WO2019214656A1 - 一类含氟异噁唑类化合物及其制备方法、药物组合物和用途 - Google Patents

一类含氟异噁唑类化合物及其制备方法、药物组合物和用途 Download PDF

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WO2019214656A1
WO2019214656A1 PCT/CN2019/086068 CN2019086068W WO2019214656A1 WO 2019214656 A1 WO2019214656 A1 WO 2019214656A1 CN 2019086068 W CN2019086068 W CN 2019086068W WO 2019214656 A1 WO2019214656 A1 WO 2019214656A1
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methyl
substituted
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柳红
李佳
周宇
臧奕
李亚洲
孙丹丹
李俊友
茹弘博
高立信
蒋华良
陈凯先
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中国科学院上海药物研究所
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    • A—HUMAN NECESSITIES
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    • A61K—PREPARATIONS FOR MEDICAL, DENTAL OR TOILETRY PURPOSES
    • A61K31/00—Medicinal preparations containing organic active ingredients
    • A61K31/33—Heterocyclic compounds
    • A61K31/395—Heterocyclic compounds having nitrogen as a ring hetero atom, e.g. guanethidine or rifamycins
    • A61K31/435—Heterocyclic compounds having nitrogen as a ring hetero atom, e.g. guanethidine or rifamycins having six-membered rings with one nitrogen as the only ring hetero atom
    • A61K31/4353—Heterocyclic compounds having nitrogen as a ring hetero atom, e.g. guanethidine or rifamycins having six-membered rings with one nitrogen as the only ring hetero atom ortho- or peri-condensed with heterocyclic ring systems
    • A61K31/437—Heterocyclic compounds having nitrogen as a ring hetero atom, e.g. guanethidine or rifamycins having six-membered rings with one nitrogen as the only ring hetero atom ortho- or peri-condensed with heterocyclic ring systems the heterocyclic ring system containing a five-membered ring having nitrogen as a ring hetero atom, e.g. indolizine, beta-carboline
    • A—HUMAN NECESSITIES
    • A61—MEDICAL OR VETERINARY SCIENCE; HYGIENE
    • A61K—PREPARATIONS FOR MEDICAL, DENTAL OR TOILETRY PURPOSES
    • A61K31/00—Medicinal preparations containing organic active ingredients
    • A61K31/33—Heterocyclic compounds
    • A61K31/395—Heterocyclic compounds having nitrogen as a ring hetero atom, e.g. guanethidine or rifamycins
    • A61K31/435—Heterocyclic compounds having nitrogen as a ring hetero atom, e.g. guanethidine or rifamycins having six-membered rings with one nitrogen as the only ring hetero atom
    • A61K31/44—Non condensed pyridines; Hydrogenated derivatives thereof
    • A61K31/445—Non condensed piperidines, e.g. piperocaine
    • A61K31/4523—Non condensed piperidines, e.g. piperocaine containing further heterocyclic ring systems
    • A—HUMAN NECESSITIES
    • A61—MEDICAL OR VETERINARY SCIENCE; HYGIENE
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    • A61K31/00—Medicinal preparations containing organic active ingredients
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    • A61K31/435—Heterocyclic compounds having nitrogen as a ring hetero atom, e.g. guanethidine or rifamycins having six-membered rings with one nitrogen as the only ring hetero atom
    • A61K31/44—Non condensed pyridines; Hydrogenated derivatives thereof
    • A61K31/445—Non condensed piperidines, e.g. piperocaine
    • A61K31/4523—Non condensed piperidines, e.g. piperocaine containing further heterocyclic ring systems
    • A61K31/4535—Non condensed piperidines, e.g. piperocaine containing further heterocyclic ring systems containing a heterocyclic ring having sulfur as a ring hetero atom, e.g. pizotifen
    • A—HUMAN NECESSITIES
    • A61—MEDICAL OR VETERINARY SCIENCE; HYGIENE
    • A61K—PREPARATIONS FOR MEDICAL, DENTAL OR TOILETRY PURPOSES
    • A61K31/00—Medicinal preparations containing organic active ingredients
    • A61K31/33—Heterocyclic compounds
    • A61K31/395—Heterocyclic compounds having nitrogen as a ring hetero atom, e.g. guanethidine or rifamycins
    • A61K31/435—Heterocyclic compounds having nitrogen as a ring hetero atom, e.g. guanethidine or rifamycins having six-membered rings with one nitrogen as the only ring hetero atom
    • A61K31/44—Non condensed pyridines; Hydrogenated derivatives thereof
    • A61K31/445—Non condensed piperidines, e.g. piperocaine
    • A61K31/4523—Non condensed piperidines, e.g. piperocaine containing further heterocyclic ring systems
    • A61K31/454—Non condensed piperidines, e.g. piperocaine containing further heterocyclic ring systems containing a five-membered ring with nitrogen as a ring hetero atom, e.g. pimozide, domperidone
    • A—HUMAN NECESSITIES
    • A61—MEDICAL OR VETERINARY SCIENCE; HYGIENE
    • A61K—PREPARATIONS FOR MEDICAL, DENTAL OR TOILETRY PURPOSES
    • A61K31/00—Medicinal preparations containing organic active ingredients
    • A61K31/33—Heterocyclic compounds
    • A61K31/395—Heterocyclic compounds having nitrogen as a ring hetero atom, e.g. guanethidine or rifamycins
    • A61K31/435—Heterocyclic compounds having nitrogen as a ring hetero atom, e.g. guanethidine or rifamycins having six-membered rings with one nitrogen as the only ring hetero atom
    • A61K31/44—Non condensed pyridines; Hydrogenated derivatives thereof
    • A61K31/445—Non condensed piperidines, e.g. piperocaine
    • A61K31/4523—Non condensed piperidines, e.g. piperocaine containing further heterocyclic ring systems
    • A61K31/4545—Non condensed piperidines, e.g. piperocaine containing further heterocyclic ring systems containing a six-membered ring with nitrogen as a ring hetero atom, e.g. pipamperone, anabasine
    • A—HUMAN NECESSITIES
    • A61—MEDICAL OR VETERINARY SCIENCE; HYGIENE
    • A61K—PREPARATIONS FOR MEDICAL, DENTAL OR TOILETRY PURPOSES
    • A61K31/00—Medicinal preparations containing organic active ingredients
    • A61K31/33—Heterocyclic compounds
    • A61K31/395—Heterocyclic compounds having nitrogen as a ring hetero atom, e.g. guanethidine or rifamycins
    • A61K31/435—Heterocyclic compounds having nitrogen as a ring hetero atom, e.g. guanethidine or rifamycins having six-membered rings with one nitrogen as the only ring hetero atom
    • A61K31/46—8-Azabicyclo [3.2.1] octane; Derivatives thereof, e.g. atropine, cocaine
    • A—HUMAN NECESSITIES
    • A61—MEDICAL OR VETERINARY SCIENCE; HYGIENE
    • A61P—SPECIFIC THERAPEUTIC ACTIVITY OF CHEMICAL COMPOUNDS OR MEDICINAL PREPARATIONS
    • A61P1/00—Drugs for disorders of the alimentary tract or the digestive system
    • A61P1/16—Drugs for disorders of the alimentary tract or the digestive system for liver or gallbladder disorders, e.g. hepatoprotective agents, cholagogues, litholytics
    • A—HUMAN NECESSITIES
    • A61—MEDICAL OR VETERINARY SCIENCE; HYGIENE
    • A61P—SPECIFIC THERAPEUTIC ACTIVITY OF CHEMICAL COMPOUNDS OR MEDICINAL PREPARATIONS
    • A61P3/00—Drugs for disorders of the metabolism
    • A61P3/06—Antihyperlipidemics
    • C—CHEMISTRY; METALLURGY
    • C07—ORGANIC CHEMISTRY
    • C07D—HETEROCYCLIC COMPOUNDS
    • C07D413/00—Heterocyclic compounds containing two or more hetero rings, at least one ring having nitrogen and oxygen atoms as the only ring hetero atoms
    • C07D413/02—Heterocyclic compounds containing two or more hetero rings, at least one ring having nitrogen and oxygen atoms as the only ring hetero atoms containing two hetero rings
    • C07D413/12—Heterocyclic compounds containing two or more hetero rings, at least one ring having nitrogen and oxygen atoms as the only ring hetero atoms containing two hetero rings linked by a chain containing hetero atoms as chain links
    • C—CHEMISTRY; METALLURGY
    • C07—ORGANIC CHEMISTRY
    • C07D—HETEROCYCLIC COMPOUNDS
    • C07D413/00—Heterocyclic compounds containing two or more hetero rings, at least one ring having nitrogen and oxygen atoms as the only ring hetero atoms
    • C07D413/14—Heterocyclic compounds containing two or more hetero rings, at least one ring having nitrogen and oxygen atoms as the only ring hetero atoms containing three or more hetero rings
    • C—CHEMISTRY; METALLURGY
    • C07—ORGANIC CHEMISTRY
    • C07D—HETEROCYCLIC COMPOUNDS
    • C07D417/00—Heterocyclic compounds containing two or more hetero rings, at least one ring having nitrogen and sulfur atoms as the only ring hetero atoms, not provided for by group C07D415/00
    • C07D417/14—Heterocyclic compounds containing two or more hetero rings, at least one ring having nitrogen and sulfur atoms as the only ring hetero atoms, not provided for by group C07D415/00 containing three or more hetero rings
    • C—CHEMISTRY; METALLURGY
    • C07—ORGANIC CHEMISTRY
    • C07D—HETEROCYCLIC COMPOUNDS
    • C07D451/00—Heterocyclic compounds containing 8-azabicyclo [3.2.1] octane, 9-azabicyclo [3.3.1] nonane, or 3-oxa-9-azatricyclo [3.3.1.0<2,4>] nonane ring systems, e.g. tropane or granatane alkaloids, scopolamine; Cyclic acetals thereof
    • C07D451/02—Heterocyclic compounds containing 8-azabicyclo [3.2.1] octane, 9-azabicyclo [3.3.1] nonane, or 3-oxa-9-azatricyclo [3.3.1.0<2,4>] nonane ring systems, e.g. tropane or granatane alkaloids, scopolamine; Cyclic acetals thereof containing not further condensed 8-azabicyclo [3.2.1] octane or 3-oxa-9-azatricyclo [3.3.1.0<2,4>] nonane ring systems, e.g. tropane; Cyclic acetals thereof
    • C07D451/04—Heterocyclic compounds containing 8-azabicyclo [3.2.1] octane, 9-azabicyclo [3.3.1] nonane, or 3-oxa-9-azatricyclo [3.3.1.0<2,4>] nonane ring systems, e.g. tropane or granatane alkaloids, scopolamine; Cyclic acetals thereof containing not further condensed 8-azabicyclo [3.2.1] octane or 3-oxa-9-azatricyclo [3.3.1.0<2,4>] nonane ring systems, e.g. tropane; Cyclic acetals thereof with hetero atoms directly attached in position 3 of the 8-azabicyclo [3.2.1] octane or in position 7 of the 3-oxa-9-azatricyclo [3.3.1.0<2,4>] nonane ring system
    • C—CHEMISTRY; METALLURGY
    • C07—ORGANIC CHEMISTRY
    • C07D—HETEROCYCLIC COMPOUNDS
    • C07D471/00—Heterocyclic compounds containing nitrogen atoms as the only ring hetero atoms in the condensed system, at least one ring being a six-membered ring with one nitrogen atom, not provided for by groups C07D451/00 - C07D463/00
    • C07D471/02—Heterocyclic compounds containing nitrogen atoms as the only ring hetero atoms in the condensed system, at least one ring being a six-membered ring with one nitrogen atom, not provided for by groups C07D451/00 - C07D463/00 in which the condensed system contains two hetero rings
    • C07D471/04—Ortho-condensed systems
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    • C07D—HETEROCYCLIC COMPOUNDS
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Definitions

  • the present invention relates to the field of medicinal chemistry and pharmacotherapy, and in particular to a class of fluoroisoxazole compounds, a process for the preparation thereof, a pharmaceutical composition containing the same, and as an FXR agonist, in particular for the treatment of nonalcoholic
  • a pharmaceutical composition containing the same and as an FXR agonist
  • Non-alcoholic fatty liver disease refers to the clinical pathological syndrome characterized by excessive alcohol deposition in the liver, in addition to alcohol and other well-defined liver damage factors. It is an acquired metabolic stress. Liver damage is closely related to insulin resistance and genetic susceptibility. In developed countries, the prevalence of NAFLD is about 2% per year in people with risk factors, and the onset of disease is getting younger and younger. It has become the “first major liver disease” in Western countries, and it is also a common type of chronic liver disease in China.
  • NAFLD hepatocellular carcinoma and transplanted liver recurrence
  • NAFLD can also affect the progression of other chronic liver diseases and participate in the development of type 2 diabetes and atherosclerosis, which is considered to be systemic metabolic syndrome. Local manifestations of the liver.
  • NAFLD mainly includes simple fatty liver (SFL) and nonalcoholic steatohepatitis (NASH), of which NASH is the most important type.
  • SFL simple fatty liver
  • NASH nonalcoholic steatohepatitis
  • the incidence of cirrhosis in NASH patients is as high as 25% within 10 years.
  • NASH drug research and treatment has been slow.
  • NASH was considered a benign complication of diabetes or obesity. This misconception led to the neglect of timely diagnosis and treatment.
  • NASH therapeutic drugs mainly include: (1) selective peripheral cannabinoid receptor (CB) inhibitor; (2) peroxisome proliferator-activated receptor (PPAR) agonist; (3) caspase Aspartate (Caspase) inhibitor; (4) phosphodiesterase 4 (PDE4) inhibitor; (5) farnesyl ester derivative X receptor (FXR) agonist; (6) chemokine receptor-5 / chemokine receptor-2 (CCR5/CCR2) dual target inhibitor; (7) apoptosis signal-regulated kinase-1 (ASK1) inhibitor and the like.
  • CB selective peripheral cannabinoid receptor
  • PPAR peroxisome proliferator-activated receptor
  • Caspase caspase Aspartate
  • PDE4 phosphodiesterase 4
  • FXR farnesyl ester derivative X receptor
  • CCR5/CCR2 chemokine receptor-5 / chemokine receptor-2
  • ASK1 apoptosis signal-regulated kinase
  • FXR farnesoid derivative X receptor
  • the FXR receptor is a bile acid receptor, and the activation of FXR controls the synthesis and transport of bile acids.
  • FXR is a member of the hormone nuclear receptor superfamily and has a typical nuclear receptor structure. Currently, there are four subtypes of FXR ⁇ 1, FXR ⁇ 2, FXR ⁇ 3 and FXR ⁇ 4.
  • FXR ligand inhibits hepatic stellate cell (HSC) expression, reduces extracellular matrix (ECM) production and increases ECM clearance by activating FXR and FXR target gene small heterodimeric chaperone receptor (SHP), thereby reducing ECM Deposition; FXR ligand can also increase the sensitivity of HSC to apoptosis, increase the apoptosis of HSC, and reverse the liver fibrosis by interacting with other nuclear factors that reverse liver fibrosis, thus playing a role in the treatment of liver fibrosis.
  • HSC hepatic stellate cell
  • ECM extracellular matrix
  • SHP small heterodimeric chaperone receptor
  • FXR agonists have entered clinical research.
  • the compound LJN-452 discovered by Novatis has good biological activity and selectivity. Its EC50 of FXR reaches 0.2nM, and it has entered Phase II clinical trials.
  • Treatment of NASH and primary biliary cholangitis (PBC); FXR agonist PX-104 developed by Gilead has entered Phase II clinical studies for NASH treatment, but unfortunately failed to reach the expected clinical endpoint .
  • LY-2562175 and INT-767 have also entered clinical research and are expected to alleviate the huge clinical needs that NASH has not yet met.
  • OCA olbecholic acid
  • the first aspect of the present invention provides a fluorine-containing isoxazole compound having the structure represented by the following Formula I, or an enantiomer or diastereomer thereof, or a pharmaceutically acceptable salt thereof, or a pharmaceutically acceptable salt thereof mixture:
  • n 0, 1, 2 or 3;
  • n 0, 1 or 2;
  • p 0, 1 or 2;
  • the ring is selected from the group consisting of a substituted or unsubstituted 6-20 membered heterocyclic group, a substituted or unsubstituted 6-20 membered aromatic ring group, a substituted or unsubstituted 6-20 membered aromatic heterocyclic group;
  • the substitution means that the hydrogen atom on the group is substituted by 1, 2, 3 or 4 substituents selected from the group consisting of hydrazine (D), hydrazine (T), halogen, C 1 -C 6 alkyl, halogenated C 1- C 6 alkyl, C 1 -C 6 alkoxy, halogenated C 1 -C 6 alkoxy, -O[(CH 2 ) q O] r R 5 , -O(CH 2 ) s C 6 -C 10 aryl, C 3 -C 8 cycloalkoxy, halogenated C 3 -C 8 cycloalkoxy, cyano, nitro, amino, amine (
  • R 1 is selected from the group consisting of a substituted or unsubstituted 6-20 membered heterocyclic group, a substituted or unsubstituted 6-20 membered aromatic ring group, a substituted or unsubstituted 6-20 membered aromatic heterocyclic group;
  • the substitution means that the hydrogen atom on the group is substituted by 1, 2, 3 or 4 substituents selected from the group consisting of hydrazine, hydrazine, halogen, C 1 -C 6 alkyl, C 1 -C 6 alkoxy , halogenated C 1 -C 6 alkoxy, -O[(CH 2 ) q O] r R 5 , -O(CH 2 ) s C 6 -C 10 aryl, C 3 -C 8 cycloalkoxy , halogenated C 3 -C 8 cycloalkoxy, cyano, nitro, amino, amine (preferably C 1 -C 6 amine), hydroxy, hydroxy
  • R 2 may be independently selected from 1, 2, 3 or 4 of the following groups of substituents: hydrogen, halogen, substituted or unsubstituted C 1 -C 6 alkyl, substituted or unsubstituted C 1 -C 6 alkoxy Base, -O[(CH 2 ) q O] r R 5 , -O(CH 2 ) s C 6 -C 10 aryl, C 3 -C 8 cycloalkoxy, halogenated C 3 -C 8 naphthenic An oxy group, a cyano group, a nitro group, an amino group, an amine group (preferably a C 1 -C 6 amine group), a hydroxyl group, a hydroxymethyl group, a carboxyl group; wherein the substituent refers to a hydrogen atom on the group 2, 3 or 4 substituents selected from the group consisting of hydrogen, halogen, nitro, cyano, trifluoromethyl, trifluoroethyl, trifluor
  • R 3 and R 4 are each independently selected from the group consisting of hydrogen, halogen, C 1 -C 6 alkyl, halo C 1 -C 6 alkyl, C 1 -C 6 alkoxy, halo C 1 -C 6 alkoxy, -O[(CH 2 ) q O] r R 5 , -O(CH 2 ) s C 6 -C 10 aryl, C 3 -C 8 cycloalkoxy, halogenated C 3 -C 8- cycloalkoxy, cyano, nitro, amino, amine (preferably C 1 -C 6 amine), hydroxy, hydroxymethyl and carboxy; or R 3 and R 4 joined together to form C 1 -C 6 alkylene;
  • m is 0, 1, or 2; preferably 0 or 1.
  • n is 0 or 1.
  • the ring is selected from the group consisting of phenyl, pyridyl, pyrimidinyl, pyridazinyl, thiazolyl, benzothiazolyl, benzo[d]isothiazolyl, imidazo[1,2-a]pyridinyl, quinoline 1,1H-fluorenyl, pyrrolo[1,2-b]pyridazinyl, benzofuranyl, benzo[b]phenylthio, 1H-carbazolyl, benzo[d]isoxazolyl , thiazoline group, 1H-pyrrolo[3,2-c]pyridyl, pyrazolo[1,5-a]pyrimidinyl, imidazo[1,2-b]pyridazinyl, pyrazolo[1, 5-a]pyrrolyl; the above group is substituted or unsubstituted; wherein said substituent means that the hydrogen atom on the group is substituted by
  • q, r, s, y1 and y2 may be 1, 2, 3 or 4, respectively;
  • the aromatic heterocyclic ring, the aryl group or the heterocyclic group each independently contains 1 to 4 hetero atoms selected from the group consisting of oxygen, sulfur and nitrogen;
  • X 4 is a bond, a C1-C6 alkylene group, a C 1-2 alkene group, a cyclopropyl group, an epoxy ethyl group;
  • R 5 and R 6 are each independently hydrogen, C 1 -C 6 alkyl, halo C 1 -C 6 alkyl, C 2 -C 6 alkenyl, hydroxymethyl or 5-7 membered heterocyclic.
  • R 8 is selected from the group consisting of hydrogen, halogen, C 1 -C 6 alkyl, halogenated C 1 -C 6 alkyl, C 1 -C 6 alkoxy, C 1 -C 6 alkoxycarbonyl, halogenated C 1 -C 6 alkoxy, C 2 -C 6 alkenyl, C 2 -C 6 alkynyl, C 3 -C 8 cycloalkyl, cyano, nitro, amino, hydroxy, hydroxymethyl, carboxy, a mercapto group, a sulfonyl group, a C 6 -C 10 aryl group and a 3-12 membered heterocyclic group;
  • t 0, 1, 2, 3, 4 or 5;
  • X l and X 2 are each independently N, CH or CR 8 ;
  • X 3 is O or S
  • R 9 is -X 4 -CO 2 R 5 , C 1 -C 6 alkyl, halogenated C 1 -C 6 alkyl, C 1 -C 6 hydroxyl, -X 4 -CONR 5 R 6 , -X 4 - CONR 5 (CH 2 ) y1 CO 2 R 6 , -X 4 -CONR 5 (CH 2 ) y2 SO 3 R 6 ; y1 and y2 are each independently 1, 2, 3 or 4; X 4 is a bond, C1- C6 alkylene, C 1-2 alkene, cyclopropyl or epoxyethyl;
  • R 5 and R 6 are each independently hydrogen, C 1-6 alkyl, halo C 1 -C 6 alkyl, C 2 -C 6 alkenyl, hydroxymethyl or 5-7 membered heterocyclyl;
  • R 10 is hydrogen, C 1 -C 6 alkylhydroxy, C 1 -C 6 alkyl, halo C 1 -C 6 alkyl, C 1 -C 6 alkoxy, C 1 -C 6 alkoxycarbonyl Halogenated C 1 -C 6 alkoxy, C 2 -C 6 alkenyl, C 2 -C 6 alkynyl or C 3 -C 8 cycloalkyl.
  • R 1 is selected from the group consisting of phenyl, pyridyl, 1-oxopyridyl, cyclohexyl, cyclopentyl, cyclopropyl; the above groups are substituted or unsubstituted;
  • the substitution means that the hydrogen atom on the group is substituted by 1, 2, 3 or 4 substituents selected from the group consisting of hydrogen, halogen, nitro, cyano, trifluoromethyl, trifluoroethyl, trifluoro propyl, trifluoromethoxy, trifluoroethoxy, phenyl, methoxycarbonyl, ethoxycarbonyl, propoxycarbonyl, C 1-6 alkyl, C l-6 cycloalkyl -alkyl, C 1 -6 alkoxy, C 6-6 cycloalkoxy, cyclopropyl.
  • the compound is selected from the group consisting of:
  • a second aspect of the invention provides a pharmaceutical composition
  • a pharmaceutical composition comprising a therapeutically effective amount of the fluoroisoxazole compound of the first aspect, or an enantiomer or diastereomer thereof, or Pharmaceutically acceptable salts, or mixtures thereof, and pharmaceutically acceptable carriers.
  • a third aspect of the invention provides an FXR agonist comprising the fluoroisoxazole compound of the first aspect, or an enantiomer or diastereomer thereof, or a pharmaceutically acceptable salt thereof, Or a mixture of them.
  • the fourth aspect of the present invention provides the use of the fluorine-containing isoxazole compound of the first aspect, or an enantiomer or diastereomer thereof, or a pharmaceutically acceptable salt thereof, or a mixture thereof, for use For the preparation of a medicament for preventing or treating FXR-mediated diseases.
  • the disease is selected from the group consisting of nonalcoholic fatty liver disease, nonalcoholic steatohepatitis, fatty liver, liver fibrosis, primary biliary cirrhosis, hyperlipidemia, dyslipidemia.
  • R 1 , R 2 , R 3 , R 4 , n, m and p have the same definitions as defined in the above formula I;
  • X is halogen or nitro;
  • Step j intermediate 12 and intermediate 9 are reacted in an organic solvent in the presence of 18-crown-6 and a base to form intermediate 13;
  • Step k intermediate 13 is deprotected in an organic solvent to form intermediate 14;
  • Step 1 reacting intermediate 14 and intermediate 15 in the presence of a base, a palladium catalyst and a ligand in an organic solvent under nitrogen to form the final product I;
  • Step m under nitrogen protection, in an organic solvent, in the presence of a base, intermediate 14 and intermediate 15 react to form the final product I;
  • R 1 , R 2 , R 3 , R 4 , n, m and p have the same definitions as defined in the above formula I;
  • X is halogen or nitro;
  • Step j intermediate 12 and intermediate 9 are reacted in an organic solvent in the presence of 18-crown-6 and a base to form intermediate 13;
  • Step k intermediate 13 is deprotected in an organic solvent to form intermediate 14;
  • Step 1 intermediate 14 and intermediate 15 are reacted in an organic solvent in the presence of a base, a palladium catalyst and a ligand under nitrogen to form an intermediate I';
  • Step m intermediate 14 and intermediate 15 are reacted in an organic solvent in the presence of a base under nitrogen to form intermediate I';
  • Step e in an organic solvent, the intermediate I' and HNR 5 R 6 , HNR 5 (CH 2 ) y1 CO 2 R 6 , or HNR 5 (CH 2 ) y2 SO 3 R 6 reaction to form the final product I;
  • the ring is selected from the group consisting of a substituted or unsubstituted 6-20 membered heterocyclic group, a substituted or unsubstituted 6-20 membered aromatic ring group, a substituted or unsubstituted 6-20 membered aromatic heterocyclic group;
  • the substitution means that the hydrogen atom on the group is substituted by 1, 2, 3 or 4 substituents selected from the group consisting of -X 4 -CO 2 R 5 ; wherein X 4 and R 5 are as defined above
  • the ring is selected from the group consisting of a substituted or unsubstituted 6-20 membered heterocyclic group, a substituted or unsubstituted 6-20 membered aromatic ring group, a substituted or unsubstituted 6-20 membered aromatic heterocyclic group;
  • the substitution means that the hydrogen atom on the group is substituted by 1, 2, 3 or 4 substituents selected from the group consisting of: -X 4 -CONR 5 R 6 , -X 4 -CONR 5 (CH 2 ) y1 CO 2 R 6 , -X 4 -CONR 5 (CH 2 ) y2 SO 3 R 6 ; wherein X 4 , R 5 , R 6 , y1, y2 are as defined above.
  • the present invention provides a method of treating a fluoroisoxazole compound of the present invention or an enantiomer or diastereomer thereof, or a pharmaceutically acceptable salt thereof, or a pharmaceutically acceptable salt thereof, to a subject in need thereof A mixture or a pharmaceutical composition or FXR agonist of the invention.
  • Figure 1 shows the effect of the compounds of the invention on serum ALP.
  • Figure 2 shows the effect of the compounds of the invention on ⁇ -SMA and Col1 ⁇ 1 mRNA.
  • Figure 3 shows the effect of the compounds of the invention on collagen content.
  • Figure 4 shows the effect of the compounds of the invention on serum concentrations of TC, TG, HDL, LDL.
  • Figure 5 shows the effect of the compounds of the invention on liver coefficient.
  • the inventors designed and prepared a novel class of fluorine-containing isoxazole compounds.
  • the compounds are effective to target agonize FXR. Based on the above findings, the inventors completed the present invention.
  • the term "6-20 membered heterocyclic group” means a saturated or partially saturated heterocyclic group having 6 to 20 ring atoms containing one or more hetero atoms selected from nitrogen and sulfur; It is not limited to morpholinyl, piperidinyl, piperazinyl, cyclohexylpiperidinyl and the like. Other "heterocyclyl” terms have similar meanings.
  • 6-20 membered heterocyclic oxy group means a 6-20 membered heterocyclic group-oxygen group.
  • Other "heterocyclicoxy” terms have similar meanings.
  • 6-20 membered aromatic ring group means an aromatic ring group having 6 to 20 ring atoms; including but not limited to, phenyl, naphthyl, phenanthryl, anthracenyl and the like.
  • aromatic ring terms have similar meanings.
  • the term "6-20 membered aromatic heterocyclic group” means an aromatic heterocyclic group having 6 to 20 ring atoms containing one or more hetero atoms selected from nitrogen and sulfur; including but not limited to : tetrazolyl, pyridyl, pyrimidinyl, pyridazinyl, thiazolyl, benzothiazolyl, benzo[d]isothiazolyl, imidazo[1,2-a]pyridyl, quinolyl, 1H- Mercapto, pyrrolo[1,2-b]pyridazinyl, benzofuranyl, benzo[b]phenylthio, 1H-carbazolyl, benzo[d]isoxazolyl, thiazolinyl , 1H-pyrrolo[3,2-c]pyridyl, pyrazolo[1,5-a]pyrimidinyl, imidazo[1,2-b]pyridazinyl, pyr
  • C 1 -C 6 alkyl means a straight or branched alkyl group having 1 to 6 carbon atoms, and includes, without limitation, methyl, ethyl, propyl, isopropyl. , butyl, isobutyl, sec-butyl, tert-butyl, pentyl and hexyl, etc.; preferably ethyl, propyl, isopropyl, butyl, isobutyl, sec-butyl and tert-butyl.
