fxr receptor agonist

专利摘要:
The present invention belongs to the technical area of pharmaceutical products, and relates in particular to a compound of formula (I), its pharmaceutically acceptable salt, an ester thereof or a stereoisomer of the compound, the salt or ester, wherein R1, X1 , X2, M, Ar, ring A, ring B and L are as defined in the specification. The present invention also relates to: a method of preparation for the compound, a pharmaceutically acceptable salt thereof, an ester thereof or a stereoisomer of the compound, the salt or ester; a pharmaceutical composition and pharmaceutical formulation containing the compound, the pharmaceutically acceptable salt, the ester or the stereoisomer of the compound, the salt or the ester; and a use of the compound, the pharmaceutically acceptable salt, the ester or the stereoisomer of the compound, the salt or the ester in the preparation of a medicament for treating and / or preventing FXR-mediated diseases.
公开号:BR112020000180A2
申请号:R112020000180-5
申请日:2018-07-06
公开日:2020-07-14
发明作者:Wenkui Ken Fang；Bo Chen；Tingzhong Wang；Jing Cheng
申请人:Xuanzhu (Hainan) Biopharmaceutical Co., Ltd.；
IPC主号:

专利说明:

[001] [001] The present invention belongs to the technical field of pharmaceuticals and relates particularly to an FXR agonist, a method of preparing the FXR agonist, a pharmaceutical formulation containing the FXR agonist and a use of the FXR agonist. CONTEXT OF THE INVENTION
[002] [002] The farnesoid X receptor (FXR) is a member of the family of nuclear receptors for ligand-activated transcription factors and has a typical nuclear receptor structure, namely a highly amino-terminal DNA binding domain (DBD) conserved, a carboxy-terminal ligand-binding domain (LBD), an amino-terminal ligand-independent transcriptional activation (AF1) domain, a carboxy-terminal ligand-dependent (AF2) transcriptional activation function domain and a region hinge. FXR and a retinoid X receptor (RXR) can form a heterodimer. After a linker binds to the LBD region of the FXR, the conformation of the FXR can be altered and the DNA binding domain of the FXR binds to the FXR response element (IR-1) of a target gene promoter to release co-compressors (for example, NCOR) and recruit co-activators, thus playing the role of transcriptional regulation.
[003] [003] FXR is expressed in several tissues and organs, including adipose tissue, liver, gastrointestinal tract and kidneys; of these, the liver has the most abundant FXR expression. The FXR signaling pathway can regulate the expression of several downstream genes, such as BSEP, SHP, CYP7A1, FGFR4, OSTα / β and SREBP-1C, in order to regulate several metabolic pathways, such as triglyceride, cholesterol, sugar metabolism in blood and cholic acid for energy stability of the metabolism, and therefore FXR has the function of treating cancers, non-alcoholic fatty liver diseases (NAFLD), metabolic disorders, inflammation and other diseases. As the main regulator of cholic acid homeostasis, FXR can regulate cholic acid metabolism by inhibiting its synthesis, binding and transport.
[004] [004] Some natural compounds of cholic acid, such as chenodeoxycholic acid (CDCA), deoxycholic acid (DCA) and lithocholic acid (LCA), as well as their taurine and glycine conjugates, can activate FXR. In addition to natural compounds, FXR agonists currently developed in the world can be divided into two main categories. One category is comprised of steroids represented by Intercept's obeticolic acid (OCA), which was approved in May 2016 for the treatment of primary cholangetic cirrhosis and non-alcoholic fatty liver disease and is in Phase III clinical trials. However, itching and other adverse reactions have been observed in clinical studies of steroids. The other category is the new molecular entities, including compounds developed initially, such as GW4604 (WO2000 / 037077). While GW4604 has a strong agonist activity, it has photolability and low bioavailability. PX-104 (WO2011020615A1) from Phenex has been transferred to Gilead and is in Phase II clinical trials.
[005] [005] In addition, GS-9674, developed by Gilead, and LJN-452, developed by Novartis, whose structures are unknown, are in Phase II clinical trials, and their indications include primary cholangetic cirrhosis, primary sclerosing cholangitis and non-alcoholic fatty liver disease.
[006] [006] A category of FXR agonists (see patent application WO2012087519A1) has been disclosed by Tully et al and, specifically compounds 30-70, have been disclosed.
[007] [007] Currently, it is of great clinical relevance the development of new FXR agonists that have high efficiency, low toxicity and good stability, thus enriching the varieties of drugs. SUMMARY OF THE INVENTION
[008] [008] The present invention relates to a compound with a new molecular entity, which can effectively agitate the FXR, increase the expression levels of the BSEP and SHP genes and, at the same time, efficiently suppress the expression of the CYP7A1 gene.
[009] [009] In one aspect, the present invention provides a compound in the form of an FXR agonist.
[010] [010] The present invention provides a compound of general formula (I), a pharmaceutically acceptable salt thereof, an ester thereof or a stereoisomer of the compound, the salt or ester, wherein R1 is selected from the group consisting of halogen, hydroxyl, amine, cyano, C1-6 alkyl, haloC1-6 alkyl, C1-6 hydroxy alkyl, amino C1-6 alkyl, C1-6 alkoxy, C1-6 alkylamine, C1-6 alkylcarbonyl, C1-6 alkoxy C1-6 alkyl, 3-8 membered cycloalkyl, 3-8 membered cycloalkyl C1-6 alkyl, 3-8 membered cycloalkyl C1-6 alkoxy, 3-8 membered heterocyclic, 3-8 membered heterocyclylC1- alkyl 6, or 3-8 membered C 1-6 alkoxy heterocyclyl;
[011] [011] In some embodiments, R1 is selected from a group consisting of halogen, hydroxyl, amine, cyano, C1-4 alkyl, haloC1-4 alkyl, hydroxy C1-4 alkyl, amino C1-4 alkyl, C1-4 alkoxy, C1-4 alkylamine, C1-4 alkylcarbonyl, C1-4 alkoxy C1-4 alkyl, 3-6 membered cycloalkyl, 3-6 membered cycloalkyl C1-4alkyl, 3-6 membered cycloalkyl C1 alkoxy -4, 3-6 membered monoheterocyclic, 3-6 membered monoheterocyclic C1-4 alkyl or 3-6 membered C1-4 alkoxy monoheterocyclyl.
[012] [012] In some embodiments, X1 and X2 are selected individually and independently from a group consisting of N, NR2, O, S or CR3R4; R2, R3 and R4 are selected individually and independently from a group consisting of hydrogen, halogen, hydroxyl, amine, cyano, C1-4 alkyl, haloC1-4 alkyl, C1-4 alkoxy or C1-4 alkylamine.
[013] [013] In some embodiments, M is C1-4 alkylene, one or more carbon atoms in C1-4 alkylene are optionally replaced by a heteroatom or a group, and the heteroatom or group is selected from a group consisting of N, NH, O, CO, S, SO or SO2.
[014] [014] In some embodiments, ring A is selected from 7-membered bridged cyclyl or 7-membered bridged nitrogen heterocyclic.
[015] [015] In some embodiments, ring B is selected from fused 8-10 membered heteroaryl and 7-14 membered fused heterocyclic which are optionally substituted by 1 to 2 Q1 and contain 1 to 3 heteroatoms or groups, and the heteroatom or group is independently selected from a group consisting of N, NH, O, S, SO or SO2.
[016] [016] In some embodiments, each Q1 is independently selected from a group consisting of halogen, hydroxyl, amine, cyano, C1-4 alkyl, haloC1-4 alkyl, hydroxy C1-4 alkyl, amino C1-4 alkyl, C1-4 alkoxy, C1-4 alkylamine, C1-4 alkylcarbonyl, C1-4 alkylsulfonyl, C1-4 alkylsulfinyl, 3-6 membered cycloalkyl, 3-6 membered cycloalkyl C1-4alkyl, 3-6 cycloalkyl C1-4 alkoxy members, 3-6-membered mono-heterocyclic, 3-6-membered mono-heterocyclyl or C1-4-alkoxy 3-6 membered mono-heterocyclyl.
[017] [017] In some embodiments, L is absent or is C1-4 alkylene, where one or more carbon atoms in C1-4 alkylene are optionally replaced by a heteroatom or a group, and the heteroatom or group is selected from a group consisting of NH, O, CO, S, SO or SO2.
[018] [018] In some embodiments, Ar is selected from a group consisting of 6-8 membered monocycloaryl, 8-10 membered fused aryl, 6-8 membered 6-8 membered monocycloaryl, 8-10 membered fused aryl -10 members C1-4 alkyl, 6-8 member monocycloaryl C1-4 alkoxy, 8-10 membered fused aryl, C1-4 alkoxy, 5-7 membered monocyclo-heteroaryl, 8-10 membered fused heteroaryl, monocyclic 5-7 membered C1-4 alkyl heteroaryl, 8-10 membered C1-4 alkyl heteroaryl, 5-7 membered C1-4 alkoxy heteroaryl, 8-10 membered C1-4 alkoxy heteroaryl, cycloalkyl 3-8 members and 3-8 membered heterocyclic, which are optionally substituted by 1 to 3 Q2.
[019] [019] In some embodiments, each Q2 is independently selected from a group consisting of halogen, hydroxyl, amine,
[020] [020] The present invention further provides a compound of general formula (II), a pharmaceutically acceptable salt thereof, an ester thereof or a stereoisomer of the compound, the salt or ester, wherein R1, X1, X2, Q1 and Ar are as described in any of the above mentioned embodiments.
[021] [021] In some embodiments, a compound of the general formula (I) or (II), a pharmaceutically acceptable salt thereof, an ester thereof or a stereoisomer of the compound, the salt or ester, wherein R1 is selected from from a group consisting of halogen, hydroxyl, amine, cyano, C1-4 alkyl, haloC1-4 alkyl, C1-4 hydroxy alkyl, amino C1-4 alkyl, C1-4 alkoxy, C1-4 alkylamine, C1- alkylcarbonyl 4, C1-4 alkoxy C1-4 alkyl, 3-4 membered cycloalkyl, 3-4 membered cycloalkyl C1-4 alkyl, 3-4 membered cycloalkyl C1-4 alkoxy, 3-4 membered monoheterocyclyl, mono -3-4 membered heterocyclic C1-4 alkyl or 3-4 membered monoheterocyclic C1-4 alkoxy.
[022] [022] In some embodiments, a compound of the general formula (I) or (II), a pharmaceutically acceptable salt thereof, an ester thereof or a stereoisomer of the compound, the salt or ester, wherein R1 is selected from a group consisting of halogen, methyl, ethyl, propyl, isopropyl, trifluoromethyl, trifluoroethyl, methoxy, ethoxy, propoxy, isopropoxy, methylamine, ethylamine, methoxymethyl, methoxyethyl, ethoxymethyl, cyclopropyl, cyclopropylmethyl, cyclopropylmethyl, cyclopropylethyl, cyclopropylethyl, cyclopropylethyl, cyclopropylethyl , cyclobutylmethoxy, epoxyethyl, epoxyethylmethyl, azacyclopropyl, azacyclopropylmethyl, oxacyclobutyl or azacyclobutyl.
[023] [023] In some embodiments, a compound of the general formula (I) or (II), a pharmaceutically acceptable salt thereof, an ester thereof or a stereoisomer of the compound, the salt or ester, wherein R1 is selected from a group consisting of cyclopropyl, cyclopropylmethyl, cyclopropylethyl, cyclopropylmethoxy, cyclobutyl, cyclobutylmethyl, cyclobutylethyl, cyclobutylmethoxy, epoxyethyl, epoxyethylmethyl, azacyclopropyl, azacyclopropylmethyl, oxacyclobutyl or azacyclobutyl; and preferably, R1 is selected from cyclopropyl or cyclobutyl.
[024] [024] In some embodiments, a compound of the general formula (I) or (II), a pharmaceutically acceptable salt thereof, an ester thereof or a stereoisomer of the compound, the salt or ester, wherein X1 and X2 are selected individually and independently from a group consisting of N, NR2, O or S; R2 is selected from a group consisting of hydrogen, halogen, hydroxyl, amine, methyl, ethyl, propyl, isopropyl or trifluoromethyl; preferably, X1 and X2 are selected individually and independently from a group consisting of N, NH, O and S; and most preferably, X1 and X2 are both O.
[025] [025] In some embodiments, a compound of the general formula (I), a pharmaceutically acceptable salt thereof, an ester thereof or a stereoisomer of the compound, the salt or ester, wherein M is selected from the group consisting of in -CH2-, -CH2-CH2-, -CH2-CH2-CH2-, -CH2-NH-, -CH2-CH2-O- or -CH2-NH-CO-; and preferably, M is selected from the group consisting of -CH2-, - CH2-CH2- or -CH2-CH2-CH2-.
[026] [026] In some embodiments, a compound of general formula (I), a pharmaceutically acceptable salt thereof, an ester thereof or a stereoisomer of the compound, the salt or ester, in which ring A is selected from saturated cyclyl 7-membered bridge or 7-membered saturated nitrogenous heterocyclic, and, if ring A is a saturated nitrogenous heterocyclic, ring A will preferably be linked to L or ring B by a ring nitrogen atom.
[027] [027] In some embodiments, a compound of general formula (I), a pharmaceutically acceptable salt thereof, an ester thereof or a stereoisomer of the compound, the salt or ester, in which ring A is selected from a 7-membered saturated bridged cyclyl or 7-membered saturated heterocyclic containing 1 nitrogen atom and 0 to 1 additional heteroatom or group, the heteroatom or group is selected from a group consisting of N, NH, O , S, CO, SO or SO2, and, when ring A is a saturated 7-membered heterocyclic containing a nitrogen atom and 0 to 1 additional heteroatom or group, ring A is preferably attached to L or ring B by a nitrogen atom in the ring; preferably, ring A is selected from the following groups:,,,,,,,
[028] [028] In some embodiments, a compound of general formula (I), a pharmaceutically acceptable salt thereof, an ester thereof or a stereoisomer of the compound, the salt or ester, in which ring B is 9-molten heteroaryl -10 members containing 1 to 2 heteroatoms or groups and optionally substituted by 1 to 2 Q1, and the heteroatom or group is selected independently from a group consisting of N, NH, O, S, SO or SO2; ring B is preferably attached to L or ring A by a ring carbon atom; each Q1 is independently selected from a group consisting of halogen, hydroxyl, amine, cyano, C1-4 alkyl, haloC1-4 alkyl, C1-4 alkoxy, 3-6 membered cycloalkyl, 3-6 membered cycloalkyl alkyl C1-4, 3-6 membered cycloalkyl C1-4 alkoxy, 3-6 membered monoheterocyclic, 3-6 membered monoheterocyclic C1-4alkyl or 3-6 membered monoheterocyclic; L is absent or is C1-2 alkylene, in which one or more carbon atoms in C1-2 alkylene are optionally replaced by a heteroatom or a group, and the heteroatom or group is selected from a group consisting of NH , O, S or CO.
[029] [029] In some embodiments, a compound of general formula (I), a pharmaceutically acceptable salt thereof, an ester thereof or a stereoisomer of the compound, the salt or ester, in which ring B is selected from groups below, optionally replaced by 1 Q1:,,,,,,
[030] [030] In some embodiments, a compound of general formula (II), a pharmaceutically acceptable salt thereof, an ester thereof or a stereoisomer of the compound, the salt or ester, wherein Q1 is selected from the group consisting of in hydrogen, halogen, hydroxyl, amine, cyano, C1-4 alkyl, haloC1-4 alkyl, C1-4 alkoxy, 3-6 membered cycloalkyl, 3-6 membered cycloalkyl C1-4alkyl, 3-6 membered cycloalkyl C1-4 alkoxy, 3-6 membered mono-heterocyclic, 3-6 membered mono-heterocyclyl C1-4 alkyl or 3-6 membered mono-heterocyclic C1-4 alkoxy; preferably, Q1 is selected from a group consisting of hydrogen, fluorine, chlorine, bromine, hydroxyl, amine, cyano, methyl, ethyl, propyl, isopropyl, fluoromethyl, difluoromethyl, trifluoromethyl, 1-fluoroethyl, 2-fluoroethyl, 1,1 -difluoroethyl, 1,2-difluoroethyl, 2,2-difluoroethyl, 2,2,2-trifluoroethyl, 3,3,3-trifluoropropyl, 1-
[031] [031] In some embodiments, a compound of the general formula (I) or (II), a pharmaceutically acceptable salt thereof, an ester thereof or a stereoisomer of the compound, the salt or ester, in which Ar is selected from a group consisting of phenyl, phenyl C1-4 alkyl, phenyl C1-4 alkoxy, monocycle-heteroaryl of 5-6 members, monocycle-heteroaryl of 5-6 members C1-4 alkyl and monocycle-heteroaryl of 5-6 members -4 alkoxy which are optionally substituted by 1 to 2 Q2, and each Q2 is independently selected from a group consisting of halogen, hydroxyl, amine, cyano, C1-4 alkyl, haloC1-4 alkyl, hydroxyC1-4 alkyl, aminoC1- 4 alkyl, C1-4 alkoxy, C1-4 alkoxy C1-4 alkyl or C1-4 alkylamine; and, preferably, Ar is selected from a group consisting of phenyl, phenylmethyl, phenylethyl, phenylmethoxy, furyl, pyrrilla, thienyl, pyrazolyl, imidazolyl, pyridyl and pyrimidinyl, which are optionally substituted by 1 to 2 Q2, and each Q2 is selected regardless of a group consisting of fluorine, chlorine, bromine, hydroxyl, amine, cyano, methyl, ethyl, propyl, isopropyl, trifluoromethyl, trifluoroethyl, methoxy, ethoxy, propoxy, isopropoxy, methylamine, ethylamine, methoxymethyl, methoxyethyl or ethoxymethyl.
[032] [032] In some embodiments, a compound of the general formula (I) or (II), a pharmaceutically acceptable salt thereof, an ester thereof or a stereoisomer of the compound, the salt or ester, where Ar is selected from among 6-membered phenyl and monocycloheteroaryl, which are optionally substituted by 1 to 2 Q2; preferably, Ar is selected from a group consisting of phenyl, pyridyl and pyrimidinyl, which are optionally substituted by 1 to 2 Q2; where each Q2 is independently selected from a group consisting of fluorine, chlorine, bromine, hydroxyl, amine, cyano, methyl, ethyl, propyl, isopropyl, trifluoromethyl, trifluoroethyl, methoxy, ethoxy, propoxy, isopropoxy, methylamine, ethylamine, methoxymethyl , methoxyethyl or ethoxymethyl, and preferably, each Q2 is independently selected from a group consisting of fluorine, chlorine, bromine, methyl, ethyl, propyl, isopropyl, trifluoromethyl, methoxy or ethoxy.
[033] [033] In some embodiments, a compound of the general formula (II), a pharmaceutically acceptable salt thereof, an ester thereof or a stereoisomer of the compound, the salt or ester, wherein R1 is selected from cyclopropyl or cyclobutyl; X1 and X2 are both O; Ar is selected from a group consisting of phenyl, pyridyl and pyrimidinyl, which are optionally substituted by 1 to 2 Q2; wherein each Q2 is independently selected from a group consisting of fluorine, chlorine, bromine, methyl, ethyl, propyl, isopropyl, methoxy or ethoxy; and Q1 is selected from a group consisting of hydrogen, fluorine, chlorine, bromine, methyl, ethyl, propyl, isopropyl, trifluoromethyl, methoxy, ethoxy, propoxy or isopropoxy.
[034] [034] In some embodiments, a compound of general formula (II), a pharmaceutically acceptable salt thereof, an ester thereof or a stereoisomer of the compound, the salt or ester, wherein R1 is selected from the group consisting of in cyclopropyl, cyclopropylmethyl, cyclopropylethyl, cyclopropylmethoxy, cyclobutyl, cyclobutylmethyl, cyclobutylethyl, cyclobutylmethoxy, epoxyethyl, epoxyethylmethyl, azacyclopropyl, azacyclopropylmethyl, oxacyclobutyl or azacyclobutyl; X1 and X2 are selected individually and independently from a group consisting of N, NH, O or S; Q1 is selected from a group consisting of fluorine, chlorine, bromine, methyl, ethyl, propyl, isopropyl, trifluoromethyl, methoxy, ethoxy, propoxy or isopropoxy; Ar is selected from a group consisting of phenyl, pyridyl and pyrimidinyl, which are optionally substituted by 1 to 2 Q2; and each Q2 is independently selected from a group consisting of methyl, ethyl, propyl, isopropyl, ethoxy or trifluoromethoxy.
[035] [035] In some embodiments, a compound of the general formula (II), a pharmaceutically acceptable salt thereof, an ester thereof or a stereoisomer of the compound, the salt or ester, wherein R1 is cyclopropyl; X1 and X2 are both O; Ar is phenyl optionally substituted by 1 to 2 Q2; each Q2 is independently selected from a group consisting of chlorine, methoxy and trifluoromethoxy; and Q1 is selected from the group consisting of hydrogen, fluorine, methyl and methoxy.
