化合物浓度。The compound was weighed accurately, and the compound was formulated into a 4 mg/mL working solution using assay buffers of different pH, shaken at 1000 rpm for 1 hour, and equilibrated at room temperature overnight. The sample was placed in a centrifuge at 12000rmp for 10min to remove undissolved particles. The supernatant was transferred to a new centrifuge tube, and the compound concentration in the supernatant was detected by LCMSMS.
实验结果表明化合物具有很好的热力学溶解度。The experimental results show that the compound has good thermodynamic solubility.
测试例5：放射性配体EP4受体结合测定Test Example 5: Radioligand EP4 Receptor Binding Assay
使用重组人EP4受体膜蛋白(从过表达人EP4受体的293细胞制备)进行放射性配体EP4结合测定。使用DMSO试剂将待测化合物和PGE 2配制成10μM储备液，以2μM为起始浓度，4倍梯度稀释至8个浓度点工作液。然后使用缓冲液(50mM HBSS,0.1％BSA,500mM NaCl)将EP4受体膜蛋白和放射性配体[ 3H]-PGE 2(PerkinElmer,Cat:NET428250UC,Lot:2469552)配制成工作液浓度。将1μL化合物工作液、DMSO、PGE 2工作液分别加到测定板中，加入100μL的EP4受体膜蛋白(20μg/孔)和100μL放射性配体[ 3H]-PGE 2(PerkinElmer,Cat:NET428250UC,Lot:2469552)(终浓度1.5nM)，密封室温下孵育1小时。在室温下，用0.5％BSA，50μL/每孔浸泡Unifilter-96GF/C滤板(Perkin Elmer)至少30min。结合完成后，使用Perkin Elmer Filtermate Harvester通过GF/C板过滤反应混合物，然后清洗滤板，在50℃下将滤板干燥1小时。干燥后，使用Perkin Elmer Unifilter-96密封胶带密封滤板孔的底部，加入50μL的MicroScint TM-20cocktail(Perkin Elmer)，密封滤板顶部。使用Perkin Elmer MicroBeta2 Reader读取过滤器上捕获的 3H计数。 Radioligand EP4 binding assays were performed using recombinant human EP4 receptor membrane protein (prepared from 293 cells overexpressing human EP4 receptor). The compound to be tested and PGE 2 were prepared into a 10 μM stock solution using DMSO reagent, and the initial concentration was 2 μM, and the 4-fold serial dilution was made to a working solution of 8 concentration points. EP4 receptor membrane protein and radioligand [ 3 H]-PGE 2 (PerkinElmer, Cat: NET428250UC, Lot: 2469552) were then formulated into working solution concentrations using buffer (50 mM HBSS, 0.1% BSA, 500 mM NaCl). Add 1 μL compound working solution, DMSO, and PGE 2 working solution to the assay plate respectively, add 100 μL EP4 receptor membrane protein (20 μg/well) and 100 μL radioligand [ 3 H]-PGE 2 (PerkinElmer, Cat: NET428250UC , Lot:2469552) (final concentration 1.5nM), sealed and incubated at room temperature for 1 hour. Soak the Unifilter-96GF/C filter plate (Perkin Elmer) with 0.5% BSA, 50 μL/well for at least 30 min at room temperature. After conjugation was complete, the reaction mixture was filtered through a GF/C plate using a Perkin Elmer Filtermate Harvester, the filter plate was washed, and the filter plate was dried at 50° C. for 1 hour. After drying, Perkin Elmer Unifilter-96 sealing tape was used to seal the bottom of the filter plate wells, and 50 μL of MicroScint ™ -20cocktail (Perkin Elmer) was added to seal the top of the filter plate. 3 H counts captured on the filter were read using a Perkin Elmer MicroBeta2 Reader.
使用GraphPad Prism 5分析数据，按以下公式计算抑制率：The data were analyzed using GraphPad Prism 5, and the inhibition rate was calculated according to the following formula:
抑制率(％)＝100-(测试组-PGE 2组)/(DMSO组-PGE 2组)*100 Inhibition rate (%)=100-(test group-PGE 2 group)/(DMSO group-PGE 2 group)*100
根据化合物不同浓度的抑制率，计算化合物通过放射性配体EP4结合测定的IC 50和Ki值。 According to the inhibition rate of different concentrations of the compound, calculate the IC 50 and Ki value of the compound through radioligand EP4 binding assay.
实验结果表明：本发明化合物与EP4受体具有很好的亲和力。The experimental results show that the compound of the present invention has a good affinity with the EP4 receptor.