  • C 1 -C 6 alkoxy means a straight or branched alkoxy group having 1 to 6 carbon atoms, and includes, without limitation, a methoxy group, an ethoxy group, a propoxy group. Base, isopropoxy and butoxy groups.
  • C 2 -C 6 alkenyl means a straight or branched alkenyl group having 2 to 6 carbon atoms and having one double bond, and includes, without limitation, a vinyl group, a propenyl group, and a butyl group. Alkenyl, isobutenyl, pentenyl, hexenyl and the like.
  • C 2 -C 6 alkynyl means a straight or branched alkynyl group having one triple bond having 2 to 6 carbon atoms, and includes, without limitation, an ethynyl group, a propynyl group, Butynyl, isobutynyl, pentynyl and hexynyl.
  • C 3 -C 8 cycloalkyl means a cyclic alkyl group having 3 to 8 carbon atoms in the ring, and includes, without limitation, a cyclopropyl group, a cyclobutyl group, a cyclopentyl group. , cyclohexyl, cycloheptyl, cyclooctyl and cyclodecyl.
  • Other "cycloalkyl” terms have similar meanings.
  • C 3 -C 8 cycloalkoxy means a cyclic alkoxy group having 3 to 8 carbon atoms in the ring, and includes, without limitation, a cyclopropoxy group or a cyclobutoxy group. , cyclopentyloxy, cyclohexyloxy, cycloheptyloxy, cyclooctyloxy, cyclodecyloxy and the like.
  • Other "cycloalkoxy” terms have similar meanings.
  • C 6 -C 10 aryl means an aromatic ring group having 6 to 10 carbon atoms which does not contain a hetero atom in the ring, such as a phenyl group, a naphthyl group or the like.
  • C 6 -C 12 aryl has a similar meaning.
  • C 1 -C 6 amine group means a group having 1 to 6 carbon atoms and having an amino group as a terminal group.
  • the halogen is F, Cl, Br or I.
  • Halogen means substituted by halogen.
  • the compound of the present invention is a compound of the formula I or an enantiomer or diastereomer thereof, or a pharmaceutically acceptable salt thereof, or a mixture thereof.
  • the compounds of the invention are preferably the following specific compounds:
  • the compounds of the invention also comprise a pharmaceutically acceptable salt of a compound of formula I, in particular a compound of formula I which is reacted with an inorganic or organic acid to form a conventional pharmaceutically acceptable salt.
  • conventional pharmaceutically acceptable salts can be prepared by reacting a compound of formula I with an inorganic or organic acid, including hydrochloric acid, hydrobromic acid, sulfuric acid, nitric acid, aminosulfonic acid, phosphoric acid, and the like, and Organic acids include citric acid, tartaric acid, lactic acid, pyruvic acid, acetic acid, benzenesulfonic acid, p-toluenesulfonic acid, methanesulfonic acid, naphthalenesulfonic acid, ethanesulfonic acid, naphthalene disulfonic acid, maleic acid, malic acid, and acrylic acid.
  • Acid fumaric acid, succinic acid, propionic acid, oxalic acid, trifluoroacetic acid, stearic acid, pamoic acid, hydroxymaleic acid, phenylacetic acid, benzoic acid, salicylic acid, glutamic acid, ascorbic acid, p-aminobenzene Sulfonic acid, 2-acetoxybenzoic acid, isethionate, etc.; or a sodium, potassium, calcium, aluminum or ammonium salt of a compound of formula I with an inorganic base; or a compound of formula I and an organic base A methylamine salt, ethylamine salt or ethanolamine salt formed.
  • the compounds of the invention also include the enantiomers (e.g., R-isomers or S-isomers) or diastereomers of the compounds of Formula I, or the racemates or meso-forms thereof.
  • enantiomers e.g., R-isomers or S-isomers
  • diastereomers of the compounds of Formula I or the racemates or meso-forms thereof.
  • the compounds of formula I of the present invention can be prepared as follows.
  • R 2 and p are the same as defined in the above formula I;
  • Step a Dissolve 1 in an organic solvent, add an oxidizing agent dropwise at 0 ° C, and stir the reaction at 0 ° C. After the reaction, the organic layer is extracted with diethyl ether, and then the aqueous layer is extracted with sodium hydroxide solution, and the aqueous layer is acidified to pH with sulfuric acid. ⁇ 1.
  • the organic layer was extracted with diethyl ether, washed with brine, dried and evaporated
  • the organic solvent is acetone, tetrahydrofuran, diethyl ether, dimethylformamide, ethylene glycol dimethyl ether, ethylene glycol diethyl ether, dioxane, ethanol, methanol, ethyl acetate, dichloromethane or a mixture thereof;
  • the oxidizing agent is selected from the group consisting of PCC, PDC, Dess-Martin oxidizing agent, Swern oxidizing agent, H 2 O 2 , potassium permanganate, and manganese dioxide;
  • Step b The intermediate 2 is dissolved in an organic solvent, N,N'-carbonyldiimidazole is added, and the reaction is allowed to stand at room temperature for one hour under nitrogen. Then, potassium monoethyl malonate and magnesium chloride are added, and the reaction is carried out at room temperature overnight. The organic layer was extracted with ethyl acetate. The organic layer was washed with brine, dried and evaporated, The organic solvent is acetone, tetrahydrofuran, diethyl ether, dimethylformamide, ethylene glycol dimethyl ether, ethylene glycol diethyl ether, dioxane, ethanol, methanol, ethyl acetate, dichloromethane or a mixture thereof;
  • R 1 , R 2 and p have the same definitions as defined in the above formula I;
  • Step c Hydroxylamine hydrochloride was dissolved in water, and sodium hydroxide was added dropwise with stirring at 0 °C. 4 After adding an organic solvent to dissolve, the above mixed solution was added dropwise, and then the temperature was raised to 90 ° C and stirred overnight. After the reaction is completed, it is cooled to room temperature, concentrated in a vacuum to a large amount of solids, and filtered to give a solid, which is dried, intermediate 5; the organic solvent is tetrahydrofuran, diethyl ether, dimethylformamide, ethylene glycol dimethyl ether, ethylene glycol Diethyl ether, dioxane, ethanol, methanol, ethyl acetate, dichloromethane or a mixture thereof.
  • the organic solvent is tetrahydrofuran, diethyl ether, dimethylformamide, ethylene glycol dimethyl ether, ethylene glycol Diethyl ether, dioxane, ethanol, methanol, eth
  • Step d Intermediate 5 is dissolved in an organic solvent, and the chlorosuccinimide is dissolved in an organic solvent and dropped into the above 5 solution at 40 ° C, and then reacted at 40 ° C, and then cooled to room temperature after the reaction, and added. A large amount of water and ethyl acetate (EA) were extracted, the organic layer was washed three times with water, and the organic layer was washed with brine, dried over anhydrous Na 2 SO 4 , dimethylformamide, ethylene glycol dimethyl ether, ethylene glycol diethyl ether, dioxane, ethanol, methanol, ethyl acetate, dichloromethane or a mixture thereof;
  • EA ethyl acetate
  • Step e adding intermediate 3 to the base, stirring at room temperature, cooling to 10 ° C, slowly adding dropwise intermediate 6 to obtain intermediate 7;
  • the organic solvent is tetrahydrofuran, diethyl ether, dimethylformamide , ethylene glycol dimethyl ether, ethylene glycol diethyl ether, dioxane, ethanol, methanol or ethyl acetate, dichloromethane;
  • the base is sodium acetate, NaOH, KOH, sodium ethoxide, sodium methoxide, sodium carbonate , potassium carbonate, triethylamine or diisopropylamine.
  • Step f The intermediate 7 is dissolved in an organic solvent, and diisobutylaluminum hydride 1 M/toluene is added dropwise under a nitrogen atmosphere at 0 ° C, and the reaction is stirred at room temperature. After the reaction is completed, methanol and a small amount of water are added dropwise at 0 ° C. After quenching and stirring for 10 minutes, the precipitate was filtered with celite and filtered with organic solvent.
  • Step g Dissolve the intermediate 8 in an organic solvent, add triphenylphosphorus, cool to 0 ° C, add carbon tetrabromide, and then react at room temperature. After the reaction, the reaction solution is concentrated in vacuo to give an orange oil. The column was silica gel and dried to give Intermediate 9.
  • the organic solvent is tetrahydrofuran, diethyl ether, dimethylformamide, ethylene glycol dimethyl ether, ethylene glycol diethyl ether, dioxane, ethanol, methanol, ethyl acetate, dichloromethane or a mixture thereof;
  • Step h Trimethylsulfonium iodide is added to the organic solvent, and the compound 10 is added under stirring to continue stirring, and then potassium t-butoxide is further added; the epoxy compound 2 is obtained.
  • the organic solvent may be dimethyl sulfoxide, tetrahydrofuran, diethyl ether, dimethylformamide, ethylene glycol dimethyl ether, ethylene glycol diethyl ether, dioxane, ethanol, methanol, ethyl acetate, dichloromethane. Or a mixture thereof.
  • Step i dissolving the intermediate 11 in an organic solvent, cooling to -10 ° C to -40 ° C, adding 1-10 equivalents of a hydrogen fluoride pyridine solution, continuing the reaction until the disappearance of the starting material, separating and purifying the intermediate 12;
  • the organic solvent It may be tetrahydrofuran, diethyl ether, dimethylformamide, ethylene glycol dimethyl ether, ethylene glycol diethyl ether, dioxane, ethanol, methanol, ethyl acetate, dichloromethane or a mixture thereof.
  • R 1 , R 2 , R 3 , R 4 , n, m and p have the same meanings as defined in the above formula I;
  • X is halogen or nitro;
  • Step j Dissolve the intermediate 12 in an organic solvent, add the compound 18-crown-6 (12.8 g, 48.6 mmol) and a strong base at 0 ° C, react for 30 minutes, transfer to room temperature, and add the intermediate 9 dropwise.
  • the reaction is stirred at room temperature, and after the reaction is completed, the intermediate 13 is isolated and purified;
  • the organic solvent is tetrahydrofuran, diethyl ether, dimethylformamide, ethylene glycol dimethyl ether, ethylene glycol diethyl ether, Dioxane, ethanol, methanol, ethyl acetate, dichloromethane or a mixture thereof;
  • the strong base is NaOH, KOH, potassium t-butoxide, sodium ethoxide or sodium methoxide;
  • Step k dissolving the intermediate 13 in an organic solvent, adding an organic solvent of trifluoroacetic acid (TFA) or hydrochloric acid (HCl), and removing the Boc protecting group to obtain the intermediate 14;
  • the organic solvent may be tetrahydrofuran or diethyl ether.
  • Step 1 Under the protection of nitrogen, the intermediates 14, 15 are dissolved in an organic solvent, and cesium carbonate, tris(dibenzylideneacetone) dipalladium catalyst and 2-dicyclohexylphosphorus-2,4,6-three are added.
  • Isopropyl biphenyl ligand after the reaction, separation and purification to obtain the final product I;
  • the organic solvent is toluene, tetrahydrofuran, diethyl ether, dimethylformamide, ethylene glycol dimethyl ether, ethylene glycol diethyl ether, two Oxycyclohexane, ethanol, methanol, ethyl acetate, dichloromethane or a mixture thereof;
  • step m dissolving the intermediates 14, 15 in an organic solvent under nitrogen, adding N,N-diisopropylethylamine, and after separation, purifying and finally obtaining the final product I;
  • the organic solvent is toluene , tetrahydrofuran, diethyl ether, dimethylformamide, ethylene glycol dimethyl ether, ethylene glycol diethyl ether, dioxane, ethanol, methanol, ethyl acetate, nitromethylpyrrolidone, dichloromethane or a mixture thereof.
  • R 1 , R 2 , R 3 , R 4 , n, m and p are as defined above;
  • X is halogen or nitro;
  • Step j intermediate 12 and intermediate 9 are reacted in an organic solvent in the presence of 18-crown-6 and a base to form intermediate 13;
  • Step k intermediate 13 is deprotected in an organic solvent to form intermediate 14;
  • Step 1 intermediate 14 and intermediate 15 are reacted in an organic solvent in the presence of a base, a palladium catalyst and a ligand under nitrogen to form an intermediate I';
  • Step m intermediate 14 and intermediate 15 are reacted in an organic solvent in the presence of a base under nitrogen to form intermediate I';
  • Step e in an organic solvent, the intermediate I' and HNR 5 R 6 , HNR 5 (CH 2 ) y1 CO 2 R 6 , or HNR 5 (CH 2 ) y2 SO 3 R 6 reaction to form the final product I;
  • the ring is selected from the group consisting of a substituted or unsubstituted 6-20 membered heterocyclic group, a substituted or unsubstituted 6-20 membered aromatic ring group, a substituted or unsubstituted 6-20 membered aromatic heterocyclic group;
  • the substitution means that the hydrogen atom on the group is substituted by 1, 2, 3 or 4 substituents selected from the group consisting of -X 4 -CO 2 R 5 ; wherein X 4 and R 5 are as defined above
  • the ring is selected from the group consisting of a substituted or unsubstituted 6-20 membered heterocyclic group, a substituted or unsubstituted 6-20 membered aromatic ring group, a substituted or unsubstituted 6-20 membered aromatic heterocyclic group;
  • the substitution means that the hydrogen atom on the group is substituted by 1, 2, 3 or 4 substituents selected from the group consisting of: -X 4 -CONR 5 R 6 , -X 4 -CONR 5 (CH 2 ) y1 CO 2 R 6 , -X 4 -CONR 5 (CH 2 ) y2 SO 3 R 6 ; wherein X 4 , R 5 , R 6 , y1, y2 are as defined above.
  • the compounds of the present invention have excellent FXR agonism and are useful in the preparation of pharmaceutical compositions for the treatment or prevention of FXR mediated diseases.
  • the present invention provides a pharmaceutical composition
  • a pharmaceutical composition comprising a therapeutically effective amount of a compound selected from the above formula (I) or an enantiomer or diastereomer thereof, or a pharmaceutically acceptable salt thereof Or one or more of the mixtures, and optionally one or more pharmaceutically acceptable carriers, excipients, adjuvants, adjuvants, and/or diluents.
  • the excipient is, for example, an odorant, a flavoring agent, a sweetener or the like.
  • the pharmaceutical composition provided by the present invention preferably contains the active ingredient in a weight ratio of 1-99%, preferably in a ratio of 65 wt% to 99 wt% of the total weight of the compound of the formula I as the active ingredient, and the remainder is pharmaceutically acceptable.
  • the compounds and pharmaceutical compositions provided by the present invention may be in various forms such as tablets, capsules, powders, syrups, solutions, suspensions, aerosols, and the like, and may be present in suitable solid or liquid carriers or diluents. Neutralizes a suitable sterilizing device for injection or drip.
  • compositions of the present invention can be prepared according to conventional methods of preparation in the pharmaceutical arts.
  • the formulation of the formulation comprises from 0.05 to 200 mg of the compound of the invention, preferably from 0.1 mg to 100 mg of the compound of the invention per unit dosage of the formulation.
  • the compounds and pharmaceutical compositions of the present invention can be used clinically in mammals, including humans and animals, by routes of administration to the mouth, nose, skin, lungs or the gastrointestinal tract. Most preferably oral. Most preferably, the daily dose is from 0.01 to 200 mg/kg body weight, taken once, or from 0.01 to 100 mg/kg body weight in divided doses. Regardless of the method of administration, the optimal dosage for the individual should be based on the particular treatment. Usually starting with a small dose, gradually increase the dose until the most suitable dose is found.
  • the present invention also provides a farnesoid X receptor (FXR) agonist comprising a compound selected from the above formula I or an enantiomer or diastereomer thereof, or a pharmaceutically acceptable drug thereof
  • FXR farnesoid X receptor
  • the compounds and compositions of the present invention are useful for the treatment and prevention of liver system and dyslipidemia associated with farnesoid X receptor (FXR), including, but not limited to, nonalcoholic steatohepatitis, liver fibrosis, cirrhosis , fatty liver, primary bile acid cirrhosis, hyperlipidemia and other diseases.
  • FXR farnesoid X receptor
  • the present invention also provides a compound represented by the above Formula I, or an enantiomer or diastereomer thereof, or a pharmaceutically acceptable salt thereof, or a mixture thereof, for use in the treatment of Fani Alcohol X receptor (FXR) related liver system and dyslipidemia, such as nonalcoholic steatohepatitis, liver fibrosis, cirrhosis, fatty liver, primary biliary cirrhosis, hyperlipidemia and other diseases the use of.
  • FXR Fani Alcohol X receptor
  • the invention also provides a treatment for liver system and dyslipidemia associated with farnesol X receptor (FXR), such as nonalcoholic steatohepatitis, liver fibrosis, cirrhosis, fatty liver, primary biliary cirrhosis
  • FXR farnesol X receptor
  • a method of a disease such as hyperlipemia which comprises administering to a patient in need of such treatment a compound selected from the above formula I or an enantiomer or diastereomer thereof, or a pharmaceutically acceptable salt thereof , or one or more of their mixtures.
  • the compounds of the present invention have excellent FXR agonism, and have good intracellular release and are safe.
  • 2,6-Dichloro-benzaldehyde oxime (100 g, 0.526 mol) was dissolved in DMF (N,N-dimethylformamide, 200 mL), and chlorosuccinimide (70.3 g, 0.526 mol) was dissolved in 100 mL DMF.
  • the above 2,6-dichloro-benzaldehyde oxime solution was added dropwise at 40 ° C, and then reacted at 40 ° C for 1 hour, and the reaction was monitored by thin layer chromatography (TLC). After the reaction was completed, it was cooled to room temperature, and a large amount of water was added.
  • Methyl 3-cyclopropyl-3-oxo-propionate (31.7 g, 0.223 mol) was dissolved in triethylamine (45.1 g, 62 mL, 0.446 mol), stirred at room temperature, and then cooled to 10 ° C after 30 min. Slowly add 2,6-dichloro-benzaldehyde chloro-hydrazine (50.0g, 0.223mol, dissolved in 300mL of ethanol) during the dropwise addition, the temperature does not exceed 24 ° C, after the completion of the dropwise addition, stirring at room temperature overnight, with a thin layer The reaction was monitored by (TLC).
  • Methyl 5-cyclopropyl-3-(2,6-dichloro-phenyl)-isoxazole-4-carboxylate (20.0 g, 64.1 mmol) was dissolved in tetrahydrofuran (100 mL).
  • Diisobutylaluminum hydride 1M/toluene (160.0 mL, 160.8 mmol) was added dropwise and stirred at room temperature for 2 hr. The reaction was monitored by thin layer chromatography (TLC). After the reaction, methanol (10 mL) and a small amount of water were added dropwise at 0 ° C and stirred for 10 minutes, then ethyl acetate (500 mL) was added.
  • DMSO dimethyl sulfoxide
  • reaction was monitored by thin layer chromatography (TLC). After the reaction, the mixture was cooled to room temperature celite and filtered, and water (100mL) and ethyl acetate (3x 100mL). The organic layer was washed with saturated brine (200 mL), dried over anhydrous Na.sub.2SO.sub.sub.sub. 4-(((5-Cyclopropyl-3-(2,6-dichlorophenyl)isoxazol-4-yl)methoxy)methyl)-4-fluoropiperidin-1-yl)benzoic acid Ethyl ester, yield 62%, ESI-MS m/z: 548 (M + ).
  • Methyl 3-cyclopropyl-3-oxo-propionate was exchanged for methyl 4-(2,2-difluorocyclopropane)-3-oxobutanoate and 2-bromobenzo[d]
  • the methyl thiazole-6-carboxylate was replaced with methyl 2-bromo-4-fluorobenzo[d]thiazole-6-carboxylate, and the remaining starting materials, reagents and preparation methods were the same as those in Example 14, to obtain the target product FL45.
  • Methyl 3-cyclopropyl-3-oxo-propionate was changed to methyl 4-(2,2-difluorocyclopropane)-3-oxobutanoate, (4-fluoro-1-tert Replacement of butoxycarbonyl substituted piperidin-4-yl)methanol with tert-butyl 3-fluoro-3-(hydroxymethyl)-8-azabicyclo[3.2.1]oct-8-carboxylate and 2-bromobenzene
  • [d] thiazole-6-formic acid methyl ester is replaced by 2-bromo-4-fluorobenzo[d]thiazole-6-carboxylic acid methyl ester, and the remaining required raw materials, reagents and preparation methods are the same as those in the example 14 to obtain the target product.
  • Methyl 3-cyclopropyl-3-oxo-propionate was changed to methyl 4-(2,2-difluorocyclopropane)-3-oxobutanoate and (4-fluoro-1-tert Replacement of butoxycarbonyl substituted piperidin-4-yl)methanol with tert-butyl 3-fluoro-3-(hydroxymethyl)-8-azabicyclo[3.2.1]oct-8-carboxylate, the remaining raw materials
  • the reagent and the preparation method were the same as those in Example 32 to obtain the target product FL49.
  • Methyl 3-cyclopropyl-3-oxo-propionate was changed to methyl 4-(2,2-difluorocyclopropane)-3-oxobutanoate, (4-fluoro-1-tert Replacement of butoxycarbonyl substituted piperidin-4-yl)methanol with tert-butyl 3-fluoro-3-(hydroxymethyl)-8-azabicyclo[3.2.1]oct-8-carboxylate and 4-iodo-
  • the ethyl benzoate was replaced with methyl 6-bromo-1-methyl-1H-indole-3-carboxylate, and the remaining starting materials, reagents and preparation methods were the same as those in Example 1, to obtain the target product FL50.
  • Methyl 3-cyclopropyl-3-oxo-propionate was exchanged for methyl 4-(2,2-difluorocyclopropane)-3-oxobutanoate and ethyl 4-iodo-benzoate
  • the product was replaced with 6-bromo-1-ethyl-1H-indole-3-carboxylic acid methyl ester, and the other desired starting materials, reagents and preparation methods were the same as those in Example 1, to obtain the target product FL51.
  • Methyl 3-cyclopropyl-3-oxo-propionate was changed to methyl 4-(2,2-difluorocyclopropane)-3-oxobutanoate, (4-fluoro-1-tert Replacement of butoxycarbonyl substituted piperidin-4-yl)methanol with tert-butyl 3-fluoro-3-(hydroxymethyl)-8-azabicyclo[3.2.1]oct-8-carboxylate and 4-iodo-
  • the ethyl benzoate was replaced with methyl 6-bromo-1-ethyl-1H-indole-3-carboxylate, and the remaining starting materials, reagents and preparation methods were the same as those in Example 1, to obtain the target product FL52.
  • Example 66 7-(3-(((5-((2,2-Difluorocyclopropyl)methyl)-3-(2-(trifluoromethoxy)phenyl)isoxazole-4- Preparation of methoxy)methyl)-3-fluoro-8-azabicyclo[3.2.1]oct-8-yl)imidazo[1,2-a]pyridine-3-carboxylic acid (FL66)
  • Example 58 Replace 6-bromo-1-methyl-1H-indole-3-carboxylic acid methyl ester with 7-bromoimidazo[1,2-a]pyridine-3-carboxylic acid methyl ester, the remaining raw materials, reagents and preparation The same procedure as in Example 58 was carried out to obtain the objective product FL66.
  • Test kit Fluorescent fluorescence reporter gene detection kit (Britelite plus model)
  • the FXR expression plasmid and the FXRE luciferase reporter plasmid were co-transfected into 293T cells, and the cells were cultured in 96-well flat-bottomed microplates to ensure expression of the plasmid, and FXR agonists were added.
  • the degree of agonist activation was calculated and quantified by measuring the luciferase fluorescent signal intensity.
  • the test compound and the positive compound OCA take 10 ⁇ M to act on the cells, and the relative activity of the test compound to the positive compound is determined, and the compound having a relative activity higher than 50% of the positive compound enters the re-screening, and the dose-dependent relationship is calculated. That is, the EC 50 value.
  • the sample was dissolved in DMSO, stored at low temperature, and the concentration of DMSO in the final system was controlled to a range that did not affect the activity of the assay.
  • the activity of the sample was tested by initial screening at a single concentration, for example 20 ⁇ g/ml.
  • the inhibition rate % inhibition is greater than 50
  • the test activity dose dependence ie, the IC 50 /EC 50 value
  • the calculation software is Graphpad Prism 4
  • the model used for fitting is sigmoidaldose-response (variable slope)
  • the bottom and top of the fitted curve are set to 0 and 100.
  • each sample is provided with a duplicate hole (n ⁇ 2) in the test, and the result is expressed by Standard Deviation (SD) or Standard Error (SE).
  • SD Standard Deviation
  • SE Standard Error
  • the reported compounds are used as a reference for each test.
  • a plurality of compounds of the present invention have excellent pharmacological activities by activity tests.
  • a plurality of compounds such as compounds FL3, FL10 and the like have an EC 50 of less than 10 nM at the cellular level, and are superior to the positive compounds OCA (Obecholic acid) and FL4-13, exhibiting stronger pharmacological activities.
  • the modeling method was as follows: SD male rats of 6-8 weeks old were intraperitoneally injected with 150 mg/kg TAA physiological saline solution three times a week for 4 weeks, and the compounds FL4-14 (FL10) and FL4-15 were started. FL11) (20 mg/kg PO qd), the same doses of OCA and FL4-13 (LY-2562175) were used as positive controls for four weeks, and TAA was continued to be administered during the administration.
  • the model can observe an increase in liver coefficient, elevated serum alkaline phosphatase (ALP) levels, up-regulation of ⁇ -smooth muscle actin ( ⁇ -SMA), type I collagen (Col1 ⁇ 1 mRNA) expression in liver and pathological sections ( Sirius red staining) increased liver collagen content.
  • ALP serum alkaline phosphatase
  • ⁇ -SMA ⁇ -smooth muscle actin
  • Col1 ⁇ 1 mRNA type I collagen
  • pathological sections Sirius red staining
  • FL4-14 in the compound significantly reduced serum ALP levels (Fig. 1) and down-regulated Col1 ⁇ 1 mRNA expression (Fig. 2).
  • Compound FL4-14 (FL10) reduced the amount of collagen in liver pathological sections and showed an anti-fibrosis effect comparable to positive OCA, but superior to the control compound FL4-13 (LY-2562175) (Fig. 3).
  • FL4-14 had no significant effect on serum cholesterol (TC), high-density ester protein (HDL) and low-density lipoprotein (LDL) levels, while positive drug OCA significantly increased serum TC, HDL and LDL levels.
  • TC serum cholesterol
  • HDL high-density ester protein
  • LDL low-density lipoprotein
  • FL10 The role of FL4-14 (FL10) may be able to avoid the side effects of clinically elevated OCL caused by OCL, and is also superior to the positive control compound FL4-13 ( Figure 4). In addition, there was no significant change in the liver coefficient of the FL4-14 (FL10) group, which also prevented the significant increase in the liver coefficient caused by the positive drug OCA (Fig. 5).
  • mice Female C57 mice, 6-8 weeks old, 6 rats, SPF grade.
  • mice Six female ICR mice were taken, and 12 hours after fasting, the rats were orally administered with FL4-14 500 mg/kg, the dosage volume was 10 mL/kg, and the solvent was 10% DMSO + 10% Tween 80 + 80%. 0.5% MC. The toxicity of the mice was observed immediately after administration, and the observation was continued for 7 days.
  • the preliminary acute toxicity test is mainly to observe whether the compound dose of 500mg/kg has obvious poisoning reaction or death.
  • the experimental results show that the compound of the present application has good safety.