[036] [036] In some embodiments, a compound of general formula (I), a pharmaceutically acceptable salt thereof, an ester thereof or a stereoisomer of the compound, the salt or ester, wherein R1 is selected from the group consisting of in cyclopropyl, cyclopropylmethyl, cyclopropylethyl, cyclopropylmethoxy, cyclobutyl, cyclobutylmethyl, cyclobutylethyl, cyclobutylmethoxy, epoxyethyl, epoxyethylmethyl, azacyclopropyl, azacyclopropylmethyl, oxacyclobutyl or azacyclobutyl; X1 and X2 are selected individually and independently from a group consisting of N, NH, O or S; M is -CH2-; ring A is; ring B is selected from the following groups:, and; L is absent; Ar is selected from a group consisting of phenyl, pyridyl and pyrimidinyl, which are optionally substituted by 1 to 2 Q2; and each Q2 is independently selected from a group consisting of fluorine, chlorine, bromine, methyl, ethyl, propyl, isopropyl, methoxy, ethoxy, trifluoromethyl or trifluoromethoxy.
[037] [037] In some embodiments, a compound of formula (I), a pharmaceutically acceptable salt thereof, an ester thereof or a stereoisomer of the compound, the salt or ester, wherein R1 is cyclopropyl; X1 and X2 are both O; M is -CH2-; ring A is;
[038] [038] Any combination can be achieved between the above mentioned embodiments and between the characteristics involved in the embodiments, and all the resulting technical solutions are listed here and belong to the technical solutions of the present invention.
[039] [039] Some compounds of the present invention are as follows: No. Structural Formula No. Structural Formula 1 2 2- 2-1 2 3 4
[040] [040] In another aspect, the present invention relates to a pharmaceutical composition containing a compound of general formula (I), a pharmaceutically acceptable salt thereof, an ester thereof or a stereoisomer of the compound, the salt or the ester.
[041] [041] In another aspect, the present invention relates to a pharmaceutical formulation containing a compound of general formula (I), a pharmaceutically acceptable salt thereof, an ester thereof or a stereoisomer of the compound, the salt or the ester, in addition to one or more pharmaceutically acceptable carriers and / or diluents, and the pharmaceutical formulation can be of any pharmaceutically acceptable dosage form. The pharmaceutical formulation can be administered to a patient in need of such treatment via oral, parenteral, rectal or pulmonary administration.
[042] [042] In another aspect, the present invention also relates to the use of a compound of general formula (I), a pharmaceutically acceptable salt thereof, an ester thereof or a stereoisomer of the compound, the salt or ester, in the preparation of drugs for the prevention and / or treatment of FXR-mediated diseases and related diseases in subjects.
[043] [043] The present invention further provides a method for the treatment and / or prevention of FXR-mediated diseases and related diseases in subjects, and the method comprises administering to a subject who needs it in an amount therapeutically and / or preventively effective amount of the compound, a pharmaceutically acceptable salt thereof, an ester thereof or a stereoisomer of the compound, the salt or ester, in the present invention or a pharmaceutical composition in the present invention.
[044] [044] As used herein, the term "effective amount" refers to an amount sufficient to fully or at least partially achieve the desired effect. For example, a preventively effective amount for a disease refers to an amount sufficient to prevent, stop or delay the occurrence of a disease; and a therapeutically effective amount for a disease refers to an amount sufficient to cure or at least partially arrest a disease and its complications in a patient suffering from the disease. Determining such an effective amount is completely within the skill of those skilled in the art. For example, an effective amount for treatment depends on the severity of a disease to be treated, the general condition of a patient's immune system, the patient's profile (such as age, body weight and sex), the method of administering the drug, other therapies administered simultaneously and so on.
[045] [045] In another aspect, the present invention also relates to the use of a compound of general formula (I), a pharmaceutically acceptable salt thereof, an ester thereof or a stereoisomer of the compound, the salt or ester, for prevention and / or treatment of FXR-mediated diseases and related diseases in subjects.
[046] [046] In the present invention, FXR-mediated diseases and related diseases include, among others: (1) disorders of lipid or lipoprotein metabolism, such as atherosclerosis, bile acid metabolism disorder, primary sclerosing cholangitis, cholesterol calculus, related diseases to fibrosis, fat in the liver (alcoholic fatty liver disease, non-alcoholic fatty liver disease, etc.), cirrhosis (primary biliary cirrhosis, primary cholangetic cirrhosis etc.), hepatitis (chronic hepatitis, non-viral hepatitis, alcoholic steatohepatitis, steato- non-alcoholic hepatitis etc.), liver failure, cholestasis (benign intrahepatic cholestasis, progressive familial intrahepatic cholestasis, extrahepatic cholestasis etc.), cholelithiasis, myocardial infarction, stroke, thrombus etc., acute liver failure , cholelithiasis and / or inflammatory bowel diseases.
[047] [047] In the present invention, subjects or patients can be any animals and, preferably, mammals, such as cattle, equines, swine, canids, felids, rodents and primates. In particular, the subjects are preferably human.
[048] [048] The present invention also provides a kit that is used to agitate FXR in cells, increase expression of BSEP and expression of SHP in cells and / or suppress expression of CYP7A1 in cells. The kit comprises the compound of the present invention, a pharmaceutically acceptable salt thereof, an ester thereof or a stereoisomer of the compound, the salt or ester and, optionally, includes a manual.
[049] [049] The present invention also provides a use of the compound of the present invention, a pharmaceutically acceptable salt thereof, an ester thereof or a stereoisomer of the compound, the salt or ester, in the preparation of a formulation to agitate the FXR in cells, increasing BSEP expression and SHP expression in cells and / or suppressing CYP7A1 expression in cells. In some embodiments, the formulation is used for in vivo or in vitro administration. For example, the formulation can be administered to a subject's body (for example, a mammal, such as cattle, horses, swine, canines, felids, rodents and primates (such as humans)); or the formulation can be administered to cells in vitro (for example, cell lines or cells derived from a subject).
[050] [050] The present invention also provides a method for agitating FXR in cells, increasing expression of BSEP and expression of PCH in cells and / or suppressing expression of CYP7A1 in cells, and the method comprises administering to cells an effective amount of the compound of the present invention, a pharmaceutically acceptable salt thereof, an ester thereof or a stereoisomer of the compound, the salt or ester. In some embodiments, the method is adopted in vivo, for example, the cells are a subject's in vivo cells (for example, a mammal, such as bovine, equine, swine, canine, feline, rodent, and primate (as a human )); or the method is adopted in vitro, for example, the cells are cells in vitro (for example, cell lines or cells derived from a subject).
[051] [051] The present invention also provides the compound of the present invention, a pharmaceutically acceptable salt thereof, an ester thereof or a stereoisomer of the compound, the salt or ester, used to agitate the FXR in cells, increasing the expression of BSEP and expression of SHP in cells and / or suppressing expression of CYP7A1 in cells.
[052] [052] In some embodiments, cells are cells derived from the liver (e.g., liver cells derived from a subject). In some embodiments, the cells are liver cancer cells or hepatocytes. In some embodiments, the cells are HepG2 cells or AML12 cells.
[053] [053] In the specifications and claims of this application, all compounds are named according to their structural chemical formulas and, if a compound name is not consistent with its structural chemical formula for the same compound, the structural chemical formula or an equation chemistry will prevail. Terms Definitions
[054] [054] In the present application, unless otherwise specified, the scientific and technological terms used herein have meanings generally understood by those skilled in the art. However, definitions and explanations for some of the related terms are provided below for a better understanding of the present invention. In addition, if the definitions and explanations of the terms provided in this application are not consistent with the meanings generally understood by those skilled in the art, the definitions and explanations of the terms provided in the present application shall prevail.
[055] [055] "Halo-", described in the present invention, refers to being replaced by a "halogen atom", and the "halogen atom", described in the present invention, includes fluorine, chlorine, bromine and iodine atoms.
[056] [056] "C1-6 alkyl" described in the present invention, refers to a straight or branched alkyl containing 1 to 6 carbon atoms, including, for example, "C1-5 alkyl", "C1-4 alkyl "," C1-3 alkyl "," C1-2 alkyl "," C2-4 alkyl "," C2-3 alkyl "and" C3-4 alkyl ", and specific examples include, but are not limited to, methyl, ethyl, n-propyl, isopropyl, n-butyl, isobutyl, sec-butyl, tert-butyl, n-pentyl, isopentyl, 2-methylbutyl, neo-pentyl, 1-ethylpropyl, n-hexyl, isohexyl, 3-methylpentyl, 2-methylpentyl, 1-methylpentyl, 3,3-dimethylbutyl, 2,2-dimethylbutyl, 1,1-dimethylbutyl, 1,2-dimethylbutyl, 1,3-dimethylbutyl, 2,3-dimethylbutyl, 2-ethylbutyl, 1, 2-dimethylpropyl etc. "C1-4alkyl", described in the present invention, refers to a specific example containing 1 to 4 carbon atoms for C1-6alkyl.
[057] [057] "HaloC1-6 alkyl", described in the present invention, refers to a group derived from the replacement of one or more hydrogen atoms in C1-6 alkyl by one or more halogen atoms and the "halogen atom "and" C1-6alkyl "are as defined above. "HaloC1-4 alkyl", described in the present invention, refers to a specific example, containing 1 to 4 carbon atoms for haloC1-6 alkyl.
[058] [058] "HydroxyC1-6 alkyl", described in the present invention, refers to a group derived from the substitution of one or more hydrogen atoms in C1-6 alkyl, by one or more hydroxyls, and "C1- alkyl 6 "is as defined above. "C1-4 alkyl hydroxy", described in the present invention, refers to a specific example, containing 1 to 4 carbon atoms for the C 1-6 alkyl hydroxy.
[059] [059] "AminoC1-6 alkyl", described in the present invention, refers to a group derived from the replacement of one or more hydrogen atoms in C1-6 alkyl by one or more amino groups and "C1-6 alkyl "is as defined above. "AminoC1-4 alkyl",
[060] [060] "C1-6 alkoxy, C1-6 alkylamine, C1-6 alkylcarbonyl, C1-6 alkylsulfonyl and C1-6 alkylsulfinyl" described in the present invention refer to groups having C1-6-O- alkyl, C1 alkyl -6-NH-, C1-6 alkyl -C (O) -, C1-6-S (O) 2- alkyl and C1-6-S (O) alkyl - where "C1-6 alkyl" is as defined above. "C1-4 alkoxy C1-4 alkylamine, C1-4 alkylcarbonyl, C1-4 alkylsulfonyl and C1-4 alkylsulfinyl" described in the present invention refer to specific examples containing 1 to 4 carbon atoms in the alkyl groups for the above examples .
[061] [061] "C1-6 alkoxy C1-6 alkyl", described in the present invention, refers to a group derived from the substitution of one or more hydrogen atoms in a C1-6 alkyl by one or more C1-6 alkoxy groups and "C1-6alkyl" is as defined above. "C1-4 alkoxy C1-4 alkyl", described in the present invention, refers to a specific example, containing 1 to 4 carbon atoms for the C1-6 alkoxy C1-6 alkyl.
[062] [062] "CR3R4", described in the present invention, refers to a group formed by the replacement of two hydrogen atoms of a methylene group with R3 and R4, respectively, and the specific bonding form is as.
[063] [063] "C1-6 alkylene", described in the present invention, refers to a group derived from the removal of two hydrogen atoms on carbon atoms other than a linear alkane group containing 1 to 6 carbon atoms, including "alkylene C1-5 "," C1-4 alkylene "," C1-3 alkylene "and" C1-2 alkylene "and specific examples include, but are not limited to, -CH2-, -CH2CH2-, -CH2CH2CH2-, -CH2CH2CH2CH2-, - CH2CH2CH2CH2CH2-, -CH2CH2CH2CH2CH2CH2- etc.
[064] [064] "Any one or more carbon atoms in C1-6 alkylene is optionally replaced by a heteroatom or a group", described in the present invention, means that any one or more carbon atoms in "C1-6 alkylene" can be optionally replaced by one or more heteroatoms or groups, that is, the carbon atoms in C1-6 alkylene cannot be replaced by any heteroatom or group, a carbon atom in C1-6 alkylene can be replaced by a heteroatom or a group or any two carbon atoms in C1-6 alkylene can be replaced by two same or different heteroatoms or groups, or any multiple carbon atoms in C1-6 alkylene can be replaced by the corresponding multiple heteroatoms or the same or different groups; and the heteroatom or group selected from a group consisting of N, NH, O, CO, S, SO or SO2.
[065] [065] "3-8 membered cycloalkyl" described in the present invention refers to a saturated monocyclic alkyl group containing 3 to 8 carbon atoms, including, for example, "3-6 membered cycloalkyl", " 3-5 membered cycloalkyl "," 3-4 membered cycloalkyl "," 4-5 membered cycloalkyl "," 4-6 membered cycloalkyl "," 4-7 membered cycloalkyl "," 4-8 membered cycloalkyl " members "etc.
[066] [066] "Heterocyclic 3-14 members" described in the present invention refers to a group obtained by removing a hydrogen atom from a monocyclic compound or a fused cyclic compound that is saturated or partially saturated and contains 3 to 14 ring atoms and at least one hetero atom (for example, 1, 2, 3, 4 or 5 hetero atoms). The "fused rings" refer to a group formed by two or more cyclic structures that share two atoms adjacent to each other. The "3-14 membered heterocyclic" includes, for example, "3-12 membered heterocyclic", "3-10 membered heterocyclic", "3-8 membered heterocyclic", "3-7 membered heterocyclic", "3-6 membered mono-heterocyclic", "3-4 membered mono-heterocyclic", "4-7 membered mono-heterocyclic", "4-6 membered mono-heterocyclic", "5-membered mono-heterocyclic" -6 members "," fused heterocyclic from 7 to 10 members "and" fused heterocyclic from 7 to 14 members ". "3-14 membered partially saturated heterocyclic" refers to a cyclic group containing double bonds and heteroatoms. "3-14-membered saturated heterocyclic" refers to a cyclic group containing heteroatoms and no unsaturated bonds. Among the specific examples are epoxietila, azaciclopropila, diazaciclopropila, oxaciclobutila, azaciclobutila, 1,4 dioxaciclo-hexyl, 1,3-dioxaciclo-hexyl, 1,3-dioxaciclopentila, tetraidrofurila, tetraidrotienila, pirrolidila, imidazolidinila, pirazolidinila, piperazinyl, morpholinyl , benzodihydrofuril, benzodihydropyranyl, benzo1,4-dioxacyclohexenyl, benzo1,3-dioxacyclohexenyl, benzotetrahydropyridyl, benzodihydrooxazinyl, benzotetrahydropyrazinyl, 1,2,3,4-tetrahydroquinazolinyl, 1,2,3,4-tetrahydrocinolin or tetrahydrocinolin. "3-6 membered heterocyclic" refers to a specific example containing 3 to 6 ring atoms for the 3-14 membered heterocyclic. "3-4 membered mono-heterocyclic" refers to a specific example of a monocyclic heterocyclic containing 3 to 4 ring atoms for the 3-14 membered heterocyclic.
[067] [067] "3-8 membered cycloalkyl C1-6 alkyl" and "3-8 membered cycloalkyl C1-6 alkoxy" described in the present invention refer to groups obtained by substituting hydrogen atoms in a C1-6 alkyl group and a C1-6 alkoxy group by 3-8 membered cycloalkyl groups, and "3-8 membered cycloalkyl", "C1-6 alkyl" and "C1-6 alkoxy" are as defined above.
[068] [068] "Heterocyclic 3-8 member C1-6 alkyl" and "Heterocyclic 3-8 member C1-6 alkoxy" described in the present invention refer to groups obtained by substituting hydrogen atoms in a C1-6 alkyl group and a C1-6 alkoxy group by 3-8 membered heterocyclic groups, and the "3-8 membered heterocyclic", "C1-6 alkyl" and "C1-6 alkoxy" are as defined above.
[069] [069] "3-6 membered C1-4 alkyl cycloalkyl", "3-4 membered C1-4 alkyl cycloalkyl", "3-6 membered C1-4 alkyl mono-heterocyclic" and "3-membered mono-heterocyclic -4 C1-4 alkyl members "described in the present invention refer to groups obtained by replacing a hydrogen atom in a C1-4 alkyl group with a 3-6 membered cycloalkyl group, a 3-4 membered cycloalkyl group, one 3-6 membered mono-heterocyclic group and 3-4 membered mono-heterocyclic group, respectively, and "3-6 membered cycloalkyl", "3-4 membered cycloalkyl", "3-- membered mono-heterocyclic" 6 members, "" 3-4 membered monoheterocyclic "and" C1-4alkyl "are as defined above.
[070] [070] "C1-4 alkoxy 3-6 membered cycloalkyl", "C1-4 alkoxy 3-4 membered cycloalkyl", "C1-4 alkoxy 3-6 membered mono-heterocyclic" and "3-membered mono-heterocyclic -4 members C1-4 alkoxy "described in the present invention refer to groups obtained by replacing a hydrogen atom in a C1-4 alkoxy group with a 3-6 membered cycloalkyl group, a 3-4 membered cycloalkyl group, one 3-6 membered mono-heterocyclic group and 3-4 membered mono-heterocyclic group, and "3-6 membered cycloalkyl", "3-4 membered cycloalkyl", "3-6 membered mono-heterocyclic" "," 3-4 membered monoheterocyclic "and" C1-4 alkoxy "are as defined above.
[071] [071] "6 to 10 membered aryl" described in the present invention refers to an aromatic monocyclic or polycyclic group containing 6 to 10 ring carbon atoms. For example, "6 to 10 membered aryl" includes "6 to 8 membered monocycloaryl", "6 to 7 membered monocycloaryl", "8-10 membered fused aryl" etc. Specific examples include, but are not limited to, phenyl, cyclooctanotetraenyl, naphthyl, etc.
[072] [072] "5-10 membered heteroaryl" described in the present invention refers to a monocyclic or polycyclic aromatic group that contains 5 to 10 ring atoms, where at least one of the ring atoms is a heteroatom, and the heteroatom is a nitrogen atom, an oxygen atom and / or a sulfur atom. "5- to 10-membered heteroaryl" includes, for example, "5-7-membered monocycle-heteroaryl", "5-6-membered monocycle-heteroaryl", "6-membered mono-heteroaryl", "7 to 7-membered heteroaryl 10 members "," 8-10 member fused heteroaryl "and" 9-10 member fused heteroaryl ". Specific examples include furyl, pyrrilla, thienyl, pyrazolyl, imidazolyl, thiazolyl, isothiazolyl, thiadiazolyl, oxazolyl, isooxazolyl, oxadiazolyl, 1,2,3-triazolyl, 1,2,4-triazolyl, 1,2,3-oxadiazolyl , 1,2,4- oxadiazolyl, 1,2,5-oxadiazolyl, 1,3,4-oxadiazolyl, pyridyl, pyrimidinyl, pyridazinyl, pyrazinyl, benzofuryl, benzoisofuryl, benzothienyl, indolyl, isoindolyl, benzooxazolyl, benzoimidazolyl, benzoimidazolyl, benzoimidazolyl, benzoimidazolyl benzotriazoyl, quinolyl, isoquinolyl etc.