测试例6：人PBMC单核细胞分化Test Example 6: Human PBMC mononuclear cell differentiation
用DMSO制备将化合物配为3000μM、1000μM、300μM和100μM的储备液。再用RPMI 1640完全培养基进行250倍稀释，制备的稀释液浓度分别为：12000nM、4000nM、1200nM和400nM。复苏商品化的人源PBMC，DPBS洗涤后测定细胞活率，用EasySep TM Human Monocyte Isolation Kit从PBMC中分离得到单核细胞，并用RPMI 1640完全培养基重悬细胞调整密度为1.5×10 6/mL。铺1.5×10 5单核细胞于96孔细胞培养板中(体积为100μL/孔)，将配制好的50μL化合物稀释液加入细胞中，同时加入50μL cytokines mix(10nM PGE2,50ng/mL GM-CSF and 100ng/mL IL-4)。化合物终浓度分别为3000nM、1000nM、300nM和100nM。37℃，5％CO 2培养箱中孵育7天，第4天时，半量换液后继续培养。培养7天后，收集细胞，进行流式检测。收获所有细胞，用死活染料(Fixable Viability Stain 780，1：1000在PBS中稀释)常温避光染色15分钟，然后用PBS洗涤两次。每个样本孔中加入细胞染色缓冲液稀释的Anti-human CD1a/CD16/CD206/CD86和CD163抗体(染色体积为100μL)，4℃避光染色40分钟，使用细胞染色缓冲液洗涤一次，400g离心5分钟。染色结束后，使用200μL的细胞染色缓冲液重悬细胞沉淀，在Thermo Attune NxT流式细胞仪上进行上机分析。所有数据使用Thermo Attune分析软件和Graphpad Prism6.0进行处理分析。数据结果显示为平均值加标准误差(SEM)。 Compounds were formulated as 3000 μM, 1000 μM, 300 μM and 100 μM stock solutions in DMSO. Then use RPMI 1640 complete medium to carry out 250-fold dilution, and the concentrations of the prepared dilutions are respectively: 12000nM, 4000nM, 1200nM and 400nM. Resuscitate commercially available human PBMC, measure cell viability after washing with DPBS, use EasySep TM Human Monocyte Isolation Kit to isolate monocytes from PBMC, and resuspend cells in RPMI 1640 complete medium to adjust the density to 1.5×10 6 /mL . Spread 1.5×10 5 monocytes in a 96-well cell culture plate (volume 100 μL/well), add 50 μL of the prepared compound dilution to the cells, and add 50 μL cytokines mix (10 nM PGE2, 50 ng/mL GM-CSF and 100ng/mL IL-4). The final compound concentrations were 3000 nM, 1000 nM, 300 nM and 100 nM, respectively. Incubate in a 37°C, 5% CO 2 incubator for 7 days. On the 4th day, half of the medium is changed and the culture is continued. After 7 days of culture, the cells were collected for flow cytometric detection. All cells were harvested, stained with a dead-viability dye (Fixable Viability Stain 780, diluted 1:1000 in PBS) for 15 minutes at room temperature in the dark, and then washed twice with PBS. Add Anti-human CD1a/CD16/CD206/CD86 and CD163 antibodies diluted in cell staining buffer to each sample well (staining volume: 100 μL), stain in the dark at 4°C for 40 minutes, wash once with cell staining buffer, and centrifuge at 400g 5 minutes. After staining, use 200 μL of cell staining buffer to resuspend the cell pellet and perform analysis on a Thermo Attune NxT flow cytometer. All data were processed and analyzed using Thermo Attune analysis software and Graphpad Prism6.0. Data results are shown as mean plus standard error (SEM).
实验结果如图1、图2和图3所示，结果表明：本发明中的化合物对PGE2介导的免疫抑制作用都有较好的拮抗作用，可提高DC细胞和M1型细胞的比例，同时可以抑制M2的比例。The experimental results are shown in Figure 1, Figure 2 and Figure 3, and the results show that the compounds of the present invention have a good antagonism to the PGE2-mediated immunosuppressive effect, can increase the ratio of DC cells and M1 cells, and at the same time The ratio of M2 can be suppressed.
测试例7：药代动力学试验Test Example 7: Pharmacokinetic Test
小鼠药代动力学试验，使用雄性ICR小鼠，20-25g，禁食过夜。取3只小鼠，口服灌胃给药5mg/kg。在给药前和在给药后15、30分钟以及1、2、4、8、24小时采血，取另外3只小鼠，静脉注射给药1mg/kg，在给药前和在给药后15、30分钟以及1、2、4、8、24小时采血。血液样品6800g，2-8℃离心6分钟，收集血浆，于-80℃保存。取各时间点血浆，加入3-5倍量含内标的乙腈溶液混合，涡旋混合1分钟，13000转/分钟4℃离心10分钟，取上清液加入3倍量水混合，取适量混合液进行LC-MS/MS分析。主要药代动力学参数用WinNonlin7.0软件非房室模型分析。Mouse pharmacokinetics test, using male ICR mice, 20-25g, fasted overnight. Take 3 mice, orally administer 5 mg/kg by gavage. Blood was collected before administration and at 15, 30 minutes and 1, 2, 4, 8, and 24 hours after administration, and the other 3 mice were given intravenous injection of 1 mg/kg, before administration and after administration 15, 30 minutes and 1, 2, 4, 8, 24 hours blood collection. The blood sample was 6800g, centrifuged at 2-8°C for 6 minutes, the plasma was collected and stored at -80°C. Take the plasma at each time point, add 3-5 times the amount of acetonitrile solution containing the internal standard and mix, vortex for 1 minute, centrifuge at 13000 rpm at 4°C for 10 minutes, take the supernatant and add 3 times the amount of water to mix, and take an appropriate amount of the mixture LC-MS/MS analysis was performed. The main pharmacokinetic parameters were analyzed by WinNonlin7.0 software non-compartmental model.
大鼠药代动力学试验，使用雄性SD大鼠，180-240g，禁食过夜。取3只大鼠，口服灌胃给药5mg/kg。取另外3只大鼠，静脉注射给药1mg/kg。其余操作同小鼠药代动力学试验。Rat pharmacokinetic test, using male SD rats, 180-240g, fasted overnight. Take 3 rats, orally administer 5 mg/kg by gavage. Take another 3 rats and administer 1mg/kg intravenously. The rest of the operations were the same as the mouse pharmacokinetic test.
