CN110452235B - Fluorine-containing isoxazole compound and preparation method, pharmaceutical composition and application thereof - Google Patents

Fluorine-containing isoxazole compound and preparation method, pharmaceutical composition and application thereof Download PDF

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CN110452235B
CN110452235B CN201810431514.0A CN201810431514A CN110452235B CN 110452235 B CN110452235 B CN 110452235B CN 201810431514 A CN201810431514 A CN 201810431514A CN 110452235 B CN110452235 B CN 110452235B
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CN110452235A (en
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柳红
李佳
周宇
臧奕
李亚洲
孙丹丹
李俊友
茹弘博
高立信
蒋华良
陈凯先
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Shanghai Institute of Materia Medica of CAS
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Abstract

The invention relates to a fluorine-containing isoxazole compound, a preparation method thereof, a pharmaceutical composition and application thereof. Specifically, the invention discloses a compound shown as a general formula I or an enantiomer or a diastereoisomer thereof, or a pharmaceutically acceptable salt thereof, or a mixture of the enantiomer and the diastereoisomer. And discloses that the compound is a small molecule agonist targeting FXR and can be used for treating FXR mediated diseases.

Description

Fluorine-containing isoxazole compound and preparation method, pharmaceutical composition and application thereof
Technical Field
The invention relates to the field of medicinal chemistry and pharmacotherapeutics, in particular to a fluorine-containing isoxazole compound, a preparation method thereof, a pharmaceutical composition containing the compound and application of the fluorine-containing isoxazole compound as an FXR agonist, especially application of the fluorine-containing isoxazole compound in preparing medicaments for treating non-alcoholic fatty liver diseases and hepatic fibrosis.
Background
Non-alcoholic fatty liver disease (NAFLD) is a clinical pathological syndrome which is mainly characterized by excessive deposition of fat in liver cells, is caused by alcohol and other definite liver damage factors, is acquired metabolic stress liver injury, and is closely related to insulin resistance and genetic susceptibility. The prevalence rate of NAFLD in population with risk factors is increased by about 2% every year in developed countries, and the disease is gradually getting younger, which has become the first major liver disease in western countries and is also a common chronic liver disease type in China. NAFLD, in addition to directly causing decompensated liver cirrhosis, hepatocellular carcinoma and relapse of transplanted liver, can also affect the progression of other chronic liver diseases and be involved in the onset of type 2 diabetes and atherosclerosis, considered as a local manifestation of the liver of systemic metabolic syndrome.
NAFLD mainly includes Simple Fatty Liver (SFL) and nonalcoholic steatohepatitis (NASH), of which NASH is the most predominant type, with incidence of cirrhosis in NASH patients as high as 25% within 10 years. Unfortunately, the slow progress in research and treatment of NASH drugs has led to the erroneous belief that NASH was considered a benign complication of diabetes or obesity, leading to an overlooking of its timely diagnosis and treatment. Currently, the development of NASH therapeutic drugs mainly includes: (1) selective peripheral cannabinoid receptor (CB) blockers; (2) Peroxisome proliferator-activated receptor (PPAR) agonists; (3) Caspase (Caspase) inhibitors; (4) phosphodiesterase 4 (PDE 4) inhibitors; (5) Farnesoid X Receptor (FXR) agonists; (6) A chemokine receptor-5/chemokine receptor-2 (CCR 5/CCR 2) dual-target inhibitor; (7) apoptosis signal-regulating kinase-1 (ASK 1) inhibitors and the like. In the research and development of the targets and the medicines thereof, the farnesoid derivative X receptor (FXR) is found to be used as a bile acid receptor, not only controls the in-vivo bile acid homeostasis, but also has an important regulation effect on lipid and sugar metabolism, and is an important target for regulating the metabolic disorder of patients with metabolic syndrome.
The FXR receptor is a bile acid receptor, and the activation of FXR controls the processes of synthesis, transportation and the like of bile acid. FXR is a member of the nuclear receptor superfamily of hormones, with a typical nuclear receptor structure, and 4 subtypes FXR α 1, FXR α 2, FXR α 3 and FXR α 4 are now found. FXR ligands inhibit Hepatic Stellate Cell (HSC) expression, reduce extracellular matrix (ECM) production and increase ECM clearance, primarily by activating FXR and FXR target gene small heterodimer partner receptors (SHPs), thereby reducing ECM deposition; the FXR ligand can also increase the sensitivity of HSC to apoptosis, increase the apoptosis of HSC, and reverse hepatic fibrosis by interacting with other nuclear factors for reversing hepatic fibrosis, thereby playing a role in treating hepatic fibrosis.
In recent years, a plurality of FXR agonists are sequentially in clinical research, such as the compound LJN-452 found by Novatis has good biological activity and selectivity, the EC50 of the FXR reaches 0.2nM, and the FXR agonist enters a phase II clinical test for treating NASH and Primary Biliary Cholangitis (PBC); the FXR agonist PX-104 developed by Gilead corporation has entered phase II clinical studies for treatment of NASH, but unfortunately has been forced to terminate short of the desired clinical endpoint. Several other compounds, such as LY-2562175 and INT-767, have also been introduced into clinical studies with the promise of alleviating the unmet enormous clinical need for NASH. Of these, obeticholic acid (OCA) was approved for marketing as the first FXR agonist, a new drug candidate for the treatment of PBC and NASH, mainly in patients who do not respond adequately or are intolerant to ursodeoxycholic acid. The mechanism mainly influences the synthesis, secretion, transportation and absorption of bile acid by exciting FXR receptor and regulating related genes. The FDA awards breakthrough drug eligibility for OCA to treat NASH with liver fibrosis, the motorway status for treating PBC, the orphan drug status for treating PBC and PSC by year 1 of 2015, and has been approved by the FDA in the united states for marketing by day 27 of 2016. Global pharmaceutical market forecasting agency, evaluatePharma, issued a bang dan "global biopharmaceutical late pipeline heavy pound drug TOP 15," forecasts that global sales of OCA in 2020 will be as high as $ 30 billion. However, although obeticholic acid plays a positive role in reducing liver fibrosis, it raises low-density lipoprotein cholesterol (LDL) and lowers high-density lipoprotein cholesterol (HDL), suggesting that the drug may increase the risk of cardiovascular diseases, and may cause side effects such as itching of the patient's body, which limits its widespread use to some extent.
Therefore, in the face of the unmet clinical needs of NASH, the development of innovative NASH therapeutic drugs aiming at the novel NASH target is urgently needed, the market vacancy is filled, and the clinical drug requirements are met.
Disclosure of Invention
The invention aims to provide a novel FXR-targeting small molecule agonist and a preparation method and application thereof.