[073] [073] "6-10 membered aryl, C1-6 alkyl" and "5-10 membered C1-6 alkyl heteroaryl" described in the present invention refer to groups obtained by substituting a hydrogen atom in an alkyl group C1-6 by a 6-10 membered aryl group and a 5-10 membered heteroaryl group, respectively. "6-10 member aryl", "5-10 member heteroaryl" and "C1-6 alkyl" are as described above.
[074] [074] "6-10 membered aryl, C1-6 alkoxy" and "5-10 membered C1-6 alkoxy heteroaryl" described in the present invention refer to groups obtained by substituting a hydrogen atom in an alkoxy group C1-6 by a 6-10 membered aryl group and a 5-10 membered heteroaryl group, respectively. "6-10 membered aryl", "5-10 membered heteroaryl" and "C1-6 alkoxy" are as described above.
[075] [075] "6-8 membered C1-4alkyl monocycloaryl", "8-10 membered C1-4alkyl fused aryl", "6-1 membered C1-4alkyl monocycle", "5-7 membered monocycloaryl alkyl C1-4 "and" fused heteroaryl of 8-10 members C1-4 alkyl "described in the present invention refer to groups obtained by substituting a hydrogen atom in a C1-4 alkyl group with a monocycloaryl group of 6- 8 members, a fused aryl group of 8-10 members fused aryl, a monocycloaryl group of 6 members, a monocycloheteroaryl group of 5-7 members and a fused heteroaryl group of 8-10 members, respectively.
[076] [076] "6-8-membered C1-4 alkoxy monocycle", "8-10-membered C1-4 alkoxy fused aryl", "6-membered C1-4 alkoxy monocycle", "5-7 membered monocycloaryl alkoxy C1-4 "and" fused heteroaryl of 8-10 members C1-4 alkoxy "described in the present invention refer to groups obtained by substituting a hydrogen atom in a C1-4 alkoxy group with a monocycloaryl group of 6- 8 members, a fused aryl group of 8-10 members fused aryl, a monocycloaryl group of 6 members, a monocycloheteroaryl group of 5-7 members and a fused heteroaryl group of 8-10 members, respectively. "6-8 membered monocycloaryl", "8-10 membered fused aryl", "6 membered monocycloaryl",
[077] [077] "L is absent", described in the present invention, means that ring A and ring B are directly linked by a chemical bond when L is absent.
[078] [078] A broken link in a structural formula or a group of the present invention represents presence or absence, for example, for the group, it encompasses and.
[079] [079] "7-membered bridged cyclyl" described in the present invention refers to a saturated or partially saturated ring cyclic structure that contains 7 carbon atoms in the ring and is formed by two or more cyclic structures that share two non-adjacent atoms in the ring. For example, "bridged 7-membered cyclyl" includes "saturated 7-membered cyclyl" and specific examples include, but are not limited to,,,, etc. "7-membered bridged saturated cyclyl" refers to a specific example of a 7-membered bridged saturated cyclyl.
[080] [080] "7-membered bridged heterocyclic" described in the present invention refers to a saturated or partially saturated cyclic structure that contains 7 ring atoms (where at least one of the ring atoms is a heteroatom or group, such as N, NH, O, S, CO, SO, SO2) and is formed by two or more cyclic structures sharing two non-adjacent atoms of the ring, and the number of heteroatoms or groups is preferably 1, 2, 3, 4 or 5 , and more preferably 1 or 2. For example, "7-membered bridged heterocyclic" includes "7-membered saturated heterocyclic", "7-membered saturated nitrogenous heterocyclic" and the like. Specific examples include, but are not limited to,,
[081] [081] "Cys" or "trans" shown in the structural formula of the compound of the present invention means the positional relationship between the main bridge (a bridge having the minimum carbon atoms) in the bridge ring A and the corresponding substituent on the structure. Taking compound 1 as an example, compound 1 has a cis structure and has a specific structural formula shown as follows: Cis- where the structural formula means that compound 1 is a racemate containing two enantiomers with the following structural formulas, respectively : and .
[082] [082] In the structural name of the compound of the present invention, "(1RS, 4RS, 5SR)" represents that the compound is a racemate comprising two enantiomers, where the absolute configurations of the two enantiomers are "1R, 4R, 5S" and "1S, 4S, 5R", respectively.
[083] [083] "Optionally", described in the present invention, means that it may or may not exist. For example, "ring B is selected from ... optionally substituted by one or more Q1", described in the present invention, includes the case where ring B is not replaced by any Q1, and the case where ring B is replaced by one or more Q1.
[084] [084] "Partially saturated", described in the present invention, means that the related group contains at least one double bond or triple bond.
[085] [085] The present invention also provides a method of preparation for the compound of formula (I), comprising, but not limited to, the process route to follow.
[086] [086] Each acronym is defined as follows: DMA: N, N-dimethylacetamide; THF: tetrahydrofuran; DCM: dichloromethane; TFA: trifluoroacetic acid; EA: ethyl acetate; PE: petroleum ether; MeOH: methanol.
[087] [087] The specific exemplary steps are as follows: (1) preparation of intermediate 1
[088] [088] Raw material 2, potassium tert-butoxide, 18-crown-6 and KI are added to an organic solvent, and raw material 1 is added next; the mixing solution reacts at 25-60 ° C; and after the reaction is complete, the reaction solution is concentrated and purified by column chromatography to obtain intermediate 1. The organic solvent is preferably tetrahydrofuran.
[089] [089] Intermediate 1 is added slowly to a solution containing an acidic reaction substance; after the reaction is complete, the pH of the reaction solution is adjusted to 7 to 8 with an alkaline solution; and the system is dried by centrifugation and concentrated to obtain intermediate 2. The solution containing an acidic substance is preferably a solution of hydrochloric acid in ethanol, a solution of trifluoroacetic acid in dichloromethane etc., and the alkaline solution is preferably a solution saturated sodium bicarbonate.
[090] [090] Intermediate 2, intermediate 3 and cesium carbonate are added to an organic solvent for microwave reaction or heating reaction; and after the reaction is complete, the reaction solution is purified by column chromatography to obtain intermediate 4. The organic solvent is preferably DMA.
[091] [091] Intermediate 4 is added to an organic solvent, and then an aqueous solution containing an alkaline substance is added for the heating reaction; after the reaction is complete, the pH of the reaction solution is adjusted to 4 to 6 with a solution of acidic substance; and the reaction solution is dried by centrifugation and purified by column chromatography to obtain a compound of formula (I). The organic solvent is preferably a mixed solution of methanol and tetrahydrofuran; the alkaline substance is preferably lithium hydroxide, sodium hydroxide, etc .; and the acidic solution is preferably hydrochloric acid.
[092] [092] The raw material 1 and raw material 2 of the present invention can be homemade or purchased.
[093] [093] The "pharmaceutically acceptable salt" of the compound of formula (I) in the present invention refers to: a salt obtained by an acid functional group in the compound of formula (I) that binds a suitable inorganic or organic (alkaline) cation , as a salt obtained by the acid functional group that binds an alkali metal or an alkaline earth metal, an ammonium salt, a salt obtained by the acid functional group that binds a nitrogenous organic base; and a salt obtained by an alkaline functional group (such as -NH2) in the compound of formula (I) linking a suitable inorganic or organic anion (acid) including an inorganic acid and organic carboxylic acid.
[094] [094] The "ester" of the compound of formula (I) in the present invention refers to: an ester formed by the esterification reaction between a compound of formula (I) and alcohol when there is a carboxyl group in the compound of formula (I ); and an ester formed by the esterification reaction between the compound of formula (I) and an organic acid, an inorganic acid, an organic acid salt or the like when there is a hydroxyl group in the compound of formula (I). In the presence of acid or alkali, the ester can be hydrolyzed to produce a corresponding acid or alcohol. It can serve as a pharmaceutically acceptable ester of the compound of general formula (I), such as alkylacyloxyalkyl ester, alkoxycarbonyloxyalkyl ester, alkoxymethyl ester, alkylacylaminamethyl ester, cycloalkylacyloxyalkyl ester and cycloalkyloxyalkylalkyl ester.
[095] [095] "Stereoisomerism" is divided into conformational isomerism and configuration isomerism, and configuration isomerism can also be divided into cis-trans and optical isomerism. Conformational isomerism refers to a phenomenon in which atoms or radicals of an organic molecule with a certain configuration are spatially organized in a different way due to the rotation or distortion of simple carbon-carbon bonds, and is common in the structures of alkane and cycloalkane compounds , such as chair forming and boat forming, appeared in the cyclohexane structure. A "cis-isomer
[096] [096] The present invention also relates to the following embodiments:
[097] [097] Embodiment 1: a compound of general formula (I), a pharmaceutically acceptable salt thereof, an ester thereof or a stereoisomer of the compound, the salt or ester,
[098] [098] Embodiment 2: the compound, a pharmaceutically acceptable salt thereof, its ester or the stereoisomer of the compound, the salt or ester of embodiment 1, wherein R1 is selected from the group consisting of halogen, hydroxyl, amine, cyano, C1-4 alkyl, haloC1-4 alkyl, hydroxy C1-4 alkyl, amino C1-4 alkyl, alkoxy
[099] [099] Embodiment 3: the compound, a pharmaceutically acceptable salt thereof, its ester or the stereoisomer of the compound, the salt or ester of embodiments 1 to 2, where M is selected from the group consisting of -CH2-, -CH2-CH2-, -CH2-CH2- CH2-, -CH2-NH-, -CH2-CH2-O- and -CH2-NH-CO-; ring A is selected from saturated 7-membered bridged cyclyl or saturated 7-membered bridged nitrogen heterocyclic, and when ring A is a saturated 7-membered bridged nitrogen heterocyclic, preferably the ring
[0100] [0100] Embodiment 4: the compound, a pharmaceutically acceptable salt thereof, its ester or the stereoisomer of the compound, the salt or ester of embodiments 1 to 3, wherein R1 is selected from a group that consists of halogen, hydroxyl, amine, cyano, C1-4 alkyl, haloC1-4 alkyl, hydroxy C1-4 alkyl, amino C1-4 alkyl, C1-4 alkoxy, C1-4 alkylamine, C1-4 alkylcarbonyl, C1- alkoxy 4 C1-4 alkyl, 3-4-membered cycloalkyl, 3-4-membered cycloalkyl, C1-4-membered cycloalkyl, 3-4-membered cycloalkyl, 3-4-membered mono-heterocyclic, 3--membered mono-heterocyclic 4 members C1-4 alkyl or 3-4 membered monoheterocyclic C1-4 alkoxy; ring A is selected from the following groups:,,,,,,,,,,,,,,,,,, and,.
[0101] [0101] Embodiment 5: the compound, a pharmaceutically acceptable salt thereof, its ester or the stereoisomer of the compound, the salt or ester of embodiments 1 to 4, wherein ring B is selected from fused heteroaryl of 9-10 members containing 1 to 2 heteroatoms or groups and is optionally substituted by 1 to 2 Q1, and the heteroatom or group is selected independently from a group consisting of N, NH, O, S, SO or SO2; preferably, ring B is attached to L or ring A by a ring carbon atom; each Q1 is independently selected from a group consisting of halogen, hydroxyl, amine, cyano, C1-4 alkyl, haloC1-4 alkyl, C1-4 alkoxy, 3-6 membered cycloalkyl, 3-6 membered cycloalkyl alkyl C1-4, 3-6 membered cycloalkyl C1-4 alkoxy, 3-6 membered monoheterocyclic, 3-6 membered monoheterocyclic C1-4alkyl or 3-6 membered monoheterocyclic; L is absent or is C1-2 alkylene, where one of the carbon atoms in C1-2 alkylene is optionally replaced by a heteroatom or a group, and the heteroatom or group is selected from a group consisting of NH, O, S or CO.
[0102] [0102] Embodiment 6: the compound, a pharmaceutically acceptable salt thereof, its ester or the stereoisomer of the compound, the salt or ester of embodiments 1 to 5, wherein Ar is selected from a group consisting of in phenyl, phenylC1-4 alkyl, phenylC1-4 alkoxy, 5-6 membered monocyclic heteroaryl, 5-6 membered C1-4alkyl heteroaryl and 5-6 membered C1-4 alkoxy heterocyclyl which are optionally substituted for 1 to 2 Q2; and each Q2 is independently selected from a group consisting of halogen, hydroxyl, amine, cyano, C1-4 alkyl, haloC1-4 alkyl, hydroxyC1-4 alkyl, aminoC1-4 alkyl, C1-4 alkoxy, C1-4 alkoxy alkyl or C1-4 alkylamine.
[0103] [0103] Embodiment 7: the compound, a pharmaceutically acceptable salt thereof, its ester or the stereoisomer of the compound, the salt or ester of embodiments 1 to 6, wherein R1 is selected from a group consisting of halogen, methyl, ethyl, propyl, isopropyl, trifluoromethyl, trifluoroethyl, methoxy, ethoxy, propoxy, isopropoxy, methylamine, ethylamine, methoxymethyl, methoxyethyl, ethoxymethyl, cyclopropyl, cyclopropylmethyl, cyclopropylethyl, cyclopropylethyl, cyclopropylethyl, cyclopropylethyl
[0104] [0104] Embodiment 8: the compound, a pharmaceutically acceptable salt thereof, its ester or the stereoisomer of the compound, the salt or ester of embodiment 7, wherein R1 is selected from the group consisting of cyclopropyl, cyclopropylmethyl, cyclopropylethyl, cyclopropylmethoxy, cyclobutyl, cyclobutylmethyl, cyclobutylethyl, cyclobutylmethoxy, epoxyethyl, epoxyethylmethyl, azacyclopropyl, azacyclopropylmethyl, oxacyclobutyl or azacyclobutyl; ring A is selected from a group consisting of, and; ring B is selected from the following groups, optionally substituted by 1 Q1:, e; Q1 is selected from a group consisting of fluorine, chlorine, bromine, methyl, ethyl, propyl, isopropyl, trifluoromethyl, methoxy, ethoxy, propoxy or isopropoxy; Ar is selected from a group consisting of phenyl, pyridyl and pyrimidinyl, which are optionally substituted by 1 to 2 Q2; and each Q2 is independently selected from a group consisting of fluorine, chlorine, bromine, methyl, ethyl, propyl, isopropyl, methoxy or ethoxy.
[0105] [0105] Embodiment 9: the compound, a pharmaceutically acceptable salt thereof, its ester or the stereoisomer of the compound, the salt or ester of embodiments 1 to 8, having the following structure as a general formula (II ),.
[0106] [0106] Embodiment 10: the compound, a pharmaceutically acceptable salt thereof, its ester or the stereoisomer of the compound, the salt or ester of embodiment 1, wherein the compound is selected from: THE
[0107] [0107] Embodiment 11: a pharmaceutical formulation containing the compound, a pharmaceutically acceptable salt thereof, an ester thereof or a stereoisomer of the compound, the salt or ester of any one of embodiments 1 to 10, wherein the formulation The pharmaceutical product contains one or more pharmaceutically acceptable carriers and / or diluents and can be of any pharmaceutically acceptable dosage form.
[0108] [0108] Embodiment 12: a use of the compound, a pharmaceutically acceptable salt thereof, an ester thereof or a stereoisomer of the compound, the salt or ester of any of embodiments 1-10 in the preparation of medicaments for the prevention and / or treatment of FXR-mediated diseases, in which the diseases comprise atherosclerosis, bile acid metabolism disorder, primary sclerosing cholangitis, cholesterol calculation, fibrosis-related diseases, fatty liver disease, cirrhosis, hepatitis, liver failure, cholestasis , cholelithiasis, myocardial infarction, stroke, thrombus, clinical complications of type I or II diabetes, hyperproliferative diseases and inflammatory bowel diseases.
[0109] [0109] Embodiment 13: the use of Embodiment 12, in which diseases are selected from alcoholic fatty liver disease, non-alcoholic fatty liver disease, primary biliary cirrhosis, primary cholangetic cirrhosis, chronic hepatitis, non-viral hepatitis , alcoholic steatohepatitis, non-alcoholic steatohepatitis, benign intrahepatic cholestasis, progressive familial intrahepatic cholestasis, drug-induced cholestasis, cholestasis related to gastrointestinal nutrition, extrahepatic cholestasis, hypercholesterolemia, neonatal jaundice, kernicterus , diabetic nephropathy, diabetic neuropathy, diabetic retinopathy, other observed results of chronic clinical manifest diabetes, hepatocellular carcinoma, colon adenoma, polyposis, colon adenocarcinoma, breast cancer, pancreatic cancer, esophageal carcinoma and other forms of gastrointestinal neoplastic diseases and liver.
[0110] [0110] The compound of the present invention has one or more of the following advantages: (1) The compound of the present invention, a pharmaceutically acceptable salt, an ester thereof or a stereoisomer of the compound, the salt or ester, has excellent stirring activity of FXR and can be used safely to treat a / or prevent non-alcoholic fatty liver disease, primary biliary cirrhosis, lipid metabolism disorder, diabetic complications, malignant tumors and related diseases.
[0111] [0111] Exemplary experimental schemes for some compounds of the present invention are provided below to show the beneficial activity and the beneficial technical effect of the compound of the present invention. However, it should be understood that the following experimental schemes are only illustrations for the present invention, and do not limit the scope of the present invention. According to the teachings of the specification, those skilled in the art can appropriately modify or alter the technical solutions of the present invention without departing from the spirit and scope of the present invention.
[0112] [0112] The cells were digested with pancreatin and collected, and the cell concentration was determined; according to the result of the count, the cells were resuspended to a density of 7.5 105 cells / ml; 2 ml of the cell suspension was inoculated into each well of a 6-well cell culture plate; the culture plate was placed in an incubator and cultured under conditions of 37 ° C and 5% CO2 for 24 hours.
[0113] [0113] The compounds to be tested were diluted to 0.3 mM and 3 mM with DMSO; 5 μL of each stock solution diluted in the previous step was added to 5 ml of medium individually. The concentrations of the working solutions obtained were 0.3 μM and 3 μM, respectively. In the control group, the medium was prepared using equivolumetric DMSO instead of stock solutions; the cell culture plate was removed from the incubator; the medium was removed from the cell culture plate and the working solutions and the control medium were added correspondingly; then, the culture plate was placed back in the incubator and cultured under conditions of 37 ° C and 5% CO2 for 24 hours.
[0114] [0114] after treatment for 24 hours, the cell culture plate was removed, and the medium was removed from the plate; the cells were washed three times with pre-cooled PBS (4 ° C); 200 μL of pancreatin (preheated to 37 ° C) was added to each well, and the plate was gently shaken so that pancreatin could evenly cover the bottom of the plate. Then, the culture plate was placed back in the incubator and incubated until the cells separate from the bottom of the plate. 1 ml of medium was added to stop digestion. The solution was gently pipetted and blown several times with a pipettor, all substances in each well were pipetted into a 1.5 mL centrifuge tube without RNAse and centrifuged at 200 × g for 5 minutes; the supernatants were removed to collect cell samples.