Abstract

一类含氟异噁唑类化合物及其制备方法、药物组合物和用途。具体地，公开了通式I所示的化合物或其对映异构体或非对映异构体，或其可药用盐，或它们的混合物。以及公开了上述化合物为靶向FXR的小分子激动剂，可用于治疗FXR介导的疾病。

Description

一类含氟异噁唑类化合物及其制备方法、药物组合物和用途 技术领域
本发明涉及药物化学和药物治疗学领域，具体涉及一类含氟异恶唑类化合物、其制备方法、含此类化合物的药物组合物及作为FXR激动剂，特别是制备用于治疗非酒精性脂肪性肝病、肝纤维化的药物的用途。
背景技术
非酒精性脂肪性肝病(NAFLD)是指除酒精和其他明确的损肝因素外所致的，以肝细胞内脂肪过度沉积为主要特征的临床病理综合征，是一种获得性代谢应激性肝损伤，与胰岛素抵抗和遗传易感性密切相关。发达国家在有风险因素人群中NAFLD患病率每年约增加2％，且起病渐趋低龄化，已成为西方国家“第一大肝病”，在我国也是常见的慢性肝病类型。NAFLD除可直接导致失代偿期肝硬化、肝细胞癌和移植肝复发外，还可影响其他慢性肝病的进展，并参与2型糖尿病和动脉粥样硬化的发病，被认为是全身代谢综合征的肝脏局部表现。
NAFLD主要包括单纯性脂肪肝(SFL)和非酒精性脂肪性肝炎(NASH)，其中NASH是最主要的一种类型，NASH病人在10年内肝硬化发生率高达25％。但遗憾的是有关NASH药物研究和治疗进展缓慢，过去NASH被认为是糖尿病或肥胖的一种良性并发症，这种错误的观念导致人们忽视了对其及时的诊断和治疗。目前，NASH治疗药物的研发主要包括：(1)选择性外周大麻素受体(CB)阻制剂；(2)过氧化物酶体增殖物激活受体(PPAR)激动剂；(3)半胱天冬酶(Caspase)抑制剂；(4)磷酸二脂酶4(PDE4)抑制剂；(5)法尼酯衍生物X受体(FXR)激动剂；(6)趋化因子受体-5/趋化因子受体-2(CCR5/CCR2)双靶点抑制剂；(7)凋亡信号调节激酶-1(ASK1)抑制剂等。在这些靶点和其药物研发中，人们发现法尼酯衍生物X受体(FXR)作为胆汁酸感受器，不但控制着体内胆汁酸稳态，而且对脂类及糖代谢也具有重要的调控作用，是调控代谢综合征患者代谢紊乱的重要靶点。
FXR受体是一种胆汁酸受体，FXR的激活控制着胆汁酸的合成、转运等过程。FXR是激素核受体超家族的一员，具有典型的核受体结构，目前发现存在FXRα1、FXRα2、FXRα3和FXRα4共4个亚型。FXR配体主要通过激活FXR及FXR靶基因小异源二聚体伴侣受体(SHP)，抑制肝星状细胞(HSC)表达，减少细胞外基质(ECM)产生并增加ECM清除，从而减少ECM沉积；FXR配体还可增加HSC对凋亡的敏感性，使HSC凋亡增多，并与其他逆转肝纤维化的核因子相互作用逆转肝纤维化，从而发挥治疗肝纤维化作用。
近年来，有多个FXR激动剂相继进入了临床研究，如Novatis发现的化合物LJN-452具有良好的生物活性和选择性，其对FXR的EC50达到0.2nM，目前已经进入II期临床试验用于NASH和原发性胆汁性胆管炎(PBC)的治疗；由Gilead公司开发的FXR激动剂PX-104曾进入II期临床研究用于NASH的治疗，但遗憾的是未达到预期临床终点被迫终止。其他几个化合物，如LY-2562175和INT-767也进入了临床研究，有望缓解NASH尚未满足的巨大临床需求。其中，奥贝胆酸(OCA)作为第一个FXR激动剂被批准上市，用于治疗PBC和NASH的新型候选药物，主要针对熊去氧胆酸没有充分应答或不能耐受的患者。其机制主要是通过激动FXR受体，调节相关基因，影响胆汁酸的合成、分泌、转运和吸收。2015年1月，FDA授予OCA治疗伴有肝纤维化的NASH 的突破性药物资格、治疗PBC的快车道地位、治疗PBC和PSC的孤儿药地位，并且已于2016年5月27日经美国FDA批准上市。全球医药市场预测机构EvaluatePharma发布《全球生物制药后期管线重磅药物TOP 15》榜单，预测OCA在2020年的全球销售额将高达30亿美元。然而，尽管奥贝胆酸在减少肝脏纤维化方面发挥了积极作用，但是它却会提升低密度脂蛋白胆固醇(LDL)和降低高密度脂蛋白胆固醇(HDL)，预示该药物可能存在增加患心血管病的风险，同时还会引起患者身体的瘙痒等副作用，这在一定程度上限制了其广泛使用。
因此，面对NASH尚未满足的临床需求，迫切需要针对NASH新型靶点开发创新的NASH治疗药物，填补市场空缺，满足临床用药需求。
发明内容
本发明的目的是提供一类新颖的靶向FXR的小分子激动剂及其制备方法和应用。
本发明第一方面提供了一种具有如下通式I所示结构的含氟异恶唑类化合物或其对映异构体或非对映异构体，或其可药用盐，或它们的混合物：
Figure PCTCN2019086068-appb-000001
其中：
m为0、1、2或3；
n为0、1或2；
p为0、1或2；
Figure PCTCN2019086068-appb-000002
环选自下组：取代或未取代的6-20元杂环基、取代或未取代的6-20元芳环基、取代或未取代的6-20元芳杂环基；其中，所述的取代指基团上的氢原子被1、2、3或4个选自下组的取代基取代：氘(D)、氚(T)、卤素、C 1-C 6烷基、卤代C 1-C 6烷基、C 1-C 6烷氧基、卤代C 1-C 6烷氧基、-O[(CH 2) qO] rR 5、-O(CH 2) sC 6-C 10芳基、C 3-C 8环烷氧基、卤代C 3-C 8环烷氧基、氰基、硝基、氨基、胺基(较佳地C 1-C 6胺基)、羟基、羧基、C 1-C 6酯基、C 6-C 10芳基、C 6-C 10芳基氧基、-X 4-CO 2R 5、C 1-C 6烷基羟基、-X 4-CONR 5R 6、-X 4-CONR 5(CH 2) y1CO 2R 6、-X 4-CONR 5(CH 2) y2SO 3R 6、3-12元杂环基和3-12元杂环基氧基；其中，所述的q、r、s、y1和y2可分别为1、2、3或4；所述的芳杂环基、芳环基或杂环基各自独立地含有1～4个选自氧、硫和氮中的杂原子；X 4为键、C1-C6亚烷基、C 1-2烯烃基、环丙基或环氧乙基；
R 1选自下组：取代或未取代的6-20元杂环基、取代或未取代的6-20元芳环基、取代或未取代的6-20元芳杂环基；其中，所述的取代指基团上的氢原子被1、2、3或4个选自下组的取代基取代：氘、氚、卤素、C 1-C 6烷基、C 1-C 6烷氧基、卤代C 1-C 6烷氧基、-O[(CH 2) qO] rR 5、-O(CH 2) sC 6-C 10芳基、C 3-C 8环烷氧基、卤代C 3-C 8环烷氧基、氰基、硝基、氨基、胺基(较佳地C 1-C 6胺基)、羟基、羟甲基、羧基、C 6-C 10芳基、C 6-C 10芳基氧基、3-12元杂环基和3-12元杂环基氧基；其中，所述的q、r和s可分别为1、2、3或4；所述的芳杂环基、芳基或杂环基各自独立地含有1～4个选自氧、硫和氮中的杂原子；
R 2可独立地选自1、2、3或4个下组的取代基：氢、卤素、取代或未取代的C 1-C 6烷基、取代或未取代的C 1-C 6烷氧基、-O[(CH 2) qO] rR 5、-O(CH 2) sC 6-C 10芳基、C 3-C 8环烷氧基、卤代C 3-C 8环烷氧基、氰基、硝基、氨基、胺基(较佳地C 1-C 6胺基)、羟基、羟甲基、羧基；其中，所述的取代指基团上的氢原子被1、2、3或4个选自下组的取代基取代：氢、卤素、硝基、氰基、三氟甲基、三氟乙基、三氟丙基、三氟甲氧基、三氟乙氧基、氨基、胺基(较佳地C 1-C 6胺基)、羟基、羟甲基、羧基、巯基和磺酰基；
R 3和R 4各自独立地选自下组：氢、卤素、C 1-C 6烷基、卤代C 1-C 6烷基、C 1-C 6烷氧基、卤代C 1-C 6烷氧基、-O[(CH 2) qO] rR 5、-O(CH 2) sC 6-C 10芳基、C 3-C 8环烷氧基、卤代C 3-C 8环烷氧基、氰基、硝基、氨基、胺基(较佳地C 1-C 6胺基)、羟基、羟甲基和羧基；或者R 3和R 4连接在一起共同形成C 1-C 6亚烷基；
R 5和R 6为自独立地氢、C 1-C 6烷基、卤代C 1-C 6烷基、C 2-C 6烯基、羟甲基或5-7元杂环基。
在另一优选例中，m为0、1或2；优选为0或1。
在另一优选例中，n为0或1。
在另一优选例中，p为0或1。
在另一优选例中，
Figure PCTCN2019086068-appb-000003
环选自下组：苯基、吡啶基、嘧啶基、哒嗪基、噻唑基、苯并噻唑基、苯并[d]异噻唑基、咪唑并[1,2-a]吡啶基、喹啉基、1H-吲哚基、吡咯并[1,2-b]哒嗪基、苯并呋喃基、苯并[b]苯硫基、1H-吲唑基、苯并[d]异恶唑基、噻唑啉基、1H-吡咯并[3,2-c]吡啶基、吡唑并[1,5-a]嘧啶基、咪唑[1,2-b]哒嗪基、吡唑并[1,5-a]吡咯基；上述基团为取代或未取代的；其中，所述的取代指基团上的氢原子被1、2、3或4个选自下组的取代基取代：D、T、卤素、C 1-C 6烷基、卤代C 1-C 6烷基、C 1-C 6烷氧基、卤代C 1-C 6烷氧基、-O[(CH 2) qO] rR 5、-O(CH 2) sC 6-C 10芳基、C 3-C 8环烷氧基、卤代C 3-C 8环烷氧基、氰基、硝基、氨基、胺基(较佳地C 1-C 6胺基)、羟基、羧基、C 1-C 6酯基、C 6-C 10芳基、C 6-C 10芳基氧基、-X 4-CO 2R 5、C 1-C 6烷基羟基、-X 4-CONR 5R 6、-X 4-CONR 5(CH 2) y1CO 2R 6、-X 4-CONR 5(CH 2) y2SO 3R 6、3-12元杂环基和3-12元杂环基氧基；
其中，所述的q、r、s、y1和y2可分别为1、2、3或4；
芳杂环、芳基或杂环基各自独立地含有1～4个选自氧、硫和氮中的杂原子；
X 4为键、C1-C6亚烷基、C 1-2烯烃基、环丙基、环氧乙基；
R 5和R 6各自独立地为氢、C 1-C 6烷基、卤代C 1-C 6烷基、C 2-C 6烯基、羟甲基或5-7元杂环基。
在另一优选例中，
Figure PCTCN2019086068-appb-000004
选自下组：
Figure PCTCN2019086068-appb-000005
其中，
R 8选自下组：氢、卤素、C 1-C 6烷基、卤代C 1-C 6烷基、C 1-C 6烷氧基、C 1-C 6烷氧基羰基、卤代C 1-C 6烷氧基、C 2-C 6烯基、C 2-C 6炔基、C 3-C 8环烷基、氰基、硝基、氨基、羟基、羟甲基、羧基、巯基、磺酰基、C 6-C 10芳基和3-12元杂环基；
t为0、1、2、3、4或5；
X l和X 2各自独立地为N、CH或CR 8；
X 3为O或S；
R 9为-X 4-CO 2R 5、C 1-C 6烷基、卤代C 1-C 6烷基、C 1-C 6羟基、-X 4-CONR 5R 6、-X 4-CONR 5(CH 2) y1CO 2R 6、-X 4-CONR 5(CH 2) y2SO 3R 6；y1和y2各自独立地为1、2、3或4；X 4为键、C1-C6亚烷基、C 1-2烯烃基、环丙基或环氧乙基；
R 5和R 6各自独立地为氢、C 1-6烷基、卤代C 1-C 6烷基、C 2-C 6烯基、羟甲基或5-7元杂环基；
R 10为氢、C 1-C 6烷基羟基，C 1-C 6烷基、卤代C 1-C 6烷基、C 1-C 6烷氧基、C 1-C 6烷氧基羰基、卤代C 1-C 6烷氧基、C 2-C 6烯基、C 2-C 6炔基或C 3-C 8环烷基。
在另一优选例中，
Figure PCTCN2019086068-appb-000006
选自下组：
Figure PCTCN2019086068-appb-000007
其中，R 8、R 9、R 10和t定义同前。
在另一优选例中，R 1选自下组：苯基、吡啶基、1-氧吡啶基、环己基、环戊基、环丙基；上述基团为取代或未取代的；其中，所述的取代指基团上的氢原子被1、2、3或4个选自下组的取代基取代：氢、卤素、硝基、氰基、三氟甲基、三氟乙基、三氟丙基、三氟甲氧基、三氟乙氧基、苯基、甲氧基羰基、乙氧基羰基、丙氧基羰基、C 1-6烷基、C l-6环烷基、C 1-6烷氧基、C l-6环烷氧基、环丙基。
在另一优选例中，所述化合物选自下组：
Figure PCTCN2019086068-appb-000008
Figure PCTCN2019086068-appb-000009
Figure PCTCN2019086068-appb-000010
Figure PCTCN2019086068-appb-000011
本发明第二方面提供了一种药物组合物，其包含治疗有效量的第一方面所述的含氟异恶唑类化合物或其对映异构体或非对映异构体，或其可药用盐，或它们的混合物，以及药学上可接受的载体。
本发明第三方面提供了一种FXR激动剂，其包含第一方面所述的含氟异恶唑类化合物或其对映异构体或非对映异构体，或其可药用盐，或它们的混合物。
本发明第四方面提供了第一方面所述的含氟异恶唑类化合物或其对映异构体或非对映异构体，或其可药用盐，或它们的混合物的用途，用于制备预防或治疗FXR介导的的疾病的药物。
在另一优选例中，所述疾病选自下组：非酒精性脂肪肝病、非酒精性脂肪性肝炎、 脂肪肝、肝纤维化、原发性胆汁性肝硬化、高血脂症、血脂异常。
本发明第五方面提供了第一方面所述的含氟异恶唑类化合物的制备方法，
(a)所述方法包括步骤：
Figure PCTCN2019086068-appb-000012
其中，
Figure PCTCN2019086068-appb-000013
R 1、R 2、R 3、R 4、n、m和p的定义与前述的通式I中的定义相同；X为卤素或硝基；
步骤j：在有机溶剂中，在18-冠醚-6和碱存在下，中间体12和中间体9进行反应，形成中间体13；
步骤k：在有机溶剂中，将中间体13进行脱保护反应，形成中间体14；
步骤l：在氮气保护下，在有机溶剂中，在碱、钯催化剂和配体的存在下，中间体14和中间体15反应，形成终产物I；或
步骤m：在氮气保护下，在有机溶剂中，在碱存在下，中间体14和中间体15反应，形成终产物I；
或
(b)所述方法包括步骤：
Figure PCTCN2019086068-appb-000014
其中，R 1、R 2、R 3、R 4、n、m和p的定义与前述的通式I中的定义相同；X为卤素或硝基；
步骤j：在有机溶剂中，在18-冠醚-6和碱存在下，中间体12和中间体9进行反应，形成中间体13；
步骤k：在有机溶剂中，将中间体13进行脱保护反应，形成中间体14；
步骤l：在氮气保护下，在有机溶剂中，在碱、钯催化剂和配体的存在下，中间体14和中间体15反应，形成中间体I’；或
步骤m：在氮气保护下，在有机溶剂中，在碱存在下，中间体14和中间体15反应，形成中间体I’；和
步骤e：在有机溶剂中，中间体I’和HNR 5R 6、HNR 5(CH 2) y1CO 2R 6、或HNR 5(CH 2) y2SO 3R 6进行反应，形成终产物I；
其中，
Figure PCTCN2019086068-appb-000015
环选自下组：取代或未取代的6-20元杂环基、取代或未取代的6-20元芳环基、取代或未取代的6-20元芳杂环基；其中，所述的取代指基团上的氢原子被1、2、3或4个选自下组的取代基取代：-X 4-CO 2R 5；其中，X 4、R 5定义同前
Figure PCTCN2019086068-appb-000016
环选自下组：取代或未取代的6-20元杂环基、取代或未取代的6-20元芳环基、取代或未取代的6-20元芳杂环基；其中，所述的取代指基团上的氢原子被1、2、3或4个选自下组的取代基取代：-X 4-CONR 5R 6、-X 4-CONR 5(CH 2) y1CO 2R 6、-X 4-CONR 5(CH 2) y2SO 3R 6；其中，X 4、R 5、R 6、y1、y2定义同前。
本发明提供了一种治疗方法，给需要的对象施用本发明所述的含氟异恶唑类化合物或其对映异构体或非对映异构体，或其可药用盐，或它们的混合物或本发明的药物组合物或FXR激动剂。
应理解，在本发明范围内中，本发明的上述各技术特征和在下文(如实施例)中具体描述的各技术特征之间都可以互相组合，从而构成新的或优选的技术方案。限于篇幅，在此不再一一累述。
附图说明
图1显示了本发明化合物对血清ALP的影响。
图2显示了本发明化合物对α-SMA和Col1α1mRNA的影响。
图3显示了本发明化合物对胶原含量的影响。
图4显示了本发明化合物对血清内TC、TG、HDL、LDL浓度的影响。
图5显示了本发明化合物对肝脏系数的影响。
具体实施方式
本发明人经过长期而深入的研究，设计制备了一类结构新颖的含氟异恶唑类化合物。所述的化合物可以有效地靶向地激动FXR。基于上述发现，发明人完成了本发明。
术语
在本发明中，除非特别指出，所用术语具有本领域技术人员公知的一般含义。
在本发明中，术语“6-20元杂环基”是指具有6-20个环原子的含有一个或多个选自氮、和硫的杂原子的饱和或部分饱和杂环基；包括但不限于：吗啉基、哌啶基、哌嗪基、环己基并哌啶基等。其他“杂环基”术语具有类似的含义。
在本发明中，术语“6-20元杂环氧基”是指6-20元杂环基-氧-。其他“杂环氧基”术语具有类似的含义。
在本发明中，术语“6-20元芳环基”是指具有6-20个环原子的芳环基；包括但不限于：苯基、萘基、菲基、蒽基等。其他“芳环基”术语具有类似的含义。
在本发明中，术语“6-20元芳杂环基”是指具有6-20个环原子的含有一个或多个选自氮、和硫的杂原子的芳杂环基；包括但不限于：四唑基、吡啶基、嘧啶基、哒嗪基、噻 唑基、苯并噻唑基、苯并[d]异噻唑基、咪唑并[1,2-a]吡啶基、喹啉基、1H-吲哚基、吡咯并[1,2-b]哒嗪基、苯并呋喃基、苯并[b]苯硫基、1H-吲唑基、苯并[d]异恶唑基、噻唑啉基、1H-吡咯并[3,2-c]吡啶基、吡唑并[1,5-a]嘧啶基、咪唑[1,2-b]哒嗪基、吡唑并[1,5-a]吡咯基等。其他“芳杂环基”术语具有类似的含义。
在本发明中，术语“C 1-C 6烷基”是指具有1至6个碳原子的直链或支链烷基，非限制性地包括甲基、乙基、丙基、异丙基、丁基、异丁基、仲丁基、叔丁基、戊基和已基等；优选乙基、丙基、异丙基、丁基、异丁基、仲丁基和叔丁基。
在本发明中，术语“C 1-C 6烷氧基”是指具有1至6个碳原子的直链或支链烷氧基，非限制性地包括甲氧基、乙氧基、丙氧基、异丙氧基和丁氧基等。
在本发明中，术语“C 2-C 6烯基”是指具有2至6个碳原子的含有一个双键的直链或支链烯基，非限制性地包括乙烯基、丙烯基、丁烯基、异丁烯基、戊烯基和己烯基等。
在本发明中，术语“C 2-C 6炔基”是指具有2至6个碳原子的含有一个三键的直链或支链炔基，非限制性地包括乙炔基、丙炔基、丁炔基、异丁炔基、戊炔基和己炔基等。
在本发明中，术语“C 3-C 8环烷基”是指在环上具有3至8个碳原子的环状烷基，非限制性地包括环丙基、环丁基、环戊基、环己基、环庚基、环辛基和环癸基等。其他“环烷基”术语具有类似的含义。
在本发明中，术语“C 3-C 8环烷氧基”是指在环上具有3至8个碳原子的环状烷氧基，非限制性地包括环丙氧基、环丁氧基、环戊氧基、环己氧基、环庚氧基、环辛氧基和环癸氧基等。其他“环烷氧基”术语具有类似的含义。
在本发明中，术语“C 6-C 10芳基”是指在环上不含杂原子的具有6至10个碳原子的芳香族环基，如苯基、萘基等。术语“C 6-C 12芳基”具有类似的含义。
在本发明中，术语“C 1-C 6酯基”是指具有1-6个碳原子且具有酯基(-O(C＝O)-)的基团。
在本发明中，术语“C 1-C 6胺基”是指具有1-6个碳原子且具有氨基作为端基的基团。
在本发明中，所述卤素为F、Cl、Br或I。卤代表示被卤素取代的。
活性成分
本发明的化合物为通式I所示的化合物或其对映异构体或非对映异构体，或其可药用盐，或它们的混合物。
本发明化合物优选为如下具体化合物：
Figure PCTCN2019086068-appb-000017
Figure PCTCN2019086068-appb-000018
Figure PCTCN2019086068-appb-000019
Figure PCTCN2019086068-appb-000020
Figure PCTCN2019086068-appb-000021
Figure PCTCN2019086068-appb-000022
Figure PCTCN2019086068-appb-000023
Figure PCTCN2019086068-appb-000024
Figure PCTCN2019086068-appb-000025
Figure PCTCN2019086068-appb-000026
Figure PCTCN2019086068-appb-000027
本发明化合物还包含通式I化合物的可药用的盐，具体地为通式I化合物与无机酸或有机酸反应形成常规的可药用盐。例如，常规的可药用盐可通过通式I化合物与无机酸或有机酸反应制得，所述无机酸包括盐酸、氢溴酸、硫酸、硝酸、胺基磺酸和磷酸等，以及所述有机酸包括柠檬酸、酒石酸、乳酸、丙酮酸、乙酸、苯磺酸、对甲苯磺酸、甲磺酸、萘磺酸、乙磺酸、萘二磺酸、马来酸、苹果酸、丙二酸、富马酸、琥珀酸、丙酸、草酸、三氟乙酸、硬酯酸、扑酸、羟基马来酸、苯乙酸、苯甲酸、水杨酸、谷氨酸、抗坏血酸、对胺基苯磺酸、2-乙酰氧基苯甲酸和羟乙磺酸等；或者通式I化合物与无机碱形成的钠盐、钾盐、钙盐、铝盐或铵盐；或者通式I化合物与有机碱形成的甲胺盐、乙胺盐或乙醇胺盐。
本发明的化合物还包括通式I化合物的对映异构体(例如R-异构体或S-异构体)或非对映异构体，或其外消旋体或内消旋体。
制备方法
本发明的通式I化合物可按照如下方案进行制备。
制备通式中间体3方案如下
Figure PCTCN2019086068-appb-000028
其中，R 2、p的定义与前述的通式I中的定义相同；
步骤a：将1溶于有机溶剂中，0℃下滴加氧化剂，在0℃搅拌反应，反应完后乙醚提取有机层，然后用氢氧化钠溶液提取合并水层，水层用硫酸酸化至PH<1。然后乙醚提取有机层，饱和食盐水洗，干燥，旋干得到中间体2。所述有机溶剂为丙酮、四氢呋喃、乙醚、二甲基甲酰胺、乙二醇二甲醚、乙二醇二乙醚、二氧六环、乙醇、甲醇、 乙酸乙酯、二氯甲烷或其混合物；所述氧化剂选自PCC、PDC、戴斯-马丁氧化剂、Swern氧化剂、H 2O 2、高锰酸钾和二氧化锰中；
步骤b：将中间体2溶入有机溶剂中，加入N,N'-羰基二咪唑，氮气保护室温反应一小时后，加入丙二酸单乙酯钾盐和氯化镁，室温反应过夜，反应完后盐酸淬灭，乙酸乙酯提取有机层，然后有机层用饱和食盐水洗，干燥，旋干，过柱子得到中间体3。所述有机溶剂为丙酮、四氢呋喃、乙醚、二甲基甲酰胺、乙二醇二甲醚、乙二醇二乙醚、二氧六环、乙醇、甲醇、乙酸乙酯、二氯甲烷或其混合物；
制备通式中间体9方案如下
Figure PCTCN2019086068-appb-000029
其中，R 1、R 2和p的定义与前述的通式I中的定义相同；
步骤c：将盐酸羟胺溶解于水中，在0℃搅拌下滴加氢氧化钠。4加入有机溶剂溶解后滴入上述混合溶液，然后升温至90℃搅拌过夜。反应完后冷却至室温，真空浓缩至大量固体析出，抽滤得固体，干燥，中间体5；所述有机溶剂为四氢呋喃、乙醚、二甲基甲酰胺、乙二醇二甲醚、乙二醇二乙醚、二氧六环、乙醇、甲醇、乙酸乙酯、二氯甲烷或其混合物。
步骤d：中间体5加入有机溶剂溶解，将氯代丁二酰亚胺溶于有机溶剂在40℃下滴入上述5溶液中，后在40℃下反应，，反应完后冷却至室温，加入大量水、乙酸乙酯(EA)萃取，用水洗涤有机层三次，饱和食盐水洗涤有机层，无水Na 2SO 4干燥，旋干有机层，得中间体6；所述有机溶剂为四氢呋喃、乙醚、二甲基甲酰胺、乙二醇二甲醚、乙二醇二乙醚、二氧六环、乙醇、甲醇、乙酸乙酯、二氯甲烷或其混合物；
步骤e：将中间体3加入碱中，在室温搅拌后，降温至10℃，下缓慢滴加中间体6反应完后得中间体7；所述有机溶剂为四氢呋喃、乙醚、二甲基甲酰胺、乙二醇二甲醚、乙二醇二乙醚、二氧六环、乙醇、甲醇或乙酸乙酯、二氯甲烷；所述碱为醋酸钠、NaOH、KOH、乙醇钠、甲醇钠、碳酸钠、碳酸钾、三乙胺或二异丙胺。
步骤f：将中间体7溶入有机溶剂中，在0℃氮气保护下，滴加二异丁基氢化铝1M/甲苯,室温搅拌反应，反应完后,在0℃下滴加甲醇和少量水淬灭并搅拌10分钟，后经硅藻土抽滤、有机溶剂洗涤滤出沉淀物，滤液真空浓缩、干燥得中间体8；所述有机溶剂为四氢呋喃、乙醚、二甲基甲酰胺、乙二醇二甲醚、乙二醇二乙醚、二氧六环、乙醇、甲醇、乙酸乙酯、二氯甲烷或其混合物；
步骤g：将中间体8溶入有机溶剂中，加入三苯基磷,冷却至0℃，加入四溴化碳，后在室温反应，反应完后，将反应液真空浓缩得橙色油状物，过硅胶柱，旋干得中间体9。所述有机溶剂为四氢呋喃、乙醚、二甲基甲酰胺、乙二醇二甲醚、乙二醇二乙醚、二氧六环、乙醇、甲醇、乙酸乙酯、二氯甲烷或其混合物；
制备通式中间体12方案如下
Figure PCTCN2019086068-appb-000030
其中，m、R 4和R 5的定义与前述的通式I中的定义相同；
步骤h：将三甲基碘化硫加溶于有机溶剂中，搅拌下加入化合物10继续搅拌，然后再加入叔丁醇钾；得环氧化合物2。所述有机溶剂可为二甲亚砜、四氢呋喃、乙醚、二甲基甲酰胺、乙二醇二甲醚、乙二醇二乙醚、二氧六环、乙醇、甲醇、乙酸乙酯、二氯甲烷或其混合物。
步骤i：将中间体11溶入有机溶剂中，降温至－10℃～－40℃，加入1-10当量的氟化氢吡啶溶液，继续反应至原料消失，分离纯化得中间体12；所述有机溶剂可为四氢呋喃、乙醚、二甲基甲酰胺、乙二醇二甲醚、乙二醇二乙醚、二氧六环、乙醇、甲醇、乙酸乙酯、二氯甲烷或其混合物。
合成通式I方案如下：
Figure PCTCN2019086068-appb-000031
其中，
Figure PCTCN2019086068-appb-000032
R 1、R 2、R 3、R 4、n、m和p的定义与前述通式I中的定义相同；X为卤素或硝基；
步骤j：将中间体12溶入有机溶剂中，在0℃下，加入化合物18-冠醚-6(12.8g,48.6mmol)和强碱，反应30分钟，移至室温，滴加中间体9(溶于有机溶剂)，在室温搅拌反应，反应完后分离纯化得中间体13；所述有机溶剂为四氢呋喃、乙醚、二甲基甲酰胺、乙二醇二甲醚、乙二醇二乙醚、二氧六环、乙醇、甲醇、乙酸乙酯、二氯甲烷或其混合物；所述强碱为NaOH、KOH、叔丁醇钾、乙醇钠或甲醇钠；
步骤k：将中间体13溶入有机溶剂中，加入三氟醋酸(TFA)或者盐酸(HCl)的有机溶剂，脱去Boc保护基，得中间体14；所述有机溶剂可为四氢呋喃、乙醚、二甲基甲酰胺、乙二醇二甲醚、乙二醇二乙醚、二氧六环、乙醇、甲醇、乙酸乙酯、二氯甲烷或其混合物。
步骤l：在氮气保护下，将中间体14、15溶入有机溶剂中，加入碳酸铯,三(二亚苄基丙酮)二钯催化剂和2-二环己基磷-2,4,6-三异丙基联苯配体，反应完后，分离纯化得终产物I；所述有机溶剂为甲苯、四氢呋喃、乙醚、二甲基甲酰胺、乙二醇二甲醚、乙二醇二乙醚、二氧六环、乙醇、甲醇、乙酸乙酯、二氯甲烷或其混合物；
或步骤m：在氮气保护下，将中间体14、15溶入有机溶剂中，加入N,N-二异丙基乙胺，反应完后，分离纯化得终产物I；所述有机溶剂为甲苯、四氢呋喃、乙醚、二甲基甲酰胺、乙二醇二甲醚、乙二醇二乙醚、二氧六环、乙醇、甲醇、乙酸乙酯、氮甲基 吡咯烷酮、二氯甲烷或其混合物。
或者合成通式I的方法如下：
Figure PCTCN2019086068-appb-000033
其中，R 1、R 2、R 3、R 4、n、m和p的定义与同前；X为卤素或硝基；
步骤j：在有机溶剂中，在18-冠醚-6和碱存在下，中间体12和中间体9进行反应，形成中间体13；