In a first aspect, the present invention provides a fluorine-containing isoxazole compound having the structure shown in the following general formula I, or an enantiomer or a diastereomer thereof, or a pharmaceutically acceptable salt thereof, or a mixture thereof:
Figure BDA0001653521430000021
wherein:
m is 0, 1,2 or 3;
n is 0, 1 or 2;
p is 0, 1 or 2;
Figure BDA0001653521430000031
the ring is selected from the group consisting of: substituted or unsubstituted 6-20 membered heterocyclic group, substituted or unsubstituted 6-20 membered aromatic ring group, substituted or unsubstituted 6-20 membered aromatic heterocyclic group; wherein, the substituted refers to that the hydrogen atom on the group is substituted by 1,2, 3 or 4 substituents selected from the following group: deuterium (D), tritium (T), halogen, C 1 -C 6 Alkyl, halo C 1 -C 6 Alkyl radical, C 1 -C 6 Alkoxy, halo C 1 -C 6 Alkoxy, -O [ (CH) 2 ) q O] r R 5 、-O(CH 2 ) s C 6 -C 10 Aryl radical, C 3 -C 8 Cycloalkoxy, halo C 3 -C 8 Cycloalkoxy, cyano, nitro, amino (preferably C) 1 -C 6 Amino), hydroxy, carboxyl, C 1 -C 6 Ester group, C 6 -C 10 Aryl radical, C 6 -C 10 Aryloxy, -X 4 -CO 2 R 5 、C 1 -C 6 Alkylhydroxy, -X 4 -CONR 5 R 6 、-X 4 -CONR 5 (CH 2 ) y1 CO 2 R 6 、-X 4 -CONR 5 (CH 2 ) y2 SO 3 R 6 3-12 membered heterocyclyl and 3-12 membered heterocyclyloxy; wherein q, r, s, y1 and y2 are 1,2, 3 or 4 respectively; the aromatic heterocyclic group, the aromatic ring group or the heterocyclic group independently contain 1 to 4 heteroatoms selected from oxygen, sulfur and nitrogen; x 4 Is a bond, C1-C6 alkylene, C 1-2 Alkylene, cyclopropyl or epoxyethyl;
R 1 selected from the group consisting of: a substituted or unsubstituted 6-20 membered heterocyclic group, a substituted or unsubstituted 6-20 membered aromatic ring group, a substituted or unsubstituted 6-20 membered aromatic heterocyclic group; wherein, theThe substitution means that hydrogen atoms on the group are substituted with 1,2, 3 or 4 substituents selected from the group consisting of: deuterium, tritium, halogen, C 1 -C 6 Alkyl radical, C 1 -C 6 Alkoxy, halo C 1 -C 6 Alkoxy, -O [ (CH) 2 ) q O] r R 5 、-O(CH 2 ) s C 6 -C 10 Aryl radical, C 3 -C 8 Cycloalkoxy, halo C 3 -C 8 Cycloalkoxy, cyano, nitro, amino (preferably C) 1 -C 6 Amino), hydroxy, hydroxymethyl, carboxy, C 6 -C 10 Aryl radical, C 6 -C 10 Aryloxy, 3-12 membered heterocyclyl and 3-12 membered heterocyclyloxy; wherein q, r and s are each 1,2, 3 or 4; the aromatic heterocyclic group, the aryl group or the heterocyclic group respectively and independently contain 1 to 4 heteroatoms selected from oxygen, sulfur and nitrogen;
R 2 substituents which may be independently selected from 1,2, 3 or 4 of the following groups: hydrogen, halogen, substituted or unsubstituted C 1 -C 6 Alkyl, substituted or unsubstituted C 1 -C 6 Alkoxy, -O [ (CH) 2 ) q O] r R 5 、-O(CH 2 ) s C 6 -C 10 Aryl radical, C 3 -C 8 Cycloalkoxy, halo C 3 -C 8 Cycloalkoxy, cyano, nitro, amino (preferably C) 1 -C 6 Amino), hydroxyl, hydroxymethyl, carboxyl; wherein, the substituted refers to that the hydrogen atom on the group is substituted by 1,2, 3 or 4 substituents selected from the following group: hydrogen, halogen, nitro, cyano, trifluoromethyl, trifluoroethyl, trifluoropropyl, trifluoromethoxy, trifluoroethoxy, amino (preferably C) 1 -C 6 Amino), hydroxyl, hydroxymethyl, carboxyl, mercapto and sulfonyl;
R 3 and R 4 Each independently selected from the group consisting of: hydrogen, halogen, C 1 -C 6 Alkyl, halo C 1 -C 6 Alkyl radical, C 1 -C 6 Alkoxy, halo C 1 -C 6 Alkoxy, -O [ (CH) 2 ) q O] r R 5 、-O(CH 2 ) s C 6 -C 10 Aryl radical, C 3 -C 8 Cycloalkoxy, halo C 3 -C 8 Cycloalkoxy, cyano, nitro, amino (preferably C) 1 -C 6 Amino), hydroxyl, hydroxymethyl, and carboxyl; or R 3 And R 4 Are connected together to form C 1 -C 6 An alkylene group;
R 5 and R 6 Is independently hydrogen, C 1 -C 6 Alkyl, halo C 1 -C 6 Alkyl radical, C 2 -C 6 Alkenyl, hydroxymethyl, or 5-7 membered heterocyclyl.
In another preferred embodiment, m is 0, 1 or 2; preferably 0 or 1.
In another preferred embodiment, n is 0 or 1.
In another preferred embodiment, p is 0 or 1.
In a further preferred embodiment of the method,
Figure BDA0001653521430000041
the ring is selected from the group consisting of: phenyl, pyridyl, pyrimidinyl, pyridazinyl, thiazolyl, benzothiazolyl, benzo [ d ]]Isothiazolyl, imidazo [1,2-a ] s]Pyridyl, quinolyl, 1H-indolyl, pyrrolo [1,2-b ]]Pyridazinyl, benzofuranyl, benzo [ b ]]Thiophenyl, 1H-indazolyl, benzo [ d ]]Isoxazolyl, thiazolinyl, 1H-pyrrolo [3,2-c ]]Pyridyl, pyrazolo [1,5-a ]]Pyrimidinyl, imidazo [1,2-b ]]Pyridazinyl, pyrazolo [1,5-a ]]A pyrrolyl group; the above groups are substituted or unsubstituted; wherein, the substituted refers to that the hydrogen atom on the group is substituted by 1,2, 3 or 4 substituents selected from the following group: D. t, halogen, C 1 -C 6 Alkyl, halo C 1 -C 6 Alkyl radical, C 1 -C 6 Alkoxy, halo C 1 -C 6 Alkoxy, -O [ (CH) 2 ) q O] r R 5 、-O(CH 2 ) s C 6 -C 10 Aryl radical, C 3 -C 8 Cycloalkoxy, halo C 3 -C 8 Cycloalkoxy, cyano, nitro, amino (preferably C) 1 -C 6 Amino), hydroxy, carboxyl, C 1 -C 6 Ester group, C 6 -C 10 Aryl radical, C 6 -C 10 Aryloxy, -X 4 -CO 2 R 5 、C 1 -C 6 Alkylhydroxy, -X 4 -CONR 5 R 6 、-X 4 -CONR 5 (CH 2 ) y1 CO 2 R 6 、-X 4 -CONR 5 (CH 2 ) y2 SO 3 R 6 3-12 membered heterocyclyl and 3-12 membered heterocyclyloxy;
wherein q, r, s, y1 and y2 are 1,2, 3 or 4 respectively;
each of the aromatic heterocyclic group, the aryl group or the heterocyclic group independently contains 1 to 4 hetero atoms selected from oxygen, sulfur and nitrogen;
X 4 is a bond, C1-C6 alkylene, C 1-2 Alkylene, cyclopropyl, epoxyethyl;
R 5 and R 6 Each independently is hydrogen, C 1 -C 6 Alkyl, halo C 1 -C 6 Alkyl radical, C 2 -C 6 Alkenyl, hydroxymethyl, or 5-7 membered heterocyclyl.