[0115] [0115] Cell lysis: Fresh RNA lysis buffer was prepared (to which 10 μL of 2-mercaptoethanol were added to 1 ml of lysis buffer); 600 μL of lysis buffer was added to each cell sample; the cells were completely lysed by vigorous vortexing for 1 to 2 minutes; cell lysates were centrifuged at 12,000 × g for 5 minutes; and the supernatants were transferred to 1.5 ml centrifuge tubes without RNAse.
[0116] [0116] RNA extraction and purification: An equal amount of 70% ethanol was added to each cell lysate; then, the centrifuge tubes were shaken vigorously, and the buffer was mixed sufficiently so that the particulate precipitates that could form after the addition of ethanol were dispersed as evenly as possible; adsorption columns were placed in collecting tubes and the mixtures were transferred to the adsorption columns. At most, 700 μL were transferred at a time; and the solutions were centrifuged at
[0117] [0117] The extracted RNA concentration and weight were measured. The RNA was stored at -80 ° C.
[0118] [0118] The RNA extracted in the second stage was incubated at 70 ° C for 5 minutes, so that the RNA was denatured. The treated sample was placed on ice;
[0119] [0119] The RNA sample was diluted to 200 ng / mL with water without RNAse; and 10 μL of the reverse transcription solution were prepared according to the following table and were mixed with 10 μL of denatured RNA. In the reverse transcription reaction, the total amount of RNA was 2 μg. In the process of the experiment, all reagents were placed on ice.
[0120] [0120] Reverse transcription is performed on a G-Storm GS1 PCR thermal cycler. The reverse transcription process was defined as follows: 25 ° C, 10 min → 37 ° C, 120 min → 85 ° C, 5 min → 4 ° C, ∞. The reverse transcription product (cDNA) was stored at -20 ° C.
[0121] [0121] According to the qPCR amplification efficiency, an appropriate cDNA concentration was selected for the qPCR experiment for a sample.
[0122] [0122] 80 μL of the reaction mixture was prepared according to the following table, and 20 μL of the reaction mixture was pipetted into a 96-well PCR reaction plate, in which there were three replicates for the cDNA sample ( 7 μL of 100 ng were added to each reaction well).
[0123] [0123] qPCR was performed in an ABI7500 real-time quantitative PCR amplifier, and the procedure was defined as follows: 50 ° C, 2 min → 95 ° C, 10 min → 95 ° C, 15 s → 60 ° C , 60 s, in which 40 cycles were determined between 95 ° C, 15 s and 60 ° C, 60 s.
[0124] [0124] Note: the relative expression data in Table 1 and Table 2 are shown with the expression below 3 μM PX-104A as 100%, where the relative expression (%) under a concentration of a compound is shown as the ratio of expression under the concentration of the compound to the expression below 3 μM of PX-104A.
[0125] [0125] It can be known from the statistical results of table 1 and table 2, the compounds of the present invention have a good shaking effect on BSEP mRNA in HepG2 cells. BSEP is a direct gene downstream of FXR and regulates the discharge of bile acid from the liver. BSEP is a good index for preliminary screening of the compound's FXR shaking activities, and has great significance in the treatment of non-alcoholic fatty liver disease.
[0126] [0126] Test samples: home compounds 1, 2, 4 and 5 of the present invention (for their chemical names and preparation methods, see the preparation example for each compound).
[0127] [0127] Experimental materials: a mixed hepatic microsome of SD and human mice, both purchased from XenoTech, with lot numbers: 1410271 (SD mouse) and 1410013 (human), respectively, where both microsomal protein concentrations were 20 mg · mL-1; Experimental promoter β-NADPH purchased from Roche (lot No. 524F0231); and a phosphate buffer solution (pH 7.4) made in the laboratory.
[0128] [0128] Preparation of test sample solutions: An adequate amount of the test sample powder was accurately weighed and dissolved in an appropriate amount of dimethyl sulfoxide (DMSO) to a concentration of 1 mM; then, the solution was diluted 20 times with methanol to produce a 50 μM working solution.
[0129] [0129] Experimental method: Table 3: Composition of the incubation system for the hepatic microsome metabolism stability experiment Materials Initial Concentration Proportion (%) Final Concentration Required Buffer solution of 100 μM 50 50 μM phosphate Magnesium chloride 20 μM 5 1 μM anhydrous Liver microsome 20 mg protein / mL 2.5 0.5 mg protein / mL Water required —— 30.5 —— additionally Test sample 50 μM 2 1 μM β-NADPH 10 μM 10 1 μM
[0130] [0130] Operational steps for the experiment: (1) According to the proportions in table 3 "Composition of the incubation system for the hepatic microsome metabolism stability experiment" above, for each compound, 6 mL of PBS (100 mM ), 0.6 mL of MgCl2 solution (20 mM) and 3.66 mL of H2O were made in a mixed solution 1 for the incubation system (not containing the microsomes, the test sample and β-NADPH).
[0131] [0131] Data analysis: Percentages of residual contents were calculated according to the following formula.
[0132] [0132] Experimental result: Table 4: Stability of compounds of the present invention in hepatic microsome metabolism in vitro Residual content (%) after 60 minutes of incubation Compo Compos Compo Compos Compos Compo sto 30- to 1 sto 2 to 4 to 5 sto 8 sto 2-1 Species 70 Hepatic microsome 49 55 54 31 96 69 20 human Hepatic microsome 53 94 69 49 73 77 - of rat
[0133] [0133] Experimental conclusion: The compounds of the present invention have good stability in the metabolism of liver microsomes, facilitating the best pharmacological effect in vivo, and have a high clinical test value and good drainage capacity.
[0134] [0134] The cells were digested with pancreatin and collected, and the concentration of the cells was determined; according to the result of the count, the cells were resuspended at a density of 7.5 105 cells / ml; 2 ml of the cell suspension was inoculated into each well of a 6-well cell culture plate; and the culture plate was placed in an incubator and grown under conditions of 37 ° C and 5% CO2 for 24 hours.
[0135] [0135] The compounds were diluted to 3000, 1000, 200, 8, 1.6, 0.32, 0.0128, 0.00256 and 0.000512 μM with DMSO. The solutions composed of different concentrations obtained in the previous step were subsequently diluted 1,000 times with the medium to obtain working solutions, and a medium containing 0.1% DMSO was adopted as a control group. The cell culture plate was removed from the incubator, the medium was removed and the working solutions and the control medium were added. The culture plate was returned to the incubator and cultured under conditions of 37 ° C and 5% CO2 for 24 hours.
[0136] [0136] after treatment for 24 hours, the cell culture plate was removed from the incubator, and the medium was removed from the plate; the cells were washed three times with pre-cooled PBS (phosphate buffer solution) (4 ° C); 200 μL of pancreatin (preheated to 37 ° C) was added to each well, and the plate was gently shaken so that pancreatin could evenly cover the bottom of the plate. Then, the culture plate was placed back in the incubator and incubated until the cells separate from the bottom of the plate. 1 ml of medium was added to stop digestion. The solution was gently pipetted and blown several times with a pipettor, all substances in each well were pipetted into a 1.5 mL centrifuge tube without RNAse and centrifuged at 200 × g for 5 minutes; the supernatants were removed to collect cell samples.
[0137] [0137] Cell lysis: Fresh RNA lysis buffer was prepared (to which 10 μL of 2-mercaptoethanol were added to 1 ml of lysis buffer); 600 μL of lysis buffer was added to each cell sample; the cells were completely lysed by vigorous vortexing for 1 to 2 minutes; cell lysates were centrifuged at 12,000 × g for 5 minutes; and the supernatants were transferred to 1.5 ml centrifuge tubes without RNAse.
[0138] [0138] RNA extraction and purification: An equal amount of 70% ethanol was added to each cell lysate; then, the centrifuge tubes were shaken vigorously, and the buffer was mixed sufficiently so that the particulate precipitates that could form after the addition of ethanol were dispersed as evenly as possible; adsorption columns were placed in collecting tubes and the mixtures were transferred to the adsorption columns. At most, 700 μL were transferred at a time; and the solutions were centrifuged at
[0139] [0139] The concentration and weight of the extracted RNA was measured. The RNA was stored at -80 ° C.
[0140] [0140] The RNA extracted in the last step was incubated at 70 ° C for 5 minutes, so that the RNA was denatured. The treated sample was placed on ice; the RNA sample was diluted to 200 ng / μL with water without RNAse; and 10 μL of the reverse transcription solution were prepared according to the following table and were mixed with 10 μL of denatured RNA. In the reverse transcription reaction, the total amount of RNA was 2 μg. In the process of the experiment, all reagents were placed on ice.
[0141] [0141] Reverse transcription is performed on a G-Storm GS1 PCR thermal cycler. The reverse transcription process was defined as follows: 25 ° C, 10 min → 37 ° C, 120 min → 85 ° C, 5 min → 4 ° C, ∞. The reverse transcription product (cDNA) was stored at -20 ° C.
[0142] [0142] According to the qPCR amplification efficiency, an appropriate cDNA concentration was selected for the qPCR experiment for a sample.
[0143] [0143] 80 μL of the reaction mixture was prepared according to the following table, and 20 μL of the reaction mixture was pipetted into a 96-well PCR reaction plate, in which there were three replicates for the cDNA sample ( 7 μL of 100 ng were added to each reaction well).
[0144] [0144] qPCR was performed in an ABI7500 real-time quantitative PCR amplifier, and the procedure was defined as follows: 50 ° C, 2 min → 95 ° C, 10 min → 95 ° C, 15 s → 60 ° C , 60 s, in which 40 cycles were determined between 95 ° C,
[0145] [0145] Note: EC50 refers to the median effective concentration, and IC50 refers to the median inhibitory concentration.
[0146] [0146] It can be known from the experimental result that the compounds of the present invention have a good stirring effect on BSEP mRNA and SHP mRNA on HepG2 cells, and the corresponding EC50 values are significantly lower than those of the compound of control 1-1B; and the compounds of the present invention have a good suppression effect of CYP7A1 on the mRNA, and the corresponding IC50 values are significantly lower than those of the control compound 1-1B. Since BSEP and SHP are the direct genes downstream of FXR, their expression agitation effects directly reflect FXR agitation activity; and since CYP7A1 is the secondary gene downstream of FXR, its expression suppression effect indirectly reflects the agitating activity of FXR. BSEP, SHP and CYP7A1 are good indexes for preliminary screening of the compound's FXR stirring activities.
[0147] [0147] The cells were digested with pancreatin and collected, and the concentration of the cells was determined; according to the result of the count, the cells were resuspended at a density of 7.5 × 104 cells / ml; 1 ml of the cell suspension was inoculated into each well of a 12-well cell culture plate; and the culture plate was placed in an incubator and grown under conditions of 37 ° C and 5% CO2 for 24 hours.
[0148] [0148] The compounds were diluted to 1000, 200, 8, 1.6, 0.32, 0.128, 0.0256 and 0.00512 μM with DMSO. The solutions composed of different concentrations obtained in the previous step were subsequently diluted 1000 times with the medium to obtain working solutions, and a medium containing 0.1% DMSO was adopted as a control group. The cell culture plate was removed from the incubator, the medium was removed and the working solutions and the control medium were added. The culture plate was returned to the incubator and cultured under conditions of 37 ° C and 5% CO2 for 24 hours.
[0149] [0149] after treatment for 24 hours, the cell culture plate was removed from the incubator, and the medium was removed from the plate; the cells were washed three times with pre-cooled PBS (phosphate buffer solution) (4 ° C); 200 μL of pancreatin (preheated to 37 ° C) was added to each well, and the plate was gently shaken so that pancreatin could evenly cover the bottom of the plate. Then, the culture plate was placed back in the incubator and incubated until the cells separate from the bottom of the plate. 1 ml of medium was added to stop digestion. The solution was gently pipetted and blown several times with a pipettor, all substances in each well were pipetted into a 1.5 mL centrifuge tube without RNAse and centrifuged at 200 × g for 5 minutes; the supernatants were removed to collect cell samples.
[0150] [0150] Cell lysis: Fresh RNA lysis buffer was prepared (to which 10 μL of 2-mercaptoethanol were added to 1 ml of lysis buffer); 600 μL of lysis buffer was added to each cell sample; the cells were completely lysed by vigorous vortexing for 1 to 2 minutes; cell lysates were centrifuged at 12,000 × g for 5 minutes; and the supernatants were transferred to 1.5 ml centrifuge tubes without RNAse.
[0151] [0151] RNA extraction and purification: An equal amount of 70% ethanol was added to the cell lysate; then, the centrifuge tubes were shaken vigorously, the buffer was mixed sufficiently, so that the precipitates of particles that may form after the addition of ethanol were dispersed as evenly as possible; adsorption columns were placed in collecting tubes and the mixtures were transferred to the adsorption columns. At most, 700 μL were transferred at a time; and the solutions were centrifuged at 12,000 × g at room temperature for 15 sec. The solutions in the collection tubes were discarded and the adsorption columns were replaced in the collection tubes; and all remaining mixtures were then transferred to the adsorption columns. 700 μL of eluent I was added to the adsorption column; and the solutions were centrifuged at 12,000 × g at room temperature for 15 sec. The adsorption columns were placed in new collection tubes; 500 μL of eluent II was added to the adsorption column; and the solutions were centrifuged at 12,000 × g at room temperature for 15 seconds. The solutions in the collection tubes were discarded and the adsorption columns were placed again in the collection tubes; 500 μL of eluent II was added to the adsorption column; the solutions were centrifuged at 12,000 × g at room temperature for 1 to 2 minutes, and the adsorption columns were placed in RNA collecting tubes; 50 μL of water without RNAse was added to the center of the adsorption column, and the resulting solution was incubated at room temperature for 1 min; the RNA was eluted in the collection tubes by centrifugation at 14,000 × g at room temperature for 2 minutes.
[0152] [0152] The extracted RNA concentration and weight were measured. The RNA was stored at -80 ° C.
[0153] [0153] The RNA extracted in the last step was incubated at 70 ° C for 5 minutes, so that the RNA was denatured. The treated sample was placed on ice; the RNA sample was diluted to 100 ng / μL with water without RNAse; and 10 μL of the reverse transcription solution were prepared according to the following table and were mixed with 10 μL of denatured RNA. In the reverse transcription reaction, the total amount of RNA was 1 μg. In the process of the experiment, all reagents were placed on ice.
[0154] [0154] Reverse transcription is performed on a G-Storm GS1 PCR thermal cycler. The reverse transcription process was defined as follows: 25 ° C, 10 min → 37 ° C, 120 min → 85 ° C, 5 min → 4 ° C, ∞. The reverse transcription product (cDNA) was stored at -20 ° C.
[0155] [0155] According to the qPCR amplification efficiency, an appropriate cDNA concentration was selected for the qPCR experiment for a sample.
[0156] [0156] qPCR was performed on an ABI7500 real-time quantitative PCR amplifier, and the procedure was defined as follows: 50 ° C, 2 min → 95 ° C, 10 min → 95 ° C, 15 s → 60 ° C , 60 s, in which 45 cycles were determined between 95 ° C, 15 s and 60 ° C, 60 s.
[0157] [0157] Results of the EC50 assay for the influence of compounds of the present invention on the relative expression of SHP mRNA in AML12 cells Compound 1-1B Compound 2-1 Compound 8 EC50 (nM) 1.51 0.76 0.89
[0158] [0158] It can be known from the experimental result that the compounds of the present invention have a good shaking effect on the SHP mRNA in AML12 cells, and the corresponding EC50 values are significantly lower than those of the control compound 1-1B . Since SHP is a direct gene downstream of FXR, the expression of SHP mRNA directly reflects the agitating activity of FXR. SHP is a good index for preliminary screening of the compound's FXR shaking activities, and the FXR agonist is expected to become a preference for the treatment of non-alcoholic fatty liver disease.
[0159] [0159] Experimental example 5: influence of compounds of the present invention in mice with NAFLD induced by high fat content
[0160] [0160] Construction of mouse models with NAFLD / NAHNA (non-alcoholic fatty liver disease / non-alcoholic steatohepatitis): six-week-old male C57BL / 6J mice were fed a high-fat diet and were weighed weekly to record the body weight of the mice; and after a high-fat diet for 8 weeks, the mice were weighed to determine if the modeling is successful.
[0161] [0161] According to body weight values (40 g was generally considered to be a critical screening point), the modeled mice were successfully divided into a blank control group, a model control group and the compound group 2-1 (1 mg / kg), in which the average body weights of all groups were consistent, with no significant difference. The mice were administered once a day (QD) (intragastric syringe # 8) for 3 weeks.
[0162] [0162] 24 hours after the last administration, blood was sampled from the inner corner; the blood sampled remained at room temperature for 1 to 2 hours and was centrifuged at 4,000 rpm for 15 minutes, and the serum was collected; the mice were then euthanized, and a large hepatic lobe and a small hepatic lobe were cut and placed in 10% neutral formalin, and the remains were stored in a refrigerator at -80 ° C. For the detailed method of administration, dosage and route of administration, see the table below.
[0163] [0163] Route, dosage and regime of administration in experiments for the influence of the compounds of the present invention in mice with DHGNA / EHNA induced by high fat content Volume group administered Route of group Dosage (mg / kg) administration (mL / 10 g) Group administration of 1 control in - 0.1 * po
[0164] [0164] The data were represented by mean ± S.E.M., and the one-way analysis of variance (ANOVA), followed by LSD or independent sample T test, was performed on the statistical data with the statistical software SPSS 16.0. Statistical differences were tested, and p <0.05 was defined as a significant difference.