犬药代动力学试验，使用雄性Beagle犬，8-10kg，禁食过夜。取3只Beagle犬，口服灌胃给药3mg/kg。取另外3只Beagle犬，静脉注射给药1mg/kg。其余操作同小鼠药代动力学试验。Canine pharmacokinetic test, using male Beagle dogs, 8-10kg, fasted overnight. Take 3 Beagle dogs, orally administer 3 mg/kg by gavage. Take another 3 Beagle dogs and give 1 mg/kg intravenously. The rest of the operations were the same as the mouse pharmacokinetic test.
猴药代动力学实验，使用雄性食蟹猴，5-7kg，禁食过夜。取3只食蟹猴，口服灌胃给药 5mg/kg。取另外3只食蟹猴，静脉注射给药3mg/kg。其余操作同小鼠药代动力学试验。For monkey pharmacokinetic experiments, male cynomolgus monkeys, 5-7kg, were fasted overnight. Get 3 cynomolgus monkeys, orally administer 5mg/kg by gavage. Take another 3 cynomolgus monkeys and administer 3 mg/kg intravenously. The rest of the operations were the same as the mouse pharmacokinetic test.
表5小鼠口服灌胃给药药代动力学实验结果Table 5 Mouse oral gavage administration pharmacokinetic experimental results
Figure PCTCN2022097611-appb-000117
Figure PCTCN2022097611-appb-000117
表6大鼠口服灌胃给药药代动力学实验结果Table 6 rat oral gavage administration pharmacokinetic experimental results
Figure PCTCN2022097611-appb-000118
Figure PCTCN2022097611-appb-000118
表7犬口服灌胃给药药代动力学实验Table 7 Dog oral gavage administration pharmacokinetic experiment
Figure PCTCN2022097611-appb-000119
Figure PCTCN2022097611-appb-000119
实验结果表明，本发明化合物的静脉给药清除率低，口服给药暴露量高，表现出优良的药代动力学性质，成药性好。Experimental results show that the compound of the present invention has low clearance rate by intravenous administration, high exposure by oral administration, excellent pharmacokinetic properties and good druggability.
测试例8：测试化合物联合anti-mouse PD-1抗体法在CT-26鼠源结肠癌肿瘤模型中的抗肿瘤作用Test Example 8: Anti-tumor effect of test compound combined with anti-mouse PD-1 antibody in CT-26 mouse colon cancer tumor model
本实验中，在CT-26鼠源结肠癌肿瘤模型中测试化合物I-18a和I-23联合anti-mouse PD-1抗体法的抗肿瘤作用。小鼠适应性饲养一周后，将处于对数期的CT-26细胞重悬于PBS， 按100μL/只将3×10 5CT-26细胞接种于右侧后部处皮下，定期观察肿瘤生长情况，待肿瘤生长至平均体积50-70mm 3时，根据肿瘤体积大小采用随机分组法将荷瘤小鼠分为4组，每组8只。试验分为溶媒组、单anti-mouse PD-1抗体组、化合物I-18a(150mg/kg)、化合物I-23(150mg/kg)分别联合anti-mouse PD-1抗体(7.5mg/kg)联合给药组，口服灌胃给药，每天两次，共给药18天；anti-mouse PD-1抗体为一周两次使用。每周测量2次肿瘤体积和小鼠体重，实验结束时称取瘤重。 In this experiment, the anti-tumor effect of compounds I-18a and I-23 combined with anti-mouse PD-1 antibody was tested in the CT-26 mouse colon cancer tumor model. After the mouse was adaptively fed for one week, the CT-26 cells in the logarithmic phase were resuspended in PBS, and 3×10 5 CT-26 cells were subcutaneously inoculated on the right rear at 100 μL/mouse, and the tumor growth was observed regularly , when the tumor grew to an average volume of 50-70 mm 3 , the tumor-bearing mice were randomly divided into 4 groups according to the size of the tumor, with 8 mice in each group. The test was divided into vehicle group, single anti-mouse PD-1 antibody group, compound I-18a (150mg/kg), compound I-23 (150mg/kg) respectively combined with anti-mouse PD-1 antibody (7.5mg/kg) The combination administration group was administered orally orally, twice a day, for a total of 18 days; the anti-mouse PD-1 antibody was used twice a week. The tumor volume and mouse body weight were measured twice a week, and the tumor weight was weighed at the end of the experiment.
肿瘤体积和相对肿瘤抑制率计算公式如下：肿瘤体积(tumor volume，TV)＝1/2×a×b 2，其中a、b分别为肿瘤测量的长和宽。化合物的抑瘤疗效TGI(％)＝(TWc-TWt/TWc)×100％，其中TWc为溶媒对照组平均瘤重，TWt为治疗组平均瘤重。实验结果如图4所示，结果表明，化合物I-18a(150mg/kg)、化合物I-23(150mg/kg)分别联合anti-mouse PD-1抗体在开始给药后第18天时表现出显著的抑瘤作用，相对肿瘤抑制率TGI(％)分别为78.48％和70.86％，相对溶媒对照组统计学上均有显著性差异(p值均小于0.05)。 The calculation formulas of tumor volume and relative tumor inhibition rate are as follows: tumor volume (tumor volume, TV)=1/2×a×b 2 , where a and b are the measured length and width of the tumor, respectively. Antitumor curative effect of compound TGI(%)=(TWc-TWt/TWc)×100%, where TWc is the average tumor weight of the vehicle control group, and TWt is the average tumor weight of the treatment group. The experimental results are shown in Figure 4. The results showed that compound I-18a (150mg/kg) and compound I-23 (150mg/kg) respectively combined with anti-mouse PD-1 antibody showed a significant effect on the 18th day after the start of administration. The relative tumor inhibition rates TGI (%) were 78.48% and 70.86%, respectively, and there were statistically significant differences compared with the vehicle control group (all p values were less than 0.05).