In a further preferred embodiment of the method,
Figure BDA0001653521430000042
selected from the group consisting of:
Figure BDA0001653521430000043
wherein the content of the first and second substances,
R 8 selected from the group consisting of: hydrogen, halogen, C 1 -C 6 Alkyl, halo C 1 -C 6 Alkyl radical, C 1 -C 6 Alkoxy radical, C 1 -C 6 Alkoxycarbonyl, halo C 1 -C 6 Alkoxy radical, C 2 -C 6 Alkenyl radical, C 2 -C 6 Alkynyl, C 3 -C 8 Cycloalkyl, cyano, nitro, amino, hydroxy, hydroxymethyl, carboxyl, mercapto, sulfonyl, C 6 -C 10 Aryl and 3-12 membered heterocyclyl;
t is 0, 1,2, 3, 4 or 5;
X l and X 2 Each independently is N, CH or CR 8 ；
X 3 Is O or S;
R 9 is-X 4 -CO 2 R 5 、C 1 -C 6 Alkyl, halo C 1 -C 6 Alkyl radical, C 1 -C 6 Hydroxy, -X 4 -CONR 5 R 6 、-X 4 -CONR 5 (CH 2 ) y1 CO 2 R 6 、-X 4 -CONR 5 (CH 2 ) y2 SO 3 R 6 (ii) a y1 and y2 are each independently 1,2, 3 or 4; x 4 Is a bond, C1-C6 alkylene, C 1-2 Alkylene, cyclopropyl or epoxyethyl;
R 5 and R 6 Each independently is hydrogen, C 1-6 Alkyl, halo C 1 -C 6 Alkyl radical, C 2 -C 6 Alkenyl, hydroxymethyl, or 5-7 membered heterocyclyl;
R 10 is hydrogen, C 1 -C 6 Alkyl hydroxy, C 1 -C 6 Alkyl, halo C 1 -C 6 Alkyl radical, C 1 -C 6 Alkoxy radical, C 1 -C 6 Alkoxycarbonyl, halo C 1 -C 6 Alkoxy radical, C 2 -C 6 Alkenyl radical, C 2 -C 6 Alkynyl or C 3 -C 8 A cycloalkyl group.
In a further preferred embodiment of the method,
Figure BDA0001653521430000051
selected from the group consisting of:
Figure BDA0001653521430000052
wherein R is 8 、R 9 、R 10 And t are as defined above.
In another preferred embodiment, R 1 Selected from the group consisting of: phenyl, pyridyl, 1-oxopyridyl, cyclohexyl, cyclopentyl,A cyclopropyl group; the above groups are substituted or unsubstituted; wherein, the substituted refers to that the hydrogen atom on the group is substituted by 1,2, 3 or 4 substituents selected from the following group: hydrogen, halogen, nitro, cyano, trifluoromethyl, trifluoroethyl, trifluoropropyl, trifluoromethoxy, trifluoroethoxy, phenyl, methoxycarbonyl, ethoxycarbonyl, propoxycarbonyl, C 1-6 Alkyl radical, C l-6 Cycloalkyl radical, C 1-6 Alkoxy radical, C l-6 Cycloalkoxy, cyclopropyl.
In another preferred embodiment, the compound is selected from the group consisting of:
Figure BDA0001653521430000053
Figure BDA0001653521430000061
Figure BDA0001653521430000071
Figure BDA0001653521430000081
in a second aspect, the present invention provides a pharmaceutical composition comprising a therapeutically effective amount of the fluoroisoxazole compound of the first aspect or its enantiomers or diastereomers, or its pharmaceutically acceptable salts, or mixtures thereof, and a pharmaceutically acceptable carrier.
In a third aspect, the invention provides an FXR agonist comprising the fluoroisoxazole compound of the first aspect, or an enantiomer or a diastereoisomer thereof, or a pharmaceutically acceptable salt thereof, or a mixture thereof.
In a fourth aspect, the present invention provides the use of the fluoroisoxazole compound of the first aspect or its enantiomers or diastereomers, or its pharmaceutically acceptable salts, or mixtures thereof, for the manufacture of a medicament for the prevention or treatment of FXR mediated diseases.
In another preferred embodiment, the disease is selected from the group consisting of: non-alcoholic fatty liver disease, non-alcoholic steatohepatitis, fatty liver, hepatic fibrosis, primary biliary cirrhosis, hyperlipidemia, and dyslipidemia.
The fifth aspect of the invention provides a method for preparing the fluorine-containing isoxazole compound according to the first aspect,
(a) The method comprises the following steps:
Figure BDA0001653521430000091
wherein the content of the first and second substances,
Figure BDA0001653521430000092
R 1 、R 2 、R 3 、R 4 n, m and p are as defined in formula I above; x is halogen or nitro;
step j: reacting intermediate 12 with intermediate 9 in an organic solvent in the presence of 18-crown-6 and a base to form intermediate 13;
step k: deprotecting intermediate 13 in an organic solvent to form intermediate 14;
step l: under the protection of nitrogen, in an organic solvent, in the presence of alkali, a palladium catalyst and a ligand, reacting an intermediate 14 with an intermediate 15 to form a final product I; or
Step m: under the protection of nitrogen, in an organic solvent and in the presence of alkali, reacting the intermediate 14 with the intermediate 15 to form a final product I;
or
(b) The method comprises the following steps:
Figure BDA0001653521430000101
wherein R is 1 、R 2 、R 3 、R 4 N, m and p andthe foregoing general formula I is as defined above; x is halogen or nitro;
step j: reacting intermediate 12 and intermediate 9 in an organic solvent in the presence of 18-crown-6 and a base to form intermediate 13;
step k: deprotecting intermediate 13 in an organic solvent to form intermediate 14;
step l: under the protection of nitrogen, reacting the intermediate 14 with the intermediate 15 in an organic solvent in the presence of a base, a palladium catalyst and a ligand to form an intermediate I'; or
Step m: under the protection of nitrogen, reacting the intermediate 14 with the intermediate 15 in an organic solvent in the presence of a base to form an intermediate I'; and
step e: in organic solvent, intermediate I' and HNR 5 R 6 、HNR 5 (CH 2 ) y1 CO 2 R 6 Or HNR 5 (CH 2 ) y2 SO 3 R 6 Carrying out reaction to form a final product I;
wherein, the first and the second end of the pipe are connected with each other,
Figure BDA0001653521430000102
the ring is selected from the group consisting of: a substituted or unsubstituted 6-20 membered heterocyclic group, a substituted or unsubstituted 6-20 membered aromatic ring group, a substituted or unsubstituted 6-20 membered aromatic heterocyclic group; wherein, the substitution refers to that the hydrogen atoms on the group are substituted by 1,2, 3 or 4 substituents selected from the following group: -X 4 -CO 2 R 5 (ii) a Wherein, X 4 、R 5 Definition in the same way as before
Figure BDA0001653521430000103
The ring is selected from the group consisting of: a substituted or unsubstituted 6-20 membered heterocyclic group, a substituted or unsubstituted 6-20 membered aromatic ring group, a substituted or unsubstituted 6-20 membered aromatic heterocyclic group; wherein, the substituted refers to that the hydrogen atom on the group is substituted by 1,2, 3 or 4 substituents selected from the following group: -X 4 -CONR 5 R 6 、-X 4 -CONR 5 (CH 2 ) y1 CO 2 R 6 、-X 4 -CONR 5 (CH 2 ) y2 SO 3 R 6 (ii) a Wherein X 4 、R 5 、R 6 Y1 and y2 are as defined above.
The invention provides a method of treatment by administering to a subject in need thereof a fluoroisoxazole compound of the invention or an enantiomer or diastereomer thereof, or a pharmaceutically acceptable salt thereof, or a mixture thereof or a pharmaceutical composition of the invention or an FXR agonist.
It is to be understood that within the scope of the present invention, the above-described features of the present invention and those specifically described below (e.g., in the examples) may be combined with each other to form new or preferred embodiments. Not to be reiterated herein, but to the extent of space.
Drawings
FIG. 1 shows the effect of compounds of the present invention on serum ALP.
FIG. 2 shows the effect of compounds of the invention on α -SMA and Col1 α 1 mRNA.
FIG. 3 shows the effect of the compounds of the invention on collagen content.
FIG. 4 shows the effect of compounds of the present invention on serum TC, TG, HDL, LDL concentrations.
FIG. 5 shows the effect of compounds of the invention on liver coefficients.
Detailed Description
The inventor designs and prepares a fluorine-containing isoxazole compound with a novel structure through long-term and intensive research. The compounds can effectively target and stimulate FXR. Based on the above findings, the inventors have completed the present invention.
Term(s) for
In the present invention, unless otherwise specified, the terms used have the ordinary meaning known to those skilled in the art.