步骤k：在有机溶剂中，将中间体13进行脱保护反应，形成中间体14；
步骤l：在氮气保护下，在有机溶剂中，在碱、钯催化剂和配体的存在下，中间体14和中间体15反应，形成中间体I’；或
步骤m：在氮气保护下，在有机溶剂中，在碱存在下，中间体14和中间体15反应，形成中间体I’；和
步骤e：在有机溶剂中，中间体I’和HNR 5R 6、HNR 5(CH 2) y1CO 2R 6、或HNR 5(CH 2) y2SO 3R 6进行反应，形成终产物I；
其中，
Figure PCTCN2019086068-appb-000034
环选自下组：取代或未取代的6-20元杂环基、取代或未取代的6-20元芳环基、取代或未取代的6-20元芳杂环基；其中，所述的取代指基团上的氢原子被1、2、3或4个选自下组的取代基取代：-X 4-CO 2R 5；其中，X 4、R 5定义同前
Figure PCTCN2019086068-appb-000035
环选自下组：取代或未取代的6-20元杂环基、取代或未取代的6-20元芳环基、取代或未取代的6-20元芳杂环基；其中，所述的取代指基团上的氢原子被1、2、3或4个选自下组的取代基取代：-X 4-CONR 5R 6、-X 4-CONR 5(CH 2) y1CO 2R 6、-X 4-CONR 5(CH 2) y2SO 3R 6；其中，X 4、R 5、R 6、y1、y2定义同前。
药物组合物和施用方法
本发明化合物具有优异的FXR激动作用，可用于制备用于治疗或预防FXR介导的疾病的药物组合物。
本发明提供了一种药物组合物，其含有治疗有效量的选自上述通式(I)所示的化合物或其对映异构体或非对映异构体，或其可药用的盐，或它们的混合物中的一种或多种，以及任选地，一种或多种可药用的载体、赋形剂、佐剂、辅料和/或稀释剂。所述辅料例如为气味剂、香味剂、甜味剂等。
本发明所提供的药物组合物优选含有重量比为1-99％的活性成份，其优选的比例是，通式I化合物作为活性成分占总重量的65wt％～99wt％，其余部分为药学可接受的载体、稀释液或溶液或盐溶液。
本发明所提供的化合物和药物组合物可以是多种形式，如片剂、胶囊、粉剂、糖浆、溶液状、悬浮液和气雾剂等，并可以存在于适宜的固体或液体的载体或稀释液中和适宜的用于注射或滴注的消毒器具中。
本发明的药物组合物的各种剂型可按照药学领域的常规制备方法制备。其制剂配方的单位计量中包含0.05-200mg本发明化合物，优选地，制剂配方的单位计量中包含0.1mg-100mg本发明化合物。
本发明化合物和药物组合物可对哺乳动物临床使用，包括人和动物，可以通过口、鼻、皮肤、肺或者胃肠道等的给药途径。最优选为口服。最优选日剂量为0.01-200mg/kg体重，一次性服用，或0.01-100mg/kg体重分次服用。不管用何种服用方法，个人的最佳剂量应依据具体的治疗而定。通常情况下是从小剂量开始，逐渐增加剂量一直到找到最适合的剂量。
本发明还提供了一种法尼醇X受体(FXR)激动剂，其包含选自上述通式I所示的化合物或其对映异构体或非对映异构体，或其可药用的盐，或它们的混合物中的一种或多种，以及任选地一种或多种可药用的载体、赋形剂、佐剂、辅料和/或稀释剂。
本发明化合物和组合物用于治疗和预防与法尼醇X受体(FXR)相关的肝脏系统和血脂疾病，所述疾病包括，但不限于，非酒精性脂肪肝炎、肝纤维化、肝硬化、脂肪肝、原发性胆汁酸肝硬化、高血脂症等疾病。
因此，本发明还提供了上述通式I所示的化合物或其对映异构体或非对映异构体，或其可药用的盐，或它们的混合物在制备用于治疗与法尼醇X受体(FXR)相关的相关的肝脏系统和血脂疾病，例如非酒精性脂肪肝炎、肝纤维化、肝硬化、脂肪肝、原发性胆汁酸肝硬化、高血脂症等疾病的药物中的用途。
本发明还提供了一种治疗与法尼醇X受体(FXR)相关的肝脏系统和血脂疾病，例如非酒精性脂肪肝炎、肝纤维化、肝硬化、脂肪肝、原发性胆汁酸肝硬化、高血脂症等疾病的方法，其包括向需要该治疗的患者施用选自上述通式I所示的化合物或其对映异构体或非对映异构体，或其可药用的盐，或它们的混合物中的一种或多种。
本发明的主要优点在于：
本发明化合物具有优异的FXR激动作用，且细胞内释放良好，且很安全。
下面结合具体实施例，进一步阐述本发明。应理解，这些实施例仅用于说明本发明而不用于限制本发明的范围。下列实施例中未注明具体条件的实验方法，通常按照常规条件，或按照制造厂商所建议的条件。除非另外说明，否则百分比和份数是重量百分比和重量份数。以下实施例中所用的实验材料和试剂如无特别说明均可从市售渠道获得。
实施例1 4-(4-(((5-环丙基-3-(2,6-二氯苯基)异恶唑-4-基)甲氧)甲基)-4-氟哌啶-1-基)苯甲酸的制备(FL1)
Figure PCTCN2019086068-appb-000036
1.1 2,6-二氯-苯甲醛肟的合成
Figure PCTCN2019086068-appb-000037
将盐酸羟胺(67.6g，0.943mol)溶解于100mL水中，在0℃搅拌下滴加氢氧化钠3N(314mL，9.43mol。2,6-二氯苯甲醛(150g，0.857mol)加入乙醇(300mL)溶解后滴入上述混合溶液，然后升温至90℃搅拌过夜。用薄层层析(TLC)监测反应，反应完后冷却至室温，真空浓缩至10％溶液，大量固体析出，抽滤得固体，干燥，得白色固体162克，产率98％。ESI-MS m/z:190(M+1)。
1.2 2,6-二氯-苯甲醛氯-肟
Figure PCTCN2019086068-appb-000038
2,6-二氯-苯甲醛肟(100g,0.526mol)加入DMF(N,N-二甲酰胺，200mL)溶解，将氯代丁二酰亚胺(70.3g,0.526mol)溶于100mL DMF在40℃下滴入上述2,6-二氯-苯甲醛肟溶液中，后在40℃下反应1小时，用薄层层析(TLC)监测反应，反应完后冷却至室温，加入大量水、乙酸乙酯(EA)萃取，用水200mL洗涤有机层三次，饱和食盐水200mL洗涤有机层，无水Na 2SO 4干燥，旋干有机层，得粗产品，加入己烷100ml研磨，抽滤，干燥，得固体88.6克，为2,6-二氯-苯甲醛氯-肟。ESI-MS m/z:224(M+1)
1.3 5-环丙基-3-(2,6-二氯-苯基)-异恶唑-4-甲酸甲酯
Figure PCTCN2019086068-appb-000039
3-环丙基-3-氧代-丙酸甲酯(31.7g,0.223mol)加入三乙胺(45.1g,62mL，0.446mol)溶解，在室温搅拌，30分钟后，降温至10℃，下缓慢滴加2,6-二氯-苯甲醛氯-肟(50.0g,0.223mol，溶于300mL乙醇中)滴加期间温度不超过24℃，滴完后，室温搅拌过夜，用薄层层析(TLC)监测反应，反应完后加水200mL、乙酸乙酯(EA)500mL萃取，水层用乙酸乙酯200mL萃取三次，合并有机层，饱和食盐水200mL洗涤有机层，无水Na 2SO 4干燥，真空浓缩至10％溶液，大量固体析出，抽滤得粗产品，加入己烷100ml研磨，抽滤，干燥，得白色固体45.0克，产率65％。ESI-MS m/z:312(M +)。
1.4 [5-环丙基-3-(2,6-二氯-苯基)-异恶唑-4-基]-甲醇
Figure PCTCN2019086068-appb-000040
5-环丙基-3-(2,6-二氯-苯基)-异恶唑-4-甲酸甲酯(20.0g,64.1mmol)溶于四氢呋喃中(100mL),在0℃氮气保护下，滴加二异丁基氢化铝1M/甲苯(160.0mL,160.8mmol),室温搅拌2小时。用薄层层析(TLC)监测反应，反应完后,在0℃下滴加甲醇(10mL)和少量水淬灭并搅拌10分钟，后加入乙酸乙酯(500mL)。经硅藻土抽滤、乙酸乙酯洗涤 滤出沉淀物，滤液真空浓缩、干燥得白色固体15.2克,产率83％，ESI-MS m/z:285(M +)。
1.5 4-溴甲基-5-环丙基-3-(2,6-二氯-苯基)-异恶唑
Figure PCTCN2019086068-appb-000041
[5-环丙基-3-(2,6-二氯-苯基)-异恶唑-4-基]-甲醇(15.0g,52.8mmol)溶于二氯甲烷100mL中，加入三苯基磷(20.8g,79.2mmol),冷却至0℃，加入四溴化碳(26.3g,79.2mmol)，后在室温反应2h，用薄层层析(TLC)监测反应，反应完后，将反应液真空浓缩得橙色油状物，石油醚(PE)：乙酸乙酯(EA)＝5：1过柱，旋干得白色固体产物16.3克，产率89％，ESI-MS m/z:346(M +).
1.6 6-叔丁氧羰基-1-氧杂-6-氮杂螺[2.5]辛烷的合成
Figure PCTCN2019086068-appb-000042
向250mL茄形瓶中加入130mL二甲亚砜(DMSO)，搅拌加热，称取NaH固体5g加入到DMSO溶液中，继续搅拌12小时，停止加热，冷却至室温。加入三甲基碘化亚砜25g，室温搅拌24小时。将预先溶于DMSO中的25g N-叔丁氧羰基哌啶酮加入反应液中，再继续搅拌反应12小时，用薄层层析(TLC)监测反应。反应结束加入100-200mL水，乙酸乙酯(EA)100mL提取3次，饱和食盐水30mL洗涤有机层三次，旋干有机层，得无色液体28g，为6-叔丁氧羰基-1-氧杂-6-氮杂螺[2.5]辛烷。
1.7 (4-氟-1-叔丁氧羰基取代哌啶-4-基)甲醇的合成
Figure PCTCN2019086068-appb-000043
将28g 6-叔丁氧羰基-1-氧杂-6-氮杂螺[2.5]辛烷溶于100mL二氯甲烷(DCM)中，冷却，加入70％氟化氢吡啶溶液120mL，再反应一个12小时，反应结束，加入100mL水，DCM 100mL萃取三次，石油醚(PE)：乙酸乙酯(EA)＝4：1过柱，旋干得基本无色液体产物21克，为(4-氟-1-Boc取代哌啶-4-基)甲醇。
1.8 4-(((5-环丙基-3-(2,6-二氯苯基)异恶唑-4-基)甲氧)甲基)-4-氟哌啶-1-羧酸叔丁酯的合成
Figure PCTCN2019086068-appb-000044
(4-氟-1-叔丁氧羰基取代哌啶-4-基)甲醇(10.0g,42.8mmol)溶于四氢呋喃50mL中，在0℃下，加入18-冠醚-6(12.8g,48.6mmol)和叔丁醇钾(5.5g,48.6mmol)，反应30分钟，移至室温，滴加4-溴甲基-5-环丙基-3-(2,6-二氯-苯基)-异恶唑(9.9g,28.6mmol，溶于50mL四氢呋喃)，在室温搅拌反应14h。用薄层层析(TLC)监测反应，反应完后加水200mL、乙酸乙酯(EA，3x 200mL)萃取，合并有机层，饱和食盐水200mL洗涤有机层，无水Na2SO4干燥，真空浓缩，石油醚(PE)：乙酸乙酯(EA)＝4：1过柱，旋干得油状液体10.1克，产率71％，ESI-MS m/z:500(M +)。
1.9 5-环丙基-3-(2,6-二氯苯基)-4-(((4-氟哌啶-4-基)甲氧)甲基)异恶唑
Figure PCTCN2019086068-appb-000045
在冰浴和氮气保护下，向250mL茄形瓶加入4-(((5-环丙基-3-(2,6-二氯苯基)异恶唑-4-基)甲氧)甲基)-4-氟哌啶-1-羧酸叔丁酯(10.0g,20.0mmol，溶于二氯甲烷50mL)，然后滴加50mL三氟乙酸(滴加大约20分钟)，在室温搅拌反应1.5小时。用薄层层析(TLC)监测反应，反应完后真空旋干溶液,加300mL乙酸乙酯和2N NaOH(2x 200mL)，饱和食盐水200mL洗涤有机层，无水Na 2SO 4干燥，真空浓缩得油状液体7.2克，产率90％，ESI-MS m/z:400(M +)。
1.10 4-(4-(((5-环丙基-3-(2,6-二氯苯基)异恶唑-4-基)甲氧)甲基)-4-氟哌啶-1-基)苯甲酸乙酯
Figure PCTCN2019086068-appb-000046
在氮气保护下，向250mL两颈瓶加入5-环丙基-3-(2,6-二氯苯基)-4-(((4-氟哌啶-4-基)甲氧)甲基)异恶唑(2.0g,5.01mmol)，4-碘-苯甲酸乙酯(1.26g,4.55mmol),碳酸铯(2.97g,9.11mmol),三(二亚苄基丙酮)二钯(208.5mg,0.228mmol)和2-二环己基磷-2,4,6-三异丙基联苯(217.1mg,0.455mmol)，后加入100mL甲苯溶解，120℃下搅拌16h。用薄层层析(TLC)监测反应，反应完后，冷却至室温硅藻土抽滤，滤液中加入水(100mL)和乙酸乙酯(3x 100mL)。饱和食盐水200mL洗涤有机层，无水Na2SO4干燥，真空浓缩，石油醚(PE)：乙酸乙酯(EA)＝4：1至1:1过柱，旋干得固体1.55克即为4-(4-(((5-环丙基-3-(2,6-二氯苯基)异恶唑-4-基)甲氧)甲基)-4-氟哌啶-1-基)苯甲酸乙酯，产率62％，ESI-MS m/z:548(M +)。
1.11终产物FL1的合成
Figure PCTCN2019086068-appb-000047
在氮气保护下，向250mL两颈瓶加入4-(4-(((5-环丙基-3-(2,6-二氯苯基)异恶唑-4-基)甲氧)甲基)-4-氟哌啶-1-基)苯甲酸乙酯(1.55g，2.82mmol),后加入30mL甲醇、30mL四氢呋喃和10mL 1，4-二氧六环溶解，然后加入2N KOH(30mL,61.0mmol)。70℃下搅拌过夜。用薄层层析(TLC)监测反应，反应完后真空旋干溶液,加入100mL水和甲基叔丁基醚50mL。水层用2N HCl调至pH 5，然后加入二氯甲烷CH2Cl2(3x 100mL)萃取，饱和食盐水200mL洗涤有机层，无水Na2SO4干燥，真空浓缩的粗产品，用MeOH/CH3CN重结晶得白色固体956.0mg,产率62％， 1H NMR(400MHz,DMSO-d 6)δ12.28(s,1H),7.77(d,J＝8.9Hz,2H),7.70-7.50(m,3H),6.96(d,J＝9.0Hz,2H),4.32(s,2H),3.61(d,J＝13.2Hz,2H),3.40(s,2H),3.04(t,J＝10.9Hz,2H),2.37-2.30(m,1H),1.71-1.38(m,4H),1.22-1.02(m,4H)；ESI-MS m/z:520(M +)。
实施例2 4-(4-(((3-(2-氯-6-氟苯基)-5-环丙基异恶唑-4-基)甲氧)甲基)-4-氟哌啶-1-基)苯甲酸的制备(FL2)
将2,6-二氯苯甲醛替换成2-氯-6-氟苯甲醛，其余所需原料、试剂及制备方法同实施例1，得产物FL2。 1H NMR(400MHz,DMSO-d 6)δ12.28(s,1H),7.77(d,J＝8.9Hz,2H),7.62(td,J＝8.3,6.2Hz,1H),7.51(d,J＝8.1Hz,1H),7.41(t,J＝8.7Hz,1H),6.95(d,J＝9.0Hz,2H),4.35(s,2H),3.61(d,J＝13.2Hz,2H),3.39(s,2H),3.03(t,J＝10.8Hz,2H),2.37-2.30(m,1H),1.70-1.35(m,4H),1.19-1.04(m,4H).LRMS(EI)m/z 504(M +)。
实施例3 6-(4-(((5-环丙基-3-(2,6-二氯苯基)异恶唑-4-基)甲氧)甲基)-4-氟哌啶-1-基)-1-乙基-1H-吲哚-3-甲酸的制备(FL3)
将4-碘-苯甲酸乙酯换成6-溴-1-乙基-1H-吲哚-3-甲酸甲酯，其余所需原料、试剂及制备方法同实施例1，得产物FL3。 1H NMR(400MHz,DMSO-d 6)δ11.81(s,1H),7.89(s,1H),7.82(d,J＝8.7Hz,1H),7.64(d,J＝7.5Hz,2H),7.57(dd,J＝9.1,6.9Hz,1H),7.01-6.91(m,2H),4.35(s,2H),4.21(q,J＝7.0Hz,2H),3.50-3.36(m,4H),2.90(t,J＝12.3Hz,2H),2.44-2.30(m,1H),1.78-1.47(m,4H),1.37(t,J＝7.2Hz,3H),1.20-1.06(m,4H).；LRMS(EI)m/z 587(M +)。
实施例4 6-(4-(((5-环丙基-3-(2,6-二氯苯基)异恶唑-4-基)甲氧)甲基)-4-氟哌啶-1-基)苯并[b]噻吩-2-甲酸的制备(FL4)
将4-碘-苯甲酸乙酯替换成6-溴-苯并[b]噻吩-2-羧酸甲酯，其余所需原料、试剂及制备方法同实施例1，得产物FL4。 1H NMR(400MHz,DMSO-d 6)δ13.08(s,1H),7.91(s,1H),7.78(d,J＝9.0Hz,1H),7.64-7.58(m,1H),7.50(d,J＝8.1Hz,1H),7.43-7.38(m,2H),7.17(dd,J＝9.0,2.1Hz,1H),4.35(s,2H),3.56(d,J＝13.1Hz,2H),3.39(s,2H),3.10-2.93(m,2H),2.36-2.30(m,1H),1.68-1.37(m,4H),1.18-1.02(m,4H)；LRMS(EI)m/z 576(M +)。
实施例5 6-(4-(((5-环丙基-3-(2,6-二氯苯基)异恶唑-4-基)甲氧)甲基)-4-氟哌啶-1-基)-2-萘甲酸(FL5)
将4-碘-苯甲酸乙酯替换成6-溴-2-萘甲酸甲酯，其余所需原料、试剂及制备方法同实施例1，得产物FL5。 1H NMR(400MHz,DMSO-d 6)δ12.73(s,1H),8.40(s,1H),7.91(d,J＝9.1Hz,1H),7.84(dd,J＝8.6,1.5Hz,1H),7.74(d,J＝8.5Hz,1H),7.65-7.60(m,2H),7.56(dd,J＝9.3,6.6Hz,1H),7.43(dd,J＝9.1,1.9Hz,1H),7.22(d,J＝1.7Hz,1H),4.33(s,2H),3.65(d,J＝13.0Hz,2H),3.39(d,J＝20.2Hz,2H),3.05(t,J＝10.7Hz,2H),2.36-2.31(m,1H),1.75-1.43(m,4H),1.20-0.97(m,4H)；LRMS(EI)m/z 570(M +).
实施例6 6-(4-(((3-(2-氯-6-氟苯基)-5-环丙基异恶唑-4-基)甲氧)甲基)-4-氟哌啶-1-基)萘甲酸的制备(FL6)
将2,6-二氯苯甲醛替换成2-氯-6-氟苯甲醛和4-碘-苯甲酸乙酯替换成6-溴-2-萘甲酸甲酯，其余所需原料、试剂及制备方法同实施例1，得产物FL6。 1H NMR(400MHz,DMSO-d 6)δ12.74(s,1H),8.41(s,1H),7.92(d,J＝9.2Hz,1H),7.85(dd,J＝8.6,1.7Hz,1H),7.75(d,J＝8.7Hz,1H),7.62(td,J＝8.3,6.2Hz,1H),7.51(d,J＝8.1Hz,1H),7.48-7.37(m,2H),7.23(d,J＝2.0Hz,1H),4.37(s,2H),3.66(d,J＝13.0Hz,2H),3.39(d,J＝20.4Hz,2H),3.05(t,J＝10.5Hz,2H),2.37-2.32(m,1H),1.78-1.42(m,4H),1.19-1.00(m,4H)；LRMS(EI)m/z 554(M +)。
实施例7 5-(4-(((5-环丙基-3-(2,6-二氯苯基)异恶唑-4-基)甲氧)甲基)-4-氟哌啶-1-基)吡啶甲酸的制备(FL7)
将4-碘-苯甲酸乙酯替换成5-溴吡啶-2-羧酸甲酯，其余所需原料、试剂及制备方法同实施例1，得产物FL7。 1H NMR(400MHz,DMSO-d 6)δ8.13-7.81(m,2H),7.80-7.40(m,4H),4.33(s,2H),3.69-3.25(m,4H),3.20-2.85(m,2H),2.42-2.24(m,1H),1.76-1.37(m,4H),1.19-0.93(m,4H)；LRMS(EI)m/z 521(M +)。
实施例8 5-(4-(((3-(2-氯-6-氟苯基)-5-环丙基异恶唑-4-基)甲氧)甲基)-4-氟哌啶-1-基)吡啶甲酸的制备(FL8)
将2,6-二氯苯甲醛替换成2-氯-6-氟苯甲醛和4-碘-苯甲酸乙酯替换成5-溴吡啶-2-羧酸甲酯，其余所需原料、试剂及制备方法同实施例1，得产物FL8。 1H NMR(400MHz,DMSO-d 6)δ8.09-7.90(m,2H),7.73-7.62(m,2H),7.56-7.26(m,2H),4.36(s,2H),3.64-3.34(m,4H),3.15-2.90(m,2H),2.41-2.20(m,1H),1.75-1.37(m,4H),1.18-0.95(m,4H)；LRMS(EI)m/z505(M +)。
实施例9 6-(4-(((5-环丙基-3-(2,6-二氯苯基)异恶唑-4-基)甲氧)甲基)-4-氟哌啶-1-基)-1-甲基-1H-吲哚-3-甲酸甲酯的制备(FL9)
将4-碘-苯甲酸乙酯换成6-溴-1-甲基-1H-吲哚-3-甲酸甲酯，其余所需原料、试剂及制备方法同实施例1，得产物FL9。 1H NMR(400MHz,CDCl 3)δ8.01(d,J＝8.7Hz,1H),7.67(s,1H),7.40(d,J＝7.8Hz,2H),7.34-7.30(m,1H),7.26(s,1H),7.02(d,J＝8.4Hz,1H),6.79(s,1H),4.39(s,3H),3.89(s,3H),3.77(s,3H),3.43-3.37(m,4H),3.05(t,J＝10.6Hz,2H),2.22-2.14(m,1H),1.91-1.71(m,4H),1.32-1.19(m,4H),1.16-1.11(m,4H)；LRMS(EI)m/z 587(M +)。
实施例10 6-(4-(((5-环丙基-3-(2,6-二氯苯基)异恶唑-4-基)甲氧)甲基)-4-氟哌啶-1-基)-1-甲基-1H-吲哚-3-甲酸的制备(FL10)
将4-碘-苯甲酸乙酯换成6-溴-1-甲基-1H-吲哚-3-甲酸甲酯，其余所需原料、试剂及制备方法同实施例1，得产物FL10。 1H NMR(400MHz,DMSO-d 6)δ11.81(s,1H),7.89-7.76(m,2H),7.72-7.49(m,3H),6.96(d,J＝8.5Hz,2H),4.35(s,2H),3.78(s,3H),3.43-3.38(m,4H),2.90(t,J＝13.0Hz,2H),2.40-2.33(m,1H),1.79-1.54(m,4H),1.21-1.06(m,4H)；LRMS(EI)m/z 573(M +)。
实施例11 6-(4-(((3-(2-氯-6-氟苯基)-5-环丙基异恶唑-4-基)甲氧)甲基)-4-氟哌啶-1-基)-1-甲基-1H-吲哚-3-甲酸的制备(FL11)
将2,6-二氯苯甲醛替换成2-氯-6-氟苯甲醛和4-碘-苯甲酸乙酯换成6-溴-1-甲基-1H-吲哚-3-甲酸甲酯，其余所需原料、试剂及制备方法同实施例1，得目标产物FL11。 1H NMR(400MHz,DMSO-d 6)δ11.79(s,1H),7.88-7.76(m,2H),7.66-7.60(m,1H),7.52(d,J＝8.1Hz,1H),7.47-7.36(m,1H),7.00-6.88(m,2H),4.38(s,2H),3.78(s,3H),3.44-3.35(m,4H),2.90(t,J＝11.4Hz,2H),2.39-2.32(m,1H),1.75-1.38(m,4H),1.21-1.04(m,4H)；LRMS(EI)m/z 557(M +)。
实施例12 6-(4-(((3-(2-氯-6-氟苯基)-5-环丙基异恶唑-4-基)甲氧)甲基)-4-氟哌啶-1-基)-1-乙基-1H-吲哚-3-甲酸的制备(FL12)
将2,6-二氯苯甲醛替换成2-氯-6-氟苯甲醛和4-碘-苯甲酸乙酯换成6-溴-1-乙基-1H-吲哚-3-甲酸甲酯，其余所需原料、试剂及制备方法同实施例1，得目标产物FL12。 1H NMR(400MHz,DMSO-d 6)δ11.82(s,1H),7.89(s,1H),7.82(d,J＝8.7Hz,1H),7.67-7.58(m,1H),7.52(d,J＝8.1Hz,1H),7.42(t,J＝8.7Hz,1H),7.02-6.86(m,2H),4.38(s,2H),4.21(q,J＝7.2Hz,2H),3.45-3.35(m,3H),2.89(t,J＝10.9Hz,2H),2.89(t,J＝10.9Hz,2H),2.39-2.33(m,1H),1.72-1.47(m,4H),1.37(t,J＝7.2Hz,2H),1.20-1.07(m,4H)；LRMS(EI)m/z 571(M +)。
实施例13 2-(4-(((5-环丙基-3-(2,6-二氯苯基)异恶唑-4-基)甲氧)甲基)-4-氟哌啶-1-基)苯并[d]噻唑-6-甲酸甲酯的制备(FL13)
在100mL茄型瓶加入5-环丙基-3-(2,6-二氯苯基)-4-(((4-氟哌啶-4-基)甲氧)甲基)异恶唑(2.0g,5.01mmol)，2-溴苯并[d]噻唑-6-甲酸甲酯(1.09g,4.01mmol),N,N-二异丙基乙胺(776.8mg,6.01mmol),后加入50mL氮甲基吡咯烷酮溶解，60℃下搅拌过夜。用薄层层析(TLC) 监测反应，反应完后，加入水(100mL)和乙酸乙酯(3x 100mL)。饱和食盐水200mL洗涤有机层，无水Na2SO4干燥，真空浓缩，石油醚(PE)：乙酸乙酯(EA)＝5：1至1:1过柱，旋干得固体2.04克即为2-(4-(((5-环丙基-3-(2,6-二氯苯基)异恶唑-4-基)甲氧)甲基)-4-氟哌啶-1-基)苯并[d]噻唑-6-甲酸甲酯，收率69％， 1H NMR(500MHz,CDCl 3)δ8.41(d,J＝2.9Hz,1H),7.91(dd,J＝15.0,2.9Hz,1H),7.64(dd,J＝16.1,13.7Hz,1H),7.57(d,J＝15.0Hz,1H),7.45(dd,J＝14.9,1.0Hz,2H),4.80(s,2H),4.41(dt,J＝24.8,11.4Hz,2H),3.96(dt,J＝24.8,11.4Hz,2H),3.90(s,3H),3.76(d,J＝50.4Hz,2H),2.73(p,J＝20.2Hz,1H),1.96(ddt,J＝50.4,24.9,11.4Hz,2H),1.66(ddt,J＝50.3,24.9,11.4Hz,2H),1.45-0.88(m,4H)；ESI-MS m/z:591(M+1)。
实施例14 2-(4-(((5-环丙基-3-(2,6-二氯苯基)异恶唑-4-基)甲氧)甲基)-4-氟哌啶-1-基)苯并[d]噻唑-6-甲酸的制备(FL14)
在氮气保护下，向250mL两颈瓶加入2-(4-(((5-环丙基-3-(2,6-二氯苯基)异恶唑-4-基)甲氧)甲基)-4-氟哌啶-1-基)苯并[d]噻唑-6-甲酸甲酯(FL13,2.05g，3.56mmol),后加入30mL甲醇、30mL四氢呋喃和10mL 1，4-二氧六环溶解，然后加入2N KOH(30mL,61.0mmol)。70℃下搅拌过夜。用薄层层析(TLC)监测反应，反应完后真空旋干溶液,加入100mL水和甲基叔丁基醚50mL。水层用2N HCl调至pH 5，然后加入二氯甲烷CH2Cl2(3x 100mL)萃取，饱和食盐水200mL洗涤有机层，无水Na2SO4干燥，真空浓缩的粗产品，用MeOH/CH3CN重结晶得白色固体1.4g,产率70％。 1H NMR(400MHz,DMSO-d 6)δ12.71(s,1H),8.38(d,J＝1.7Hz,1H),7.86(dd,J＝8.4,1.8Hz,1H),7.66-7.52(m,3H),7.48(d,J＝8.5Hz,1H),4.34(s,2H),3.87(d,J＝12.7Hz,2H),3.41(d,J＝19.8Hz,2H),3.32-3.29(m,2H),2.38-2.31(m,1H),1.79-1.44(m,4H),1.20-1.03(m,4H)；LRMS(EI)m/z 577(M +)。