尽管上面已经示出和描述了本发明的实施例，可以理解的是，上述实施例是示例性的，不能理解为对本发明的限制，本领域的普通技术人员在本发明的范围内可以对上述实施例进行变化、修改、替换和变型。Although the embodiments of the present invention have been shown and described above, it can be understood that the above embodiments are exemplary and should not be construed as limiting the present invention, those skilled in the art can make the above-mentioned The embodiments are subject to changes, modifications, substitutions and variations.

Claims (22)

  1. 式I所示苯并杂环化合物、其互变异构体、立体异构体、水合物、溶剂化物、药学上可接受的盐或前药：Benzoheterocyclic compounds represented by formula I, tautomers, stereoisomers, hydrates, solvates, pharmaceutically acceptable salts or prodrugs thereof:
    Figure PCTCN2022097611-appb-100001
    Figure PCTCN2022097611-appb-100001
    其中，R 1、R 2分别为氢或-C≡C-R 11，且R 1、R 2不同； Wherein, R 1 and R 2 are respectively hydrogen or -C≡CR 11 , and R 1 and R 2 are different;
    所述R 11为氢或选自：C 1-C 5烷基、3-6元环烷基； The R 11 is hydrogen or selected from: C 1 -C 5 alkyl, 3-6 membered cycloalkyl;
    所述R 11任选地被一个或多个选自下列的取代基取代：羟基、卤素、C 1-C 5烷基；当取代基为多个时，所述取代基相同或不同； The R 11 is optionally substituted by one or more substituents selected from the following: hydroxyl, halogen, C 1 -C 5 alkyl; when there are multiple substituents, the substituents are the same or different;
    L 1为C 1-C 5亚烷基； L 1 is C 1 -C 5 alkylene;
    L 2不存在或为未取代或被Rb取代的C 1-C 3亚烷基； L 2 does not exist or is unsubstituted or substituted by Rb C 1 -C 3 alkylene;
    所述L 1任选地被Ra 1和/或Ra 2取代； The L 1 is optionally substituted by Ra 1 and/or Ra 2 ;
    所述Ra 1、Ra 2各自独立地为C 1-C 5烷基、卤素、羟基、氨基、C 1-C 5烷氧基、C 1-C 5卤代烷基、C 1-C 5卤代烷氧基； The Ra 1 and Ra 2 are each independently C 1 -C 5 alkyl, halogen, hydroxyl, amino, C 1 -C 5 alkoxy, C 1 -C 5 haloalkyl, C 1 -C 5 haloalkoxy ;
    或所述Ra 1、Ra 2与它们共同连接的C原子一起形成3-6元环烷基或4-6元杂环烷基； Or the Ra 1 and Ra 2 form a 3-6-membered cycloalkyl group or a 4-6-membered heterocycloalkyl group together with the C atoms they are connected to;
    所述Rb为C 1-C 3烷基或卤素； The Rb is C 1 -C 3 alkyl or halogen;
    环A为苯基或5-6元杂芳基；Ring A is phenyl or 5-6 membered heteroaryl;
    环B不存在或为3-8元环烷基、4-8元杂环烷基、苯基、5-6元杂芳基；Ring B does not exist or is 3-8 membered cycloalkyl, 4-8 membered heterocycloalkyl, phenyl, 5-6 membered heteroaryl;
    R 3、R 4各自独立地选自：羟基、卤素、氨基、氰基、C 1-C 5烷基、C 1-C 5烷氧基、3-6元环烷基、4-6元杂环烷基、3-6元环烷氧基；所述C 1-C 5烷基、C 1-C 5烷氧基、3-6元环烷基、4-6元杂环烷基、3-6元环烷氧基任选地被一个或多个选自下列的取代基取代：羟基、卤素、氨基、氰基、3-6元环烷基； R 3 and R 4 are each independently selected from: hydroxyl, halogen, amino, cyano, C 1 -C 5 alkyl, C 1 -C 5 alkoxy, 3-6 membered cycloalkyl, 4-6 membered hetero Cycloalkyl, 3-6 membered cycloalkoxy; said C 1 -C 5 alkyl, C 1 -C 5 alkoxy, 3-6 membered cycloalkyl, 4-6 membered heterocycloalkyl, 3 -6-membered cycloalkoxy is optionally substituted by one or more substituents selected from the group consisting of hydroxyl, halogen, amino, cyano, 3-6-membered cycloalkyl;
    m为0、1、2或3；m is 0, 1, 2 or 3;
    n为0、1、2或3。n is 0, 1, 2 or 3.