In the present invention, the term "6-20 membered heterocyclic group" means a saturated or partially saturated heterocyclic group having 6 to 20 ring atoms and containing one or more heteroatoms selected from nitrogen, and sulfur; including but not limited to: morpholinyl, piperidinyl, piperazinyl, cyclohexylpiperidinyl, and the like. Other "heterocyclyl" terms have similar meanings.
In the present invention, the term "6-20 membered heterocyclyloxy" refers to 6-20 membered heterocyclyl-oxy-. Other "heterocyclyloxy" terms have similar meanings.
In the present invention, the term "6-20 membered aromatic ring group" means an aromatic ring group having 6 to 20 ring atoms; including but not limited to: phenyl, naphthyl, phenanthryl, anthracyl, and the like. Other "aromatic ring" terms have similar meanings.
In the present invention, the term "6-to 20-membered aromatic heterocyclic group" means an aromatic heterocyclic group having 6 to 20 ring atoms and containing one or more heteroatoms selected from nitrogen, and sulfur; including but not limited to: tetrazolyl, pyridyl, pyrimidinyl, pyridazinyl, thiazolyl, benzothiazolyl, benzo [ d ] isothiazolyl, imidazo [1,2-a ] pyridyl, quinolinyl, 1H-indolyl, pyrrolo [1,2-b ] pyridazinyl, benzofuranyl, benzo [ b ] thiophenyl, 1H-indazolyl, benzo [ d ] isoxazolyl, thiazolinyl, 1H-pyrrolo [3,2-c ] pyridyl, pyrazolo [1,5-a ] pyrimidinyl, imidazo [1,2-b ] pyridazinyl, pyrazolo [1,5-a ] pyrrolyl, and the like. Other "arylheterocyclyl" terms have similar meanings.
In the present invention, the term "C 1 -C 6 Alkyl "means a straight or branched chain alkyl group having 1 to 6 carbon atoms, including, but not limited to, methyl, ethyl, propyl, isopropyl, butyl, isobutyl, sec-butyl, tert-butyl, pentyl, hexyl, and the like; ethyl, propyl, isopropyl, butyl, isobutyl, sec-butyl and tert-butyl are preferred.
In the present invention, the term "C 1 -C 6 Alkoxy "means a straight or branched chain alkoxy group having 1 to 6 carbon atoms, including without limitation methoxy, ethoxy, propoxy, isopropoxy, butoxy, and the like.
In the present invention, the term "C 2 -C 6 The alkenyl group "means a straight or branched alkenyl group having 2 to 6 carbon atoms and containing one double bond, and includes, but is not limited to, ethenyl, propenyl, butenyl, isobutenyl, pentenyl, hexenyl, and the like.
In the inventionIn, the term "C 2 -C 6 Alkynyl "refers to a straight or branched chain alkynyl group having 2 to 6 carbon atoms containing one triple bond and includes, without limitation, ethynyl, propynyl, butynyl, isobutynyl, pentynyl, hexynyl, and the like.
In the present invention, the term "C 3 -C 8 Cycloalkyl "refers to cyclic alkyl groups having 3 to 8 carbon atoms in the ring and includes, without limitation, cyclopropyl, cyclobutyl, cyclopentyl, cyclohexyl, cycloheptyl, cyclooctyl, cyclodecyl, and the like. Other "cycloalkyl" terms have similar meanings.
In the present invention, the term "C 3 -C 8 Cycloalkoxy "means a cyclic alkoxy group having 3 to 8 carbon atoms in the ring, and includes, without limitation, cyclopropoxy, cyclobutoxy, cyclopentoxy, cyclohexyloxy, cycloheptyloxy, cyclooctyloxy, cyclodecyloxy, and the like. Other "cycloalkoxy" terms have similar meanings.
In the present invention, the term "C 6 -C 10 Aryl "refers to an aromatic ring group having 6 to 10 carbon atoms, such as phenyl, naphthyl, and the like, which does not contain heteroatoms in the ring. The term "C 6 -C 12 Aryl "has a similar meaning.
In the present invention, the term "C 1 -C 6 An ester group "refers to a group having 1 to 6 carbon atoms and having an ester group (-O (C = O) -).
In the present invention, the term "C 1 -C 6 The amino group "means a group having 1 to 6 carbon atoms and having an amino group as a terminal group.
In the present invention, the halogen is F, cl, br or I. Halo represents substitution by halogen.
Active ingredient
The compound of the invention is a compound shown in a general formula I or an enantiomer or a diastereoisomer thereof, or a pharmaceutically acceptable salt thereof, or a mixture of the enantiomer and the diastereoisomer.
The compounds of the invention are preferably the following specific compounds:
Figure BDA0001653521430000121
Figure BDA0001653521430000131
Figure BDA0001653521430000141
Figure BDA0001653521430000151
Figure BDA0001653521430000161
Figure BDA0001653521430000171
Figure BDA0001653521430000181
Figure BDA0001653521430000191
Figure BDA0001653521430000201
Figure BDA0001653521430000211
Figure BDA0001653521430000221
Figure BDA0001653521430000231
the compound also comprises a pharmaceutically acceptable salt of the compound shown in the general formula I, and particularly the compound shown in the general formula I reacts with inorganic acid or organic acid to form a conventional pharmaceutically acceptable salt. For example, conventional pharmaceutically acceptable salts can be prepared by reacting a compound of formula I with inorganic acids including hydrochloric acid, hydrobromic acid, sulfuric acid, nitric acid, sulfamic acid, phosphoric acid and the like, or organic acids including citric acid, tartaric acid, lactic acid, pyruvic acid, acetic acid, benzenesulfonic acid, p-toluenesulfonic acid, methanesulfonic acid, naphthalenesulfonic acid, ethanesulfonic acid, naphthalenedisulfonic acid, maleic acid, malic acid, malonic acid, fumaric acid, succinic acid, propionic acid, oxalic acid, trifluoroacetic acid, stearic acid, pamoic acid, hydroxymaleic acid, phenylacetic acid, benzoic acid, salicylic acid, glutamic acid, ascorbic acid, p-aminobenzenesulfonic acid, 2-acetoxybenzoic acid, isethionic acid and the like; or sodium salt, potassium salt, calcium salt, aluminum salt or ammonium salt formed by the compound of the general formula I and inorganic base; or methylamine salt, ethylamine salt or ethanolamine salt formed by the compound in the general formula I and organic base.
The compounds of the invention also include enantiomers (e.g., R-or S-isomers) or diastereomers of the compounds of formula I, or racemates or meso forms thereof.
Preparation method
The compounds of formula I of the present invention may be prepared according to the following scheme.
Scheme for preparing intermediate 3 of general formula
Figure BDA0001653521430000232
Wherein R is 2 P is as defined in formula I above;
step a: dissolving 1 in organic solvent, dripping oxidant at 0 deg.C, stirring at 0 deg.C for reaction, extracting organic layer with diethyl ether, extracting with sodium hydroxide solution, mixing water layers, acidifying with sulfuric acid to pH<1. Then extracting with diethyl etherThe organic layer was washed with saturated brine, dried and spin-dried to give intermediate 2. The organic solvent is acetone, tetrahydrofuran, diethyl ether, dimethylformamide, ethylene glycol dimethyl ether, ethylene glycol diethyl ether, dioxane, ethanol, methanol, ethyl acetate, dichloromethane or a mixture thereof; said oxidant is selected from the group consisting of PCC, PDC, dess-Martin oxidant, swern oxidant, H 2 O 2 Potassium permanganate and manganese dioxide;
step b: dissolving the intermediate 2 in an organic solvent, adding N, N' -carbonyldiimidazole, reacting for one hour at room temperature under the protection of nitrogen, adding potassium monoethyl malonate and magnesium chloride, reacting overnight at room temperature, quenching with hydrochloric acid after the reaction is finished, extracting an organic layer with ethyl acetate, washing the organic layer with saturated salt water, drying, spin-drying, and passing through a column to obtain an intermediate 3. The organic solvent is acetone, tetrahydrofuran, diethyl ether, dimethylformamide, ethylene glycol dimethyl ether, ethylene glycol diethyl ether, dioxane, ethanol, methanol, ethyl acetate, dichloromethane or a mixture thereof;
preparation of intermediate of general formula 9 the scheme is as follows
Figure BDA0001653521430000241
Wherein R is 1 、R 2 And p is as defined in formula I above;
step c: hydroxylamine hydrochloride was dissolved in water and sodium hydroxide was added dropwise with stirring at 0 ℃.4 adding organic solvent to dissolve, dripping the mixed solution, then heating to 90 ℃, and stirring overnight. After the reaction is finished, cooling to room temperature, concentrating in vacuum until a large amount of solid is separated out, and performing suction filtration to obtain solid, and drying to obtain an intermediate 5; the organic solvent is tetrahydrofuran, diethyl ether, dimethylformamide, ethylene glycol dimethyl ether, ethylene glycol diethyl ether, dioxane, ethanol, methanol, ethyl acetate, dichloromethane or a mixture thereof.