实施例15 6-(4-(((3-(2-氯-6-氟苯基)-5-环丙基异恶唑-4-基)甲氧)甲基)-4-氟哌啶-1-基)-1-乙基-1H-吲哚-3-甲酸甲酯的制备(FL15)
将2,6-二氯苯甲醛替换成2-氯-6-氟苯甲醛和4-碘-苯甲酸乙酯换成6-溴-1-乙基-1H-吲哚-3-甲酸甲酯，其余所需原料、试剂及制备方法同实施例1，得目标产物FL15。 1H NMR(400MHz,CDCl 3)δ8.02(d,J＝8.0Hz,1H),7.74(s,1H),7.41-7.36m,1H),7.31(d,J＝8.0Hz,1H),7.11(t,J＝8.4Hz,1H),7.01(d,J＝6.8Hz,1H),6.82(s,1H),4.42(s,2H),4.14(q,J＝7.3Hz,2H),3.89(s,3H),3.38(d,J＝19.1Hz,4H),3.05(t,J＝12.9Hz,2H),2.20-2.13(m,1H),1.92-1.65(m,4H),1.50(t,J＝7.3Hz,3H),1.30-1.22(m,2H),1.16-1.11(m,2H)；LRMS(EI)m/z585(M +)。
实施例16 6-(4-(((5-环丙基-3-(2,6-二氯苯基)异恶唑-4-基)甲氧)甲基)-4-氟哌啶-1-基)-1-(2-羟乙基)-1H-吲哚-3-甲酸的制备(FL16)
将4-碘-苯甲酸乙酯替换成6-溴-1-(2-((三甲基硅基)氧)乙基)-1H-吲哚-3-甲酸，其余所需原料、试剂及制备方法同实施例1，得目标产物FL16。 1H NMR(400MHz,DMSO-d 6)δ8.17-7.96(m,3H),7.69-7.69(m,3H),7.59-7.55(m,1H),4.37(s,2H),4.30(s,2H),3.79-3.70(m,4H),3.50(s,4H),2.41-2.35(m,1H),2.08-1.80(m,4H),1.20-1.06(m,4H)；LRMS(EI)m/z603(M +)。
实施例17 5-(4-(((5-环丙基-3-(2,6-二氯苯基)异恶唑-4-基)甲氧)甲基)-4-氟哌啶-1-基)吡啶甲酸甲酯的制备(FL17)
将4-碘-苯甲酸乙酯替换成5-溴吡啶-2-羧酸甲酯，其余所需原料、试剂及制备方法同实施例1，得目标产物FL17。 1H NMR(400MHz,CDCl 3)δ8.35(d,J＝2.9Hz,1H),8.01(d,J＝8.8Hz,1H),7.40(d,J＝1.9Hz,1H),7.38(s,1H),7.33(dd,J＝9.3,6.6Hz,1H),7.17(dd,J＝8.9,2.9 Hz,1H),4.37(s,2H),3.97(s,3H),3.67(d,J＝13.1Hz,2H),3.36(d,J＝17.9Hz,2H),3.22(td,J＝12.8,2.6Hz,2H),2.17-2.10(m,1H),1.83(t,J＝11.6Hz,2H),1.75-1.50(m,2H),1.33-1.26(m,2H),1.16-1.11(m,2H)；LRMS(EI)m/z 535(M +)。
实施例18 6-(4-(((5-环丙基-3-(2,6-二氯苯基)异恶唑-4-基)甲氧)甲基)-4-氟哌啶-1-基)-2-萘甲酸甲酯的制备(FL18)
将2,6-二氯苯甲醛替换成2-氯-6-氟苯甲醛和4-碘-苯甲酸乙酯替换成6-溴-2-萘甲酸甲酯，其余所需原料、试剂及制备方法同实施例1，得目标产物FL18。 1H NMR(400MHz,CDCl 3)δ8.48(s,1H),7.99(d,J＝7.7Hz,1H),7.82(d,J＝8.1Hz,1H),7.68(d,J＝5.3Hz,1H),7.37(td,J＝8.2,5.9Hz,1H),7.31(s,1H),7.29(s,1H),7.09(t,J＝8.4Hz,2H),4.41(s,2H),3.96(s,3H),3.63(d,J＝12.6Hz,2H),3.37(d,J＝19.1Hz,2H),3.20(s,2H),2.20-2.10(m,1H),1.95-1.80(m,2H),1.57(s,2H),1.31-1.24(m,2H),1.15-1.10(m,2H)；LRMS(EI)m/z 568(M +)。
实施例19 2-(4-(((3-(2-氯-6-氟苯基)-5-环丙基异恶唑-4-基)甲氧)甲基)-4-氟哌啶-1-基)苯并[d]噻唑-6-甲酸的制备(FL19)
将2,6-二氯苯甲醛替换成2-氯-6-氟苯甲醛，其余所需原料、试剂及制备方法同实施例14，得目标产物FL19。 1H NMR(400MHz,DMSO-d 6)δ12.69(s,1H),8.38(d,J＝1.7Hz,1H),7.86(dd,J＝8.5,1.8Hz,1H),7.61(td,J＝8.3,6.2Hz,1H),7.49(t,J＝8.0Hz,2H),7.44-7.36(m,1H),4.36(s,2H),3.86(d,J＝12.7Hz,2H),3.53-3.17(m,4H),2.37-2.30(m,1H),1.88-1.43(m,4H),1.20-1.04(m,4H)；LRMS(EI)m/z 561(M +)。
实施例20 3-((4-(((5-环丙基-3-(2,6-二氯苯基)异恶唑-4-基)甲氧)甲基)-4-氟哌啶-1-基)-1H-吲唑-6-甲酸的制备(FL20)
将4-碘-苯甲酸乙酯替换成3-溴-1-(四氢-2H-吡喃-2-基)-1H-吲唑-6-甲酸甲酯，其余所需原料、试剂及制备方法同实施例1，得目标产物FL20。 1H NMR(400MHz,DMSO-d 6)δ13.00(s,1H),12.31(s,1H),7.95(s,1H),7.82(d,J＝8.6Hz,1H),7.65-7.60(m,2H),7.56-7.52(m,2H),4.35(s,2H),3.58(d,J＝12.7Hz,2H),3.40(d,J＝20.4Hz,2H),3.09-2.96(m,2H),2.39-2.33(m,1H),1.80-1.57(m,4H),1.20-1.06(m,4H)；LRMS(EI)m/z 560(M +)。
实施例21 3-(4-(((3-(2-氯-6-氟苯基)-5-环丙基异恶唑-4-基)甲氧)甲基)-4-氟哌啶-1-基)-1H-吲唑-6-甲酸的制备(FL21)
将2,6-二氯苯甲醛替换成2-氯-6-氟苯甲醛和4-碘-苯甲酸乙酯替换成3-溴-1-(四氢-2H-吡喃-2-基)-1H-吲唑-6-甲酸甲酯，其余所需原料、试剂及制备方法同实施例1，得目标产物FL21。 1H NMR(400MHz,DMSO-d 6)δ12.97(s,1H),12.34(s,1H),7.95(s,1H),7.82(d,J＝8.6Hz,1H),7.63-7.57(m,1H),7.54(dd,J＝8.6,1.3Hz,1H),7.50(d,J＝8.6Hz,1H),7.44-7.36(m,1H),4.38(s,2H),3.61-3.54(m,2H),3.39(d,J＝20.6Hz,2H),3.08-2.99(m,2H),2.39-2.32(m,1H),1.78-1.56(m,4H),1.21-1.07(m,4H)；LRMS(EI)m/z 544(M +)。
实施例22 3-((4-(((5-环丙基-3-(2,6-二氯苯基)异恶唑-4-基)甲氧)甲基)-4-氟哌啶-1-基)-1H-吲唑-5-甲酸的制备(FL22)
将4-碘-苯甲酸乙酯替换成3-溴-1-(四氢-2H-吡喃-2-基)-1H-吲唑-5-甲酸甲酯，其余所需原料、试剂及制备方法同实施例1，得目标产物FL22。 1H NMR(400MHz,DMSO-d 6)δ12.71(s,1H),12.36(s,1H),8.35(s,1H),7.86(dd,J＝8.8,1.2Hz,1H),7.65-7.60(m,2H),7.55(dd,J＝9.2,6.8Hz,1H),7.41(d,J＝8.8Hz,1H),4.35(s,2H),3.59(d,J＝15.5Hz,2H),3.41(d,J＝20.1Hz,2H),3.17-2.88(m,2H),2.39-2.33(m,1H),1.82-1.54(m,4H),1.21-1.00(m,4H)；LRMS(EI)m/z 560(M +)。
实施例23 3-(4-(((3-(2-氯-6-氟苯基)-5-环丙基异恶唑-4-基)甲氧)甲基)-4-氟哌啶-1-基)-1H-吲唑-5-甲酸的制备(FL23)
将2,6-二氯苯甲醛替换成2-氯-6-氟苯甲醛和4-碘-苯甲酸乙酯替换成3-溴-1-(四氢-2H-吡喃-2-基)-1H-吲唑-6-甲酸甲酯，其余所需原料、试剂及制备方法同实施例1，得目标产物FL23。 1H NMR(400MHz,DMSO-d 6)δ12.71(s,1H),12.36(s,1H),8.34(s,1H),7.86(dd,J＝8.8,1.3Hz,1H),7.61(td,J＝8.3,6.2Hz,1H),7.51(d,J＝8.1Hz,1H),7.43-7.43(m,2H),4.38(s,2H),3.59(d,J＝14.2Hz,2H),3.40(d,J＝20.3Hz,2H),3.16-3.03(m,2H),2.39-2.32(m,1H),1.88-1.49(m,4H),1.20-0.98(m,4H)；LRMS(EI)m/z 544(M +)。
实施例24 5-环丙基-3-(2,6-二氯苯基)-4-(((4-氟-1-(4-(三氟甲基)苯基)哌啶-4-基)甲氧)甲基)异恶唑的制备(FL24)
将4-碘-苯甲酸乙酯替换成1-溴-4-(三氟甲基)苯，其余所需原料、试剂及制备方法同实施例1，得目标产物FL24。 1H NMR(400MHz,CD3OD)δ7.63-7.59(m,2H),7.58-7.52(m,1H),7.49(d,J＝8.8Hz,2H),7.04(d,J＝8.8Hz,2H),4.32(s,2H),3.58(d,J＝13.1Hz,2H),3.39(s,2H),3.10-2.94(m,2H),2.37-2.30(m,1H),1.73-1.36(m,4H),1.22-1.00(m,4H)；LRMS(EI)m/z 544(M +)。
实施例25 3-(2-氯-6-氟苯基)-5-环丙基-4-(((4-氟-1-(4-(三氟甲基)苯基)哌啶-4-基)甲氧)甲基)异恶唑的制备(FL25)
将2,6-二氯苯甲醛替换成2-氯-6-氟苯甲醛和4-碘-苯甲酸乙酯替换成1-溴-4-(三氟甲基)苯，其余所需原料、试剂及制备方法同实施例1，得目标产物FL25。 1H NMR(400MHz,DMSO-d 6)δ7.62(td,J＝8.3,6.2Hz,1H),7.51-7.48(m,3H),7.46-7.37(m,1H),7.05(d,J＝8.8Hz,2H),4.35(s,2H),3.86-3.50(m,2H),3.36(d,J＝20.2Hz,2H),3.10-2.89(m,2H),2.37-2.30(m,1H),1.67-1.38(m,4H),1.21-1.06(m,4H)；LRMS(EI)m/z 527(M +)。
实施例26 6-(4-(((5-环丙基-3-(2,6-二氟苯基)异恶唑-4-基)甲氧)甲基)-4-氟哌啶-1-基)-1-甲基-1H-吲哚-3-甲酸的制备(FL26)
将2,6-二氯苯甲醛替换成2,6-二氟苯甲醛和4-碘-苯甲酸乙酯换成6-溴-1-甲基-1H-吲哚-3-甲酸甲酯，其余所需原料、试剂及制备方法同实施例1，得目标产物FL26。 1H NMR(400MHz,DMSO-d 6)δ11.76(s,1H),7.84(s,1H),7.81(d,J＝9.5Hz,1H),7.74-7.59(m,1H),7.40-7.21(m,2H),7.01-6.92(m,2H),4.41(s,2H),3.77(s,3H),3.42-3.36(m,4H),3.03-2.77(m,2H),2.38-2.31(m,1H),1.82-1.42(m,4H),1.19-1.03(m,4H)；LRMS(EI)m/z 540(M +).
实施例27 6-(4-(((5-环丙基-3-(2,6-二氟苯基)异恶唑-4-基)甲氧)甲基)-4-氟哌啶-1-基)-1-乙基-1H-吲哚-3-甲酸的制备(FL27)
将2,6-二氯苯甲醛替换成2,6-二氟苯甲醛和4-碘-苯甲酸乙酯换成6-溴-1-乙基-1H-吲哚-3-甲酸甲酯，其余所需原料、试剂及制备方法同实施例1，得目标产物FL27。 1H NMR(400MHz,DMSO-d 6)δ11.81(s,1H),7.89(s,1H),7.81(d,J＝8.7Hz,1H),7.69-7.58(m,1H),7.33-7.28(m,2H),6.98(d,J＝1.5Hz,1H),6.96-6.92(m,1H),4.41(s,2H),4.20(q,J＝7.2Hz,2H),3.39-3.36(m,4H),3.06-2.77(m,2H),2.38-2.31(m,1H),1.75-1.50(m,4H),1.36(t,J＝7.2Hz,3H),1.18-1.03(m,4H)；LRMS(EI)m/z 554(M +).
实施例28 2-(3-(((3-(2-氯-6-氟苯基)-5-环丙基异恶唑-4-基)甲氧)甲基)-3-氟-8-氮杂双环[3.2.1]辛-8-基)苯并[d]噻唑-6-甲酸的制备(FL28)
将2,6-二氯苯甲醛替换成2-氯-6-氟苯甲醛和(4-氟-1-叔丁氧羰基取代哌啶-4-基)甲醇替换成叔丁基3-氟-3-(羟甲基)-8-氮杂双环[3.2.1]辛-8-羧化物，其余所需原料、试剂及制备方法同实施例13、14，得目标产物FL28。 1H NMR(400MHz,DMSO-d 6)δ12.66(s,1H),8.40(s,1H), 7.87(d,J＝8.4Hz,1H),7.65(dd,J＝14.4,7.8Hz,1H),7.59-7.43(m,3H),4.39(s,2H),4.31(s,2H),3.51(d,J＝30.0Hz,2H),2.42-2.35(m,1H),2.06-1.84(m,8H),1.33-1.04(m,4H)；LRMS(EI)m/z 587(M +).
实施例29 2-(3-(((3-(2-氯-6-氟苯基)-5-环丙基异恶唑-4-基)甲氧)甲基)-3-氟-8-氮杂双环[3.2.1]辛-8-基)-4-氟苯并[d]噻唑-6-甲酸的制备(FL29)
将2,6-二氯苯甲醛替换成2-氯-6-氟苯甲醛、2-溴苯并[d]噻唑-6-甲酸甲酯替换成2-溴-4-氟苯并[d]噻唑-6-甲酸甲酯和(4-氟-1-叔丁氧羰基取代哌啶-4-基)甲醇替换成叔丁基3-氟-3-(羟甲基)-8-氮杂双环[3.2.1]辛-8-羧化物，其余所需原料、试剂及制备方法同实施例13、14，得目标产物FL29。 1H NMR(400MHz,DMSO-d 6)δ13.02(s,1H),8.27(d,J＝1.5Hz,1H),7.68-7.61(m,2H),7.56(d,J＝8.1Hz,1H),7.51-7.43(m,1H),4.39(s,2H),4.34(s,2H),3.51(d,J＝29.8Hz,2H),2.42-2.35(m,1H),2.02-1.91(m,8H),1.20-1.07(m,4H)；LRMS(EI)m/z605(M +)。
实施例30 2-(4-(((3-(2-氯-6-氟苯基)-5-环丙基异恶唑-4-基)甲氧)甲基)-4-氟哌啶-1-基)-4-氟苯并[d]噻唑-6-甲酸的制备(FL30)
将2,6-二氯苯甲醛替换成2-氯-6-氟苯甲醛和2-溴苯并[d]噻唑-6-甲酸甲酯替换成2-溴-4-氟苯并[d]噻唑-6-甲酸甲酯，其余所需原料、试剂及制备方法同实施例14，得目标产物FL30。 1H NMR(400MHz,DMSO-d 6)δ12.97(s,1H),8.21(d,J＝1.5Hz,1H),7.64(d,J＝1.1Hz,1H),7.62(s,1H),7.59(dd,J＝11.5,1.5Hz,1H),7.54(dd,J＝9.0,7.2Hz,1H),4.34(s,2H),3.79-3.37(m,6H),2.40-2.33(m,1H),1.82-1.69(m,2H),1.54-1.37(m,2H),1.21-1.06(m,4H)；LRMS(EI)m/z 579(M +).
实施例31 2-(4-(((5-环丙基-3-(2,6-二氯苯基)异恶唑-4-基)甲氧)甲基)-4-氟哌啶-1-基)-4-氟苯并[d]噻唑-6-甲酸的制备(FL31)
将2-溴苯并[d]噻唑-6-甲酸甲酯替换成2-溴-4-氟苯并[d]噻唑-6-甲酸甲酯，其余所需原料、试剂及制备方法同实施例14，得目标产物FL31。 1H NMR(400MHz,DMSO-d 6)δ12.94(s,1H),8.25(d,J＝1.5Hz,1H),7.63(d,J＝1.9Hz,1H),7.61-7.59(m,2H),7.55(dd,J＝9.3,6.6Hz,1H),4.33(s,2H),3.88(d,J＝11.6Hz,2H),3.46-3.28(m,4H),2.38-2.31(m,1H),1.84-1.50(m,4H),1.19-1.06(m,4H)；LRMS(EI)m/z 595(M +).
实施例32 2-(4-(((3-(2-氯-6-氟苯基)-5-环丙基异恶唑-4-基)甲氧)甲基)-4-氟哌啶-1-基)-4-氟苯并[d]噻唑-6-甲酸甲酯的制备(FL32)
将2,6-二氯苯甲醛替换成2-氯-6-氟苯甲醛和2-溴苯并[d]噻唑-6-甲酸甲酯替换成2-溴-4-氟苯并[d]噻唑-6-甲酸甲酯，其余所需原料、试剂及制备方法同实施例14，得目标产物FL32。 1H NMR(400MHz,CDCl 3)δ8.08(d,J＝0.8Hz,1H),7.70(dd,J＝11.3,0.7Hz,1H),7.42(s,1H),7.40(s,1H),7.32(dd,J＝8.7,7.4Hz,1H),4.36(s,2H),3.91(s,3H),3.71-3.43(m,6H),2.17-2.11(m,1H),1.84-1.68(m,2H),1.70-1.57(m,2H),1.31-1.25(m,2H),1.17-1.12(m,2H)；LRMS(EI)m/z 593(M +).
实施例33 2-(4-(((5-环丙基-3-(2,6-二氯苯基)异恶唑-4-基)甲氧)甲基)-3-氟-8-氮杂双环[3.2.1]辛-8-基)-4-氟苯并[d]噻唑-6-甲酸(FL33)
将2-溴苯并[d]噻唑-6-甲酸甲酯替换成2-溴-4-氟苯并[d]噻唑-6-甲酸甲酯和(4-氟-1-叔丁氧羰基取代哌啶-4-基)甲醇替换成叔丁基3-氟-3-(羟甲基)-8-氮杂双环[3.2.1]辛-8-羧化物，其余所需原料、试剂及制备方法同实施例14，得目标产物FL33。 1H NMR(400MHz,DMSO-d 6)δ12.96(s,1H),8.27(d,J＝1.5Hz,1H),7.70(d,J＝1.2Hz,1H),7.68(s,1H),7.64-7.58(m,2H), 4.36(s,4H),3.56-3.29(m,4H),2.46-2.32(m,1H),2.05-1.83(m,6H),1.21-1.05(m,4H)；LRMS(EI)m/z 621(M +).
实施例34 2-(4-(((5-环丙基-3-(2,6-二氯苯基)异恶唑-4-基)甲氧)甲基)-3-氟-8-氮杂双环[3.2.1]辛-8-基)苯并[d]噻唑-6-甲酸的制备(FL34)
将(4-氟-1-叔丁氧羰基取代哌啶-4-基)甲醇替换成叔丁基3-氟-3-(羟甲基)-8-氮杂双环[3.2.1]辛-8-羧化物，其余所需原料、试剂及制备方法同实施例14。 1H NMR(400MHz,DMSO-d 6)δ8.40(d,J＝1.7Hz,1H),7.87(dd,J＝8.5,1.8Hz,1H),7.69(d,J＝1.3Hz,1H),7.67(s,1H),7.60(dd,J＝9.1,7.1Hz,1H),7.53(d,J＝8.5Hz,1H),4.35(s,2H),4.30(s,2H),3.51(d,J＝30.2Hz,2H),2.42-2.31(m,1H),2.01-1.87(m,6H),1.47-1.40(m,2H),1.21-1.07(m,4H)；LRMS(EI)m/z 603(M +).
实施例35 2-(3-(((5-环丙基-3-(2-(三氟甲基)苯基)异恶唑-4-基)甲氧)甲基)-3-氟-8-氮杂双环[3.2.1]辛-8-基)-4-氟苯并[d]噻唑-6-甲酸甲酯的制备(FL35)
将2,6-二氯苯甲醛替换成2-(三氟甲基)苯甲醛、2-溴苯并[d]噻唑-6-甲酸甲酯替换成2-溴-4-氟苯并[d]噻唑-6-甲酸甲酯和(4-氟-1-叔丁氧羰基取代哌啶-4-基)甲醇替换成叔丁基3-氟-3-(羟甲基)-8-氮杂双环[3.2.1]辛-8-羧化物，其余所需原料、试剂及制备方法同实施例14，得目标产物FL35。 1H NMR(400MHz,CDCl 3)δ8.31(d,J＝1.6Hz,1H),8.01(dd,J＝8.5,1.5Hz,1H),7.60-7.50(m,2H),7.43-7.38(m,2H),4.47(s,2H),4.44(s,2H),3.92(s,3H),3.51(d,J＝28.0Hz,2H),2.33-2.05(m,5H),1.65-1.49(m,4H),1.28-1.19(m,2H),1.15-1.08(m,2H)；LRMS(EI)m/z 650(M +).
实施例36 2-(3-(((5-环丙基-3-(2-(三氟甲基)苯基)异恶唑-4-基)甲氧)甲基)-3-氟-8-氮杂双环[3.2.1]辛-8-基)-4-氟苯并[d]噻唑-6-甲酸的制备(FL36)
将2,6-二氯苯甲醛替换成2-(三氟甲基)苯甲醛、2-溴苯并[d]噻唑-6-甲酸甲酯替换成2-溴-4-氟苯并[d]噻唑-6-甲酸甲酯和(4-氟-1-叔丁氧羰基取代哌啶-4-基)甲醇替换成叔丁基3-氟-3-(羟甲基)-8-氮杂双环[3.2.1]辛-8-羧化物，其余所需原料、试剂及制备方法同实施例14，得目标产物FL36。 1H NMR(400MHz,DMSO-d 6)δ13.00(s,1H),8.27(d,J＝1.5Hz,1H),7.73-7.69(m,1H),7.67(dd,J＝6.5,1.7Hz,1H),7.64(dd,J＝4.1,1.6Hz,1H),7.61(d,J＝1.5Hz,1H),7.58-7.55(m,1H),4.44(s,2H),4.35(s,2H),3.54(d,J＝29.6Hz,2H),2.41-2.34(m,1H),2.05-1.85(m,6H),1.51(q,J＝6.5Hz,2H),1.21-1.05(m,4H)；LRMS(EI)m/z 636(M +).
实施例37 2-(3-(((5-环丙基-3-(2-(三氟甲基)苯基)异恶唑-4-基)甲氧)甲基)-3-氟-8-氮杂双环[3.2.1]辛-8-基)苯并[d]噻唑-6-甲酸的制备(FL37)
将2,6-二氯苯甲醛替换成2-(三氟甲基)苯甲醛和(4-氟-1-叔丁氧羰基取代哌啶-4-基)甲醇替换成叔丁基3-氟-3-(羟甲基)-8-氮杂双环[3.2.1]辛-8-羧化物，其余所需原料、试剂及制备方法同实施例14，得目标产物FL37。 1H NMR(500MHz,CDCl 3)δ8.58(d,J＝3.1Hz,1H),8.43(dd,J＝14.4,3.7Hz,1H),8.07(dd,J＝14.9,3.0Hz,1H),7.68(d,J＝15.0Hz,1H),7.57-7.29(m,2H),7.12(dd,J＝14.2,3.7Hz,1H),4.80(s,2H),4.77(ddd,J＝12.9,7.4,4.2Hz,2H),4.11(d,J＝50.6Hz,2H),2.72(p,J＝20.1Hz,1H),2.36(ddd,J＝50.6,24.9,6.4Hz,2H),2.11(ddd,J＝50.4,24.9,6.4Hz,2H),2.01-1.76(m,4H),1.34-1.11(m,2H),1.11-0.89(m,2H).LRMS(EI)m/z618(M +).
实施例38 2-(4-(((5-环丙基-3-(2-(三氟甲基)苯基)异恶唑-4-基)甲氧)甲基)-4-氟哌啶-1-基)-4-氟苯并[d]噻唑-6-甲酸的制备(FL38)
将2,6-二氯苯甲醛替换成2-(三氟甲基)苯甲醛和(4-氟-1-叔丁氧羰基取代哌啶-4-基)甲醇替换成叔丁基3-氟-3-(羟甲基)-8-氮杂双环[3.2.1]辛-8-羧化物，其余所需原料、试剂及制备方 法同实施例14，得目标产物FL38。 1H NMR(500MHz,CDCl 3)δ8.00(d,J＝8.0,2H),7.53(d,J＝8.4,2H),7.17(s,1H),6.86(s,1H),3.96(s,3H),3.91(s,6H),3.74(s,2H),3.32-3.41(m,3H),2.85-2.92(m,2H),2.52-2.71(m,1H),2.28-2.48(m,1H),2.19-2.30(m,1H),2.07-2.14(m,4H),1.70-1.82(m,4H)；LRMS(EI)m/z 610(M +).
实施例39 2-(4-(((5-环丙基-3-(2-(三氟甲基)苯基)异恶唑-4-基)甲氧)甲基)-4-氟哌啶-1-基)苯并[d]噻唑-6-甲酸的制备(FL39)
将2,6-二氯苯甲醛替换成2-(三氟甲基)苯甲醛，其余所需原料、试剂及制备方法同实施例14，得目标产物FL39. 1H NMR(500MHz,CDCl 3)δ8.58(d,J＝3.1Hz,1H),8.43(dd,J＝14.3,3.7Hz,1H),8.07(dd,J＝15.0,2.9Hz,1H),7.68(d,J＝15.0Hz,1H),7.55-7.32(m,2H),7.12(dd,J＝14.2,3.7Hz,1H),4.80(s,2H),4.52(dt,J＝24.9,11.0Hz,2H),4.24(dt,J＝24.9,11.0Hz,2H),3.77(d,J＝50.4Hz,2H),2.89(p,J＝20.2Hz,1H),2.05(ddt,J＝50.4,24.9,11.0Hz,2H),1.74(ddt,J＝50.4,24.7,11.0Hz,2H),1.23(m,2H),1.11-0.87(m,2H).LRMS(EI)m/z592(M +).
实施例40 6-(3-(((5-环丙基-3-(2,6-二氯苯基)异恶唑-4-基)甲氧)甲基)-3-氟-8-氮杂双环[3.2.1]辛-8-基)-1-甲基-1H-吲哚-3-甲酸甲酯的制备(FL40)
将(4-氟-1-叔丁氧羰基取代哌啶-4-基)甲醇替换成叔丁基3-氟-3-(羟甲基)-8-氮杂双环[3.2.1]辛-8-羧化物和4-碘-苯甲酸乙酯换成6-溴-1-甲基-1H-吲哚-3-甲酸甲酯，其余所需原料、试剂及制备方法同实施例1，得目标产物FL40。 1H NMR(500MHz,CDCl 3)δ9.00(s,1H),8.10(d,J＝7.5Hz,1H),7.64(t,J＝7.5Hz,1H),7.46(d,J＝7.4Hz,2H),6.62(dd,J＝7.5,1.4Hz,1H),6.48(d,J＝1.4Hz,1H),4.80(s,2H),4.24-4.15(m,2H),3.93(d,J＝25.3Hz,2H),3.82(d,J＝80.0Hz,6H),2.71(p,J＝10.2Hz,1H),2.22-1.96(m,4H),1.79(ddd,J＝25.3,12.5,3.2Hz,2H),1.54(tdd,J＝5.8,3.8,1.9Hz,2H),1.35-1.13(m,2H),1.10-0.90(m,2H).LRMS(EI)m/z 613(M +).