  2. 式I所示苯并杂环化合物、其互变异构体、立体异构体、水合物、溶剂化物、药学上可接受的盐或前药：Benzoheterocyclic compounds represented by formula I, tautomers, stereoisomers, hydrates, solvates, pharmaceutically acceptable salts or prodrugs thereof:
    Figure PCTCN2022097611-appb-100002
    Figure PCTCN2022097611-appb-100002
    其中，R 1、R 2分别为氢或-C≡C-R 11，且R 1、R 2不同； Wherein, R 1 and R 2 are respectively hydrogen or -C≡CR 11 , and R 1 and R 2 are different;
    所述R 11为氢或选自：C 1-C 5烷基、3-6元环烷基； The R 11 is hydrogen or selected from: C 1 -C 5 alkyl, 3-6 membered cycloalkyl;
    所述R 11任选地被一个或多个选自下列的取代基取代：羟基、卤素、C 1-C 5烷基；当取代基为多个时，所述取代基相同或不同； The R 11 is optionally substituted by one or more substituents selected from the following: hydroxyl, halogen, C 1 -C 5 alkyl; when there are multiple substituents, the substituents are the same or different;
    L 1为C 1-C 5亚烷基； L 1 is C 1 -C 5 alkylene;
    L 2不存在或为未取代或被Rb取代的C 1-C 3亚烷基； L 2 does not exist or is unsubstituted or substituted by Rb C 1 -C 3 alkylene;
    所述L 1任选地被Ra 1和/或Ra 2取代； The L 1 is optionally substituted by Ra 1 and/or Ra 2 ;
    所述Ra 1、Ra 2各自独立地为C 1-C 5烷基、卤素、羟基、氨基、C 1-C 5烷氧基、C 1-C 5卤代烷基、C 1-C 5卤代烷氧基； The Ra 1 and Ra 2 are each independently C 1 -C 5 alkyl, halogen, hydroxyl, amino, C 1 -C 5 alkoxy, C 1 -C 5 haloalkyl, C 1 -C 5 haloalkoxy ;
    或所述Ra 1、Ra 2与它们共同连接的C原子一起形成3-6元环烷基或4-6元杂环烷基； Or the Ra 1 and Ra 2 form a 3-6-membered cycloalkyl group or a 4-6-membered heterocycloalkyl group together with the C atoms they are connected to;
    所述Rb为C 1-C 3烷基或卤素； The Rb is C 1 -C 3 alkyl or halogen;
    环A为苯基或5-6元杂芳基；Ring A is phenyl or 5-6 membered heteroaryl;
    环B不存在或为3-6元环烷基、4-6元杂环烷基、苯基、5-6元杂芳基；Ring B does not exist or is 3-6 membered cycloalkyl, 4-6 membered heterocycloalkyl, phenyl, 5-6 membered heteroaryl;
    R 3、R 4各自独立地选自：羟基、卤素、氨基、氰基、C 1-C 5烷基、C 1-C 5烷氧基、3-6元环烷基、4-6元杂环烷基、3-6元环烷氧基；所述C 1-C 5烷基、C 1-C 5烷氧基、3-6元环烷基、4-6元杂环烷基、3-6元环烷氧基任选地被一个或多个选自下列的取代基取代：羟基、卤素、氨基、氰基、3-6元环烷基； R 3 and R 4 are each independently selected from: hydroxyl, halogen, amino, cyano, C 1 -C 5 alkyl, C 1 -C 5 alkoxy, 3-6 membered cycloalkyl, 4-6 membered hetero Cycloalkyl, 3-6 membered cycloalkoxy; said C 1 -C 5 alkyl, C 1 -C 5 alkoxy, 3-6 membered cycloalkyl, 4-6 membered heterocycloalkyl, 3 -6-membered cycloalkoxy is optionally substituted by one or more substituents selected from the group consisting of hydroxyl, halogen, amino, cyano, 3-6-membered cycloalkyl;
    m为0、1、2或3；m is 0, 1, 2 or 3;
    n为0、1、2或3。n is 0, 1, 2 or 3.
  3. 如权利要求1所述的式I所示苯并杂环化合物、其互变异构体、立体异构体、水合物、溶剂化物、药学上可接受的盐或前药，其特征在于，在所述环A中，所述5-6元杂芳基含有一个或多个选自N、O或S的杂原子；当所述杂原子为多个时，所述杂原子相同或不同；The benzoheterocyclic compound shown in formula I as claimed in claim 1, its tautomer, stereoisomer, hydrate, solvate, pharmaceutically acceptable salt or prodrug, is characterized in that, in In the ring A, the 5-6 membered heteroaryl group contains one or more heteroatoms selected from N, O or S; when there are multiple heteroatoms, the heteroatoms are the same or different;
    较佳地，所述5-6元杂芳基选自：噻吩、呋喃、吡咯、吡唑、咪唑、三氮唑、噻唑、噻二唑、噁唑、吡啶、嘧啶、哒嗪、吡嗪。Preferably, the 5-6 membered heteroaryl group is selected from: thiophene, furan, pyrrole, pyrazole, imidazole, triazole, thiazole, thiadiazole, oxazole, pyridine, pyrimidine, pyridazine, pyrazine.
  4. 如权利要求1所述的式I所示苯并杂环化合物、其互变异构体、立体异构体、水合物、溶剂化物、药学上可接受的盐或前药，其特征在于，L 1为-CH 2-、-CH(CH 3)-、-CH 2CH 2-或-CH 2CH 2CH 2-；较佳地，L 1为-CH 2-、-CH(CH 3)-。 The benzoheterocyclic compound represented by formula I as claimed in claim 1, its tautomer, stereoisomer, hydrate, solvate, pharmaceutically acceptable salt or prodrug, is characterized in that, L 1 is -CH 2 -, -CH(CH 3 )-, -CH 2 CH 2 - or -CH 2 CH 2 CH 2 -; preferably, L 1 is -CH 2 -, -CH(CH 3 )- .