Step d: dissolving intermediate 5 in organic solvent, dripping chlorosuccinimide in organic solvent at 40 deg.C into the above 5 solution, reacting at 40 deg.C, cooling to room temperatureAdding water and Ethyl Acetate (EA) at room temperature, extracting, washing organic layer with water for three times, washing organic layer with saturated saline, and anhydrous Na 2 SO 4 Drying and spin-drying the organic layer to obtain an intermediate 6; the organic solvent is tetrahydrofuran, diethyl ether, dimethylformamide, ethylene glycol dimethyl ether, ethylene glycol diethyl ether, dioxane, ethanol, methanol, ethyl acetate, dichloromethane or a mixture thereof;
step e: adding the intermediate 3 into alkali, stirring at room temperature, cooling to 10 ℃, and slowly dropwise adding the intermediate 6 to react to obtain an intermediate 7; the organic solvent is tetrahydrofuran, diethyl ether, dimethylformamide, ethylene glycol dimethyl ether, ethylene glycol diethyl ether, dioxane, ethanol, methanol or ethyl acetate, and dichloromethane; the alkali is sodium acetate, naOH, KOH, sodium ethoxide, sodium methoxide, sodium carbonate, potassium carbonate, triethylamine or diisopropylamine.
Step f: dissolving the intermediate 7 in an organic solvent, dropwise adding diisobutylaluminum hydride 1M/toluene at 0 ℃ under the protection of nitrogen, stirring at room temperature for reaction, after the reaction is finished, dropwise adding methanol and a small amount of water at 0 ℃ for quenching, stirring for 10 minutes, filtering out precipitates through diatomite suction filtration and organic solvent washing, and carrying out vacuum concentration and drying on the filtrate to obtain an intermediate 8; the organic solvent is tetrahydrofuran, diethyl ether, dimethylformamide, ethylene glycol dimethyl ether, ethylene glycol diethyl ether, dioxane, ethanol, methanol, ethyl acetate, dichloromethane or a mixture thereof;
step g: dissolving the intermediate 8 in an organic solvent, adding triphenylphosphine, cooling to 0 ℃, adding carbon tetrabromide, reacting at room temperature, after the reaction is finished, concentrating the reaction solution in vacuum to obtain orange oily matter, passing through a silica gel column, and spin-drying to obtain an intermediate 9. The organic solvent is tetrahydrofuran, diethyl ether, dimethylformamide, ethylene glycol dimethyl ether, ethylene glycol diethyl ether, dioxane, ethanol, methanol, ethyl acetate, dichloromethane or a mixture thereof;
scheme for preparing intermediate 12 of general formula
Figure BDA0001653521430000251
Wherein, m and R 4 And R 5 Are as defined in formula I above;
step h: dissolving trimethyl sulfur iodide in an organic solvent, adding the compound 10 while stirring, continuing stirring, and then adding potassium tert-butoxide; to obtain the epoxy compound 2. The organic solvent can be dimethyl sulfoxide, tetrahydrofuran, diethyl ether, dimethylformamide, ethylene glycol dimethyl ether, ethylene glycol diethyl ether, dioxane, ethanol, methanol, ethyl acetate, dichloromethane or their mixture.
Step i: dissolving the intermediate 11 into an organic solvent, cooling to-10 to-40 ℃, adding 1 to 10 equivalents of pyridine hydrogen fluoride solution, continuing to react until the raw materials disappear, and separating and purifying to obtain an intermediate 12; the organic solvent may be tetrahydrofuran, diethyl ether, dimethylformamide, ethylene glycol dimethyl ether, ethylene glycol diethyl ether, dioxane, ethanol, methanol, ethyl acetate, dichloromethane or their mixture.
The scheme for synthesizing the general formula I is as follows:
Figure BDA0001653521430000252
wherein the content of the first and second substances,
Figure BDA0001653521430000253
R 1 、R 2 、R 3 、R 4 n, m and p are as defined in formula I above; x is halogen or nitro;
step j: dissolving the intermediate 12 in an organic solvent, adding a compound 18-crown-6 (12.8g, 48.6 mmol) and strong base at 0 ℃, reacting for 30 minutes, moving to room temperature, dropwise adding the intermediate 9 (dissolved in the organic solvent), stirring at room temperature for reacting, and separating and purifying after the reaction to obtain an intermediate 13; the organic solvent is tetrahydrofuran, diethyl ether, dimethylformamide, ethylene glycol dimethyl ether, ethylene glycol diethyl ether, dioxane, ethanol, methanol, ethyl acetate, dichloromethane or a mixture thereof; the strong base is NaOH, KOH, potassium tert-butoxide, sodium ethoxide or sodium methoxide;
step k: dissolving the intermediate 13 into an organic solvent, adding an organic solvent of trifluoroacetic acid (TFA) or hydrochloric acid (HCl), and removing Boc protecting groups to obtain an intermediate 14; the organic solvent may be tetrahydrofuran, diethyl ether, dimethylformamide, ethylene glycol dimethyl ether, ethylene glycol diethyl ether, dioxane, ethanol, methanol, ethyl acetate, dichloromethane or their mixture.
Step l: under the protection of nitrogen, dissolving the intermediates 14 and 15 in an organic solvent, adding cesium carbonate, a tris (dibenzylideneacetone) dipalladium catalyst and a 2-dicyclohexyl phosphorus-2, 4, 6-triisopropyl biphenyl ligand, and after the reaction is finished, separating and purifying to obtain a final product I; the organic solvent is toluene, tetrahydrofuran, diethyl ether, dimethylformamide, ethylene glycol dimethyl ether, ethylene glycol diethyl ether, dioxane, ethanol, methanol, ethyl acetate, dichloromethane or a mixture thereof;
or step m: under the protection of nitrogen, dissolving the intermediates 14 and 15 in an organic solvent, adding N, N-diisopropylethylamine, and after the reaction is finished, separating and purifying to obtain a final product I; the organic solvent is toluene, tetrahydrofuran, diethyl ether, dimethylformamide, ethylene glycol dimethyl ether, ethylene glycol diethyl ether, dioxane, ethanol, methanol, ethyl acetate, azomethylpyrrolidone, dichloromethane or a mixture thereof.