实施例41 6-(3-(((5-环丙基-3-(2-(三氟甲基)苯基)异恶唑-4-基)甲氧)甲基)-3-氟-8-氮杂双环[3.2.1]辛-8-基)-1-甲基-1H-吲哚-3-甲酸甲酯的制备(FL41)
将2,6-二氯苯甲醛替换成2-(三氟甲基)苯甲醛、(4-氟-1-叔丁氧羰基取代哌啶-4-基)甲醇替换成叔丁基3-氟-3-(羟甲基)-8-氮杂双环[3.2.1]辛-8-羧化物和4-碘-苯甲酸乙酯换成6-溴-1-甲基-1H-吲哚-3-甲酸甲酯，其余所需原料、试剂及制备方法同实施例1，得目标产物FL14。 1H NMR(500MHz,CDCl 3)δ9.00(s,1H),8.43(dd,J＝14.4,3.7Hz,1H),8.10(d,J＝15.0Hz,1H),7.65-7.35(m,2H),7.12(dd,J＝14.2,3.7Hz,1H),6.62(dd,J＝15.0,2.9Hz,1H),6.48(d,J＝2.9Hz,1H),4.80(s,2H),4.12(tt,J＝8.6,6.4Hz,2H),4.00(d,J＝50.4Hz,2H),3.82(d,J＝80.0Hz,6H),2.80(dd,J＝40.3,20.2Hz,1H),2.11(m,4H),1.86-1.39(m,4H),1.36-0.78(m,4H).LRMS(EI)m/z 628(M +).
实施例42 6-(3-(((5-环丙基-3-(2,6-二氯苯基)异恶唑-4-基)甲氧)甲基)-3-氟-8-氮杂双环[3.2.1]辛-8-基)-1-甲基-1H-吲哚-3-甲酸的制备(FL42)
将(4-氟-1-叔丁氧羰基取代哌啶-4-基)甲醇替换成叔丁基3-氟-3-(羟甲基)-8-氮杂双环[3.2.1]辛-8-羧化物和4-碘-苯甲酸乙酯换成6-溴-1-甲基-1H-吲哚-3-甲酸甲酯，其余所需原料、试剂及制备方法同实施例1，得目标产物FL42。 1H NMR(500MHz,CDCl 3)δ9.26(s,1H),7.64(dd,J＝16.1,13.7Hz,1H),7.60-7.28(m,3H),6.72(d,J＝2.9Hz,1H),6.12(dd,J＝15.0,3.1Hz,1H),4.80(s,2H),4.26(tt,J＝8.5,6.4Hz,2H),3.96(d,J＝50.4Hz,2H),3.74(s,3H),3.03(p,J＝20.2Hz,1H),2.34(ddd,J＝50.4,24.8,6.4Hz,2H),2.22-1.84(m,4H),1.81-1.40(m,2H),1.37-0.82(m,4H).LRMS(EI)m/z 599(M +).
实施例43 6-(3-(((5-环丙基-3-(2-(三氟甲基)苯基)异恶唑-4-基)甲氧)甲基)-3-氟-8-氮杂双环[3.2.1]辛-8-基)-1-甲基-1H-吲哚-3-甲酸的制备(FL43)
将2,6-二氯苯甲醛替换成2-(三氟甲基)苯甲醛、(4-氟-1-叔丁氧羰基取代哌啶-4-基)甲醇替换成叔丁基3-氟-3-(羟甲基)-8-氮杂双环[3.2.1]辛-8-羧化物和4-碘-苯甲酸乙酯换成6-溴-1-甲基-1H-吲哚-3-甲酸甲酯，其余所需原料、试剂及制备方法同实施例1，得目标产物FL43。 1H NMR(500MHz,CDCl 3)δ9.26(s,1H),8.43(dd,J＝14.4,3.7Hz,1H),7.78-7.29(m,3H),7.12(dd,J＝14.2,3.7Hz,1H),6.68(d,J＝2.9Hz,1H),6.12(dd,J＝15.0,3.1Hz,1H),4.80(s,2H),4.25(tt,J＝8.6,6.4Hz,2H),4.03(d,J＝50.4Hz,2H),3.74(s,3H),3.13-2.77(m,1H),2.52-1.96(m,6H),1.81(s,1H),1.41-0.73(m,4H).LRMS(EI)m/z 614(M +).
实施例44 6-(3-(((3-(2,6-二氯苯基)-5-((2,2-二氟环丙基)甲基)异恶唑-4-基)甲氧)甲基)-4-氟哌啶-1-基)-1-甲基-1H-吲哚-3-甲酸的制备(FL44)
将3-环丙基-3-氧代-丙酸甲酯换成甲基4-(2,2-二氟环丙烷)-3-氧代丁酸甲酯和4-碘-苯甲酸乙酯换成6-溴-1-甲基-1H-吲哚-3-甲酸甲酯，其余所需原料、试剂及制备方法同实施例1， 1H NMR(500MHz,CDCl 3)δ9.26(s,1H),7.89-7.58(m,1H),7.52-7.21(m,3H),6.79(d,J＝2.9Hz,1H),6.12(dd,J＝15.0,3.1Hz,1H),4.80(s,2H),3.76(t,J＝25.2Hz,5H),3.64-3.40(m,3H),3.39-3.18(m,2H),2.94(dd,J＝24.8,14.6Hz,1H),2.35-2.02(m,2H),1.98-1.50(m,3H),0.80(m,2H).LRMS(EI)m/z 623(M +).
实施例45 2-(4-(((3-(2,6-二氯苯基)-5-((2,2-二氟环丙基)甲基)异恶唑-4-基)甲氧)甲基)-4-氟哌啶-1-基)-4-氟苯并[d]噻唑-6-甲酸的制备(FL45)
将3-环丙基-3-氧代-丙酸甲酯换成甲基4-(2,2-二氟环丙烷)-3-氧代丁酸甲酯和2-溴苯并[d]噻唑-6-甲酸甲酯替换成2-溴-4-氟苯并[d]噻唑-6-甲酸甲酯，其余所需原料、试剂及制备方法同实施例14，得目标产物FL45。 1H NMR(500MHz,CDCl 3)δ8.35(d,J＝2.9Hz,1H),7.96(dd,J＝15.9,2.9Hz,1H),7.64(dd,J＝16.2,13.7Hz,1H),7.46(dd,J＝14.8,1.1Hz,2H),4.80(s,2H),4.37(dt,J＝24.9,10.9Hz,2H),4.03(dt,J＝24.9,10.9Hz,2H),3.63(d,J＝50.4Hz,2H),3.23(dd,J＝24.7,17.0Hz,1H),2.94(dd,J＝24.8,17.0Hz,1H),2.09(ddt,J＝50.4,24.7,10.9Hz,2H),1.90-1.37(m,3H),1.33-0.54(m,2H).LRMS(EI)m/z 645(M +).
实施例46 2-(4-(((5-环丙基-3-(2,6-二氟苯基)异恶唑-4-基)甲氧)甲基)-4-氟哌啶-1-基)苯并[d]噻唑-6-甲酸的制备(FL46)
将2,6-二氯苯甲醛替换成2,6-二氟苯甲醛，其余所需原料、试剂及制备方法同实施例14，得目标产物FL46。 1H NMR(400MHz,DMSO-d 6)δ13.02(s,1H),8.25(d,J＝1.3Hz,1H),7.63(ddd,J＝11.6,7.7,1.5Hz,2H),7.30(t,J＝8.1Hz,2H),4.40(s,2H),3.88(d,J＝10.3Hz,2H),3.62-3.26(m,6H),2.34(ddd,J＝13.4,8.3,5.3Hz,1H),1.87-1.31(m,4H),1.36-0.86(m,4H).LRMS(EI)m/z 544(M +).
实施例47 2-(4-(((5-环丙基-3-(2,6-二氟苯基)异恶唑-4-基)甲氧)甲基)-4-氟哌啶-1-基)-4-氟苯并[d]噻唑-6-甲酸的制备(FL47)
将2,6-二氯苯甲醛替换成2,6-二氟苯甲醛和2-溴苯并[d]噻唑-6-甲酸甲酯替换成2-溴-4-氟苯并[d]噻唑-6-甲酸甲酯，其余所需原料、试剂及制备方法同实施例14，得目标产物DC69，收率80％。 1H NMR(500MHz,CDCl 3)δ8.32(d,J＝2.9Hz,1H),7.64(dd,J＝16.0,3.0Hz,1H),7.53-7.19(m,1H),7.21-6.83(m,2H),4.78(s,2H),4.56(dt,J＝24.9,11.4Hz,2H),4.00-3.78(m,4H),2.91(p,J＝20.1Hz,1H),2.12-1.68(m,4H),1.41-0.71(m,4H).；LRMS(EI)m/z562(M +).
实施例48 2-(3-(((3-(2,6-二氯苯基)-5-((2,2-二氟环丙基)甲基)异恶唑-4-基)甲氧甲基)-3-氟-8-氮杂双环[3.2.1]辛-8-基)-4-氟苯并[d]噻唑-6-甲酸(FL48)
将3-环丙基-3-氧代-丙酸甲酯换成甲基4-(2,2-二氟环丙烷)-3-氧代丁酸甲酯、(4-氟-1-叔丁氧羰基取代哌啶-4-基)甲醇替换成叔丁基3-氟-3-(羟甲基)-8-氮杂双环[3.2.1]辛-8-羧化物和2-溴苯并[d]噻唑-6-甲酸甲酯替换成2-溴-4-氟苯并[d]噻唑-6-甲酸甲酯，其余所需原料、试剂及制备方法同实施例14，得目标产物FL48。 1H NMR(500MHz,CDCl 3)δ9.00-8.07(m,1H),8.05(dd,J＝14.9,3.0Hz,1H),7.63(t,J＝7.5Hz,1H),7.47(d,J＝7.4Hz,2H),4.82(s,2H),4.66(tt,J＝8.5,6.4Hz,2H),4.27(d,J＝50.4Hz,1H),3.29-2.91(m,2H),2.60-1.90(m,4H),1.89-1.31(m,5H),1.26-0.66(m,2H)；LRMS(EI)m/z 671(M +).
实施例49 2-(3-(((3-(2,6-二氯苯基)-5-((2,2-二氟环丙基)甲基)异恶唑-4-基)甲氧甲基)-3-氟-8-氮杂双环[3.2.1]辛-8-基)苯并[d]噻唑-6-甲酸的制备(FL49)
将3-环丙基-3-氧代-丙酸甲酯换成甲基4-(2,2-二氟环丙烷)-3-氧代丁酸甲酯和(4-氟-1-叔丁氧羰基取代哌啶-4-基)甲醇替换成叔丁基3-氟-3-(羟甲基)-8-氮杂双环[3.2.1]辛-8-羧化物，其余所需原料、试剂及制备方法同实施例32，得目标产物FL49。 1H NMR(500MHz,CDCl 3)δ9.01-8.08(m,1H),8.07(dd,J＝14.9,3.0Hz,1H),7.90-7.53(m,2H),7.46(dd,J＝14.9,1.0Hz,2H),4.80(s,2H),4.64(tt,J＝8.5,6.4Hz,2H),4.25(d,J＝50.4Hz,1H),3.29-2.90(m,2H),2.60-1.93(m,4H),1.89-1.32(m,5H),1.26-0.67(m,2H)；LRMS(EI)m/z 653(M +).
实施例50 2-(3-(((3-(2,6-二氯苯基)-5-((2,2-二氟环丙基)甲基)异恶唑-4-基)甲氧甲基)-3-氟-8-氮杂双环[3.2.1]辛-8-基)-1-甲基-1H-吲哚-3-甲酸(FL50)
将3-环丙基-3-氧代-丙酸甲酯换成甲基4-(2,2-二氟环丙烷)-3-氧代丁酸甲酯、(4-氟-1-叔丁氧羰基取代哌啶-4-基)甲醇替换成叔丁基3-氟-3-(羟甲基)-8-氮杂双环[3.2.1]辛-8-羧化物和4-碘-苯甲酸乙酯换成6-溴-1-甲基-1H-吲哚-3-甲酸甲酯，其余所需原料、试剂及制备方法同实施例1，得目标产物FL50。 1H NMR(500MHz,CDCl 3)δ9.26(s,1H),7.64(dd,J＝16.1,13.7Hz,1H),7.49-7.32(m,3H),6.73(d,J＝2.9Hz,1H),6.12(dd,J＝15.0,3.1Hz,1H),4.80(s,2H),4.56-4.24(m,2H),4.19(dd,J＝72.1,28.4Hz,2H),3.98-3.77(m,2H),3.74(s,3H),3.52(dd,J＝24.8,16.4Hz,1H),3.19-2.48(m,3H),2.02-1.24(m,3H),1.27-0.29(m,4H).LRMS(EI)m/z 649(M +).
实施例51 2-(3-(((3-(2,6-二氯苯基)-5-((2,2-二氟环丙基)甲基)异恶唑-4-基)甲氧甲基)-4-氟哌啶-1-基)-1-乙基-1H-吲哚-3-甲酸的制备(FL51)
将3-环丙基-3-氧代-丙酸甲酯换成甲基4-(2,2-二氟环丙烷)-3-氧代丁酸甲酯和4-碘-苯甲酸乙酯换成6-溴-1-乙基-1H-吲哚-3-甲酸甲酯，其余所需原料、试剂及制备方法同实施例1，得目标产物FL51。 1H NMR(500MHz，CDCl 3)δ9.26(s,1H),7.64(dd,J＝16.1,13.7Hz,1H),7.56-7.23(m,3H),6.79(d,J＝3.1Hz,1H),6.12(dd,J＝15.0,3.1Hz,1H),4.80(s,2H),4.24(q,J＝12.6Hz,2H),3.77(d,J＝50.4Hz,2H),3.52(dt,J＝24.9,12.9Hz,2H),3.23(ddd,J＝25.0,13.5,6.6Hz,3H),2.85(dd,J＝24.8,15.2Hz,1H),2.32-2.00(m,2H),1.94-1.48(m,3H),1.06(t,J＝12.6Hz,3H),1.24-0.39(m,2H)；LRMS(EI)m/z 637(M +).
实施例52 2-(3-(((3-(2,6-二氯苯基)-5-((2,2-二氟环丙基)甲基)异恶唑-4-基)甲氧甲基)-3-氟-8-氮杂双环[3.2.1]辛-8-基)-1-乙基-1H-吲哚-3-甲酸的制备(FL52)
将3-环丙基-3-氧代-丙酸甲酯换成甲基4-(2,2-二氟环丙烷)-3-氧代丁酸甲酯、(4-氟-1-叔丁氧羰基取代哌啶-4-基)甲醇替换成叔丁基3-氟-3-(羟甲基)-8-氮杂双环[3.2.1]辛-8-羧化物和4-碘-苯甲酸乙酯换成6-溴-1-乙基-1H-吲哚-3-甲酸甲酯，其余所需原料、试剂及制备方法同实施例1，得目标产物FL52。 1H NMR(500MHz,CDCl 3)δ9.26(s,1H),7.64(dd,J＝16.1,13.7 Hz,1H),7.57-7.33(m,3H),6.71(d,J＝3.1Hz,1H),6.12(dd,J＝15.0,3.1Hz,1H),4.80(s,2H),4.47-4.08(m,6H),4.07-3.75(m,2H),3.46(dd,J＝24.8,12.4Hz,1H),3.16-2.78(m,3H),2.15-1.47(m,3H),1.40-0.16(m,7H)；LRMS(EI)m/z 663(M +).
实施例53 2-(3-(((3-(2,6-二氯苯基)-5-((2,2-二氟环丙基)甲基)异恶唑-4-基)甲氧甲基)-4-氟哌啶-1-基)苯并[d]噻唑-6-甲酸的制备(FL53)
将3-环丙基-3-氧代-丙酸甲酯换成甲基4-(2,2-二氟环丙烷)-3-氧代丁酸甲酯，其余所需原料、试剂及制备方法同实施例14，得目标产物FL53。 1H NMR(500MHz,CDCl 3)δ8.58(d,J＝1.6Hz,1H),8.07(dd,J＝7.5,1.4Hz,1H),7.83-7.57(m,2H),7.46(d,J＝7.4Hz,2H),4.80(s,2H),4.50(dt,J＝12.3,5.5Hz,2H),4.19(dt,J＝12.4,5.5Hz,2H),3.88(dd,J＝12.4,7.6Hz,1H),3.81(d,J＝25.3Hz,2H),2.84(dd,J＝12.4,7.6Hz,1H),2.02(ddt,J＝25.2,12.3,5.4Hz,2H),1.74(ddt,J＝25.1,12.4,5.5Hz,2H),1.69-1.51(m,1H),1.42-1.20(m,1H),0.78-0.64(m,1H).LRMS(EI)m/z 627(M +).
实施例54 2-(3-(((3-(2,6-二氯苯基)-5-(2,2-二氟环丙基)异恶唑-4-基)甲氧甲基)-3-氟-8-氮杂双环[3.2.1]辛-8-基)-4-氟苯并[d]噻唑-6-甲酸的制备(FL54)
将3-环丙基-3-氧代-丙酸甲酯换成甲基4-(2,2-二氟环丙烷)-3-氧代丁酸甲酯换成3-(2,2-二氟环丙基-3-氧代丙酸、(4-氟-1-叔丁氧羰基取代哌啶-4-基)甲醇替换成叔丁基3-氟-3-(羟甲基)-8-氮杂双环[3.2.1]辛-8-羧化物和2-溴苯并[d]噻唑-6-甲酸甲酯替换成2-溴-4-氟苯并[d]噻唑-6-甲酸甲酯，其余所需原料、试剂及制备方法同实施例14，得目标产物FL54。 1H NMR(500MHz,CDCl 3)δ8.35(d,J＝3.1Hz,1H),7.74(dd,J＝15.9,3.1Hz,1H),7.64(dd,J＝16.1,13.7Hz,1H),7.46(dd,J＝14.9,1.0Hz,2H),4.77-4.71(m,2H),4.70(s,2H),4.31(d,J＝50.6Hz,2H),3.33(tt,J＝41.8,23.1Hz,1H),2.84(ddd,J＝50.4,24.8,6.7Hz,2H),2.17-1.95(m,2H),1.83(ddd,J＝50.5,24.8,6.7Hz,2H),1.71-1.48(m,2H),1.39-0.98(m,2H)；LRMS(EI)m/z657(M +).
实施例55 22-(3-(((3-(2,6-二氯苯基)-5-(2,2-二氟环丙基)异恶唑-4-基)甲氧甲基)-3-氟-8-氮杂双环[3.2.1]辛-8-基)苯并[d]噻唑-6-甲酸的制备(FL55)
将3-环丙基-3-氧代-丙酸甲酯换成甲基4-(2,2-二氟环丙烷)-3-氧代丁酸甲酯换成3-(2,2-二氟环丙基-3-氧代丙酸和(4-氟-1-叔丁氧羰基取代哌啶-4-基)甲醇替换成叔丁基3-氟-3-(羟甲基)-8-氮杂双环[3.2.1]辛-8-羧化物，其余所需原料、试剂及制备方法同实施例14，得目标产物FL55。 1H NMR(500MHz,CDCl 3)δ8.58(d,J＝3.1Hz,1H),8.07(dd,J＝14.9,3.0Hz,1H),7.83-7.53(m,2H),7.46(dd,J＝14.9,1.0Hz,2H),4.89-4.59(m,4H),4.29(d,J＝50.4Hz,2H),3.26(tt,J＝41.8,23.1Hz,1H),2.76(ddd,J＝50.5,24.8,6.8Hz,2H),2.18-1.97(m,2H),1.84(ddd,J＝50.4,24.8,6.7Hz,2H),1.66-1.42(m,2H),1.37-0.99(m,2H).LRMS(EI)m/z639(M +)。
实施例56 2-(3-(((3-(2,6-二氯苯基)-5-(2,2-二氟环丙基)异恶唑-4-基)甲氧甲基)-3-氟-8-氮杂双环[3.2.1]辛-8-基)-1-甲基-1H-吲哚-3-甲酸的制备(FL56)
将3-环丙基-3-氧代-丙酸甲酯换成甲基4-(2,2-二氟环丙烷)-3-氧代丁酸甲酯换成3-(2,2-二氟环丙基-3-氧代丙酸、(4-氟-1-叔丁氧羰基取代哌啶-4-基)甲醇替换成叔丁基3-氟-3-(羟甲基)-8-氮杂双环[3.2.1]辛-8-羧化物和4-碘-苯甲酸乙酯换成6-溴-1-甲基-1H-吲哚-3-甲酸甲酯，其余所需原料、试剂及制备方法同实施例1，得目标产物FL56。 1H NMR(500MHz,CDCl 3)δ9.26(s,1H),7.64(dd,J＝16.1,13.7Hz,1H),7.56-7.26(m,3H),6.62(d,J＝2.9Hz,1H),6.12(dd,J＝15.0,3.1Hz,1H),4.76(s,2H),4.26(s,2H),3.81(d,J＝50.4Hz,2H),3.74(s,3H),2.88(tt,J＝41.9,23.0Hz,1H),2.34-2.02(m,6H),1.72-0.90(m,4H)；LRMS(EI)m/z 635(M +)。
实施例57 2-(3-(((5-(2,2-二氟环丙基)-3-(2-(三氟甲氧基)苯基)异恶唑-4-基)甲氧)甲基)-3-氟-8-氮杂双环[3.2.1]辛-8-基)-4-氟苯并[d]噻唑-6-甲酸的制备(FL57)
将2,6-二氯苯甲醛替换成2-(三氟甲基)苯甲醛，其余所需原料、试剂及制备方法同实施例54，得目标产物FL57。 1H NMR(500MHz,CDCl 3)δ8.43(dd,J＝14.4,3.7Hz,1H),8.35(d,J＝3.1Hz,1H),7.74(dd,J＝16.0,3.0Hz,1H),7.43(ddd,J＝18.1,14.0,3.3Hz,2H),7.12(dd,J＝14.2,3.7Hz,1H),4.81(s,2H),4.73(ddd,J＝8.4,6.5,2.0Hz,2H),4.19(d,J＝50.4Hz,2H),3.01(tt,J＝41.8,22.9Hz,1H),2.52-1.65(m,8H),1.55-0.76(m,2H)；LRMS(EI)m/z 672(M +).
实施例58 2-(3-(((5-((2,2-二氟环丙基)甲基)-3-(2-(三氟甲氧基)苯基)异恶唑-4-基)甲氧)甲基)-3-氟-8-氮杂双环[3.2.1]辛-8-基)-1-甲基-1H-吲哚-3-甲酸的制备(FL58)
将2,6-二氯苯甲醛替换成2-(三氟甲基)苯甲醛，其余所需原料、试剂及制备方法同实施例50，得目标产物FL58。 1H NMR(500MHz,CDCl 3)δ9.25(s,1H),8.42(dd,J＝14.2,3.7Hz,1H),7.59-7.32(m,3H),7.11(dd,J＝14.2,3.7Hz,1H),6.69(d,J＝3.0Hz,1H),6.11(dd,J＝15.0,2.9Hz,1H),4.79(s,2H),4.55-4.10(m,4H),3.73(s,3H),3.34(dd,J＝24.7,16.3Hz,1H),2.64(dd,J＝24.7,16.3Hz,1H),2.50-2.08(m,6H),1.77-1.24(m,3H),1.12-0.83(m,2H)；LRMS(EI)m/z 664(M +).
实施例59 2-(3-(((5-((2,2-二氟环丙基)甲基)-3-(2-(三氟甲氧基)苯基)异恶唑-4-基)甲氧)甲基)-3-氟-8-氮杂双环[3.2.1]辛-8-基)-4-氟苯并[d]噻唑-6-甲酸的制备(FL59)
将2,6-二氯苯甲醛替换成2-(三氟甲基)苯甲醛，其余所需原料、试剂及制备方法同实施例48，得目标产物FL59。 1H NMR(500MHz,CDCl 3)δ8.43(dd,J＝14.4,3.7Hz,1H),8.35(d,J＝3.1Hz,1H),7.94(dd,J＝16.0,3.0Hz,1H),7.59-7.30(m,2H),7.12(dd,J＝14.2,3.7Hz,1H),4.80(s,2H),4.67-4.56(m,2H),4.41(d,J＝50.4Hz,2H),3.21(dd,J＝24.7,14.3Hz,1H),2.98(dd,J＝24.9,14.2Hz,1H),2.45(ddd,J＝50.5,24.8,6.7Hz,2H),2.08-1.84(m,2H),1.85-1.46(m,5H),1.15-0.51(m,2H).LRMS(EI)m/z 686(M +).
实施例60 2-(4-(((5-环丙基-3-(2-(三氟甲氧基)苯基)异恶唑-4-基)氧)甲基)-4-氟哌啶-1-基)-4-氟苯并[d]噻唑-6-甲酸的制备(FL60)
制备方法同实施例38，得目标产物FL60。 1H NMR(500MHz,CDCl 3)δ8.00(d,J＝8.0,2H),7.53(d,J＝8.4,2H),7.17(s,1H),6.86(s,1H),3.96(s,3H),3.91(s,6H),3.32-3.41(m,3H),2.85-2.92(m,2H),2.52-2.71(m,1H),2.28-2.48(m,1H),2.19-2.30(m,1H),2.07-2.14(m,4H),1.70-1.82(m,4H)；LRMS(EI)m/z 610(M+).；LRMS(EI)m/z 596(M +).
实施例61 2-(4-(((5-环丙基-3-(2-(三氟甲氧基)苯基)异恶唑-4-基)氧)甲基)-3-氟-8-氮杂双环[3.2.1]辛-8-基)-4-氟苯并[d]噻唑-6-甲酸的制备(FL61)
制备方法同实施例36，制得目标产物FL61。 1H NMR(CDCl 3,500MHz)8.27(d,J＝1.5Hz,1H),7.73-7.69(m,1H),7.67(dd,J＝6.5,1.7Hz,1H),7.64(dd,J＝4.1,1.6Hz,1H),7.61(d,J＝1.5Hz,1H),7.58-7.55(m,1H),4.35(s,2H),3.54(d,J＝29.6Hz,2H),2.41-2.34(m,1H),2.05-1.85(m,6H),1.51(q,J＝6.5Hz,2H),1.21-1.05(m,4H)；LRMS(EI)m/z 636(M+).LRMS(EI)m/z 622(M +).
实施例62 2-(4-(((5-环丙基-3-(2，6-二氯苯基)异恶唑-4-基)氧)甲基)-4-氟哌啶-1-基)-4-氟苯并[d]噻唑-6-甲酸的制备(FL62)
制备方法同实施例31，制得目标产物FL62。 1H NMR(CDCl 3,500MHz)8.25(d,J＝1.5Hz,1H),7.63(d,J＝1.9Hz,1H),7.61-7.59(m,2H),7.55(dd,J＝9.3,6.6Hz,1H),3.88(d,J＝11.6Hz,2H),3.46-3.28(m,4H),2.38-2.31(m,1H),1.84-1.50(m,4H),1.19-1.06(m,4H)；LRMS(EI)m/z581(M +).
实施例63 2-(4-(((5-环丙基-3-(2，6-二氯苯基)异恶唑-4-基)氧)甲基)-3-氟-8-氮杂双环[3.2.1]辛-8-基)-4-氟苯并[d]噻唑-6-甲酸的制备(FL63)
制备方法同实施例33，制得目标产物FL63。 1H NMR(CDCl 3,500MHz)8.27(d,J＝1.5Hz,1H),7.70(d,J＝1.2Hz,1H),7.68(s,1H),7.64-7.58(m,2H),4.36(s,2H),3.56-3.29(m,4H),2.46-2.32(m,1H),2.05-1.83(m,6H),1.21-1.05(m,4H)；LRMS(EI)m/z 607(M +).
实施例64 2-(4-(((5-环丙基-3-(2，6-二氯苯基)异恶唑-4-基)氧)甲基)-3-氟-8-氮杂双环[3.2.1]辛-8-基)-1-甲基-1H-吲哚-3-甲酸的制备(FL64)
制备方法同实施例42，制得目标产物FL64。 1H NMR(CDCl 3,500MHz)δ9.00(s,1H),8.43(dd,J＝14.4,3.7Hz,1H),8.10(d,J＝15.0Hz,1H),7.65-7.35(m,2H),7.12(dd,J＝14.2,3.7Hz,1H),6.62(dd,J＝15.0,2.9Hz,1H),6.48(d,J＝2.9Hz,1H),4.12(tt,J＝8.6,6.4Hz,2H),4.00(d,J＝50.4Hz,2H),3.82(d,J＝80.0Hz,6H),2.80(dd,J＝40.3,20.2Hz,1H),2.11(m,4H),1.86-1.39(m,4H),1.36-0.78(m,4H).LRMS(EI)m/z 585(M +).
实施例65 2-(4-(((5-环丙基-3-(2，6-二氯苯基)异恶唑-4-基)氧)甲基)-4-氟哌啶-1-基)-1-甲基-1H-吲哚-3-甲酸的制备(FL65)
制备方法同实施例10，制得目标产物FL65。 1H NMR(CDCl 3,500MHz)δ9.18(s,1H),7.58(dd,J＝16.1,13.6Hz,1H),7.48-7.25(m,3H),7.01(d,J＝2.9Hz,1H),6.07(dd,J＝14.8,2.9Hz,1H),4.47(d,J＝50.2Hz,2H),3.71(s,3H),3.34(ddt,J＝117.5,24.7,13.1Hz,4H),2.75(p,J＝20.1Hz,1H),2.18-1.61(m,4H),1.19-0.76(m,4H).LRMS(EI)m/z 559(M +).