  5. 如权利要求1所述的式I所示苯并杂环化合物、其互变异构体、立体异构体、水合物、溶剂化物、药学上可接受的盐或前药，其特征在于，R 3、R 4各自独立地选自：卤素、氰基、C 1-C 5烷基、C 1-C 5烷氧基、3-6元环烷基；所述C 1-C 5烷基、C 1-C 5烷氧基、3-6元环 烷基任选地被1、2或3个选自下列的取代基取代：羟基、氟、氯。 The benzoheterocyclic compound represented by formula I according to claim 1, its tautomers, stereoisomers, hydrates, solvates, pharmaceutically acceptable salts or prodrugs, is characterized in that R 3. R 4 are each independently selected from: halogen, cyano, C 1 -C 5 alkyl, C 1 -C 5 alkoxy, 3-6 membered cycloalkyl; said C 1 -C 5 alkyl, C 1 -C 5 alkoxy, 3-6 membered cycloalkyl is optionally substituted with 1, 2 or 3 substituents selected from the group consisting of hydroxyl, fluorine, chlorine.
  6. 如权利要求1所述的式I所示苯并杂环化合物、其互变异构体、立体异构体、水合物、溶剂化物、药学上可接受的盐或前药，其特征在于，具有结构Ia、Ib、Ic、Id，The benzoheterocyclic compound shown in formula I according to claim 1, its tautomers, stereoisomers, hydrates, solvates, pharmaceutically acceptable salts or prodrugs, is characterized in that it has Structure Ia, Ib, Ic, Id,
    Figure PCTCN2022097611-appb-100003
    Figure PCTCN2022097611-appb-100003
    其中，Z 1、Z 2、Z 3、Z 4、Z 5各自独立地选自CR 3或N； Wherein, Z 1 , Z 2 , Z 3 , Z 4 , Z 5 are each independently selected from CR 3 or N;
    环B、R 1、R 2、R 3、R 4、n、L 2的定义如权利要求1中所述；Ra如权利要求1中Ra 1或Ra 2所定义； Ring B, R 1 , R 2 , R 3 , R 4 , n, L 2 are as defined in claim 1; Ra is as defined in claim 1 as Ra 1 or Ra 2 ;
    较佳地，在结构Ia中，Z 1、Z 2、Z 3、Z 4、Z 5中至多含有两个N； Preferably, in structure Ia, Z 1 , Z 2 , Z 3 , Z 4 , Z 5 contain at most two Ns;
    较佳地，在结构Ib中，Z 1、Z 2、Z 3、Z 4中至多含有两个N。 Preferably, in structure Ib, Z 1 , Z 2 , Z 3 , and Z 4 contain at most two Ns.
  7. 如权利要求1所述的式I所示苯并杂环化合物、其互变异构体、立体异构体、水合物、溶剂化物、药学上可接受的盐或前药，其特征在于，具有如下结构IIa、IIb、IIc、IId、IIe、IIf、IIg、IIm、IIn：The benzoheterocyclic compound shown in formula I according to claim 1, its tautomers, stereoisomers, hydrates, solvates, pharmaceutically acceptable salts or prodrugs, is characterized in that it has The following structures IIa, IIb, IIc, IId, IIe, IIf, IIg, IIm, IIn:
    Figure PCTCN2022097611-appb-100004
    Figure PCTCN2022097611-appb-100004
    Figure PCTCN2022097611-appb-100005
    Figure PCTCN2022097611-appb-100005
  8. 如权利要求1-6任一项所述的式I所示苯并杂环化合物、其互变异构体、立体异构体、水合物、溶剂化物、药学上可接受的盐或前药，其特征在于，L 2不存在或选自-CH 2-、-CH 2CH 2-、-CH 2CH 2CH 2-、-CH(CH 3)-、-CH(CH 3)CH 2-；较佳地，L 2不存在或为-CH 2-或-CH(CH 3)-。 The benzoheterocyclic compound represented by formula I according to any one of claims 1-6, its tautomers, stereoisomers, hydrates, solvates, pharmaceutically acceptable salts or prodrugs, It is characterized in that L 2 does not exist or is selected from -CH 2 -, -CH 2 CH 2 -, -CH 2 CH 2 CH 2 -, -CH(CH 3 )-, -CH(CH 3 )CH 2 -; Preferably, L 2 is absent or is -CH 2 - or -CH(CH 3 )-.
  9. 如权利要求1-7任一项所述的式I所示苯并杂环化合物、其互变异构体、立体异构体、水合物、溶剂化物、药学上可接受的盐或前药，其特征在于，R 1、R 2分别为氢或-C≡C-R 11，且R 1、R 2不同； The benzoheterocyclic compound represented by formula I according to any one of claims 1-7, its tautomers, stereoisomers, hydrates, solvates, pharmaceutically acceptable salts or prodrugs, It is characterized in that R 1 and R 2 are hydrogen or -C≡CR 11 respectively, and R 1 and R 2 are different;
    R 11为氢、C 1-C 5烷基或3-6元环烷基； R 11 is hydrogen, C 1 -C 5 alkyl or 3-6 membered cycloalkyl;
    较佳地，所述R 11为甲基。 Preferably, the R 11 is methyl.
  10. 如权利要求6所述的式I所示苯并杂环化合物、其互变异构体、立体异构体、水合物、溶剂化物、药学上可接受的盐或前药，其特征在于，Ra为甲基、乙基、丙基。The benzoheterocyclic compound represented by formula I as claimed in claim 6, its tautomer, stereoisomer, hydrate, solvate, pharmaceutically acceptable salt or prodrug, is characterized in that, Ra For methyl, ethyl, propyl.