Or the method for synthesizing the general formula I is as follows:
Figure BDA0001653521430000261
wherein R is 1 、R 2 、R 3 、R 4 N, m and p are as defined above; x is halogen or nitro;
step j: reacting intermediate 12 and intermediate 9 in an organic solvent in the presence of 18-crown-6 and a base to form intermediate 13;
step k: deprotecting intermediate 13 in an organic solvent to form intermediate 14;
step l: under the protection of nitrogen, in an organic solvent, in the presence of alkali, a palladium catalyst and a ligand, reacting the intermediate 14 with the intermediate 15 to form an intermediate I'; or
Step m: under the protection of nitrogen, reacting the intermediate 14 with the intermediate 15 in an organic solvent in the presence of a base to form an intermediate I'; and
step e: in organic solvent, intermediate I' and HNR 5 R 6 、HNR 5 (CH 2 ) y1 CO 2 R 6 Or HNR 5 (CH 2 ) y2 SO 3 R 6 Carrying out a reaction to form a final product I;
wherein the content of the first and second substances,
Figure BDA0001653521430000271
the ring is selected from the group consisting of: a substituted or unsubstituted 6-20 membered heterocyclic group, a substituted or unsubstituted 6-20 membered aromatic ring group, a substituted or unsubstituted 6-20 membered aromatic heterocyclic group; wherein, the substituted refers to that the hydrogen atom on the group is substituted by 1,2, 3 or 4 substituents selected from the following group: -X 4 -CO 2 R 5 (ii) a Wherein, X 4 、R 5 Definition in the same way as before
Figure BDA0001653521430000272
The ring is selected from the group consisting of: substituted or unsubstituted 6-20 membered heterocyclic group, substituted or unsubstituted 6-20 membered aromatic ring group, substituted or unsubstituted 6-20 membered aromatic heterocyclic group; wherein, the substitution refers to that the hydrogen atoms on the group are substituted by 1,2, 3 or 4 substituents selected from the following group: -X 4 -CONR 5 R 6 、-X 4 -CONR 5 (CH 2 ) y1 CO 2 R 6 、-X 4 -CONR 5 (CH 2 ) y2 SO 3 R 6 (ii) a Wherein, X 4 、R 5 、R 6 Y1, y2 are as defined above.
Pharmaceutical compositions and methods of administration
The compound has excellent FXR agonism and can be used for preparing a pharmaceutical composition for treating or preventing FXR-mediated diseases.
The invention provides a pharmaceutical composition, which comprises one or more compounds selected from the compounds shown in the general formula (I) or enantiomers or diastereomers thereof, or pharmaceutically acceptable salts thereof, or mixtures thereof, in a therapeutically effective amount, and optionally, one or more pharmaceutically acceptable carriers, excipients, adjuvants and/or diluents. The auxiliary materials are, for example, odorants, sweeteners, and the like.
The pharmaceutical composition provided by the invention preferably contains 1-99% by weight of active ingredients, and the preferred proportion is that the compound of the general formula I as the active ingredient accounts for 65-99% by weight of the total weight, and the rest is pharmaceutically acceptable carriers, diluents or solutions or salt solutions.
The compounds and pharmaceutical compositions provided herein may be in a variety of forms such as tablets, capsules, powders, syrups, solutions, suspensions, and aerosols, and the like, and may be presented in suitable solid or liquid carriers or diluents and in suitable sterile devices for injection or instillation.
Various dosage forms of the pharmaceutical composition of the present invention can be prepared according to conventional preparation methods in the pharmaceutical field. The unit dosage of the preparation formula comprises 0.05-200mg of the compound of the invention, and preferably, the unit dosage of the preparation formula comprises 0.1-100 mg of the compound of the invention.
The compounds and pharmaceutical compositions of the present invention may be administered to mammals, including humans and animals, clinically, by oral, nasal, dermal, pulmonary or gastrointestinal routes of administration. Most preferably oral. Most preferably, the daily dose is 0.01-200mg/kg body weight, and is administered once or in portions of 0.01-100mg/kg body weight. Regardless of the method of administration, the optimal dosage for an individual will depend on the particular treatment. Usually, the dosage is increased gradually starting from a small dosage until the most suitable dosage is found.
The invention also provides a Farnesoid X Receptor (FXR) agonist comprising one or more selected from the group consisting of a compound represented by the general formula I above, or an enantiomer or a diastereoisomer thereof, or a pharmaceutically acceptable salt thereof, or a mixture thereof, and optionally one or more pharmaceutically acceptable carriers, excipients, adjuvants, and/or diluents.
The compounds and compositions of the present invention are useful for the treatment and prevention of diseases of the liver system and blood lipids associated with Farnesoid X Receptor (FXR), including, but not limited to, nonalcoholic steatohepatitis, liver fibrosis, cirrhosis, fatty liver, primary biliary cirrhosis, hyperlipidemia, and the like.
Therefore, the invention also provides the application of the compound shown in the general formula I, or the enantiomer or diastereoisomer thereof, or the pharmaceutically acceptable salt thereof, or the mixture thereof in preparing the medicines for treating liver system and blood fat diseases related to Farnesoid X Receptor (FXR), such as nonalcoholic steatohepatitis, liver fibrosis, cirrhosis, fatty liver, primary bile acid cirrhosis, hyperlipidemia and other diseases.
The invention also provides a method for treating diseases of the liver system and blood lipids associated with Farnesoid X Receptor (FXR), such as nonalcoholic steatohepatitis, liver fibrosis, cirrhosis, fatty liver, primary bile acid cirrhosis, hyperlipidemia, etc., comprising administering to a patient in need of such treatment one or more compounds selected from the group consisting of a compound represented by formula I above, or an enantiomer or diastereoisomer thereof, or a pharmaceutically acceptable salt thereof, or a mixture thereof.
The main advantages of the invention are:
the compound has excellent FXR agonism, good intracellular release and high safety.
The invention will be further illustrated with reference to the following specific examples. It should be understood that these examples are for illustrative purposes only and are not intended to limit the scope of the present invention. The experimental procedures, in which specific conditions are not noted in the following examples, are generally carried out under conventional conditions or conditions recommended by the manufacturers. Unless otherwise indicated, percentages and parts are by weight. The test materials and reagents used in the following examples are commercially available without specific reference.
EXAMPLE 1 preparation of 4- (4- (((5-cyclopropyl-3- (2, 6-dichlorophenyl) isoxazol-4-yl) methoxy) methyl) -4-fluoropiperidin-1-yl) benzoic acid (FL 1)
Figure BDA0001653521430000281
1.1 Synthesis of 2, 6-dichloro-benzaldoxime
Figure BDA0001653521430000291
Hydroxylamine hydrochloride (67.6 g, 0.943mol) was dissolved in 100mL of water, sodium hydroxide 3N (314mL, 9.43mol.2, 6-dichlorobenzaldehyde (150g, 0.857mol) was added dropwise with ethanol (300 mL) to dissolve it at 0 ℃ with stirring, the mixture was added dropwise to the solution, the mixture was heated to 90 ℃ with stirring overnight, the reaction was monitored by Thin Layer Chromatography (TLC), after completion of the reaction, it was cooled to room temperature, concentrated in vacuo to a 10% solution, a large amount of solid precipitated, filtered to give a solid by suction, and dried to give 162 g of a white solid, yield 98% ESI-MS M/z:190 (M + 1).