[0165] [0165] Experimental result of the influence of compounds of the present invention in mice with NAFLD / NAFLD induced by high fat content Weight of LDLp1 TG liver Scorep2 / Weight DHGNA body activity Model group 0.31 - - 3 Compound 2-1 0.21 -53% * -9% * 1
[0166] [0166] LDL represents low density lipoprotein; TG represents triglycerides; and p-value has statistical significance, where p1 ﹤ 0.01 and p2 ﹤ 0.001.
[0167] [0167] Compound 2-1 of the present invention exhibits a good improvement effect on the NAFLD symptom of mice with the administration dose. An increase in liver weight is a common side effect caused by the FXR agonist. It can be known from the experimental result that the compounds of the present invention do not cause an obvious increase in the liver weight / body weight ratio of the experimental animal, that is, they do not show obvious side effects and, therefore, the compounds of the present invention are high security. DETAILED DESCRIPTION OF THE INVENTION
[0168] [0168] The above mentioned content of the present invention is further described in detail below by the detailed description in the form of examples. However, the scope of the object of the present invention above should not be construed as being limited to the following examples. Any technique achieved based on the above mentioned content of the present invention must be within the scope of the present invention.
[0169] [0169] (1RS, 4RS, 5Sr) -5-hydroxy-2-azabicyclo [2.2.1] tert-butyl heptane-2-carboxylate (700 mg; 3.3 mol), potassium tert-butoxide (560 mg ; 5.0 mmol), 18-crown-6 (1.32 g, 5.0 mmol) and KI (830 mg; 5.0 mmol) were added to THF (15 mL); then 4- (chloromethyl) -5-cyclopropyl- 3- (2,6-dichlorophenyl) isoxazole (1.20 g, 4.0 mmol) was added, and the mixture solution was reacted at 50 ° C for 6 hours . Water (100 ml) and ethyl acetate (100 ml) were added for extraction and the organic phase was concentrated and purified by silica gel column chromatography (petroleum ether: ethyl acetate = 8: 1) to obtain the compound titer (1.38 g; yield 87.3%).
[0170] [0170] (1RS, 4RS, 5SR) -5 - ((5-cyclopropyl-3- (2,6-dichlorophenyl) isoxazol-4-yl)
[0171] [0171] Methyl 4-amino-3-methoxybenzoate (1.81 g; 10.0 mmol) and KSCN (3.88 g; 40.0 mmol) were added to glacial acetic acid (10 ml) and, 15 minutes then, bromine (1.60 g; 10.0 mmol) was added. The mixing solution was reacted at 25 ° C for 16 hours. The pH of the reaction solution was adjusted to 8 with ammonia hydroxide.
[0172] [0172] Crude methyl 2-amino-4-methoxybenzo [d] thiazol-6-carboxylate (2.40 g; 10.0 mmol) and copper bromide (3.35 g; 15.0 mmol) acetonitrile (20 ml). Tert-butyl nitrite (1.55 g, 15.0 mmol) was added dropwise at 0 ° C and, after the addition was complete, the mixing solution was stirred and reacted at 25 ° C for 0.5 hour . Water (100 mL) and ethyl acetate (100 mL) were added for extraction, and the organic phase was concentrated and purified by silica gel column chromatography (petroleum ether: ethyl acetate = 10: 1) to obtain the title compound (1.21 g; total yield in two stages: 40.1%).
[0173] [0173] Methyl 2-bromo-4-methoxybenzo [d] thiazol-6-carboxylate (157 mg; 0.52 mmol), 4 - (((((1RS, 4RS, 5SR) -2-azabicyclo [2.2.1 ] heptan-5-yl) oxy) methyl) -5-cyclopropyl-3- (2,6-dichlorophenyl) isoxazole (100 mg; 0.26 mmol) and cesium carbonate (254 mg; 0.78 mol) were added the DMA (2 ml), and the mixing solution was reacted at 110 ° C for 16 hours.
[0174] [0174] 2 - (((1RS, 4RS, 5SR) -5 - ((5-cyclopropyl-3- (2,6-dichlorophenyl) isoxazole-
[0175] [0175] Molecular formula: C28H25Cl2N3O5S Molecular weight: 585.1 LC-MS (m / z): 586.2 (M + H +)
[0176] [0176] 1H NMR (400 MHz, DMSO-d6) : 7.95 (s, 1H); 7.72-7.52 (m, 3H); 7.40 - 7.35 (m, 1H); 4.35-4.45 (m, 2H); 3.87 (s, 3H); 3.65-3.55 (m, 1H); 2.95-2.85 (m, 1H); 2.40-2.30 (m, 1H); 1.92-1.81 (m, 1H), 1.63-1.52 (m, 1H); 1.50-1.40 (m, 1H); 1.28-1.02 (m, 4H); 0.90-0.80 (m, 2H) Example 2: Preparation of 2 - ((1RS, 4RS, 5SR) -5 - ((5-cyclopropyl-3- (2,6-dichlorophenyl) isoxazol-4-yl ) methoxy) -2-azabicyclo [2.2.1] heptan-2-yl) benzo [d] thiazol-6-carboxylic acid (compound 2)
[0177] [0177] (1RS, 4RS, 5Sr) -5-hydroxy-2-azabicyclo [2.2.1] tert-butyl heptane-2-carboxylate (213 mg; 1.0 mmol), potassium tert-butoxide (168 mg ; 1.5 mmol) and 18-crown-6 (396 mg; 1.5 mmol) were added to THF (20 mL) and, 25 minutes later, 4- (bromomethyl) -5-cyclopropyl-3- (2 , 6-dichlorophenyl) isoxazole (416 mg; 1.2 mmol) was added. The mixing solution was reacted at 25 ° C for 16 hours. Water (100 mL) and ethyl acetate (100 mL) were added for extraction, and the organic phase was concentrated and purified by silica gel column chromatography (petroleum ether: ethyl acetate = 3: 1) to obtain the title compound (367 mg; yield: 76.7%).
[0178] [0178] (1RS, 4RS, 5SR) -5 - ((5-cyclopropyl-3- (2,6-dichlorophenyl) isoxazol4-yl) methoxy) -2-azabicyclo [2.2.1] heptane-2-carboxylate tert -butyl (360 mg; 0.75 mmol) was added to DCM (10 ml) and then TFA (3 ml) was added. The mixing solution was reacted under an ice water bath for 2 hours. The pH of the reaction solution was adjusted to 8 with a saturated sodium bicarbonate solution under an ice water bath, and water (100 ml) and DCM (100 ml) were added for extraction. The organic phase was dried over anhydrous sodium sulfate and concentrated to obtain the title compound (280 mg; yield: 98.3%).
[0179] [0179] 4 - ((((((1RS, 4RS, 5SR) -2-azabicyclo [2.2.1] heptan-5-yl) oxy) methyl) -5-cyclopropyl- 3- (2,6-dichlorophenyl) isoxazole ( 280 mg; 0.74 mmol), methyl 2-bromobenzo [d] thiazol-6-carboxylate (403 mg, 1.48 mmol) and cesium carbonate (724 mg; 2.22 mmol) were added to DMA (6 ml), and the mixing solution was reacted at 100 ° C under microwave for 1 hour. Water (100 mL) and ethyl acetate (100 mL) were added for extraction, and the organic phase was concentrated and purified by silica gel column chromatography (petroleum ether: ethyl acetate = 10: 1) to obtain the title compound (305 mg; yield: 72.6%).
[0180] [0180] 2 - ((1RS, 4RS, 5SR) -5 - ((5-cyclopropyl-3- (2,6-dichlorophenyl) isoxazol-4-yl) methoxy) -2-azabicyclo [2.2.1] heptan- Methyl 2-yl) benzo [d] thiazol-6-carboxylate (300 mg;
[0181] [0181] Molecular formula: C27H23Cl2N3O4S Molecular weight: 555.1 LC-MS (m / z): 556.2 (M + H +)
[0182] [0182] 1H NMR (400 MHz, DMSO-d6) : 8.31 (s, 1H); 7.90-7.82 (m, 1H); 7.75- 7.55 (m, 3H); 7.56-7.35 (m, 1H); 4.32-4.25 (m, 2H); 3.62-3.58 (m, 1H); 2.98-2.88 (m, 1H); 2.55-2.52 (m, 2H); 2.40-2.30 (m, 1H); 2.10-1.99 (m, 1H); 1.98-1.85 (m, 1H); 1.65-1.54 (m, 1H); 1.52-1.42 (m, 1H); 1.30-1.25 (m, 1H); 1.20-1.10 (m, 2H); 1.10-1.05 (m, 2H).
[0183] [0183] (1S, 4R) -2-azabicyclo [2.2.1] heptan-5-en-3-one (3.0 g; 27.5 mmol) was dissolved in THF (80 mL), and LiAlH4 (1 , 36 g; 35.8 mmol) was added at 0 ° C. The reaction solution was reacted at 25 ° C for 3 hours and then reacted at an elevated temperature of 60 ° C for 4 hours. Then, water (2 mL) was added at 0 ° C to stop the reaction. The reaction solution was filtered through celite, the filter cake was washed with ethyl acetate (50 ml) and the filtrate was concentrated to 50 ml. Boc2O (9.0 g, 41.2 mmol) was added to the concentrated filtrate and the mixing solution was reacted at 25 ° C for 16 hours. The reaction solution was concentrated and purified by silica gel column chromatography (petroleum ether: ethyl acetate = 10: 1) to obtain the product (3.0 mg; two step yield: 55.9%).
[0184] Tert-butyl [0184] (1S, 4R) -2-azabicyclo [2.2.1] heptan-5-ene-2-carboxylate (1.5 g; 7.68 mmol) and NaBH4 (0.24 g; 6 , 3 mmol) were added to THF (20 mL), and the solution of the mixture was reacted at 25 ° C and under a nitrogen atmosphere for 0.5 hour.
[0185] [0185] (1S, 4S, 5R) -5-hydroxy-2-azabicyclo [2.2.1] tert-butyl heptane-2-carboxylate (0.2 g, 0.94 mmol) was dissolved in THF (30 mL ), and potassium tert-butoxide (158 mg; 1.4 mmol) and 18-crown-6-ether (248 mg; 0.94 mmol) were added and then the solution of the mixture reacted at 25 ° C for 0.5 hour. 4- (chloromethyl) -5-cyclopropyl-3- (2,6-dichlorophenyl) isoxazole (427 mg; 1.4 mmol) and KI (232.4 mg; 1.4 mmol) were added, and the mixture solution reacted at 40 ° C for 2 hours. The reaction solution was concentrated and purified by silica gel column chromatography (petroleum ether: ethyl acetate = 2: 1) to obtain a product (300 mg; yield: 66.6%).
[0186] [0186] (1S, 4S, 5R) -5 - ((5-cyclopropyl-3- (2,6-dichlorophenyl) isoxazol-4-yl) methoxy) - 2-azabicyclo [2.2.1] heptane-2-carboxylate tert-butyl (0.25 g, 0.52 mmol) was added to DCM (8 ml), TFA (2 ml) was added and the mixture solution was reacted at 25 ° C for 2 hours. The reaction solution was concentrated to obtain a crude product (400 mg), which would be used directly in the next step without being purified.
[0187] [0187] 4 - ((((((1S, 4S, 5R) -2-azabicyclo [2.2.1] heptan-5-yl) oxy) methyl) -5-cyclopropyl - 3- (2,6-dichlorophenyl) trifluoroacetate ) isoxazole (400 mg; crude product), methyl 2-bromobenzo [d] thiazol-6-carboxylate (212 mg, 0.78 mmol) and cesium carbonate (508 mg, 1.56 mmol) were added to dimethyl adipate (8 ml), and the mixing solution was reacted at 120 ° C under microwave for 0.5 hour. Then, the reaction solution was poured into water (20 ml) and filtered. The filter cake was purified by silica gel column chromatography (dichloromethane: methanol = 20: 1) to obtain a product (200 mg; two-step yield: 67.4%).
[0188] [0188] 2 - ((1S, 4S, 5R) -5 - ((5-cyclopropyl-3- (2,6-dichlorophenyl) isoxazol-4-yl) methoxy) - 2-azabicyclo [2.2.1] heptan- 2-yl) methyl benzo [d] thiazol-6-carboxylate (200 mg; 0.35 mmol) and lithium hydroxide monohydrate (103 mg; 2.45 mmol) were dissolved in the mixed methanol solvent (8 mL), tetrahydrofuran (8 ml) and water (8 ml), and the resulting mixture was stirred and reacted at 40 ° C for 2 hours. The reaction solution was concentrated, and water (20 ml) was added to the residue. Then, the pH of the reaction solution was adjusted to 2 with 1M diluted hydrochloric acid, and ethyl acetate (50 mL x 2) was added for extraction. The organic phases were combined and concentrated and the residue was purified by C18 reverse phase silica gel column chromatography (methanol: water = 0% -70%) to obtain a product (70 mg; yield: 35.9%) .
[0189] [0189] Molecular formula: C27H23Cl2N3O4S Molecular weight: 555.1 LC-MS (m / z): 556.1 (M + H +)
[0190] [0190] [α] 20 D = -84.97 (C = 1.0; CH3OH)
[0191] [0191] 1H-NMR (400 MHz, CDCl3) δ: 8.34 (s, 1H); 8.05 (dd, J = 1.6 Hz; J = 8.4 Hz, 1H); 7.55 (d, J = 8.8 Hz; 1H); 7.32-7.45 (m, 3H), 4.26-4.33 (m, 2H); 3.61 (d, J = 7.2 Hz, 1H); 3.50 (s, 2H), 3.03 (s, 1H), 2.60 (s, 1H), 2.10-2.15 (m, 1H), 2.01-2.10 (m, 1H); 1.65-1.69 (m, 2H); 1.44 (d, J = 13.6 Hz, 1H); 1.26-1.30 (m, 2H); 1.11 - 1.17 (m, 2H).
[0192] [0192] (1R, 4S) -2-azabicyclo [2.2.1] hept-5-en-3-one (5.0 g, 45.8 mmol) was added to anhydrous THF (100 mL) and hydride aluminum and lithium (2.26 g; 59.5 mol) were added in batches at 0 ° C. The mixing solution was reacted at 25 ° C for 3 hours.
[0193] [0193] (1R, 4S) -2-azabicyclo [2.2.1] hept-5-ene (4.35 g, 45.8 mmol) was added to THF 100 mL), then (Boc) 2O (15, 0 g; 68.7 mmol) was added, and the mixing solution was reacted at 25 ° C for 1 hour. The system was dried by centrifugation directly and purified by silica gel column chromatography (PE: EA = 10: 1) to obtain a product (7.0 g; yield in two stages: 78.4%).
[0194] Tert-butyl [0194] (1R, 4S) -2-azabicyclo [2.2.1] heptan-5-ene-2-carboxylate (1.5 g; 7.68 mmol) and sodium tetrahydroborate (0.24 g ; 6.30 mmol) were added to THF (9 mL), and the mixing solution was stirred and reacted at 23 ° C and under an atmosphere of nitrogen for 0.5 hour. The solution of dimethyl sulfate (0.57 ml; 6.30 mmol) in THF (2 ml) was added at 35 ° C, and the mixture solution was reacted at 35 ° C for 4 hours with vigorous stirring. The reaction solution was cooled to 0 ° C, and stopped with water (5.0 ml). Then, 1M NaOH (15 mL) and hydrogen peroxide (30% by weight in H2O; 0.96 mL) were added in sequence, and the mixing solution was reacted at 23 ° C for 1 hour. EA (100 mL) and water (50 mL) were added for extraction. The organic phase was dried by centrifugation and purified by silica gel column chromatography (PE: EA = 2: 1) to obtain a product (600 mg; yield: 36.7%).
[0195] [0195] (1R, 4R, 5S) -5-hydroxy-2-azabicyclo [2.2.1] tert-butyl heptane-2-carboxylate (600 mg; 2.8 mmol), 18-crown-6-ether ( 739 mg; 2.8 mmol) and potassium tert-butoxide (627 mg; 5.6 mmol) were added to THF (80 mL), and the mixing solution was reacted at 25 ° C for 0.5 hour. Then, 4- (chloromethyl) -5-cyclopropyl-3- (2,6-dichlorophenyl) isoxazole (1.28 g, 4.2 mmol) and potassium iodide (697 mg, 4.2 mmol) were added and the mixing solution was reacted at 50 ° C for 4 hours. EA (100 mL) and water (50 mL) were added for extraction. The organic phase was dried by centrifugation and purified by column chromatography (PE: EA = 2: 1) to obtain a product (700 mg; yield: 52.2%).
[0196] [0196] (1R, 4R, 5S) -5 - ((5-cyclopropyl-3- (2,6-dichlorophenyl) isoxazol-4-yl) methoxy) -2-
[0197] [0197] 4 - (((((1R, 4R, 5S) -2-azabicyclo [2.2.1] heptan-5-yl) oxy) methyl) -5-cyclopropyl- 3- (2,6-dichlorophenyl) isoxazole ( 400 mg; 1.06 mmol), methyl 2-bromobenzo [d] thiazol-6-formate (431 mg; 1.58 mmol) and cesium carbonate (691 mg; 2.12 mmol) were added to DMA (10 ml), and the mixing solution was reacted at 110 ° C under microwave for 0.5 hour. After the reaction was complete, the reaction solution was cooled to 25 ° C. Ethyl acetate (100 ml) and water (50 ml) were then added for extraction. The organic phase was dried by centrifugation and purified by silica gel column chromatography (PE: EA = 5: 1) to obtain a product (450 mg; yield: 74.5%).
[0198] [0198] 2 - (((1R, 4R, 5S) -5 - ((5-cyclopropyl-3- (2,6-dichlorophenyl) isoxazol-4-yl) methoxy) -2-azabicyclo [2.2.1] benzo [ d] methyl thiazol-6-carboxylate (450 mg; 0.79 mmol) was added to the mixed solution of methanol (5 ml) and THF (5 ml), the mixing solution was then added to an aqueous solution (2 ml ) containing lithium hydroxide monohydrate (133 mg; 3.2 mmol), and the system was heated to 50 ° C and reacted for 12 hours. After the reaction was complete, the reaction solution was cooled to 25 ° C and the pH The system was adjusted to 4 with 1N HCl The solvent was distilled by centrifugation. Ethyl acetate (100 ml) and an aqueous sodium chloride solution (50 ml) were added for extraction. The organic phase was dried by centrifugation and purified by silica gel column chromatography (DCM: MeOH = 40: 1) to obtain a product (390 mg; yield: 89.1%).
[0199] [0199] Molecular formula: C27H23Cl2N3O4S Molecular weight: 555.1 LC-MS (m / z): 556.1 (M + H) +
[0200] [0200] [α] 20 D = +76.2 (C = 1.0; CH3OH)
[0201] [0201] 1H-NMR (400 MHz, CDCl3) δ: 8.25 (s, 1H); 8.08-8.04 (m, 1H); 7.62 (m, 1H); 7.47-7.34 (m, 3H); 4.35-4.28 (m, 2H); 3.71 - 3.68 (m, 2H); 2.68 (s, 1H); 2.15-2.08 (m, 2H); 1.85-1.70 (m, 2H); 1.55-1.48 (m, 1H); 1.30-1.21 (m, 3H); 1.20-1.09 (m, 2H).
[0202] [0202] Tert-butyl (1RS, 4RS, 5Rs) -5-hydroxy-2-azabicyclo [2.2.1] tert-butyl heptane-2-carboxylate (0.25 g, 1.17 mmol) was dissolved in THF (30 mL), potassium tert-butoxide (196 mg; 1.75 mmol) and 18-crown-6-ether (310 mg; 1.17 mmol) were added and the solution of the mixture reacted at 25 ° C for 0 , 5 hour. Then 4- (chloromethyl) -5-cyclopropyl-3- (2,6-dichlorophenyl) isoxazole (530 mg; 1.75 mmol) and KI (290 mg; 1.75 mmol) were added, and the solution of mixture was reacted at 30 ° C for 4 hours. The reaction solution was concentrated and the resulting crude product was purified by silica gel column chromatography (petroleum ether: ethyl acetate = 3: 1) to obtain the target product (450 mg; yield: 80%).
[0203] [0203] (1RS, 4RS, 5RS) -5 - ((5-cyclopropyl-3- (2,6-dichlorophenyl) isoxazol-4-yl) methoxy) -2-azabicyclo [2.2.1] heptane-2-carboxylate tert-butyl (0.4 g; 0.83 mmol) was added to dichloromethane (10 ml), trifluoroacetic acid (4 ml) was added and the mixture solution was reacted at 25 ° C for 2 hours. The reaction solution was concentrated to obtain a crude product (500 mg), which would be used directly in the next step.
[0204] [0204] 4 - ((((((1RS, 4RS, 5RS) -2-azabicyclo [2.2.1] heptan-5-yl) oxy) methyl) - 5-cyclopropyl-3- (2,6-dichlorophenyl) trifluoroacetate isoxazole (500 mg; crude product), methyl 2-bromobenzo [d] thiazol-6-carboxylate (338 mg, 1.24 mmol) and cesium carbonate (811 mg, 2.49 mmol) were added to DMA (6 ml), and the mixing solution was reacted at 120 ° C under microwave for 0.5 hour. Then, the reaction solution was poured into water (20 ml) and filtered. The filter cake was purified by silica gel column chromatography (dichloromethane: methanol = 20: 1) to obtain the target product (400 mg; two-step yield: 84.5%).
[0205] [0205] 2 - (((1RS, 4RS, 5RS) -5 - ((5-cyclopropyl-3- (2,6-dichlorophenyl) isoxazol-4-yl) methoxy) -2-azabicyclo [2.2.1] heptan- 2-yl) methyl benzo [d] thiazol-6-carboxylate (200 mg; 0.35 mmol) and lithium hydroxide monohydrate (80 mg; 1.9 mmol) were dissolved in the mixed methanol solvent (10 mL), tetrahydrofuran (10 ml) and water (10 ml), and the resulting mixture was stirred and reacted at 40 ° C for 2 hours. The reaction solution was concentrated, and water (10 ml) was added to the residue. Then, the pH of the solution was adjusted to 2 with hydrochloric acid diluted to 1M and ethyl acetate was added for extraction (100 mL x 2). The organic phases were combined and concentrated and the residue was purified by C18 reverse phase silica gel column chromatography (methanol: water = 0% -70%) to obtain the target product (50 mg; yield: 25.7 %).
[0206] [0206] Molecular formula: C27H23Cl2N3O4S Molecular weight: 555.1 LC-MS (m / z): 556.1 (M + H +)
[0207] [0207] 1H-NMR (400 MHz, CDCl3) δ: 8.39 (s, 1H); 8.08 (d, J = 8.0 Hz, 1H); 7.57 (d, J = 8.4 Hz, 1H); 7.35 (d, J = 8.0 Hz, 1H); 7.10-7.19 (m, 2H); 4.22-4.33 (m, 2H); 4.00 - 4.03 (m, 1H); 3.64 (s, 1H); 3.36 (s, 1H); 2.76 (s, 1H); 2.06-2.13 (m, 1H); 1.86-1.99 (m, 2H); 1.62 (d, J = 10.0 Hz, 1H); 1.20-1.47 (m, 4H); 1.07-1.12 (m, 2H). Example 4: Preparation of 2 - ((1RS, 4RS, 5SR) -5 - ((5-cyclopropyl-3- (2,6-dichlorophenyl) isoxazol-4-yl) methoxy) -2-azabicycles [2.2.1] heptan-2-yl) -4-fluorobenzo [d] thiazol-6-carboxylic acid (compound 4)
[0208] [0208] Methyl 3-fluoro-4-nitrobenzoate (1.99 g; 10.0 mol) was added to methanol (200 mL), Pd / C (200 mg) was added and the mixing solution was subjected to hydrogenation for 16 hours. After suction filtration of the reaction solution, the filtrate was concentrated to obtain the title compound (1.68 g; yield: 99.4%).
[0209] [0209] Methyl 4-amino-3-fluorobenzoate (1.68 g; 9.9 mmol) and KSCN (3.84 g; 39.6 mmol) were added to glacial acetic acid (20 ml) and, 15 minutes then, bromine (1.58 g, 9.9 mmol) was added. The mixing solution was reacted at 25 ° C for 16 hours. Water (100 mL) was added and the pH of the reaction solution was adjusted to 8 with ammonia hydroxide. After suction filtration, the solid was dried to obtain a crude title compound (2.30 g), which would be used directly in the next step.