  11. 如权利要求1-5或7任一项所述的式I所示苯并杂环化合物、其互变异构体、立体异构体、水合物、溶剂化物、药学上可接受的盐或前药，其特征在于，环B为不存在或为环丙基、环丁基、环戊基、环己基、苯基、噻吩、呋喃、吡咯、吡唑、咪唑、三氮唑、噻唑、噻二唑、噁唑、吡啶、嘧啶、哒嗪、吡嗪。The benzoheterocyclic compound represented by formula I according to any one of claims 1-5 or 7, its tautomers, stereoisomers, hydrates, solvates, pharmaceutically acceptable salts or precursors The drug is characterized in that ring B does not exist or is cyclopropyl, cyclobutyl, cyclopentyl, cyclohexyl, phenyl, thiophene, furan, pyrrole, pyrazole, imidazole, triazole, thiazole, thiadi Azole, oxazole, pyridine, pyrimidine, pyridazine, pyrazine.
  12. 如权利要求1-5或7任一项所述的式I所示苯并杂环化合物、其互变异构体、立体异构体、水合物、溶剂化物、药学上可接受的盐或前药，其特征在于，环B为氧杂环丁烷、
    Figure PCTCN2022097611-appb-100006
    或为选自下列杂芳基的饱和或部分饱和的杂环：噻吩、呋喃、吡咯、吡唑、咪唑、三氮唑、噻唑、噻二唑、噁唑、吡啶、嘧啶、哒嗪、吡嗪。     The benzoheterocyclic compound represented by formula I according to any one of claims 1-5 or 7, its tautomers, stereoisomers, hydrates, solvates, pharmaceutically acceptable salts or precursors Medicine, it is characterized in that, ring B is oxetane,
    Figure PCTCN2022097611-appb-100006
    Or a saturated or partially saturated heterocycle selected from the following heteroaryl groups: thiophene, furan, pyrrole, pyrazole, imidazole, triazole, thiazole, thiadiazole, oxazole, pyridine, pyrimidine, pyridazine, pyrazine .
  13. 如权利要求1所述的式I所示苯并杂环化合物、其互变异构体、立体异构体、水合物、溶剂化物、药学上可接受的盐或前药，其特征在于，包括：The benzoheterocyclic compound represented by formula I according to claim 1, its tautomer, stereoisomer, hydrate, solvate, pharmaceutically acceptable salt or prodrug, is characterized in that, comprising :
    Figure PCTCN2022097611-appb-100007
    Figure PCTCN2022097611-appb-100007
    Figure PCTCN2022097611-appb-100008
    Figure PCTCN2022097611-appb-100008
    Figure PCTCN2022097611-appb-100009
    Figure PCTCN2022097611-appb-100009
  14. 如权利要求1所述的式I所示苯并杂环化合物、其互变异构体、立体异构体、水合物、溶剂化物、药学上可接受的盐或前药，其特征在于，包括：The benzoheterocyclic compound represented by formula I according to claim 1, its tautomer, stereoisomer, hydrate, solvate, pharmaceutically acceptable salt or prodrug, is characterized in that, comprising :
    Figure PCTCN2022097611-appb-100010
    Figure PCTCN2022097611-appb-100010
    Figure PCTCN2022097611-appb-100011
    Figure PCTCN2022097611-appb-100011
    Figure PCTCN2022097611-appb-100012
    Figure PCTCN2022097611-appb-100012
  15. 一种中间体B-1：An intermediate B-1:
    Figure PCTCN2022097611-appb-100013
    Figure PCTCN2022097611-appb-100013
    其中，R 6为-OH、-Cl、-O-C 1-C 5烷基、-O-苄基； Wherein, R 6 is -OH, -Cl, -OC 1 -C 5 alkyl, -O-benzyl;
    环A、环B、L 1、R 1、R 2、R 3、R 4、m、n的定义如权利要求1中所述。 Ring A, Ring B, L 1 , R 1 , R 2 , R 3 , R 4 , m, n are as defined in claim 1.