1.2 2, 6-dichloro-benzaldehyde chloro-oxime
Figure BDA0001653521430000292
Dissolving 2, 6-dichloro-benzaldoxime (100g, 0.526 mol) in DMF (N, N-dimethylamide, 200 mL), dissolving chlorosuccinimide (70.3 g,0.526 mol) in 100mL DMF, dropping the solution into the above 2, 6-dichloro-benzaldoxime solution at 40 deg.C, reacting at 40 deg.C for 1 hour, monitoring the reaction by Thin Layer Chromatography (TLC), cooling to room temperature after the reaction is completed, adding a large amount of water, extracting with Ethyl Acetate (EA), washing the organic layer three times with 200mL of water, washing the organic layer with 200mL of saturated saline, and adding anhydrous Na 2 SO 4 Drying, spin-drying the organic layer to obtain a crude product, adding 100ml of hexane, grinding, filtering, and drying to obtain 88.6 g of solid which is 2, 6-dichloro-benzaldehyde chloro-oxime. ESI-MS M/z 224 (M + 1)
1.3 5-cyclopropyl-3- (2, 6-dichloro-phenyl) -isoxazole-4-carboxylic acid methyl ester
Figure BDA0001653521430000293
Dissolving 3-cyclopropyl-3-oxo-methyl propionate (31.7g, 0.223mol) in triethylamine (45.1g, 62mL,0.446 mol), stirring at room temperature, after 30 minutes, cooling to 10 deg.C, adding 2, 6-dichloro-benzaldehyde chloro-oxime (50.0 g,0.223mol, dissolved in 300mL ethanol) slowly and dropwise while the temperature does not exceed 24 deg.C, after dropwise addition, stirring at room temperature overnight, monitoring the reaction by Thin Layer Chromatography (TLC), after reaction, adding water 200mL and Ethyl Acetate (EA) 500mL, extracting the water layer three times with ethyl acetate 200mL, combining the organic layers, washing the organic layer with saturated saline 200mL, anhydrous Na 2 SO 4 Drying, vacuum concentrating to 10% solution, precipitating a large amount of solid, vacuum filtering to obtain crude product, adding hexane 100ml, grinding, vacuum filtering, and drying to obtain white solid 45.0 g, yield 65%. ESI-MS M/z 312 (M) + )。
1.4[ 2- (5-cyclopropyl) -3- (2, 6-dichloro-phenyl) -isoxazol-4-yl ] -methanol
Figure BDA0001653521430000301
5-cyclopropyl-3- (2, 6-dichloro-phenyl) -isoxazole-4-carboxylic acid methyl ester (20.0 g, 64.1mmol) was dissolved in tetrahydrofuran (100 mL), diisobutylaluminum hydride 1M/toluene (160.0 mL, 160.8mmol) was added dropwise at 0 ℃ under nitrogen, and the mixture was stirred at room temperature for 2 hours. The reaction was monitored by Thin Layer Chromatography (TLC) and after completion was quenched by addition of methanol (10 mL) and a small amount of water dropwise at 0 deg.C and stirred for 10 minutes before addition of ethyl acetate (500 mL). Filtering through diatomite, washing with ethyl acetate, filtering to obtain precipitate, vacuum concentrating the filtrate, and drying to obtain white solid 15.2 g, yield 83%, ESI-MS M/z 285 (M) + )。
1.5 4-bromomethyl-5-cyclopropyl-3- (2, 6-dichloro-phenyl) -isoxazole
Figure BDA0001653521430000302
[ 5-cyclopropyl-3- (2, 6-dichloro-phenyl) -isoxazol-4-yl]-first of allAlcohol (15.0g, 52.8mmol) was dissolved in 100mL of methylene chloride, triphenylphosphine (20.8g, 79.2mmol) was added, the mixture was cooled to 0 ℃ and carbon tetrabromide (26.3g, 79.2mmol) was added, the mixture was reacted at room temperature for 2 hours, the reaction was monitored by Thin Layer Chromatography (TLC), and after completion of the reaction, the reaction mixture was concentrated in vacuo to give an orange oil, petroleum Ether (PE): ethyl Acetate (EA) =5:1 column chromatography, spin-drying to obtain white solid product 16.3 g, yield 89%, ESI-MS M/z:346 (M) + ).
1.6 Synthesis of 6-tert-butyloxycarbonyl-1-oxa-6-azaspiro [2.5] octane
Figure BDA0001653521430000303
130mL of dimethyl sulfoxide (DMSO) was added to a 250mL eggplant-shaped bottle, and the mixture was stirred and heated, 5g of NaH solid was weighed and added to the DMSO solution, and stirring was continued for 12 hours, and the heating was stopped and the mixture was cooled to room temperature. 25g of trimethyl sulfoxide iodide was added thereto, and the mixture was stirred at room temperature for 24 hours. 25g of N-t-butoxycarbonylpiperidone dissolved in DMSO in advance was added to the reaction solution, and the reaction was further stirred for 12 hours and monitored by Thin Layer Chromatography (TLC). After the reaction, 100-200mL of water was added, 100mL of Ethyl Acetate (EA) was extracted 3 times, and 30mL of saturated brine was used to wash the organic layer three times and spin-dry the organic layer to obtain 28g of a colorless liquid, which was 6-t-butoxycarbonyl-1-oxa-6-azaspiro [2.5] octane.
1.7 Synthesis of (4-fluoro-1-tert-butoxycarbonyl-substituted-piperidin-4-yl) methanol
Figure BDA0001653521430000304
28g of 6-tert-butoxycarbonyl-1-oxa-6-azaspiro [2.5] octane was dissolved in 100mL of Dichloromethane (DCM), cooled, added with 120mL of 70% hydrogen fluoride pyridine solution, reacted for another 12 hours, and the reaction was completed, added with 100mL of water, extracted three times with 100mL of DCM, petroleum Ether (PE): ethyl Acetate (EA) =4: the product was applied to a column 1 and spin-dried to give 21 g of a substantially colorless liquid product, which was (4-fluoro-1-Boc-substituted-piperidin-4-yl) -methanol.
1.8 Synthesis of 4- (((5-cyclopropyl-3- (2, 6-dichlorophenyl) isoxazol-4-yl) methoxy) methyl) -4-fluoropiperidine-1-carboxylic acid tert-butyl ester
Figure BDA0001653521430000311
(4-fluoro-1-tert-Butoxycarbonyl-substituted-piperidin-4-yl) -methanol (10.0g, 42.8mmol) was dissolved in 50mL of tetrahydrofuran, and 18-crown-6 (12.8g, 48.6 mmol) and potassium tert-butoxide (5.5g, 48.6 mmol) were added thereto at 0 ℃ to conduct a reaction for 30 minutes, and then the mixture was allowed to cool to room temperature, and 4-bromomethyl-5-cyclopropyl-3- (2, 6-dichloro-phenyl) -isoxazole (9.9g, 28.6mmol, in 50mL of tetrahydrofuran) was added dropwise and the mixture was stirred at room temperature for 14 hours. The reaction was monitored by Thin Layer Chromatography (TLC), after completion of the reaction, 200mL of water was added and extracted with ethyl acetate (EA, 3X 200mL), the organic layers were combined, washed with 200mL of saturated brine, dried over anhydrous Na2SO4, concentrated in vacuo, and Petroleum Ether (PE): ethyl Acetate (EA) =4:1 column chromatography, spin-drying to obtain 10.1 g oily liquid, yield 71%, ESI-MS M/z:500 (M) + )。
1.9 5-cyclopropyl-3- (2, 6-dichlorophenyl) -4- (((4-fluoropiperidin-4-yl) methoxy) methyl) isoxazole