实施例66 7-(3-(((5-((2,2-二氟环丙基)甲基)-3-(2-(三氟甲氧基)苯基)异恶唑-4-基)甲氧)甲基)-3-氟-8-氮杂双环[3.2.1]辛-8-基)咪唑并[1,2-a]吡啶-3-甲酸的制备(FL66)
将6-溴-1-甲基-1H-吲哚-3-甲酸甲酯换成7-溴咪唑并[1,2-a]吡啶-3-甲酸甲酯,其余所需原料、试剂及制备方法同实施例58，制得目标产物FL66。 1H NMR(500MHz,CDCl 3)δ8.43(dd,J＝14.4,3.7Hz,1H),8.33(d,J＝15.0Hz,1H),7.66(s,1H),7.56-7.30(m,2H),7.12(dd,J＝14.2,3.7Hz,1H),6.65(d,J＝3.1Hz,1H),6.56(dd,J＝14.9,3.0Hz,1H),4.80(s,2H),4.29-4.16(m,3H),4.13(s,1H),3.57(dd,J＝24.8,14.7Hz,1H),2.80-2.39(m,3H),2.12-1.83(m,4H),1.79-1.36(m,3H),1.14-0.60(m,2H)；LRMS(EI)m/z 651(M +).
实施例67 2-(4-(((5-环丙基-3-(2-(三氟甲氧基)苯基)异恶唑-4-基)甲氧)甲基)-3-氟-8-氮杂双环[3.2.1]辛-8-基)咪唑并[1,2-a]吡啶-3-甲酸的制备(FL67)
将2,6-二氯苯甲醛替换成2-(三氟甲基)苯甲醛、(4-氟-1-叔丁氧羰基取代哌啶-4-基)甲醇替换成叔丁基3-氟-3-(羟甲基)-8-氮杂双环[3.2.1]辛-8-羧化物和4-碘-苯甲酸乙酯换成7-溴咪唑并[1,2-a]吡啶-3-甲酸甲酯,其余所需原料、试剂及制备方法同实施例1，制得目标产物FL67。 1H NMR(500MHz,CDCl 3)δ8.43(dd,J＝7.5,1.6Hz,1H),8.33(d,J＝7.5Hz,1H),7.66(s,1H),7.43(dtd,J＝30.9,7.4,1.5Hz,1H),7.12(dd,J＝7.4,1.5Hz,1H),6.70(d,J＝1.4Hz,1H),6.57(dd,J＝7.5,1.6Hz,1H),4.80(s,1H),4.35-4.05(m,1H),3.94(d,J＝25.1Hz,1H),3.15(ddd,J＝25.3,12.4,3.3Hz,1H),2.56(p,J＝10.2Hz,1H),2.21-1.95(m,2H),1.90-1.51(m,4H),1.40-1.13(m,2H),1.13-0.84(m,2H).LRMS(EI)m/z 601(M +).
实施例68 2-(4-(((5-环丙基-3-(2-(三氟甲氧基)苯基)异恶唑-4-基)甲氧)甲基)-4-氟哌啶-1-基)咪唑并[1,2-a]吡啶-3-甲酸的制备(FL68)
将2,6-二氯苯甲醛替换成2-(三氟甲基)苯甲醛和4-碘-苯甲酸乙酯换成7-溴咪唑并[1,2-a]吡啶-3-甲酸甲酯,其余所需原料、试剂及制备方法同实施例，制得目标产物FL68。 1H NMR(500MHz,CDCl 3)δ8.39(dd,J＝14.2,3.7Hz,1H),8.29(d,J＝14.8Hz,1H),7.62(s,1H),7.52-7.30(m,2H),7.09(dd,J＝14.2,3.7Hz,1H),6.75-6.50(m,2H),4.78(s,2H),3.79(d,J＝50.2 Hz,2H),3.43(dt,J＝25.0,12.9Hz,2H),3.12(dt,J＝24.8,12.9Hz,2H),2.86-2.55(m,1H),2.09(ddt,J＝50.5,25.1,12.9Hz,2H),1.71(ddt,J＝50.6,24.8,13.0Hz,2H),1.41-0.82(m,4H).LRMS(EI)m/z 575(M +).
实施例69 6-(3-(((5-环丙基-3-(2,6-二氯苯基)异恶唑-4-基)甲氧)甲基)-4-氟哌啶-1-基)咪唑并[1,2-a]吡啶-3-甲酸的制备(FL69)
将4-碘-苯甲酸乙酯换成7-溴咪唑并[1,2-a]吡啶-3-甲酸甲酯,其余所需原料、试剂及制备方法同实施例1，制得目标产物FL69。 1H NMR(500MHz,CDCl 3)δ8.33(d,J＝15.0Hz,1H),7.98-7.55(m,2H),7.46(dd,J＝14.9,1.0Hz,2H),6.93(d,J＝3.1Hz,1H),6.80(dd,J＝14.9,3.0Hz,1H),4.80(s,2H),3.83(d,J＝50.6Hz,2H),3.65-3.34(m,3H),3.14(dt,J＝24.9,13.2Hz,2H),2.26-1.57(m,4H),1.45-0.73(m,4H).LRMS(EI)m/z 560(M +).
实施例70 6-(3-(((5-环丙基-3-(2,6-二氯苯基)异恶唑-4-基)甲氧)甲基)-3-氟-8-氮杂双环[3.2.1]辛-8-基)咪唑并[1,2-a]吡啶-3-甲酸的制备(FL70)
将(4-氟-1-叔丁氧羰基取代哌啶-4-基)甲醇替换成叔丁基3-氟-3-(羟甲基)-8-氮杂双环[3.2.1]辛-8-羧化物和4-碘-苯甲酸乙酯换成7-溴咪唑并[1,2-a]吡啶-3-甲酸甲酯,其余所需原料、试剂及制备方法同实施例1，制得目标产物FL70。 1H NMR(500MHz,CDCl 3)δ8.33(d,J＝15.0Hz,1H),7.74-7.53(m,2H),7.46(dd,J＝14.9,1.0Hz,2H),6.66(d,J＝3.0Hz,1H),6.58(dd,J＝14.9,3.0Hz,1H),4.80(s,2H),4.46(d,J＝50.4Hz,2H),4.34-4.15(m,2H),3.04(p,J＝20.0Hz,1H),2.72(ddd,J＝50.4,24.8,6.7Hz,2H),2.51-2.05(m,4H),1.84-1.49(m,2H),1.45-0.58(m,4H).LRMS(EI)m/z 586(M +).
实施例71 2-(4-(((3-(2-氯-6-氟苯基)-5-环丙基异恶唑-4-基)甲氧)甲基)-4-氟哌啶-1-基)咪唑并[1,2-a]吡啶-3-甲酸的制备(FL71)
将2,6-二氯苯甲醛替换成2-氯-6-氟苯甲醛和4-碘-苯甲酸乙酯替换成7-溴咪唑并[1,2-a]吡啶-3-甲酸甲酯,其余所需原料、试剂及制备方法同实施例1，制得目标产物FL71。 1H NMR(500MHz,CDCl 3)δ8.33(d,J＝14.9Hz,1H),7.66(s,1H),7.51-7.11(m,3H),7.00-6.71(m,2H),4.80(s,2H),3.87(d,J＝50.4Hz,2H),3.30(dtd,J＝37.6,25.0,12.9Hz,4H),2.86(p,J＝20.0Hz,1H),2.32-1.51(m,4H),1.46-0.80(m,4H).LRMS(EI)m/z 544(M +).
实施例72 2-(4-(((3-(2-氯-6-氟苯基)-5-环丙基异恶唑-4-基)甲氧)甲基)-3-氟-8-氮杂双环[3.2.1]辛-8-基)-咪唑并[1,2-a]吡啶-3-甲酸的制备(FL72)
将2,6-二氯苯甲醛替换成2-氯-6-氟苯甲醛、(4-氟-1-叔丁氧羰基取代哌啶-4-基)甲醇替换成叔丁基3-氟-3-(羟甲基)-8-氮杂双环[3.2.1]辛-8-羧化物和4-碘-苯甲酸乙酯替换成7-溴咪唑并[1,2-a]吡啶-3-甲酸甲酯,其余所需原料、试剂及制备方法同实施例，制得目标产物FL72。 1H NMR(500MHz,CDCl 3)δ8.33(d,J＝7.5Hz,1H),7.66(s,1H),7.50-7.11(m,3H),6.78-6.51(m,2H),4.80(s,2H),4.42(d,J＝25.1Hz,2H),4.26-3.97(m,2H),2.66(p,J＝10.1Hz,1H),2.55-2.04(m,6H),1.40-0.81(m,6H).LRMS(EI)m/z 570(M +).
实施例73 2-(4-(((3-(2-氯-6-氟苯基)-5-环丙基异恶唑-4-基)氧)甲基)-3-氟-8-氮杂双环[3.2.1]辛-8-基)-咪唑并[1,2-a]吡啶-3-甲酸的制备(FL73)
制备方法同实施例72，制得目标产物FL73。 1H NMR(500MHz,CDCl 3)δ8.33(d,J＝15.0Hz,1H),7.66(s,1H),7.46-7.14(m,3H),6.86-6.49(m,2H),5.23(d,J＝50.4Hz,2H),4.39-4.05(m,2H),2.94-2.14(m,7H),1.86-0.73(m,6H).LRMS(EI)m/z 556(M +).
Figure PCTCN2019086068-appb-000048
实施例74 2-(6-(3-(((5-环丙基-3-(2，6-二氯苯基)异恶唑-4-基)甲氧)甲基)-4-氟哌啶-1-基)-1-甲基-1H-吲哚-3-酰胺)乙酸甲酯的制备(FL74)
将2-(6-(3-(((5-环丙基-3-(2，6-二氯苯基)异恶唑-4-基)甲氧)甲基)-4-氟哌啶-1-基)-1-甲基-1H-吲哚-3甲酸(0.06mmo1)，甘氨酸甲酯(0.06mmo1),2-(7-偶氮苯并三氮唑)-N,N,N',N'-四甲基脲六氟磷酸酯(0.065mmo1),N,N-二异丙基乙胺(0.05mI)加入二氯甲烷(2mL)中，反应搅拌2h。用薄层层析(TLC)监测反应，反应完后真空旋干溶液,然后加入饱和碳酸氢钠，乙酸乙酯萃取，有机层无水Na 2SO 4干燥，石油醚(PE)：乙酸乙酯(EA)＝(0-100％)过柱，得FL74。 1H NMR(500MHz,CDCl 3)δ8.37(s,1H),7.64(dd,J＝16.2,13.7Hz,1H),7.52-7.35(m,3H),7.04(s,1H),6.83(d,J＝2.9Hz,1H),6.12(dd,J＝15.0,2.9Hz,1H),4.80(s,2H),3.87(s,2H),3.78(d,J＝36.3Hz,4H),3.67(d,J＝40.8Hz,4H),3.34(ddt,J＝111.0,24.9,12.5Hz,4H),2.87(dd,J＝40.2,20.1Hz,1H),1.86(ddtd,J＝62.8,37.3,24.8,12.5Hz,4H),1.42-0.74(m,4H).LRMS(EI)m/z 644(M +).
实施例75 2-(6-(3-(((5-环丙基-3-(2，6-二氯苯基)异恶唑-4-基)甲氧)甲基)-4-氟哌啶-1-基)-1-甲基-1H-吲哚-3-酰胺)乙酸的制备(FL75)
在氮气保护下，向250mL两颈瓶加入2-(6-(3-(((5-环丙基-3-(2，6-二氯苯基)异恶唑-4-基)甲氧)甲基)-4-氟哌啶-1-基)-1-甲基-1H-吲哚-3-酰胺)乙酸甲酯(0.05mmol),后加入2mL甲醇和2mL 1，4-二氧六环溶解，然后加入4N LiOH(4mL)。70℃下搅拌过夜。用薄层层析(TLC)监测反应，反应完后真空旋干溶液,加入水和甲基叔丁基醚。水层用2N HCl调至pH 5，然后加入二氯甲烷CH2Cl2萃取，饱和食盐水洗涤有机层，无水Na2SO4干燥，真空浓缩的粗产品，二氯甲烷(PE)：甲醇(EA)过柱，得FL75。 1H NMR(500MHz,CDCl 3)δ8.30(s,1H),7.77(s,1H),7.58(dd,J＝16.1,13.6Hz,1H),7.53-7.32(m,3H),7.09(d,J＝3.1Hz,1H),6.07(dd,J＝14.9,3.0Hz,1H),4.76(s,2H),3.73(d,J＝20.1Hz,4H),3.61(d,J＝39.4Hz,3H),3.34(dt,J＝24.6,11.3Hz,2H),3.12(dt,J＝24.7,11.2Hz,2H),2.89(p,J＝19.9Hz,1H),2.09-1.58(m,4H),1.40-0.80(m,4H).LRMS(EI)m/z 630(M +).
实施例76 2-(6-(3-(((5-环丙基-3-(2-(三氟甲氧基)苯基)异恶唑-4-基)甲氧)甲基)-3-氟-8-氮杂双环[3.2.1]辛-8-基)-1-甲基-1H-吲哚-3-酰胺)乙酸的制备(FL76)
制备方法同实施例75，制得目标产物FL76。 1H NMR(500MHz,CDCl 3)δ8.43(dd,J＝14.2,3.7Hz,1H),8.37(s,1H),7.57-7.35(m,3H),7.12(dd,J＝14.3,3.8Hz,1H),6.59(d,J＝2.9Hz,1H),6.53(s,1H),6.12(dd,J＝15.0,3.1Hz,1H),4.80(s,2H),4.25(tt,J＝8.6,6.4Hz,2H),3.98(d,J＝50.4Hz,2H),3.74(s,3H),3.60(s,2H),3.04(p,J＝20.1Hz,1H),2.39(ddd,J＝ 50.4,24.7,6.4Hz,2H),2.26-1.93(m,4H),1.75-1.39(m,2H),1.33-0.68(m,4H).LRMS(EI)m/z 671(M +).
实施例77 2-(6-(3-(((5-((2,2-二氟环丙基)甲基)-3-(2-(三氟甲氧基)苯基)异恶唑-4-基)甲氧)甲基)-3-氟氮杂双环[3.2.1]辛-8-基)-4-氟苯并[d]噻唑-6-酰胺)乙酸的制备(FL77)
制备方法同实施例75，制得目标产物FL77。 1H NMR(500MHz,CDCl 3)δ8.43(dd,J＝14.4,3.7Hz,1H),8.17(d,J＝2.9Hz,1H),7.48-7.33(m,2H),7.30-7.06(m,2H),5.33(s,1H),4.80(s,2H),4.51-4.14(m,2H),4.04(d,J＝50.6Hz,2H),3.84(s,2H),3.12(ddd,J＝169.2,24.8,13.8Hz,2H),2.24(ddd,J＝50.4,24.7,6.6Hz,2H),1.99-1.20(m,7H),1.24-0.30(m,2H).LRMS(EI)m/z 743(M +).
实施例78 2-(6-(3-(((5-(2,2-二氟环丙基)-3-(2-(三氟甲氧基)苯基)异恶唑-4-基)甲氧)甲基)-3-氟氮杂双环[3.2.1]辛-8-基)-1-甲基-1H-吲哚-3-酰胺)乙酸的制备(FL78)
制备方法同实施例75，制得目标产物FL78。 1H NMR(500MHz,CDCl 3)δ8.43(dd,J＝14.4,3.7Hz,1H),8.37(s,1H),7.74(s,1H),7.53-7.29(m,3H),7.12(dd,J＝14.2,3.7Hz,1H),6.73(d,J＝2.9Hz,1H),6.12(dd,J＝15.0,3.1Hz,1H),4.88(s,2H),4.47-4.27(m,2H),4.21(d,J＝50.4Hz,2H),3.99-3.80(m,2H),3.74(s,3H),3.60(s,2H),3.14-2.66(m,3H),1.63(ddd,J＝50.4,24.8,13.3Hz,2H),1.41-0.94(m,4H).LRMS(EI)m/z 707(M +).
实施例79 2-(6-(3-(((5-(2,2-二氟环丙基)-3-(2-(三氟甲氧基)苯基)异恶唑-4-基)甲氧)甲基)-3-氟氮杂双环[3.2.1]辛-8-基)-4-氟苯并[d]噻唑-6-酰胺)乙酸的制备(FL79)
制备方法同实施例75，制得目标产物FL79。 1H NMR(500MHz,CDCl 3)δ8.43(dd,J＝14.4,3.7Hz,1H),8.17(d,J＝2.9Hz,1H),7.46(dddd,J＝33.1,29.1,15.5,3.2Hz,3H),7.12(dd,J＝14.2,3.7Hz,1H),6.14(s,1H),4.73(s,2H),4.66(tt,J＝8.6,6.4Hz,2H),4.14(d,J＝50.4Hz,2H),3.84(s,2H),3.08(tt,J＝41.8,23.0Hz,1H),2.46(ddd,J＝50.4,24.9,6.4Hz,2H),2.30-1.67(m,6H),1.41-0.90(m,2H).LRMS(EI)m/z 729(M +).
实施例80 1H NMR(500MHz,CDCl3)LRMS(EI)m/z(M+).2-(6-(3-(((5-((2,2-二氟环丙基)甲基)-3-(2-(三氟甲氧基)苯基)异恶唑-4-基)甲氧)甲基)-3-氟氮杂双环[3.2.1]辛-8-基)-1-甲基-1H-吲哚-3-酰胺)乙酸的制备(FL80)
制备方法同实施例75，制得目标产物FL80。 1H NMR(500MHz,CDCl 3)δ8.43(dd,J＝14.3,3.7Hz,1H),8.37(s,1H),7.61-7.35(m,3H),7.12(dd,J＝14.2,3.7Hz,1H),6.62(d,J＝3.1Hz,1H),6.30(s,1H),6.12(dd,J＝15.0,3.1Hz,1H),4.80(s,2H),4.31(tt,J＝8.6,6.4Hz,2H),4.08(d,J＝50.4Hz,2H),3.93(dd,J＝24.7,15.8Hz,1H),3.74(s,3H),3.60(s,2H),2.61-2.32(m,3H),2.28-2.02(m,4H),1.86-1.44(m,3H),1.17-0.67(m,2H).LRMS(EI)m/z 721(M +).
实施例81 2-(6-(3-(((5-环丙基-3-(2-(三氟甲氧基)苯基)异恶唑-4-基)甲氧)甲基)-3-氟-8-氮杂双环[3.2.1]辛-8-基)-4-氟苯并[d]噻唑-6-酰胺)乙酸的制备(FL81)
制备方法同实施例75，制得目标产物FL81。 1H NMR(500MHz,CDCl 3)δ8.43(dd,J＝14.4,3.7Hz,1H),8.17(d,J＝2.9Hz,1H),7.54(dd,J＝16.0,3.0Hz,1H),7.51-7.32(m,2H),7.12(dd,J＝14.2,3.7Hz,1H),6.21(s,1H),4.80(s,2H),4.55(tt,J＝8.6,6.4Hz,2H),4.01(d,J＝50.4Hz,2H),3.84(s,2H),2.76(p,J＝20.1Hz,1H),2.25(ddd,J＝50.4,24.9,6.4Hz,2H),2.04-1.47(m,6H),1.42-0.48(m,4H).LRMS(EI)m/z 693(M +).
实施例82 2-(6-(3-(((5-环丙基-3-(2，6-二氯苯基)异恶唑-4-基)甲氧)甲基)-3-氟-8-氮杂双环[3.2.1]辛-8-基)-1-甲基-1H-吲哚-3-酰胺)乙酸的制备(FL82)
制备方法同实施例75，制得目标产物FL82。 1H NMR(500MHz,CDCl 3)δ8.36(s,1H),7.63(dd,J＝16.1,13.6Hz,1H),7.52-7.35(m,3H),6.58(d,J＝3.1Hz,1H),6.29-6.01(m,2H),4.79(s,2H),4.18(tt,J＝8.6,6.4Hz,2H),3.93(d,J＝50.4Hz,2H),3.73(s,3H),3.59(s,2H),2.71(p,J＝20.2Hz,1H),2.34-1.90(m,4H),1.82(ddd,J＝50.4,24.9,6.4Hz,2H),1.66-1.38(m,2H),1.28-0.99(m,4H).LRMS(EI)m/z 656(M +).
实施例83 2-(2-(3-(((3-(2,6-二氯苯基)-5-((2,2-二氟环丙基)甲基)异恶唑-4-基)甲氧)甲基)-3-氟-8-氮杂双环[3.2.1]辛-8-基)-4-氟苯并[d]噻唑-6-酰胺)乙酸的制备(FL83)
制备方法同实施例75，制得目标产物FL83。 1H NMR(500MHz,CDCl 3)δ8.15(d,J＝2.9Hz,1H),7.63(dd,J＝16.1,13.7Hz,1H),7.51(dd,J＝16.0,3.0Hz,1H),7.44(dd,J＝14.8,1.1Hz,2H),6.12(s,1H),4.79(s,2H),4.76-4.60(m,2H),4.46(d,J＝50.2Hz,2H),3.83(s,2H),3.06(ddd,J＝80.4,24.8,14.2Hz,2H),2.72(ddd,J＝50.4,24.8,6.7Hz,2H),2.21-1.42(m,7H),1.07-0.47(m,2H).LRMS(EI)m/z 728(M +).
实施例84 2-(6-(3-(((5-环丙基-3-(2，6-二氯苯基)异恶唑-4-基)甲氧)甲基)-3-氟-8-氮杂双环[3.2.1]辛-8-基)-4-氟苯并[d]噻唑-6-酰胺)乙酸的制备(FL84)
制备方法同实施例75，制得目标产物FL84。 1H NMR(500MHz,CDCl 3)δ8.15(d,J＝2.9Hz,1H),7.63(dd,J＝16.1,13.7Hz,1H),7.52(dd,J＝15.9,2.9Hz,1H),7.44(dd,J＝14.9,1.0Hz,2H),6.20(s,1H),4.79(s,2H),4.64(tt,J＝8.4,6.4Hz,2H),4.08(d,J＝50.4Hz,2H),3.83(s,2H),2.73(dd,J＝40.3,20.1Hz,1H),2.16-1.49(m,8H),1.54-0.62(m,4H).LRMS(EI)m/z678(M +).
实施例85 2-(2-(3-(((3-(2,6-二氯苯基)-5-((2,2-二氟环丙基)甲基)异恶唑-4-基)氧)甲基)-3-氟-8-氮杂双环[3.2.1]辛-8-基)-4-氟苯并[d]噻唑-6-酰胺)乙酸的制备(FL85)
制备方法同实施例75，制得目标产物FL85。 1H NMR(500MHz,CDCl 3)δ8.17(d,J＝2.9Hz,1H),7.64(dd,J＝16.1,13.7Hz,1H),7.54(dd,J＝16.0,3.0Hz,1H),7.46(dd,J＝14.9,1.0Hz,2H),6.20(s,1H),4.94(d,J＝50.4Hz,2H),4.58(tt,J＝8.6,6.4Hz,2H),3.84(s,2H),3.33(ddd,J＝92.3,24.7,15.6Hz,2H),2.32(ddd,J＝50.6,24.9,6.4Hz,2H),1.90(ddd,J＝50.4,24.7,6.3Hz,2H),1.79-1.47(m,5H),1.14-0.64(m,2H).LRMS(EI)m/z 714(M +).
实施例86 2-(6-(3-(((5-环丙基-3-(2，6-二氯苯基)异恶唑-4-基)氧)甲基)-3-氟-8-氮杂双环[3.2.1]辛-8-基)-4-氟苯并[d]噻唑-6-酰胺)乙酸的制备(FL86)
制备方法同实施例75，制得目标产物FL86。 1H NMR(500MHz,CDCl 3)δ8.17(d,J＝2.9Hz,1H),7.73(s,1H),7.60(ddd,J＝19.0,16.0,8.3Hz,2H),7.46(dd,J＝14.9,1.0Hz,2H),5.24(d,J＝50.4Hz,2H),4.98-4.62(m,2H),3.84(s,2H),2.74(p,J＝20.2Hz,1H),2.42(ddd,J＝50.4,24.8,6.7Hz,2H),2.24-1.83(m,4H),1.79-1.51(m,2H),1.45-0.64(m,4H).LRMS(EI)m/z 634(M +).
Figure PCTCN2019086068-appb-000049
实施例87 2-(6-(3-(((5-环丙基-3-(2，6-二氯苯基)异恶唑-4-基)甲氧)甲基)-3-氟-8-氮杂双环[3.2.1]辛-8-基)-1-甲基-1H-吲哚-3-酰胺)乙磺酸的制备(FL87)
将2-(6-(3-(((5-环丙基-3-(2，6-二氯苯基)异恶唑-4-基)甲氧)甲基)-4-氟哌啶-1-基)-1-甲基-1H-吲哚-3甲酸(0.1mmol)溶于四氢呋喃(1.0mL)中,加入N-甲基马琳(0.7mmol).在室温搅拌至酸溶解。再加入2-氯-4,6-二甲氧基-1,3,5-三嗪(0.15mmol)在50℃反应 20分钟有沉淀产生。搅拌均匀下加入氨基乙磺酸(0.40mmol，溶于DMA 4mL中)。然后密封在80℃反应2小时。反应完后冷却至室温，加入水，乙酸乙酯萃取。有机层无水Na 2SO 4干燥，旋干，过柱，得纯FL87。 1H NMR(500MHz,CDCl 3)δ8.37(s,1H),7.64(dd,J＝16.1,13.7Hz,1H),7.52-7.38(m,3H),7.19(d,J＝2.9Hz,1H),6.87(s,1H),6.68(s,1H),6.12(dd,J＝15.0,3.1Hz,1H),4.80(s,2H),3.71(t,J＝25.2Hz,5H),3.59-3.05(m,8H),2.73(p,J＝20.1Hz,1H),2.49-1.84(m,4H),1.48-0.65(m,4H).LRMS(EI)m/z 680(M +).
实施例88 2-(6-(3-(((5-环丙基-3-(2，6-二氯苯基)异恶唑-4-基)甲氧)甲基)-4-氟哌啶-1-基)-1-甲基-1H-吲哚-3-酰胺)乙磺酸的制备(FL88)
制备方法同实施例87，制得目标产物FL88。 1H NMR(500MHz,CDCl 3)δ8.37(s,1H),7.61(s,1H),7.54-7.39(m,3H),6.68(s,1H),6.58(d,J＝2.9Hz,1H),6.51(s,1H),6.12(dd,J＝14.9,3.0Hz,1H),4.80(s,2H),4.18(tt,J＝8.6,6.4Hz,2H),3.93(d,J＝50.4Hz,2H),3.74(s,3H),3.56-3.11(m,4H),2.92-2.54(m,1H),2.37-1.92(m,4H),1.79(ddd,J＝50.4,24.7,6.4Hz,2H),1.67-1.41(m,2H),1.38-0.77(m,4H).LRMS(EI)m/z 706(M +).
实施例89 2-(2-(3-(((3-(2,6-二氯苯基)-5-((2,2-二氟环丙基)甲基)异恶唑-4-基)甲氧)甲基)-3-氟-8-氮杂双环[3.2.1]辛-8-基)-4-氟苯并[d]噻唑-6-酰胺)乙磺酸的制备(FL89)
制备方法同实施例87，制得目标产物FL89。 1H NMR(500MHz,CDCl 3)δ8.17(d,J＝2.9Hz,1H),7.71-7.58(m,1H),7.52(dd,J＝15.9,2.9Hz,1H),7.47(d,J＝0.6Hz,1H),7.44(d,J＝1.4Hz,1H),6.66(s,1H),6.41(s,1H),4.80(s,2H),4.79-4.70(m,2H),4.15(d,J＝50.4Hz,2H),3.68(td,J＝16.6,1.5Hz,2H),3.33(td,J＝16.8,1.5Hz,2H),3.19(dd,J＝24.7,16.1Hz,1H),3.04-2.70(m,3H),2.21-1.03(m,8H),0.85-0.66(m,1H).LRMS(EI)m/z 778(M +).
实施例90 2-(6-(3-(((5-环丙基-3-(2，6-二氯苯基)异恶唑-4-基)甲氧)甲基)-3-氟-8-氮杂双环[3.2.1]辛-8-基)-4-氟苯并[d]噻唑-6-酰胺)乙磺酸的制备(FL90)
,其余所需原料、试剂及制备方法同实施例，制得目标产物FL90。 1H NMR(500MHz,CDCl 3)δ8.37(s,1H),7.64(dd,J＝16.1,13.7Hz,1H),7.52-7.38(m,3H),7.19(d,J＝2.9Hz,1H),6.87(s,1H),6.68(s,1H),6.12(dd,J＝15.0,3.1Hz,1H),4.80(s,2H),3.71(t,J＝25.2Hz,5H),3.59-3.05(m,8H),2.73(p,J＝20.1Hz,1H),2.49-1.84(m,2H),1.48-0.65(m,4H).LRMS(EI)m/z 728(M +).
实施例91 2-(6-(3-(((5-环丙基-3-(2-(三氟甲氧基)苯基)异恶唑-4-基)甲氧)甲基)-3-氟-8-氮杂双环[3.2.1]辛-8-基)-1-甲基-1H-吲哚-3-酰胺)乙磺酸的制备(FL91)
制备方法同实施例87，制得目标产物FL91。 1H NMR(500MHz,CDCl 3)δ8.43(dd,J＝14.4,3.7Hz,1H),8.37(s,1H),7.55-7.25(m,3H),7.12(dd,J＝14.2,3.7Hz,1H),6.66(s,1H),6.60(d,J＝2.9Hz,1H),6.54(s,1H),6.12(dd,J＝15.0,3.1Hz,1H),4.80(s,2H),4.14(tt,J＝8.6,6.4Hz,2H),3.97(d,J＝50.4Hz,2H),3.74(s,3H),3.54-3.19(m,4H),2.68(p,J＝20.1Hz,1H),2.48-1.71(m,6H),1.68-0.41(m,6H).LRMS(EI)m/z 721(M +).