[0210] [0210] Crude methyl 2-amino-4-fluorobenzo [d] thiazol-6-carboxylate (2.30 g; 9.9 mmol) and copper bromide (3.31 g; 14.9 mmol) were added acetonitrile (20 ml). Tert-butyl nitrite (1.53 g; 14.9 mmol) was added dropwise at 0 ° C and, after the addition was complete, the mixing solution was stirred and reacted at 25 ° C for 1 hour. The reaction solution was concentrated directly and purified by silica gel column chromatography (petroleum ether: ethyl acetate = 10: 1) to obtain the title compound (480 mg; total yield in two stages: 16.7%).
[0211] [0211] Methyl 2-bromo-4-fluorobenzo [d] thiazol-6-carboxylate (151 mg; 0.52 mmol), 4 - (((((1RS, 4RS, 5SR) -2-azabicyclo [2.2.1 ] heptan-5-yl) oxy) methyl) -5-cyclopropyl-3- (2,6-dichlorophenyl) isoxazole (100 mg; 0.26 mmol) and cesium carbonate (254 mg; 0.78 mmol) the DMA (2 ml), and the mixing solution was reacted at 110 ° C for 16 hours.
[0212] [0212] 2 - (((1RS, 4RS, 5SR) -5 - ((5-cyclopropyl-3- (2,6-dichlorophenyl) isoxazol-4-yl) methoxy) -2-azabicyclo [2.2.1] heptan- Methyl 2-yl) -4-fluorobenzo [d] thiazol-6-carboxylate (50 mg; 0.09 mmol) was dissolved in the mixed solution of THF (2 mL) and methanol (2 mL) and then hydroxide 1M lithium (0.9 mL) was added. The mixing solution was reacted at 40 ° C for 1 hour and then the pH of the reaction solution was adjusted to 4 with 1M hydrochloric acid. Ethyl acetate (30 mL) was added for extraction and the organic phase was concentrated and purified by silica gel column chromatography (dichloromethane: methanol = 15: 1) to obtain the compound (30 mg; yield: 61.5% ).
[0213] [0213] Molecular formula: C27H22Cl2FN3O4S Molecular weight: 573.1 LC-MS (m / z): 574.1 (M + H +)
[0214] [0214] 1H NMR (400 MHz, DMSO-d6) δ: 8.18 (s, 1H); 7.76-7.48 (m, 4H); 4.32 - 4.20 (m, 2H); 3.65-3.58 (m, 1H); 3.45-3.40 (m, 2H); 2.58-2.52 (m, 1H); 2.40-2.28 (m, 1H); 1.95-1.85 (m, 1H); 1.65-1.55 (m, 1H); 1.53-1.42 (m, 1H); 1.20-1.13 (m, 2H); 1.13 - 1.09 (m, 2H); 0.90-0.78 (m, 2H). Example 5: Preparation of 2 - ((1RS, 4RS, 5SR) -5 - ((5-cyclopropyl-3- (2,6-dichlorophenyl)
[0215] [0215] Methyl 4-amino-3-methylbenzoate (33.0 g; 0.2 mol) and KSCN (68.0 g; 0.7 mol) were added to glacial acetic acid (200 mL) at 0 ° C and bromine (31.9 g; 0.2 mol) was added slowly dropwise at 0 ° C. After the dropwise addition was complete, the mixing solution was heated to 25 ° C and reacted for 10 hours. Then, the reaction solution was poured into ice water and the pH of the solution was adjusted to 8 with ammonium hydroxide. A solid was precipitated, filtered under reduced pressure and dried. The resulting solid would be used directly in the next step.
[0216] [0216] Methyl 2-Amino-4-methylbenzo [d] thiazol-6-carboxylate (44.5 g, 0.20 mol) and copper bromide (75.9 g, 0.34 mol) were dissolved in acetonitrile (300 mL) at 0 ° C, and tert-butyl nitrite (24.7 g, 0.24 mol) were added slowly dropwise. After the dropwise addition was complete, the mixing solution was stirred and reacted at 25 ° C for 3 hours. The reaction solution was poured into ice water and the solid was precipitated and filtered under reduced pressure. The filter cake was washed with a mixed solvent (PE: EA = 5: 1, 600 mL), and the filtrate was concentrated to obtain the target product (34.3 g; two-step yield: 60.0%) .
[0217] [0217] (1RS, 4RS, 5SR) -5 - ((5-cyclopropyl-3- (2,6-dichlorophenyl) isoxazol-4-yl) methoxy) -2-azabicyclo [2.2.1] heptane-2-carboxylate tert-butyl (200 mg; 0.42 mmol) was added to dichloromethane (20 mL) and then trifluoroacetic acid (10 mL) was added. The mixing solution was reacted at 25 ° C for 2 hours and then the reaction solution was concentrated and used directly in the next step.
[0218] [0218] Methyl 2-bromo-4-methylbenzo [d] thiazol-6-carboxylate (120 mg; 0.42 mmol), 4 - (((((1RS, 4RS, 5SR) -2-azabicycles [2.2.1 ] heptan-5-yl) oxy) methyl) -5-cyclopropyl-3- (2,6-dichlorophenyl)
[0219] [0219] 2 - (((1RS, 4RS, 5SR) -5 - ((5-cyclopropyl-3- (2,6-dichlorophenyl) isoxazol-4-yl) methoxy) -2-azabicyclo [2.2.1] heptan- Methyl 2-yl) -4-methylbenzo [d] thiazol-6-carboxylate (45 mg; 0.077 mmol) and lithium hydroxide monohydrate (16 mg; 0.38 mmol) were added to the mixed methanol solution (8 mL) , THF (8 ml) and water (4 ml), and the mixing solution was reacted at 25 ° C for 5 hours. The pH of the reaction solution was adjusted to 5 with hydrochloric acid, and the resulting solution was then concentrated to obtain the target product (34 mg; yield: 77.4%).
[0220] [0220] Molecular formula: C28H25Cl2N3O4S Molecular weight: 569.1 LC-MS (m / z): 570.2 (M + H) +
[0221] [0221] 1H NMR (400 MHz, CDCl3) : 8.19 (s, 1H); 7.84 (s, 1H); 7.56-7.50 (m, 3H); 4.36 (s, 2H); 3.63-3.48 (m, 2H); 3.17-3.08 (m, 1H); 2.69-2.62 (m, 1H); 2.55 (s, 3H); 2.48-2.35 (m, 1H); 2.15-2.03 (m, 2H); 1.75-1.68 (m, 1H); 1.49-1.46 (m, 1H); 1.40-1.32 (m, 2H); 1.18-1.21 (m, 2H) Example 6: Preparation of 2 - ((1RS, 4SR, 6RS) -6 - ((5-cyclopropyl-3- (2,6-dichlorophenyl)
[0222] [0222] (1RS, 4SR, 6RS) -2-azabicyclo [2.2.1] heptan-6-ol (0.5 g; 3.3 mmol) was dissolved in dichloromethane (20 mL), and di-tert-dicarbonate butyl (0.73 g; 3.3 mmol) and triethylamine (0.37 g; 3.7 mmol) were added. The mixing solution was reacted at 25 ° C for 3 hours. The solvent was distilled off by centrifugation and the residue was purified by column chromatography (petroleum ether: ethyl acetate = 2: 1) to obtain the target compound (0.65 g; yield: 91.5%).
[0223] [0223] (1RS, 4SR, 6Rs) -6-hydroxy-2-azabicyclo [2.2.1] tert-butyl heptane-2-carboxylate (0.1 g; 0.47 mmol), 4- (chloromethyl) - 5- cyclopropyl-3- (2,6-dichlorophenyl) isoxazole (0.21 g, 0.69 mmol), potassium tert-butoxide (79 mg; 0.70 mmol), 18-crown-6 (0.15 g; 0.57 mmol) and potassium iodide (0.12 g; 0.72 mmol) were dissolved in tetrahydrofuran (10 mL), and the mixture solution was heated to 60 ° C and reacted for 3 hours with stirring. Water (20 mL) and ethyl acetate (20 mL) were added for extraction and the aqueous phase was extracted with ethyl acetate (20 mL x 3). The organic phases were combined, dried over anhydrous sodium sulfate and filtered. The filtrate was concentrated and the resulting crude product was purified by silica gel column chromatography (petroleum ether: ethyl acetate = 5: 1) to obtain the compound (0.2 g; yield: 90.9%).
[0224] [0224] (1RS, 4SR, 6Rs) -6 - ((5-cyclopropyl-3- (2,6-dichlorophenyl) isoxazol-4-yl) methoxy) -2-azabicyclo [2.2.1] heptane-2-carboxylate of tert-butyl (0.20 g; 0.42 mmol) was dissolved in dichloromethane (2 ml) and a solution (2 ml) of hydrochloric acid in ethanol was added after the solution of the mixture was cooled to 0 ° C. The resulting mixing solution was reacted for 3 hours. The pH of the reaction system was adjusted to 7 to 8 with a saturated sodium bicarbonate solution, and then the reaction solution was extracted with dichloromethane (10 mL x 2). The organic phase was dried over anhydrous sodium sulfate and filtered, and the filtrate was concentrated to obtain a crude compound (0.158 g; yield: 100%) that would be used directly in the next step.
[0225] [0225] 4 - ((((((1RS, 4SR, 6RS) -2-azabicyclo [2.2.1] heptan-6-yl) oxy) methyl) -5-cyclopropyl- 3- (2,6-dichlorophenyl) isoxazole ( 0.158 g; 0.42 mmol) was dissolved in N, N-dimethylacetamide (3 mL) and then methyl 2-bromobenzo [d] thiazol-6-carboxylate (0.23 g; 0.84 mmol) and cesium carbonate (0.41 g, 1.3 mmol) was added. The mixing solution was heated to 100 ° C under microwave and reacted for 30 minutes.
[0226] [0226] 2 - (((1RS, 4SR, 6RS) -6 - ((5-cyclopropyl-3- (2,6-dichlorophenyl) isoxazol-4-yl) methoxy) -2-azabicyclo [2.2.1] heptan- Methyl 2-yl) -benzo [d] thiazol-6-carboxylate (0.15 g, 0.26 mmol) was dissolved in the mixed solution of tetrahydrofuran (2 ml) and methanol (2 ml), and a hydroxide solution sodium (52 mg; 1.3 mmol) in water (1 mL) was added. The mixing solution was stirred and reacted at 60 ° C for 3 hours. The pH of the reaction solution was adjusted to 3 to 4 with 1N hydrochloric acid. Water (10 ml) was added, and the solution was extracted with ethyl acetate (10 ml x 2). The organic phases were combined, dried over anhydrous sodium sulfate and filtered. The filtrate was concentrated and the resulting crude product was purified by silica gel column chromatography (dichloromethane: methanol = 20: 1) to obtain the compound (0.11 g; yield: 73.3%).
[0227] [0227] Molecular formula: C27H23Cl2N3O4S Molecular weight: 555.1 LC-MS (m / z): 556.1 (M + H +)
[0228] [0228] 1H-NMR (400 MHz, MeOD) δ: 8.33 (s, 1H); 7.98 (dd, J1 = 8.8 Hz, J2 = 1.6 Hz, 1H); 7.49-7.51 (m, 2H); 7.41-7.46 (m, 2H); 4.38-4.47 (m, 2H); 3.57-3.58 (m, 1H); 3.42-3.44 (m, 1H); 2.95-3.01 (m, 1H); 2.62-2.65 (m, 1H); 2.32-2.36 (m, 1H); 1.75-1.79 (m, 1H); 1.61 - 1.69 (m, 2H); 1.33-1.36 (m, 1H); 1.07-1.19 (m, 4H) Example 7: Preparation of 2 - ((1RS, 4SR, 6SR) -6 - ((5-cyclopropyl-3- (2,6-dichlorophenyl) isoxazol-4-yl ) methoxy) -2-azabicyclo [2.2.1] heptan-2-yl) benzo [d] thiazol-6-carboxylic acid (compound 7)
[0229] [0229] (1RS, 4SR, 6Rs) -6-hydroxy-2-azabicyclo [2.2.1] tert-butyl heptane-2-carboxylate (0.2 g; 0.94 mmol), p-nitrobenzoic acid (0.172 g; 1.03 mmol), diethyl azodicarboxylate (0.24 g, 1.38 mmol) and triphenylphosphine (0.37 g; 1.41 mmol) were dissolved in tetrahydrofuran (10 mL), and the mixture solution reacted at 25 ° C for 3 hours. The solvent was distilled by centrifugation and the resulting crude product was purified by silica gel column chromatography (petroleum ether: ethyl acetate = 5: 1) to obtain the target product (0.32 g; yield: 94, 1%).
[0230] [0230] (1RS, 4SR, 6Sr) -6 - ((4-nitrobenzoyl) oxy) -2-azabicyclo [2.2.1] tert-butyl heptane-2-carboxylate (0.32 g, 0.88 mmol) and potassium hydroxide (0.5 g; 8.9 mmol) was dissolved in the mixed solvent of tetrahydrofuran (30 ml) and water (3 ml), and the solution of the mixture was stirred and reacted at 25 ° C for 16 hours. The solvent was distilled by centrifugation and the resulting crude product was purified by silica gel column chromatography (petroleum ether: ethyl acetate = 2: 1) to obtain the target product (0.13 g; yield: 68, 4%).
[0231] [0231] (1RS, 4SR, 6SR) -6-hydroxy-2-azabicyclo [2.2.1] tert-butyl heptane-2-carboxylate (0.13 g; 0.61 mmol), 4- (chloromethyl) - 5- cyclopropyl-3- (2,6-dichlorophenyl) isoxazole (0.28 g, 0.92 mmol), potassium tert-butoxide (0.1 g; 0.89 mmol), 18-crown-6 (0 , 2 g; 0.76 mmol) and potassium iodide (0.15 g; 0.90 mmol) were dissolved in tetrahydrofuran (20 mL), and the mixture solution was heated to 60 ° C and reacted for 3 hours with agitation. Water (20 mL) and ethyl acetate (20 mL) were added for extraction and the aqueous phase was extracted with ethyl acetate (20 mL x 3). The organic phases were combined, dried over anhydrous sodium sulfate and filtered. The filtrate was concentrated and the resulting crude product was purified by silica gel column chromatography (petroleum ether: ethyl acetate = 5: 1) to obtain the target product (0.25 g; yield: 86.2% ).
[0232] [0232] (1RS, 4SR, 6SR) -6 - ((5-cyclopropyl-3- (2,6-dichlorophenyl) isoxazol-4-yl) methoxy) -2-azabicyclo [2.2.1] heptane-2-carboxylate tert-butyl (0.25 g, 0.52 mmol) was dissolved in dichloromethane (2 ml) and a solution (2 ml) of hydrochloric acid in ethanol was added after the solution of the mixture was cooled to 0 ° C. The resulting mixing solution was reacted for 3 hours. The pH of the reaction system was adjusted to 7 to 8 with a saturated sodium bicarbonate solution, and then the reaction solution was extracted with dichloromethane (10 mL x 2). The organic phase was dried over anhydrous sodium sulfate and filtered, and the filtrate was concentrated. The waste would be used directly in the next step.
[0233] [0233] 4 - ((((((1RS, 4SR, 6SR) -2-azabicyclo [2.2.1] heptan-6-yl) oxy) methyl) -5-cyclopropyl- 3- (2,6-dichlorophenyl) isoxazole ( crude product from the last step; 0.52 mmol) was dissolved in N, N-dimethylacetamide (3 mL) and then methyl 2-bromobenzo [d] thiazol-6-carboxylate (0.28 g; 1.0 mmol) and cesium carbonate (0.51 g; 1.6 mmol) were added.
[0234] [0234] 2 - (((1RS, 4SR, 6SR) -6 - ((5-cyclopropyl-3- (2,6-dichlorophenyl) isoxazol-4-yl) methoxy) -2-azabicyclo [2.2.1] heptan- Methyl 2-yl) -benzo [d] thiazol-6-carboxylate (0.24 g, 0.42 mmol) was dissolved in the mixed solution of tetrahydrofuran (2 ml) and methanol (2 ml), and a hydroxide solution sodium (80 mg, 2.0 mmol) in water was added. The mixing solution was stirred and reacted at 60 ° C for 3 hours. The pH of the reaction solution was adjusted to 3 to 4 with 1N hydrochloric acid, and water (10 ml) was added. The solution was extracted with ethyl acetate (10 ml x 2). The organic phases were combined, dried over anhydrous sodium sulfate and filtered. The filtrate was concentrated and the resulting crude product was purified by silica gel column chromatography (dichloromethane: methanol = 20: 1) to obtain the target product (0.13 g; yield: 56.5%).
[0235] [0235] Molecular formula: C27H23Cl2N3O4S Molecular weight: 555.1 LC-MS (m / z): 556.1 (M + H +)
[0236] [0236] 1H-NMR (400 MHz, MeOD) δ: 8.33 (s, 1H); 7.98 (dd, J1 = 8.8 Hz, J2 = 1.6 Hz, 1H); 7.49-7.51 (m, 2H); 7.41-7.46 (m, 2H); 4.39-4.47 (m, 2H); 3.57-3.59 (m, 1H); 3.43-3.45 (m, 1H); 2.95-3.01 (m, 1H); 2.62-2.65 (m, 1H); 2.32-2.36 (m, 1H); 1.75-1.79 (m, 1H); 1.61 - 1.69 (m, 2H); 1.33-1.36 (m, 1H); 1.07-1.20 (m, 4H) Example 8: Preparation of 2 - ((1S, 4S, 5R) -5 - ((5-cyclopropyl-3- (2-trifluoromethoxy) phenyl) isoxazol-4-yl ) methoxy) -2-azabicyclo [2.2.1] heptan-2-yl) -4-fluorobenzo [d] thiazol-6-formic acid (compound 8)
[0237] [0237] Methyl 3-fluoro-4-nitrobenzoate (3.0 g; 15.1 mmol) was dissolved in MeOH (50 ml), and Pd / C (1.2 g) was added to an N2 atmosphere. The mixing solution was subjected to hydrogenation at 25 ° C for 3 hours. The Pd / C was separated by filtration through celite and the residue was concentrated to obtain the product (2.5 g; yield: 98.0%).
[0238] [0238] Methyl 4-amino-3-fluorobenzoate (2.5 g; 14.8 mmol) and KSCN (5.7 g; 59.1 mmol) were added to glacial acetic acid (50 ml) and the solution was stirred at 25 ° C for 15 minutes. Then, bromine (2.4 g; 14.9 mmol) was added dropwise, and the mixing solution continued to react at 25 ° C for 16 hours. The reaction solution was filtered and the filtrate was diluted with water (50 ml). The pH of the filtrate was adjusted to 8 with ammonia hydroxide. Then, the resulting solution was filtered and the filter cake was purified by silica gel column chromatography (petroleum ether: ethyl acetate = 5: 1) to obtain the product (200 mg, yield: 6.0% ).
[0239] [0239] Methyl 2-Amino-4-fluorobenzo [d] thiazole-6-formate (200 mg; 0.88 mmol)
[0240] [0240] Hydroxylamine hydrochloride (5.88 g; 84.7 mmol) was dissolved in water (60 ml), and the solution was stirred at 0 ° C. NaOH (3.5 g; 87.6 mmol) was dissolved in water (60 mL) and then added dropwise to the reaction flask. 2- trifluoromethoxybenzaldehyde (14 g; 73.6 mmol) was dissolved in an anhydrous ethanol solution (60 mL) and then added dropwise to the reaction flask. After the addition was complete, the mixture solution was reacted at 25 ° C for 1 hour. The reaction solution was diluted with water (300 ml) and extracted with ethyl acetate (500 ml x 3).
[0241] [0241] (E) -2-trifluoromethoxybenzaldoxima (15.4 g; crude product) was dissolved in DMF (150 mL), and then NCS (11.23 g; 84.1 mmol) was added in batches at a temperature not exceeding 25 ° C. After the addition was complete, the mixture solution was reacted at 25 ° C for 1 hour. The reaction solution was diluted with water (150 ml) and extracted with ethyl acetate (500 ml x 3). The organic phases were combined,
[0242] [0242] Potassium carbonate (10.44 g; 75.52 mmol) was added to THF (100 mL), and the ethyl 3-cyclopropyl-3-oxopropionate hydrochloride solution (10.44 g; 73.4 mmol) in THF (50 mL) was added to the reaction system. The system was stirred at -10 ° C for 30 min. Then, the solution of N-hydroxy-2-trifluoromethoxy) benzimidoyl (16.6 g; crude product) in THF (50 mL) was added to the reaction system at -5 ° C, and the system reacted at 35 ° C. ° C for 6 hours. The reaction solution was diluted with water (200 ml) and extracted with ethyl acetate (500 ml x 3).
[0243] [0243] Methyl 5-cyclopropyl-3- (2- (trifluoromethoxy) phenyl) isoxazol-4-formate (2.4 g, 7.33 mmol) was dissolved in anhydrous THF (50 mL), and DIBAL-H ( 1.5 M methylbenzene solution; 15 mL) was added dropwise at 0 ° C. After the addition was complete, the mixing solution was reacted at 25 ° C for 2 hours. MeOH (2 ml) was added dropwise to the reaction system at 0 ° C to stop the reaction and then water (50 ml) and ethyl acetate (100 ml) were added. The resulting solution was filtered through celite; then, the filtrate was separated by standing and the aqueous phase was extracted with ethyl acetate (100 ml x 2). The organic phases were combined, dried, concentrated and purified by silica gel column chromatography (petroleum ether: ethyl acetate = 5: 1) to obtain a product (1.0 g; yield: 45.6%).
[0244] [0244] (5-cyclopropyl-3- (2- (trifluoromethoxy) phenyl) isoxazol-4-yl) -methanol (1 g; 3.34 mmol) was dissolved in dichloromethane (20 mL) and triphenylphosphine (1.31 g ; 5.01 mmol) was added. Carbon tetrabromide (1.66 g; 5.07 mmol) was added in batches at 0 ° C and, after completion of the addition, the mixing solution was reacted at 25 ° C for 2 hours. The reaction solution was concentrated and purified by a silica gel column (petroleum ether: ethyl acetate = 10: 1) to obtain a product (860 mg; yield: 72.0%).
[0245] [0245] (1RS, 4RS, 5Rs) -5-hydroxy-2-azabicyclo [2.2.1] heptane-2-formate (0.15 g, 0.7 mmol), potassium tert-butoxide (118 mg; 1 , 05 mmol) and 18-crown-6 (93 mg; 0.35 mmol) were added in THF (20 mL) and the solution of the mixture was reacted at 25 ° C for 5 minutes. Then 4- (bromomethyl) -5-cyclopropyl-3- (2-
[0246] [0246] (1RS, 4RS, 5SR) -5 - ((5-cyclopropyl-3- (2-dichlorophenyl) isoxazol-4-yl) methoxy) -2-azabicyclo [2.2.1] heptane-2-carboxylate tert -butyl (0.2 g, 0.4 mmol) was added to DCM (10 ml) and then TFA (4 ml) was added. The mixing solution was reacted at 25 ° C for 2 hours and the reaction solution was concentrated to obtain the crude product (300 mg).
[0247] [0247] 4 - ((((((1S, 4S, 5R) -2-azabicyclo [2.2.1] heptan-5-yl) oxy) methyl) -5-cyclopropyl-3- (2-trifluoromethoxy) phenyl) trifluoroacetate isoxazole (300 mg; crude product), 2-bromo-4-fluorobenzo
[0248] [0248] 2 - ((1S, 4S, 5R) -5 - ((5-cyclopropyl-3- (2- (trifluoromethoxy) phenyl) isoxazol-4-yl) methoxy) -2-azabicycle [2.2.1] heptan -2-yl) -4-fluorobenzo [d] thiazol-6-methyl formate (200 mg; 0.33 mmol) and lithium hydroxide monohydrate (70 mg; 1.65 mmol) were dissolved in the mixed methanol solution ( 10 ml), tetrahydrofuran (10 ml) and water (10 ml), and the mixing solution was stirred at 25 ° C for 4.6 hours. The reaction solution was concentrated and the pH of the aqueous phase was adjusted to 2 with dilute hydrochloric acid (1M), and then the aqueous phase was extracted with ethyl acetate (50 ml x 3). The organic phases were combined and concentrated and the residue was purified by TLC (dichloromethane: methanol = 10: 1) to obtain the product (120 mg; yield 61.7%).
[0249] [0249] Molecular formula: C28H23F4N3O5S Molecular weight: 589.56 LC-MS (M / e): 590.2 (M + H) +
[0250] [0250] 1H-NMR (400 MHz, CDCl3) δ: 8.09 (s, 1H); 7.72 (d, J = 6.0 Hz, 1H); 7.52 (s, 2H); 7.35-7.38 (m, 2H); 4.29-4.35 (m, 2H); 3.59 (s, 1H); 3.49 (s, 1H); 3.00 (s, 1H);
2.63 (s, 1H); 2.07 (s, 2H); 1.69 (s, 2H); 1.47-1.50 (m, 1H); 1.21 (s, 2H); 1.10 (m, 2H).