  16. 如权利要求15所述的中间体B-1，其特征在于，具有结构B-1a或B-1b：Intermediate B-1 according to claim 15, having structure B-1a or B-1b:
    Figure PCTCN2022097611-appb-100014
    Figure PCTCN2022097611-appb-100014
    其中，Z 1、Z 2、Z 3、Z 4、Z 5各自独立地选自CR 3或N； Wherein, Z 1 , Z 2 , Z 3 , Z 4 , Z 5 are each independently selected from CR 3 or N;
    环B、R 1、R 2、R 3、R 4、n的定义如权利要求1中所述；Ra如权利要求1中Ra 1或Ra 2所定义； Ring B, R 1 , R 2 , R 3 , R 4 , n are as defined in claim 1; Ra is as defined in claim 1 as Ra 1 or Ra 2 ;
    较佳地，在结构Ia中，Z 1、Z 2、Z 3、Z 4、Z 5中至多含有两个N； Preferably, in structure Ia, Z 1 , Z 2 , Z 3 , Z 4 , Z 5 contain at most two Ns;
    较佳地，在结构Ib中，Z 1、Z 2、Z 3、Z 4中至多含有两个N； Preferably, in structure Ib, Z 1 , Z 2 , Z 3 , and Z 4 contain at most two Ns;
    较佳地，所述中间体B-1具有结构B-1c、B-1d、B-1f、B-1g：Preferably, the intermediate B-1 has the structures B-1c, B-1d, B-1f, B-1g:
    Figure PCTCN2022097611-appb-100015
    Figure PCTCN2022097611-appb-100015
  17. 一种制备如权利要求1-14任一所述的式I所示苯并杂环化合物、其互变异构体、立体异构体、水合物、溶剂化物、药学上可接受的盐或前药的方法，其特征在于，包括：A preparation of the benzoheterocyclic compound represented by the formula I as described in any one of claims 1-14, its tautomer, stereoisomer, hydrate, solvate, pharmaceutically acceptable salt or precursor The method of medicine is characterized in that, comprising:
    1)将如权利要求15或16所述的中间体B-1与中间体B-2或中间体B-2的盐反应，得到所述式I所示苯并杂环化合物；1) reacting the intermediate B-1 as claimed in claim 15 or 16 with the intermediate B-2 or the salt of the intermediate B-2 to obtain the benzoheterocyclic compound shown in the formula I;
    所述中间体B-2具有结构：The intermediate B-2 has the structure:
    Figure PCTCN2022097611-appb-100016
    Figure PCTCN2022097611-appb-100016
    其中，R 5为C 1-C 6烷基、苄基； Wherein, R 5 is C 1 -C 6 alkyl, benzyl;
    较佳地，所述中间体B-2具有结构B-2a或B-2bPreferably, the intermediate B-2 has the structure B-2a or B-2b
    Figure PCTCN2022097611-appb-100017
    Figure PCTCN2022097611-appb-100017
  18. 如权利要求17所述的方法，其特征在于，所述方法还包括：The method of claim 17, further comprising:
    2)当所述中间体B-1中R 6不为-OH或-Cl时，将基团-COR 6转化为-COOH或-COCl后，再与中间体B-2反应；和/或 2) When R in the intermediate B- 1 is not -OH or -Cl, the group -COR is converted into -COOH or -COCl, and then reacted with the intermediate B- 2 ; and/or
    3)在所述中间体B-1与中间体B-2反应后，将基团-COOR 5水解为-COOH。 3) After the reaction of intermediate B-1 with intermediate B-2, the group -COOR 5 is hydrolyzed to -COOH.
  19. 一种药物组合物，其特征在于，所述药物组合物包括：如权利要求1-14中任一所述的式I所示苯并杂环化合物、其互变异构体、立体异构体、水合物、溶剂化物、药学上可接受的盐或前药；和药学上可接受的载体。A pharmaceutical composition, characterized in that the pharmaceutical composition comprises: the benzoheterocyclic compound shown in formula I according to any one of claims 1-14, its tautomers, stereoisomers , hydrate, solvate, pharmaceutically acceptable salt or prodrug; and a pharmaceutically acceptable carrier.
  20. 一种药物组合物，其特征在于，所述药物组合物包括：如权利要求1-14中任一所述的式I所示苯并杂环化合物、其互变异构体、立体异构体、水合物、溶剂化物、药学上可接受的盐或前药；和第二种药物；A pharmaceutical composition, characterized in that the pharmaceutical composition comprises: the benzoheterocyclic compound shown in formula I according to any one of claims 1-14, its tautomers, stereoisomers , hydrate, solvate, pharmaceutically acceptable salt or prodrug; and a second drug;
    较佳地，所述第二种药物包括抗体；Preferably, said second drug comprises an antibody;
    较佳地，所述抗体包括抗PD-L1抗体、抗PD-1抗体。Preferably, the antibodies include anti-PD-L1 antibodies and anti-PD-1 antibodies.
  21. 一种如权利要求1-14中任一所述的式I所示苯并杂环化合物、其互变异构体、立体异构体、水合物、溶剂化物、药学上可接受的盐或前药的用途，或如权利要求19或20所述的药物组合物的用途，所述用途包括：A benzoheterocyclic compound represented by formula I as described in any one of claims 1-14, its tautomers, stereoisomers, hydrates, solvates, pharmaceutically acceptable salts or precursors The purposes of medicine, or the purposes of pharmaceutical composition as described in claim 19 or 20, described purposes comprises:
    1)对EP2和/或EP4产生拮抗作用；1) produce antagonistic effect on EP2 and/or EP4;
    2)与EP2和/或EP4受体结合；2) Binding to EP2 and/or EP4 receptors;
    3)预防、治疗EP2和/或EP4受体介导的疾病疾病、3) Prevention and treatment of diseases mediated by EP2 and/or EP4 receptors,
    4)制备EP2和/或EP4拮抗剂、4) preparing EP2 and/or EP4 antagonists,
    5)制备预防、治疗与EP2和/或EP4受体介导的疾病的药物、药物组合物或制剂。5) Preparation of medicines, pharmaceutical compositions or preparations for preventing and treating diseases mediated by EP2 and/or EP4 receptors.
  22. 如权利要求21所述的用途，其中所述EP2和/或EP4受体介导的疾病包括炎性疾病、自身免疫疾病、神经变性疾病、心血管疾病和癌症。The use according to claim 21, wherein the diseases mediated by the EP2 and/or EP4 receptors include inflammatory diseases, autoimmune diseases, neurodegenerative diseases, cardiovascular diseases and cancers.
PCT/CN2022/097611 2021-06-08 2022-06-08 Benzoheterocyclic compound for treating ep2 and ep4 receptor-mediated diseases WO2022257961A1 (en)

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