实施例92 2-(6-(3-(((5-环丙基-3-(2-(三氟甲氧基)苯基)异恶唑-4-基)甲氧)甲基)-4-氟哌啶-1-基)-1-甲基-1H-吲哚-3-酰胺)乙磺酸的制备(FL92)
制备方法同实施例87，制得目标产物FL92。 1H NMR(500MHz,CDCl 3)δ8.43(dd,J＝14.4,3.7Hz,1H),8.37(s,1H),7.54-7.27(m,3H),7.20(d,J＝3.1Hz,1H),7.12(dd,J＝14.2,3.7Hz,1H),6.70(s,1H),6.52(s,1H),6.12(dd,J＝15.0,3.1Hz,1H),4.80(s,2H),3.76(t,J＝25.2Hz,5H),3.64-3.04(m,8H),2.78(p,J＝20.1Hz,1H),2.14-1.68(m,4H), 1.48-0.77(m,4H).LRMS(EI)m/z 695(M +).
实施例93 2-(6-(3-(((5-(2,2-二氟环丙基)-3-(2-(三氟甲氧基)苯基)异恶唑-4-基)甲氧)甲基)-3-氟氮杂双环[3.2.1]辛-8-基)-4-氟苯并[d]噻唑-6-酰胺)乙磺酸的制备(FL93)
制备方法同实施例87，制得目标产物FL93。 1H NMR(500MHz,CDCl 3)δ8.43(dd,J＝14.4,3.7Hz,1H),8.17(d,J＝2.9Hz,1H),7.54(dd,J＝16.0,3.0Hz,1H),7.51-7.35(m,2H),7.12(dd,J＝14.2,3.7Hz,1H),6.57(d,J＝72.5Hz,2H),4.80(s,2H),4.79-4.73(m,2H),4.27(d,J＝50.4Hz,2H),3.72-2.89(m,6H),2.52(ddd,J＝50.5,24.8,6.6Hz,2H),2.23-1.41(m,7H),1.32-0.40(m,2H).LRMS(EI)m/z(M +).
实施例94 2-(6-(3-(((5-环丙基-3-(2-(三氟甲氧基)苯基)异恶唑-4-基)甲氧)甲基)-3-氟-8-氮杂双环[3.2.1]辛-8-基)-4-氟苯并[d]噻唑-6-酰胺)乙磺酸的制备(FL94)
制备方法同实施例87，制得目标产物FL94。 1H NMR(500MHz,CDCl 3)δ8.37(s,1H),7.64(dd,J＝16.1,13.7Hz,1H),7.52-7.38(m,3H),7.19(d,J＝2.9Hz,1H),6.87(s,1H),6.68(s,1H),6.12(dd,J＝15.0,3.1Hz,1H),4.80(s,2H),3.71(t,J＝25.2Hz,5H),3.59-3.05(m,8H),2.73(p,J＝20.1Hz,1H),2.49-1.84(m,2H),1.48-0.65(m,4H).LRMS(EI)m/z 743(M +).
实施例95 2-(6-(3-(((5-(2,2-二氟环丙基)-3-(2-(三氟甲氧基)苯基)异恶唑-4-基))甲基)-3-氟氮杂双环[3.2.1]辛-8-基)-4-氟苯并[d]噻唑-6-酰胺)乙磺酸的制备(FL95)
制备方法同实施例87，制得目标产物FL95。 1H NMR(500MHz,CDCl 3)δ8.43(dd,J＝14.4,3.7Hz,1H),8.17(d,J＝2.9Hz,1H),7.46(dddd,J＝29.1,18.4,15.6,3.3Hz,3H),7.12(dd,J＝14.2,3.7Hz,1H),6.67(s,1H),6.39(s,1H),4.75-4.47(m,4H),3.89-3.57(m,3H),3.33(td,J＝16.8,1.5Hz,2H),2.82(dd,J＝24.8,16.6Hz,1H),2.52-1.51(m,9H),1.19-0.59(m,2H).LRMS(EI)m/z 779(M +).
实施例96 2-(6-(3-(((5-环丙基-3-(2-(三氟甲氧基)苯基)异恶唑-4-基)氧)甲基)-3-氟-8-氮杂双环[3.2.1]辛-8-基)-4-氟苯并[d]噻唑-6-酰胺)乙磺酸的制备(FL96)
制备方法同实施例87，制得目标产物FL96。 1H NMR(500MHz,CDCl 3)δ8.43(dd,J＝14.4,3.7Hz,1H),8.17(d,J＝2.9Hz,1H),7.45(dddd,J＝33.1,29.1,15.5,3.2Hz,3H),7.12(dd,J＝14.2,3.7Hz,1H),6.65(s,1H),6.50(s,1H),5.00-4.68(m,4H),3.68(td,J＝16.3,1.4Hz,4H),2.94-2.24(m,3H),2.16-1.44(m,6H),1.50-0.74(m,4H).LRMS(EI)m/z 729(M +).
实施例97 6-(4-(((3-(2,6-二氯苯基)-5-(2-(三氟甲基)环丙基)异恶唑-4-基)甲氧)甲基)-4-氟哌啶-1-基)-1-甲基-1H-吲哚-3-甲酸的制备(FL97)
制备方法同实施例1，制得目标产物FL97。 1H NMR(500MHz,CDCl 3)δ9.26(s,1H),7.64(dd,J＝16.1,13.7Hz,1H),7.47(d,J＝0.6Hz,1H),7.44(dd,J＝8.2,6.8Hz,2H),7.21(d,J＝2.9Hz,1H),6.12(dd,J＝15.0,3.1Hz,1H),4.93(s,2H),3.76(s,1H),3.74(s,3H),3.66(s,1H),3.44(dt,J＝24.9,11.3Hz,2H),3.20(dt,J＝24.9,11.3Hz,2H),2.07(ddt,J＝50.4,24.7,11.3Hz,2H),1.91-1.61(m,3H),1.57-1.17(m,2H),0.94(ddd,J＝24.3,21.7,19.5Hz,1H).LRMS(EI)m/z 641(M +).
实施例98 2-(4-(((3-(2,6-二氯苯基)-5-(2-(三氟甲基)环丙基)异恶唑-4-基)甲氧)甲基)-4-氟哌啶-1-基)-4-氟苯并[d]噻唑-6-甲酸的制备(FL98)
制备方法同实施例13，制得目标产物FL98。 1H NMR(500MHz,CDCl 3)δ8.35(d,J＝3.1Hz,1H),7.90(dd,J＝15.9,2.9Hz,1H),7.64(dd,J＝16.1,13.7Hz,1H),7.46(dd,J＝14.9,1.0Hz,2H),4.72(s,2H),4.61(dt,J＝24.9,11.4Hz,2H),4.05(dt,J＝24.7,11.4Hz, 2H),3.79(d,J＝50.4Hz,2H),2.04(ddt,J＝50.5,24.9,11.4Hz,2H),1.90-1.32(m,4H),1.18(ddd,J＝24.5,21.2,19.6Hz,1H),0.96(ddd,J＝24.6,21.7,19.9Hz,1H).LRMS(EI)m/z 663(M +).
实施例99 2-(3-(((3-(2,6-二氯苯基)-5-(2-(三氟甲基)环丙基)异恶唑-4-基)甲氧)甲基)-4-氟哌啶-1-基)-3-氟-8-氮杂双环[3.2.1]辛-8-基)-4-氟苯并[d]噻唑-6-甲酸的制备(FL99)
制备方法同实施例13，制得目标产物FL98。 1H NMR(500MHz,CDCl 3)δ8.35(d,J＝3.1Hz,1H),7.95(dd,J＝15.9,2.9Hz,1H),7.64(dd,J＝16.1,13.7Hz,1H),7.46(dd,J＝14.9,1.0Hz,2H),4.91(s,2H),4.72(tt,J＝8.4,6.4Hz,2H),4.21(d,J＝50.4Hz,2H),2.24(ddd,J＝50.4,24.7,6.4Hz,2H),2.09-1.54(m,7H),1.55-1.35(m,1H),1.24(ddd,J＝24.4,21.2,19.7Hz,1H),0.90(ddd,J＝24.5,21.6,19.7Hz,1H).LRMS(EI)m/z 689(M +).
实施例100 6-(3-氟-3-(((3-(2-(三氟甲氧基)苯基)-5-(2-(三氟甲基)环丙基)异恶唑-4-基)甲氧)甲基)-8-氮杂双环[3.2.1]辛-8-基)-1-甲基-1H-吲哚-3-甲酸的制备(FL100)
制备方法同实施例1，制得目标产物FL100。 1H NMR(500MHz,CDCl 3)δ9.26(s,1H),8.43(dd,J＝14.4,3.7Hz,1H),7.82-7.25(m,3H),7.12(dd,J＝14.2,3.7Hz,1H),6.64(d,J＝2.9Hz,1H),6.12(dd,J＝15.0,3.1Hz,1H),4.92(s,2H),4.45(s,1H),4.41-4.24(m,3H),3.74(s,3H),2.82(ddd,J＝50.4,24.8,6.7Hz,2H),2.05(dddd,J＝47.4,31.5,17.8,7.0Hz,4H),1.72(td,J＝21.4,17.9Hz,1H),1.59-1.15(m,4H),0.89(ddd,J＝24.4,21.7,19.4Hz,1H).LRMS(EI)m/z 682(M +).
实施例101 4-氟-2-(3-氟-3-(((3-(2-(三氟甲氧基)苯基)-5-(2-(三氟甲氧)环丙基)异恶唑-4-基)甲氧)甲基)-8-氮杂双环[3.2.1]辛-8-基)苯并[d]噻唑-6-甲酸的制备(FL101)
制备方法同实施例13，制得目标产物FL101。 1H NMR(500MHz,CDCl 3)δ8.40(dd,J＝14.2,3.7Hz,1H),8.32(d,J＝2.9Hz,1H),7.92(dd,J＝16.0,3.0Hz,1H),7.53-7.31(m,2H),7.09(dd,J＝14.2,3.7Hz,1H),4.75(s,2H),4.61(tt,J＝8.6,6.4Hz,2H),4.29(d,J＝50.4Hz,2H),2.65(ddd,J＝50.4,24.7,6.4Hz,2H),2.37-1.91(m,4H),1.88-1.34(m,4H),1.32-0.80(m,2H)..LRMS(EI)m/z 704M +).
实施例102 6-(4-氟-4-(((3-(2-(三氟甲氧基)苯基)-5-(2-(三氟甲基)环丙基)异恶唑-4-基)甲氧)甲基)哌啶-1-基)-1-甲基-1H-吲哚-3-甲酸的制备(FL102)
制备方法同实施例1，制得目标产物FL102。 1H NMR(500MHz,CDCl 3)δ9.26(s,1H),8.43(dd,J＝14.4,3.7Hz,1H),7.78-7.36(m,3H),7.24-7.01(m,2H),6.12(dd,J＝15.0,3.1Hz,1H),4.91(s,2H),3.74(t,J＝25.2Hz,5H),3.32(ddt,J＝105.9,24.7,11.3Hz,4H),2.22-1.56(m,5H),1.57-0.83(m,3H).LRMS(EI)m/z 656(M +).
药理活性试验实施例
实施例1
筛选方法:
编号:142
名称:FXR-荧光素酶模型激动剂筛选
仪器:
微孔板检测仪：EnVisionTM(PerkinElmer)
材料:
筛选板：ViewPlate-96具有清晰底部的白色96微孔板
检测试剂盒：萤光虫荧光报告基因检测试剂盒(Britelite plus型号)
过程:
将FXR表达质粒和FXRE荧光素酶报告质粒共转染至293T细胞，在96孔平底微孔板中培养细胞并确保质粒表达后，加入FXR激动剂。通过检测荧光素酶荧光信号强度，计算并定量判定激动剂激活程度。其中初筛时，待测化合物和阳性化合物OCA取10μM作用于细胞，分别测定待测化合物对阳性化合物的相对活性，相对活性高于阳性化合物50％的化合物进入复筛，计算其剂量依赖关系，即EC 50值。
样品处理:
样品用DMSO溶解，低温保存，DMSO在最终体系中的浓度控制在不影响检测活性的范围之内。
数据处理及结果说明:
初筛选择单浓度条件下，例如20μg/ml，对样品的活性进行测试。对于在一定条件下表现出活性的样品，例如抑制率％抑制大于50，测试活性剂量依赖关系，即IC 50/EC 50值，通过样品活性对样品浓度进行非线性拟和得到，计算所用软件为Graphpad Prism 4，拟合所使用的模型为sigmoidaldose-response(可变斜率)，对于大多数抑制剂筛选模型，将拟合曲线底部和顶部设定为0和100。一般情况下，每个样品在测试中均设置复孔(n≥2)，在结果中以标准偏差(Standard Deviation,SD)或者标准误差(Standard Error,SE)表示。一般情况下，每次测试均有已报道的化合物作为参照。
Figure PCTCN2019086068-appb-000050
通过活性测试，本发明多个化合物具有优异的药理活性。化合物FL3、FL10等多个化合物在细胞水平的EC 50低于10nM，优于阳性化合物OCA(奥贝胆酸)和FL4-13，表现出更强的药理活性。
FL4-13：
Figure PCTCN2019086068-appb-000051
实施例2.硫代乙酰胺(TAA)诱导的大鼠肝纤维化模型实验
为了进一步评价化合物的药效，我们进行了TAA诱导的大鼠肝纤维化模型实验。
造模方法为：6-8周龄的SD雄性大鼠，腹腔注射150mg/kg TAA生理盐水溶液， 每周三次，连续注射4周后，开始给予化合物FL4-14(FL10)、FL4-15(FL11)(20mg/kg P.O qd)，相同给药剂量的OCA和FL4-13(LY-2562175)作为阳性对照，给药四周，给药期间继续给予TAA造模。该模型能观察到肝脏系数增加、血清碱性磷酸酶(ALP)水平升高、肝脏内α-平滑肌肌动蛋白(α-SMA)、Ⅰ型胶原蛋白(Col1α1mRNA)表达的上调以及病理切片中(天狼猩红染色)肝脏胶原的含量增加。
实验结果表明：化合物中FL4-14(FL10)能明显降低血清中ALP水平(图1)、下调Col1α1mRNA表达(图2)。化合物FL4-14(FL10)减少肝脏病理切片中胶原的含量，表现出同阳性OCA相当的抗肝纤维化效果，但优于对照化合物FL4-13(LY-2562175)(图3)。且FL4-14对大鼠血清内胆固醇(TC)、高密酯蛋白(HDL)、低密脂蛋白(LDL)含量无明显影响，而阳性药物OCA能显著升高大鼠血清内TC、HDL、LDL含量的作用，提示FL4-14(FL10)可能可以避免临床上OCA会引起LDL升高而带来的副作用，也优于阳性对照化合物FL4-13(图4)。另外，FL4-14(FL10)组肝脏系数无明显变化，也避免了阳性药物OCA带来的肝脏系数显著增大的作用(图5)。
实施例3.药代动力学测试
对挑选化合物FL36与对照化合物FL4-13进行了初步药代动力学测试。
1.材料
1.1动物与分组
雄性SD大鼠，12只。
1.2药品与试剂
FL4-13；FL36；DMSO(二甲亚砜)；EtOH；PEG300(聚乙二醇)；NaCl；HPMC(羟丙基甲基纤维素)
2.方法
2.1给药方法
Figure PCTCN2019086068-appb-000052
2.2大鼠药代动力学参数如下表
Figure PCTCN2019086068-appb-000053
Figure PCTCN2019086068-appb-000054
分析数据可以看出：在同等给药剂量下，化合物FL36在血浆中的暴露量(AUC)远高于阳性对照化合物FL4-13，呈现出良好的药代动力学性质。
实施例4.小鼠急性毒性初步试验
1.材料
1.1动物与分组
雌性C57小鼠，6-8周龄，6只，SPF级。
1.2药品与试剂
FL4-14(FL10)(MW:572.45)
DMSO，Tween80，MC
2.方法
2.1实验过程
取雌性ICR小鼠6只，禁食不禁水12小时后，口服灌胃给予受试物FL4-14 500mg/kg，给药体积为10mL/kg，溶剂为10％DMSO+10％Tween80+80％0.5％MC。给药后立即观察小鼠的毒性反应情况，连续观察7天。
2.2动物体重及给药后观察表如下
Figure PCTCN2019086068-appb-000055
Figure PCTCN2019086068-appb-000056
结论：该初步急毒实验主要是观察化合物500mg/kg给药剂量有无明显中毒反应或死亡，实验结果表明：本申请的化合物具有良好的安全性。
在本发明提及的所有文献都在本申请中引用作为参考，就如同每一篇文献被单独引用作为参考那样。此外应理解，在阅读了本发明的上述讲授内容之后，本领域技术人员可以对本发明作各种改动或修改，这些等价形式同样落于本申请所附权利要求书所限定的范围。

Claims (11)

  1. 一种具有如下通式I所示结构的含氟异恶唑类化合物或其对映异构体或非对映异构体，或其可药用盐，或它们的混合物：
    Figure PCTCN2019086068-appb-100001
    其中：
    m为0、1、2或3；
    n为0、1或2；
    p为0、1或2；
    Figure PCTCN2019086068-appb-100002
    环选自下组：取代或未取代的6-20元杂环基、取代或未取代的6-20元芳环基、取代或未取代的6-20元芳杂环基；其中，所述的取代指基团上的氢原子被1、2、3或4个选自下组的取代基取代：氘(D)、氚(T)、卤素、C 1-C 6烷基、卤代C 1-C 6烷基、C 1-C 6烷氧基、卤代C 1-C 6烷氧基、-O[(CH 2) qO] rR 5、-O(CH 2) sC 6-C 10芳基、C 3-C 8环烷氧基、卤代C 3-C 8环烷氧基、氰基、硝基、氨基、胺基(较佳地C 1-C 6胺基)、羟基、羧基、C 1-C 6酯基、C 6-C 10芳基、C 6-C 10芳基氧基、-X 4-CO 2R 5、C 1-C 6烷基羟基、-X 4-CONR 5R 6、-X 4-CONR 5(CH 2) y1CO 2R 6、-X 4-CONR 5(CH 2) y2SO 3R 6、3-12元杂环基和3-12元杂环基氧基；其中，所述的q、r、s、y1和y2可分别为1、2、3或4；所述的芳杂环基、芳环基或杂环基各自独立地含有1～4个选自氧、硫和氮中的杂原子；X 4为键、C1-C6亚烷基、C 1-2烯烃基、环丙基或环氧乙基；
    R 1选自下组：取代或未取代的6-20元杂环基、取代或未取代的6-20元芳环基、取代或未取代的6-20元芳杂环基；其中，所述的取代指基团上的氢原子被1、2、3或4个选自下组的取代基取代：氘、氚、卤素、C 1-C 6烷基、C 1-C 6烷氧基、卤代C 1-C 6烷氧基、-O[(CH 2) qO] rR 5、-O(CH 2) sC 6-C 10芳基、C 3-C 8环烷氧基、卤代C 3-C 8环烷氧基、氰基、硝基、氨基、胺基(较佳地C 1-C 6胺基)、羟基、羟甲基、羧基、C 6-C 10芳基、C 6-C 10芳基氧基、3-12元杂环基和3-12元杂环基氧基；其中，所述的q、r和s可分别为1、2、3或4；所述的芳杂环基、芳基或杂环基各自独立地含有1～4个选自氧、硫和氮中的杂原子；
    R 2可独立地选自1、2、3或4个下组的取代基：氢、卤素、取代或未取代的C 1-C 6烷基、取代或未取代的C 1-C 6烷氧基、-O[(CH 2) qO] rR 5、-O(CH 2) sC 6-C 10芳基、C 3-C 8环烷氧基、卤代C 3-C 8环烷氧基、氰基、硝基、氨基、胺基(较佳地C 1-C 6胺基)、羟基、羟甲基、羧基；其中，所述的取代指基团上的氢原子被1、2、3或4个选自下组的取代基取代：氢、卤素、硝基、氰基、三氟甲基、三氟乙基、三氟丙基、三氟甲氧基、三氟乙氧基、氨基、胺基(较佳地C 1-C 6胺基)、羟基、羟甲基、羧基、巯基和磺酰基；
    R 3和R 4各自独立地选自下组：氢、卤素、C 1-C 6烷基、卤代C 1-C 6烷基、C 1-C 6烷氧基、卤代C 1-C 6烷氧基、-O[(CH 2) qO] rR 5、-O(CH 2) sC 6-C 10芳基、C 3-C 8环烷氧基、卤代C 3-C 8环烷氧基、氰基、硝基、氨基、胺基(较佳地C 1-C 6胺基)、羟基、羟甲基和羧基； 或者R 3和R 4连接在一起共同形成C 1-C 6亚烷基；
    R 5和R 6为自独立地氢、C 1-C 6烷基、卤代C 1-C 6烷基、C 2-C 6烯基、羟甲基或5-7元杂环基。
  2. 根据权利要求1所述的含氟异恶唑类化合物或其对映异构体或非对映异构体，或其可药用盐，或它们的混合物，其特征在于，
    Figure PCTCN2019086068-appb-100003
    环选自下组：苯基、吡啶基、嘧啶基、哒嗪基、噻唑基、苯并噻唑基、苯并[d]异噻唑基、咪唑并[1,2-a]吡啶基、喹啉基、1H-吲哚基、吡咯并[1,2-b]哒嗪基、苯并呋喃基、苯并[b]苯硫基、1H-吲唑基、苯并[d]异恶唑基、噻唑啉基、1H-吡咯并[3,2-c]吡啶基、吡唑并[1,5-a]嘧啶基、咪唑[1,2-b]哒嗪基、吡唑并[1,5-a]吡咯基；上述基团为取代或未取代的；其中，所述的取代指基团上的氢原子被1、2、3或4个选自下组的取代基取代：D、T、卤素、C 1-C 6烷基、卤代C 1-C 6烷基、C 1-C 6烷氧基、卤代C 1-C 6烷氧基、-O[(CH 2) qO] rR 5、-O(CH 2) sC 6-C 10芳基、C 3-C 8环烷氧基、卤代C 3-C 8环烷氧基、氰基、硝基、氨基、胺基(较佳地C 1-C 6胺基)、羟基、羧基、C 1-C 6酯基、C 6-C 10芳基、C 6-C 10芳基氧基、-X 4-CO 2R 5、C 1-C 6烷基羟基、-X 4-CONR 5R 6、-X 4-CONR 5(CH 2) y1CO 2R 6、-X 4-CONR 5(CH 2) y2SO 3R 6、3-12元杂环基和3-12元杂环基氧基；
    其中，所述的q、r、s、y1和y2可分别为1、2、3或4；
    芳杂环、芳基或杂环基各自独立地含有1～4个选自氧、硫和氮中的杂原子；
    X 4为键、C1-C6亚烷基、C 1-2烯烃基、环丙基、环氧乙基；
    R 5和R 6各自独立地为氢、C 1-C 6烷基、卤代C 1-C 6烷基、C 2-C 6烯基、羟甲基或5-7元杂环基。
  3. 根据权利要求1所述的含氟异恶唑类化合物或其对映异构体或非对映异构体，或其可药用盐，或它们的混合物，其特征在于，
    Figure PCTCN2019086068-appb-100004
    选自下组：
    Figure PCTCN2019086068-appb-100005
    其中，
    R 8选自下组：氢、卤素、C 1-C 6烷基、卤代C 1-C 6烷基、C 1-C 6烷氧基、C 1-C 6烷氧基羰基、卤代C 1-C 6烷氧基、C 2-C 6烯基、C 2-C 6炔基、C 3-C 8环烷基、氰基、硝基、氨基、羟基、羟甲基、羧基、巯基、磺酰基、C 6-C 10芳基和3-12元杂环基；
    t为0、1、2、3、4或5；
    X l和X 2各自独立地为N、CH或CR 8；
    X 3为O或S；
    R 9为-X 4-CO 2R 5、C 1-C 6烷基、卤代C 1-C 6烷基、C 1-C 6羟基、-X 4-CONR 5R 6、 -X 4-CONR 5(CH 2) y1CO 2R 6、-X 4-CONR 5(CH 2) y2SO 3R 6；y1和y2各自独立地为1、2、3或4；X 4为键、C1-C6亚烷基、C 1-2烯烃基、环丙基或环氧乙基；
    R 5和R 6各自独立地为氢、C 1-6烷基、卤代C 1-C 6烷基、C 2-C 6烯基、羟甲基或5-7元杂环基；
    R 10为氢、C 1-C 6烷基羟基，C 1-C 6烷基、卤代C 1-C 6烷基、C 1-C 6烷氧基、C 1-C 6烷氧基羰基、卤代C 1-C 6烷氧基、C 2-C 6烯基、C 2-C 6炔基或C 3-C 8环烷基。
  4. 根据权利要求1所述的含氟异恶唑类化合物或其对映异构体或非对映异构体，或其可药用盐，或它们的混合物，其特征在于，
    Figure PCTCN2019086068-appb-100006
    选自下组：
    Figure PCTCN2019086068-appb-100007
    其中，R 8、R 9、R 10和t定义同前。
  5. 根据权利要求1所述的含氟异恶唑类化合物或其对映异构体或非对映异构体，或其可药用盐，或它们的混合物，其特征在于，R 1选自下组：苯基、吡啶基、1-氧吡啶基、环己基、环戊基、环丙基；上述基团为取代或未取代的；其中，所述的取代指基团上的氢原子被1、2、3或4个选自下组的取代基取代：氢、卤素、硝基、氰基、三氟甲基、三氟乙基、三氟丙基、三氟甲氧基、三氟乙氧基、苯基、甲氧基羰基、乙氧基羰基、丙氧基羰基、C 1-6烷基、C l-6环烷基、C 1-6烷氧基、C l-6环烷氧基、环丙基。
  6. 根据权利要求1所述的含氟异恶唑类化合物或其对映异构体或非对映异构体，或其可药用盐，或它们的混合物，其特征在于，所述化合物选自下组：
    Figure PCTCN2019086068-appb-100008
    Figure PCTCN2019086068-appb-100009
    Figure PCTCN2019086068-appb-100010
    Figure PCTCN2019086068-appb-100011
  7. 一种药物组合物，其特征在于，其包含治疗有效量的权利要求1-6任一项所述的含氟异恶唑类化合物或其对映异构体或非对映异构体，或其可药用盐，或它们的混合物，以及药学上可接受的载体。
  8. 一种FXR激动剂，其特征在于，其包含权利要求1-6任一项所述的含氟异恶唑类化合物或其对映异构体或非对映异构体，或其可药用盐，或它们的混合物。
  9. 权利要求1-6任一项所述的含氟异恶唑类化合物或其对映异构体或非对映异构体，或其可药用盐，或它们的混合物的用途，其特征在于，用于制备预防或治疗FXR介导的的疾病的药物。
  10. 如权利要求9所述的用途，其特征在于，所述疾病选自下组：非酒精性脂肪肝病、非酒精性脂肪性肝炎、脂肪肝、肝纤维化、原发性胆汁性肝硬化、高血脂症、血脂异常。
  11. 如权利要求1-6任一项所述的含氟异恶唑类化合物的制备方法，其特征在于，
    (a)所述方法包括步骤：
    Figure PCTCN2019086068-appb-100012
    其中，
    Figure PCTCN2019086068-appb-100013
    R 1、R 2、R 3、R 4、n、m和p的定义与权利要求1所述的通式I中的定义相同；X为卤素或硝基；
    步骤j：在有机溶剂中，在18-冠醚-6和碱存在下，中间体12和中间体9进行反应，形成中间体13；
    步骤k：在有机溶剂中，将中间体13进行脱保护反应，形成中间体14；
    步骤l：在氮气保护下，在有机溶剂中，在碱、钯催化剂和配体的存在下，中间体14和中间体15反应，形成终产物I；或
    步骤m：在氮气保护下，在有机溶剂中，在碱存在下，中间体14和中间体15反应，形成终产物I；
    或
    (b)所述方法包括步骤：
    Figure PCTCN2019086068-appb-100014
    其中，R 1、R 2、R 3、R 4、n、m和p的定义与权利要求1所述的通式I中的定义相同；X为卤素或硝基；
    步骤j：在有机溶剂中，在18-冠醚-6和碱存在下，中间体12和中间体9进行反应，形成中间体13；
    步骤k：在有机溶剂中，将中间体13进行脱保护反应，形成中间体14；
    步骤l：在氮气保护下，在有机溶剂中，在碱、钯催化剂和配体的存在下，中间体14和中间体15反应，形成中间体I’；或
    步骤m：在氮气保护下，在有机溶剂中，在碱存在下，中间体14和中间体15反应，形成中间体I’；和
    步骤e：在有机溶剂中，中间体I’和HNR 5R 6、HNR 5(CH 2) y1CO 2R 6、或HNR 5(CH 2) y2SO 3R 6进行反应，形成终产物I；
    其中，
    Figure PCTCN2019086068-appb-100015
    环选自下组：取代或未取代的6-20元杂环基、取代或未取代的6-20元芳环基、取代或未取代的6-20元芳杂环基；其中，所述的取代指基团上的氢原子被1、2、3或4个选自下组的取代基取代：-X 4-CO 2R 5；其中，X 4、R 5定义同前
    Figure PCTCN2019086068-appb-100016
    环选自下组：取代或未取代的6-20元杂环基、取代或未取代的6-20元芳环基、取代或未取代的6-20元芳杂环基；其中，所述的取代指基团上的氢原子被1、2、3或4个选自下组的取代基取代：-X 4-CONR 5R 6、-X 4-CONR 5(CH 2) y1CO 2R 6、-X 4-CONR 5(CH 2) y2SO 3R 6；其中，X 4、R 5、R 6、y1、y2定义同前。
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