权利要求:
Claims (16)
[1]
1. Compound, CHARACTERIZED by the fact that it has the general formula (I), a pharmaceutically acceptable salt thereof, an ester thereof or a stereoisomer of the compound, the salt or ester, in which R1 is selected from a group that consists of halogen, hydroxyl, amine, cyano, C1-6 alkyl, haloC1-6 alkyl, C1-6 hydroxy alkyl, C1-6 alkyl, C1-6 alkoxy, C1-6 alkylamine, C1-6 alkylcarbonyl, C1- alkoxy 6 C1-6 alkyl, 3-8 member cycloalkyl, 3-8 member cycloalkyl C1-6 alkyl, 3-8 member cycloalkyl C1-6 alkoxy, 3-8 member heterocyclyl, 3-8 member heterocyclyl C1 alkyl -6, or 3-8 membered heterocyclic C1-6 alkoxy; X1 and X2 are selected individually and independently from a group consisting of N, NR2, O, S or CR3R4; R2, R3 and R4 are selected individually and independently from a group consisting of hydrogen, halogen, hydroxyl, amine, cyano, C1-6 alkyl, haloC1-6 alkyl, C1-6 alkoxy or C1-6 alkylamine; M is C1-6 alkylene, in which one or more carbon atoms in C1-6 alkylene are optionally replaced by a heteroatom or a group, and the heteroatom or group is selected from a group consisting of N, NH, O, CO, S, SO or SO2; ring A is selected from 7-membered bridged cyclyl or 7-membered bridged heterocyclic; ring B is selected from a group consisting of 6-10 membered aryl, 5-10 membered heteroaryl, 3-14 membered heterocyclic and 3-8 membered cycloalkyl that are optionally substituted by a Q1 or more; each Q1 is independently selected from a group consisting of halogen, hydroxyl, amine, cyano, C1-6 alkyl, haloC1-6 alkyl, hydroxy C1-6 alkyl, amino C1-6 alkyl, C1-6 alkoxy, C1 alkylamine -6, C1-6 alkylcarbonyl, C1-6 alkylsulfonyl, C1-6 alkylsulfinyl, 3-8 membered cycloalkyl, 3-8 membered cycloalkyl C1-6alkyl, 3-8 membered cycloalkyl C1-6 alkoxy, 3-heterocyclic -8 members C1-6 alkyl, or 3-8 membered heterocyclic C1-6 alkoxy; L is absent or is C 1-6 alkylene, where one or more carbon atoms in C 1-6 alkylene are optionally replaced by a heteroatom or a group, and the heteroatom or group is selected from a group consisting of N , NH, O, CO, S, SO or SO2; Ar is selected from a group consisting of 6-10 member aryl, 6-10 member aryl C1-6 alkyl, 6-10 member aryl C1-6 alkoxy, 5-10 member heteroaryl, 5 heteroaryl -10 members C1-6 alkyl, 5-10 members heteroaryl C1-6 alkoxy, 3-8 members cycloalkyl and 3-8 members heterocyclic which are optionally substituted by one or more Q2; and each Q2 is independently selected from a group consisting of halogen, hydroxyl, amine, cyano, C1-6 alkyl, haloC1-6 alkyl, hydroxy C1-6 alkyl, amino C1-6 alkyl, C1-6 alkoxy, alkoxy C1-6 alkyl, C1-6 alkylamine, C1-6 alkylcarbonyl, C1-6 alkylsulfonyl, C1-6 alkylsulfinyl, 3-8 membered cycloalkyl, 3-8 membered cycloalkyl C1-6alkyl, 3-8 membered cycloalkyl alkoxy C1-6, 3-8 membered heterocyclic, 3-8 membered heterocyclic C1-6 alkyl, or 3-8 membered heterocyclic C1-6 alkoxy.
[2]
2. A compound, a pharmaceutically acceptable salt thereof, an ester thereof or a stereoisomer of the compound, the salt or ester according to claim 1, CHARACTERIZED by the fact that
R1 is selected from the group consisting of halogen, hydroxyl, amine, cyano, C1-4 alkyl, haloC1-4 alkyl, hydroxy C1-4 alkyl, amino C1-4 alkyl, alkoxy
C1-4, C1-4 alkylamine, C1-4 alkylcarbonyl, C1-4 alkoxy C1-4 alkyl, 3-6 membered cycloalkyl, 3-6 membered cycloalkyl C1-4 alkyl, 3-6 membered cycloalkyl alkoxy
C1-4, 3-6 membered mono-heterocyclic, 3-6 membered mono-heterocyclic alkyl
C1-4 or 3-6 membered mono-heterocyclic C1-4 alkoxy;
Preferably, X1 and X2 are selected individually and independently from a group consisting of N, NR2, O, S or CR3R4; R2, R3 and
R4 are selected individually and independently from a group consisting of hydrogen, halogen, hydroxyl, amine, cyano, C1-4 alkyl, haloC1-4 alkyl, C1-4 alkoxy or C1-4 alkylamine;
preferably, M is C 1-4 alkylene, in which one or more carbon atoms in C 1-4 alkylene are optionally replaced by a heteroatom or group, and the heteroatom or group is selected from a group consisting of N, NH,
O, CO, S, SO or SO2;
preferably, ring A is selected from 7-membered bridged cyclyl or 7-membered bridged nitrogen heterocyclic; preferably, ring B is selected from 8-10 membered fused heteroaryl and 7-14 membered fused heterocyclic which are optionally substituted by 1 to 2 Q1 and contain 1 to 3 heteroatoms or groups, and the heteroatom or group is independently selected from from a group consisting of N, NH,
O, S, SO or SO2;
preferably, each Q1 is independently selected from a group consisting of halogen, hydroxyl, amine, cyano, C1-4 alkyl, haloC1-4 alkyl,
hydroxy C1-4 alkyl, aminoC1-4 alkyl, C1-4 alkoxy, C1-4 alkylamine, C1-4 alkylcarbonyl,
C1-4 alkylsulfonyl, C1-4 alkylsulfinyl, 3-6 membered cycloalkyl, 3-6 membered cycloalkyl C1-4alkyl, 3-6 membered cycloalkyl C1-4 alkoxy, mono-heterocyclic
3-6 member, 3-6 member mono-heterocyclic C1-4 alkyl or 3-6 member mono-heterocyclic C1-4 alkoxy; preferably, L is absent or is C 1-4 alkylene, in which one or more carbon atoms in C 1-4 alkylene are optionally replaced by a heteroatom or a group, and the heteroatom or group is selected from a group consisting of in NH, O, CO, S, SO or SO2; preferably, Ar is selected from a group consisting of 6-8 membered monocycloaryl, 8-10 membered fused aryl, 6-8 membered C1-4 alkyl monocycle, fused 8-10 membered C1-4 alkyl aryl , 6-8 membered C1-4 alkoxy monocycloyl, 8-10 membered C1-4 alkoxy fused aryl, 5-7 membered monocyclo-heteroaryl, 8-10 membered fused heteroaryl, 5-7 membered C1 heteroaryl -4 alkyl, 8-10 membered C1-4 heteroaryl fused alkyl, 5-7 membered C1-4 alkoxy heteroaryl alkoxy, 8-10 membered C1-4 alkoxy fused heteroaryl, 3-8 membered cycloalkyl and heterocyclic 3-8 members, which are optionally replaced by 1 to 3 Q2; and preferably, each Q2 is independently selected from a group consisting of halogen, hydroxyl, amine, cyano, C1-4 alkyl, haloC1-4 alkyl, hydroxy C1-4 alkyl, amino C1-4 alkyl, C1-4 alkoxy , C1-4 alkoxy alkyl, C1-4 alkylamine, C1-4 alkylcarbonyl, C1-4 alkylsulfonyl, C1-4 alkylsulfinyl, 3-6 membered cycloalkyl, 3-6 membered cycloalkyl C1-4alkyl, 3-6 cycloalkyl C1-4 alkoxy members, 3-6 membered monoheterocyclic, 3-6 membered monoheterocyclyl, or C1-4 alkoxy 3-6 membered monoheterocyclyl.
[3]
3. A compound, a pharmaceutically acceptable salt thereof, an ester thereof or a stereoisomer of the compound, the salt or ester according to any one of claims 1 to 2, CHARACTERIZED by the fact that M is selected from the group consisting of -CH2-, -CH2-CH2-, -CH2-CH2- CH2-, -CH2-NH-, -CH2-CH2-O- and -CH2-NH-CO-;
preferably, ring A is selected from saturated 7-membered bridged cyclyl or 7-membered saturated nitrogenous heterocyclic, and when ring A is a saturated nitrogenous heterocyclic bridged, preferably ring A is attached to L or to the ring B by a nitrogen atom in the ring; preferably, X1 and X2 are selected individually and independently from a group consisting of N, NR2, O or S; R2 is selected from a group consisting of hydrogen, halogen, hydroxyl, amine, methyl, ethyl, propyl, isopropyl or trifluoromethyl.
[4]
4. A compound, a pharmaceutically acceptable salt thereof, an ester thereof or the stereoisomer of the compound, the salt or ester according to any one of claims 1 to 3, CHARACTERIZED by the fact that R1 is selected from the group that consists of halogen, hydroxyl, amine, cyano, C1-4 alkyl, haloC1-4 alkyl, hydroxy C1-4 alkyl, amino C1-4 alkyl, C1-4 alkoxy, C1-4 alkylamine, C1-4 alkylcarbonyl, C1- alkoxy 4 C1-4 alkyl, 3-4-membered cycloalkyl, 3-4-membered cycloalkyl, C1-4-membered cycloalkyl, 3-4-membered cycloalkyl, 3-4-membered mono-heterocyclic, 3--membered mono-heterocyclyl 4 members C1-4 alkyl or 3-4 membered monoheterocyclic C1-4 alkoxy; and, preferably, ring A is selected from the following groups:,,,,,,,,,,,,,,,,,,,,,,,,.
[5]
5. A compound, a pharmaceutically acceptable salt thereof, an ester thereof or the stereoisomer of the compound, the salt or ester according to any one of claims 1 to 4, CHARACTERIZED by the fact that ring B is selected from fused heteroaryl of 9-10 members containing 1 to 2 heteroatoms or groups and is optionally substituted by 1 to 2 Q1, and the heteroatom or group is selected independently from a group consisting of N, NH, O, S, SO or SO2; preferably, ring B is attached to L or ring A by a ring carbon atom; preferably, each Q1 is independently selected from a group consisting of halogen, hydroxyl, amine, cyano, C1-4 alkyl, haloC1-4 alkyl, C1-4 alkoxy, 3-6 membered cycloalkyl, 3-6 cycloalkyl C1-4 alkyl members, 3-6 members C1-4 alkoxy cycloalkyl, 3-6 members mono-heterocyclic, 3-6 members C1-4 alkyl mono-heterocyclic or 3-6 members C1- alkoxy mono-heterocyclic 4; and, preferably, L is absent or is C1-2 alkylene, where one or more carbon atoms in C1-2 alkylene are optionally replaced by a heteroatom or group, and the heteroatom or group is selected from a group consisting of NH, O, S or CO.
[6]
6. A compound, a pharmaceutically acceptable salt thereof, an ester thereof or the stereoisomer of the compound, the salt or ester according to any one of claims 1 to 5, CHARACTERIZED by the fact that Ar is selected from a group consisting of in phenyl, phenylC1-4 alkyl, phenylC1-4 alkoxy, 5-6 membered monocyclic heteroaryl, 5-6 membered C1-4alkyl heteroaryl and 5-6 membered C1-4 alkoxy heterocyclyl which are optionally substituted for 1 to 2 Q2; and each Q2 is independently selected from a group consisting of halogen, hydroxyl, amine,
cyano, C1-4 alkyl, haloC1-4 alkyl, hydroxyC1-4 alkyl, aminoC1-4 alkyl, C1-4 alkoxy, C1-4 alkoxy alkyl or C1-4 alkylamine.
[7]
7. A compound, a pharmaceutically acceptable salt thereof, an ester thereof or the stereoisomer of the compound, the salt or ester according to any one of claims 1 to 6, CHARACTERIZED by the fact that R1 is selected from the group consisting of halogen, methyl, ethyl, propyl, isopropyl, trifluoromethyl, trifluoroethyl, methoxy, ethoxy, propoxy, isopropoxy, methylamine, ethylamine, methoxymethyl, methoxyethyl, ethoxymethyl, cyclopropyl, cyclopropylmethyl, cyclopropylmethyl, cyclopropylmethyl, cyclopropylmethyl, cyclopropylethyl epoxyethylmethyl, azacyclopropyl, azacyclopropylmethyl, oxacyclobutyl or azacyclobutyl; X1 and X2 are selected individually and independently from a group consisting of N, NH, O or S; M is selected from -CH2-, -CH2-CH2- or -CH2-CH2-CH2-; ring A is selected from the following groups:,,,,,,,,,,,,, and; ring B is selected from the following groups, optionally substituted by 1 Q1:,,,,,,,, and;
Q1 is selected from the group consisting of fluorine, chlorine, bromine, hydroxyl, amine, cyano, methyl, ethyl, propyl, isopropyl, fluoromethyl, difluoromethyl, trifluoromethyl, 1-fluoroethyl, 2-fluoroethyl, 1,1-difluoroethyl, 1 , 2-difluoroethyl, 2,2-difluoroethyl, 2,2,2-trifluoroethyl, 3,3,3-trifluoropropyl, 1-trifluoromethylethyl, methoxy, ethoxy, propoxy, isopropoxy, cyclopropyl, cyclopropylmethyl, cyclopropylethyl, cyclopropylmethyl, cyclopropylmethyl, cyclobutyl , cyclobutylethyl, cyclobutylmethoxy, cyclopentyl, cyclohexyl, epoxyethyl, epoxyethylmethyl, azacyclopropyl, azacyclopropylmethyl, oxacyclobutyl, azacyclobutyl, tetrahydrofuryl, pyrrolidyl, imidazolidinyl, pyrazininyl, pyridine, pyridine, pyridine, pyridine; L is absent; Ar is selected from a group consisting of phenyl, phenylmethyl, phenylethyl, phenylmethoxy, furyl, pyrrilla, thienyl, pyrazolyl, imidazolyl, pyridyl and pyrimidinyl, which are optionally substituted by 1 to 2 Q2; and each Q2 is independently selected from a group consisting of fluorine, chlorine, bromine, hydroxyl, amine, cyan, methyl, ethyl, propyl, isopropyl, trifluoromethyl, trifluoroethyl, methoxy, ethoxy, propoxy, isopropoxy, methylamine, ethylamine, methoxymethyl, methoxyethyl or ethoxymethyl.
[8]
8. A compound, a pharmaceutically acceptable salt thereof, an ester thereof or a stereoisomer of the compound, the salt or ester according to claim 7, CHARACTERIZED by the fact that R1 is selected from the group consisting of cyclopropyl, cyclopropylmethyl, cyclopropylethyl, cyclopropylmethoxy, cyclobutyl, cyclobutylmethyl, cyclobutylethyl, cyclobutylmethoxy, epoxyethyl, epoxyethylmethyl, azacyclopropyl, azacyclopropylmethyl, oxacyclobutyl or azacyclobutyl; ring A is selected from a group consisting of, and;
ring B is selected from the following groups, optionally substituted by 1 Q1:, e; Q1 is selected from a group consisting of fluorine, chlorine, bromine, methyl, ethyl, propyl, isopropyl, trifluoromethyl, methoxy, ethoxy, propoxy or isopropoxy; Ar is selected from a group consisting of phenyl, pyridyl and pyrimidinyl, which are optionally substituted by 1 to 2 Q2; each Q2 is independently selected from a group consisting of fluorine, chlorine, bromine, methyl, ethyl, propyl, isopropyl, methoxy or ethoxy; and preferably, X1 and X2 are both O.
[9]
9. The compound, a pharmaceutically acceptable salt thereof, an ester thereof or a stereoisomer of the compound, the salt or ester according to any one of claims 1 to 8, CHARACTERIZED by the fact that having the structure shown below as a general formula (II):.
[10]
10. The compound, a pharmaceutically acceptable salt thereof, an ester thereof or a stereoisomer of the compound, the salt or ester according to claim 1, CHARACTERIZED by the fact that the compound is selected from: Cis-Cis-,,
,, Trans-, Cis-, Cis- Cis-, and Trans-.
[11]
11. Compound, CHARACTERIZED by the fact that it has the general formula (II), a pharmaceutically acceptable salt of it, an ester thereof or a stereoisomer of the compound, the salt or the ester,
wherein X1, X2, R1, Q1 and Ar are as described in any one of claims 1 to 9, wherein Q2 is independently selected from a group consisting of, methyl, ethyl, propyl, isopropyl, ethoxy or trifluoromethoxy.
[12]
12. As a compound, a pharmaceutically acceptable salt thereof, an ester thereof or a stereoisomer of the compound, the salt or ester according to claim 11, CHARACTERIZED by the fact that R1 is selected from the group consisting of cyclopropyl, cyclopropylmethyl, cyclopropylethyl, cyclopropylmethoxy, cyclobutyl, cyclobutylmethyl, cyclobutylethyl, cyclobutylmethoxy, epoxyethyl, epoxyethylmethyl, azacyclopropyl, azacyclopropylmethyl, oxacyclobutyl or azacyclobutyl; X1 and X2 are selected individually and independently from a group consisting of N, NH, O or S; Q1 is selected from a group consisting of fluorine, chlorine, bromine, methyl, ethyl, propyl, isopropyl, trifluoromethyl, methoxy, ethoxy, propoxy or isopropoxy; Ar is selected from a group consisting of phenyl, pyridyl and pyrimidinyl, which are optionally substituted by 1 to 2 Q2; each Q2 is independently selected from a group consisting of methyl, ethyl, propyl, isopropyl, ethoxy or trifluoromethoxy; preferably, R1 is selected from cyclopropyl and cyclobutyl; and preferably, Ar is phenyl optionally substituted by 1 to 2 Q2.
[13]
13. The compound, a pharmaceutically acceptable salt thereof, an ester thereof or a stereoisomer of the compound, the salt or ester according to claim 12, CHARACTERIZED by the fact that
.
[14]
14. Pharmaceutical formulation, CHARACTERIZED by the fact that it contains the compound, a pharmaceutically acceptable salt thereof, an ester thereof or a stereoisomer of the compound, the salt or ester as defined in any one of claims 1 to 13, wherein the pharmaceutical formulation contains one or more pharmaceutically acceptable carriers and / or diluents and can be of any pharmaceutically acceptable dosage form.
[15]
15. Use of the compound, a pharmaceutically acceptable salt thereof, an ester thereof or a stereoisomer of the compound, the salt or ester as defined in any of claims 1-13, CHARACTERIZED by the fact that it is in the preparation of a medicament for prevent and / or treat FXR-mediated diseases, preferably diseases are selected from atherosclerosis, bile acid metabolism disorder, primary sclerosing cholangitis, cholesterol calculation, fibrosis-related diseases, fatty liver, cirrhosis, hepatitis, liver failure, cholestasis , cholelithiasis, myocardial infarction, stroke, thrombus, clinical complications of type I or type II diabetes, hyperproliferative diseases and inflammatory bowel diseases.
[16]
16. Use according to claim 15, CHARACTERIZED by the fact that the diseases are selected from alcoholic fatty liver disease, non-alcoholic fatty liver disease, primary biliary cirrhosis, primary cholangetic cirrhosis, chronic hepatitis, non-viral hepatitis, steato- alcoholic hepatitis, non-alcoholic steatohepatitis, benign intrahepatic cholestasis, progressive familial intrahepatic cholestasis, drug-induced cholestasis, cholestasis related to gastrointestinal nutrition, extrahepatic cholestasis, hypercholesterolemia,
neonatal jaundice, kernicterus, diabetic nephropathy, diabetic neuropathy, diabetic retinopathy, other observed results of clinical manifest chronic diabetes,
hepatocellular carcinoma, colon adenoma, polyposis, colon adenocarcinoma,
breast cancer, pancreatic cancer, esophageal carcinoma and other forms of gastrointestinal and hepatic neoplastic diseases.

类似技术:

---

公开号 | 公开日 | 专利标题

---

BR112020000180A2|2020-07-14|fxr receptor agonist

---

JP2021107425A|2021-07-29|SUBSTITUTED 1,2,3,4-TETRAHYDROCYCLOPENTA[b]INDOL-3-YL)ACETIC ACID DERIVATIVE USEFUL IN TREATMENT OF AUTOIMMUNE AND INFLAMMATORY DISORDERS

---

US20210228599A1|2021-07-29|Farnesoid x receptor modulators

---

JP2017537960A|2017-12-21|Azabicyclooctane derivatives as FXR agonists for use in the treatment of liver and gastrointestinal diseases

---

CN106083978A|2016-11-09|Sulfonyl amino carbonyl derivant, its pharmaceutical composition and application

---

CN103391937A|2013-11-13|Compositions and methods for modulating farnesoid x receptors

---

US10105373B2|2018-10-23|Fused triterpene compounds and uses thereof

---

BR112021010871A2|2021-08-31|THRB RECEPTOR AGONIST COMPOUND AND METHOD OF PREPARATION AND USE THEREOF

---

CN106573901A|2017-04-19|Fumarate analogs and uses thereof in the treatment of an autoimmune disease or an inflammatory disease

---

WO2020156241A1|2020-08-06|Aromatic ring or heteroaromatic ring compounds, preparation method therefor and medical use thereof

---

CN111868056A|2020-10-30|1,2, 4-oxadiazole compound and preparation method and medical application thereof

---

WO2020001304A1|2020-01-02|Fxr receptor agonist

---

WO2019149089A1|2019-08-08|Nitrogen-containing benzoheterocycle compound comprising carboxylic acid group, preparation method and use thereof

---

JP6997870B2|2022-02-10|FXR receptor stimulant

---

WO2020108485A1|2020-06-04|Solid form, crystalline form, and crystal form a of fxr agonist, and preparation method therefor and application thereof

---

US10647673B2|2020-05-12|Acetophenone compound, preparation method thereof, and application thereof in fatty liver prevention and treatment

---

TW202140009A|2021-11-01|3-carboxamido-4-hydroxynaltrexone deuterated derivative, preparation method therefor and application thereof

---

BR112015028399B1|2021-11-16|11-HYDROXYL DERIVATIVES OF BILIARY ACIDS AND AMINO ACID CONJUGATES THEREOF AS FARNESOID X RECEPTOR MODULATORS, PHARMACEUTICAL COMPOSITION AND USE OF REFERRED TO DERIVATIVES

---

JP2022518603A|2022-03-15|Aromatic ring or aromatic heterocyclic compound, its production method and pharmaceutical use

---

WO2020168152A2|2020-08-20|Substituted amide compounds useful as farnesoid x receptor modulators

---

NZ714082B2|2021-05-27|11-hydroxyl-derivatives of bile acids and amino acid conjugates thereof as farnesoid x receptor modulators

同族专利:

---

公开号 | 公开日

---

AU2018298253B2|2020-11-19|

---

JP2020527598A|2020-09-10|

---

CN112876467A|2021-06-01|

---

KR20200029509A|2020-03-18|

---

US20210024522A1|2021-01-28|

---

AU2018298253A1|2020-02-27|

---

EP3650449A4|2021-04-07|

---

CN110944997B|2021-04-09|

---

CA3068928A1|2019-01-10|

---

WO2019007418A1|2019-01-10|

---

EP3650449A1|2020-05-13|

---

CN110944997A|2020-03-31|

引用文献:

---

公开号 | 申请日 | 公开日 | 申请人 | 专利标题

---

---

ES2326850T3|1998-12-23|2009-10-20|Glaxo Group Limited|TESTS FOR NUCLEAR RECEPTORS LIGANDOS.|

---

EP2289883A1|2009-08-19|2011-03-02|Phenex Pharmaceuticals AG|Novel FXR binding and activity modulating compounds|

---

SI2655370T1|2010-12-20|2017-12-29|Novartis Ag|Compositions and methods for modulating fxr|

---

WO2012087520A1|2010-12-20|2012-06-28|Irm Llc|Compositions and methods for modulating farnesoid x receptors|

---

EP3034501A1|2014-12-17|2016-06-22|Gilead Sciences, Inc.|Hydroxy containing FXR modulating compounds|

---

CN107106555A|2014-12-18|2017-08-29|诺华股份有限公司|Application of the azabicyclo-octane derivative as FXR activators in treatment liver and gastrointestinal disease|

---

CN110177783A|2016-08-23|2019-08-27|阿德利克斯股份有限公司|For treating the hormone receptor modulator of metabolic condition and illness|CN110177783A|2016-08-23|2019-08-27|阿德利克斯股份有限公司|For treating the hormone receptor modulator of metabolic condition and illness|

---

BR112019006651A2|2016-10-04|2019-07-02|Enanta Pharm Inc|isoxazole analogues as fxr agonists and methods of using it|

---

US10597391B2|2016-10-26|2020-03-24|Enanta Pharmaceuticals, Inc.|Urea-containing isoxazole derivatives as FXR agonists and methods of use thereof|

---

US10689391B2|2017-12-12|2020-06-23|Enanta Pharmaceuticals, Inc.|Isoxazole analogs as FXR agonists and methods of use thereof|

---

CN110128432B|2018-02-02|2021-03-02|广东东阳光药业有限公司|Nitrogenous tricyclic compound and application thereof in medicine|

---

WO2019160813A1|2018-02-14|2019-08-22|Enanta Pharmaceuticals, Inc.|Isoxazole derivatives as fxr agonists and methods of use thereof|

---

AU2020312735A1|2019-07-18|2021-12-16|Enyo Pharma|Method for decreasing adverse-effects of interferon|

---

WO2021014349A1|2019-07-23|2021-01-28|Novartis Ag|Treatment comprising fxr agonists|

---

WO2021014350A1|2019-07-23|2021-01-28|Novartis Ag|Combination treatment of liver diseases using fxr agonists|

---

WO2021044287A1|2019-09-03|2021-03-11|Novartis Ag|Treatment of liver disease or disorder comprising actrii receptor antagonists|

---

WO2021053618A1|2019-09-19|2021-03-25|Novartis Ag|Treatment comprising fxr agonists|

---

WO2021064575A1|2019-09-30|2021-04-08|Novartis Ag|Treatment comprising the use of fxr agonists|

---

TW202135811A|2019-12-20|2021-10-01|瑞士商諾華公司|Combination treatment of liver diseases using integrin inhibitors|

---

WO2021144330A1|2020-01-15|2021-07-22|INSERM |Use of fxr agonists for treating an infection by hepatitis d virus|

法律状态:
2021-06-22| B25D| Requested change of name of applicant approved|Owner name: XUANZHU (SHIJIAZHUANG) BIOPHARMACEUTICAL CO., LTD. (CN) |

---

2021-07-13| B25G| Requested change of headquarter approved|Owner name: XUANZHU (SHIJIAZHUANG) BIOPHARMACEUTICAL CO., LTD. (CN) |

---

2021-08-03| B25D| Requested change of name of applicant approved|Owner name: XUANZHU BIOPHARMACEUTICAL CO., LTD. (CN) |

---

2021-11-03| B350| Update of information on the portal [chapter 15.35 patent gazette]|

优先权:

---

申请号 | 申请日 | 专利标题

---

CN201710547157.X|2017-07-06|

---

CN201710547157|2017-07-06|

---

PCT/CN2018/094813|WO2019007418A1|2017-07-06|2018-07-06|Fxr receptor agonist|

---