Figure BDA0001653521430000312
To a 250mL eggplant-shaped bottle was added tert-butyl 4- (((5-cyclopropyl-3- (2, 6-dichlorophenyl) isoxazol-4-yl) methoxy) methyl) -4-fluoropiperidine-1-carboxylate (10.0 g,20.0mmol, dissolved in dichloromethane (50 mL)) under ice-bath and nitrogen blanket, followed by dropwise addition of 50mL trifluoroacetic acid (dropwise addition for about 20 minutes), and the reaction was stirred at room temperature for 1.5 hours. The reaction was monitored by Thin Layer Chromatography (TLC), after completion of the reaction the solution was vacuum-dried, 300mL of ethyl acetate and 2N NaOH (2X 200mL) were added, the organic layer was washed with 200mL of saturated saline, and anhydrous Na 2 SO 4 Drying and concentration in vacuo gave 7.2 g of an oily liquid, 90% yield, ESI-MS M/z:400 (M) + )。
1.10 4- (4- (((5-cyclopropyl-3- (2, 6-dichlorophenyl) isoxazol-4-yl) methoxy) methyl) -4-fluoropiperidin-1-yl) benzoic acid ethyl ester
Figure BDA0001653521430000313
5-cyclopropyl-3- (2, 6-dichlorophenyl) -4- (((4-fluoropiperidin-4-yl) methoxy) methyl) isoxazole (2.0g, 5.01mmol), 4-iodo-benzoic acid ethyl ester (1.26g, 4.55mmol), cesium carbonate (2.97g, 9.11mmol), tris (dibenzylideneacetone) dipalladium (208.5mg, 0.228mmol) and 2-dicyclohexylphosphorus-2, 4, 6-triisopropylbiphenyl (217.1mg, 0.455mmol) were charged into a 250mL two-necked flask under nitrogen protection, followed by dissolution in 100mL of toluene and stirring at 120 ℃ for 16 hours. The reaction was monitored by Thin Layer Chromatography (TLC), after completion of the reaction, it was cooled to room temperature and filtered through celite, and water (100 mL) and ethyl acetate (3X 100mL) were added to the filtrate. The organic layer was washed with 200mL of saturated brine, dried over anhydrous Na2SO4, and concentrated in vacuo, petroleum Ether (PE): ethyl Acetate (EA) =4:1 to 1, passing through a column, and spin-drying to obtain 1.55g of solid, namely 4- (4- (((5-cyclopropyl-3- (2, 6-dichlorophenyl) isoxazol-4-yl) methoxy) methyl) -4-fluoropiperidine-1-yl) ethyl benzoate, wherein the yield is 62%, and ESI-MS M/z is 548 (M-Z: 4 (ESI-MS M/z) + )。
1.11 Synthesis of the end product FL1
Figure BDA0001653521430000321
To a 250mL two-necked flask was added ethyl 4- (4- (((5-cyclopropyl-3- (2, 6-dichlorophenyl) isoxazol-4-yl) methoxy) methyl) -4-fluoropiperidin-1-yl) benzoate (1.55g, 2.82mmol) under nitrogen, followed by the addition of 30mL of methanol, 30mL of tetrahydrofuran, and 10mL of 1, 4-dioxane for dissolution, followed by the addition of 2N KOH (30mL, 61.0 mmol). Stirring was carried out overnight at 70 ℃. The reaction was monitored by Thin Layer Chromatography (TLC) and after completion the solution was spun dry in vacuo and 100mL of water and 50mL of methyl tert-butyl ether were added. The aqueous layer was adjusted to pH 5 with 2N HCl, then dichloromethane CH2Cl2 (3X 100mL) was added and the organic layer was washed with saturated brine 200mL, dried over anhydrous Na2SO4, the crude product was concentrated in vacuo and recrystallized from MeOH/CH3CN to give 956.0mg of a white solid in 62% yield, 1 H NMR(400MHz,DMSO-d 6 )δ12.28(s,1H),7.77(d,J＝8.9Hz,2H),7.70-7.50(m,3H),6.96(d,J＝9.0Hz,2H),4.32(s,2H),3.61(d,J＝13.2Hz,2H),3.40(s,2H),3.04(t,J＝10.9Hz,2H),2.37-2.30(m,1H),1.71-1.38(m,4H),1.22-1.02(m,4H)；ESI-MS m/z:520(M + )。
EXAMPLE 2 preparation of 4- (4- (((3- (2-chloro-6-fluorophenyl) -5-cyclopropylisoxazol-4-yl) methoxy) methyl) -4-fluoropiperidin-1-yl) benzoic acid (FL 2)
2, 6-dichlorobenzaldehyde is replaced by 2-chloro-6-fluorobenzaldehyde, and the rest required raw materials, reagents and preparation methods are the same as those in example 1, so that a product FL2 is obtained. 1 H NMR(400MHz,DMSO-d 6 )δ12.28(s,1H),7.77(d,J＝8.9Hz,2H),7.62(td,J＝8.3,6.2Hz,1H),7.51(d,J＝8.1Hz,1H),7.41(t,J＝8.7Hz,1H),6.95(d,J＝9.0Hz,2H),4.35(s,2H),3.61(d,J＝13.2Hz,2H),3.39(s,2H),3.03(t,J＝10.8Hz,2H),2.37-2.30(m,1H),1.70-1.35(m,4H),1.19-1.04(m,4H).LRMS(EI)m/z 504(M + )。
EXAMPLE 3 preparation of 6- (4- (((5-cyclopropyl-3- (2, 6-dichlorophenyl) isoxazol-4-yl) methoxy) methyl) -4-fluoropiperidin-1-yl) -1-ethyl-1H-indole-3-carboxylic acid (FL 3)
The 4-iodo-ethyl benzoate was replaced with 6-bromo-1-ethyl-1H-indole-3-carboxylic acid methyl ester, and the remaining required raw materials, reagents and preparation methods were the same as in example 1, to obtain product FL3. 1 H NMR(400MHz,DMSO-d 6 )δ11.81(s,1H),7.89(s,1H),7.82(d,J＝8.7Hz,1H),7.64(d,J＝7.5Hz,2H),7.57(dd,J＝9.1,6.9Hz,1H),7.01-6.91(m,2H),4.35(s,2H),4.21(q,J＝7.0Hz,2H),3.50-3.36(m,4H),2.90(t,J＝12.3Hz,2H),2.44-2.30(m,1H),1.78-1.47(m,4H),1.37(t,J＝7.2Hz,3H),1.20-1.06(m,4H).；LRMS(EI)m/z587(M + )。
EXAMPLE 4 preparation of 6- (4- (((5-cyclopropyl-3- (2, 6-dichlorophenyl) isoxazol-4-yl) methoxy) methyl) -4-fluoropiperidin-1-yl) benzo [ b ] thiophene-2-carboxylic acid (FL 4)
Replacement of 4-iodo-benzoic acid ethyl ester with 6-bromo-benzo [ b]Thiophene-2-carboxylic acid methyl ester, the other required raw materials, reagents and preparation methods are the same as those of example 1, and a product FL4 is obtained. 1 H NMR(400MHz,DMSO-d 6 )δ13.08(s,1H),7.91(s,1H),7.78(d,J＝9.0Hz,1H),7.64-7.58(m,1H),7.50(d,J＝8.1Hz,1H),7.43-7.38(m,2H),7.17(dd,J＝9.0,2.1Hz,1H),4.35(s,2H),3.56(d,J＝13.1Hz,2H),3.39(s,2H),3.10-2.93(m,2H),2.36-2.30(m,1H),1.68-1.37(m,4H),1.18-1.02(m,4H)；LRMS(EI)m/z 576(M + )。
EXAMPLE 5- (4- (((5-cyclopropyl-3- (2, 6-dichlorophenyl) isoxazol-4-yl) methoxy) methyl) -4-fluoropiperidin-1-yl) -2-naphthoic acid (FL 5)
The 4-iodo-ethyl benzoate was replaced with 6-bromo-2-methyl naphthoate, and the remaining required raw materials, reagents, and preparation methods were the same as in example 1, to give product FL5. 1 H NMR(400MHz,DMSO-d 6 )δ12.73(s,1H),8.40(s,1H),7.91(d,J＝9.1Hz,1H),7.84(dd,J＝8.6,1.5Hz,1H),7.74(d,J＝8.5Hz,1H),7.65-7.60(m,2H),7.56(dd,J＝9.3,6.6Hz,1H),7.43(dd,J＝9.1,1.9Hz,1H),7.22(d,J＝1.7Hz,1H),4.33(s,2H),3.65(d,J＝13.0Hz,2H),3.39(d,J＝20.2Hz,2H),3.05(t,J＝10.7Hz,2H),2.36-2.31(m,1H),1.75-1.43(m,4H),1.20-0.97(m,4H)；LRMS(EI)m/z 570(M + ).
EXAMPLE 6 preparation of (FL 6) 4- (((3- (2-chloro-6-fluorophenyl) -5-cyclopropylisoxazol-4-yl) methoxy) methyl) -4-fluoropiperidin-1-yl) naphthoic acid
2, 6-dichlorobenzaldehyde is replaced by 2-chloro-6-fluorobenzaldehyde and 4-iodo-ethyl benzoate is replaced by 6-bromo-2-methyl naphthoate, and the rest of the required raw materials, reagents and preparation method are the same as those in example 1, so as to obtain a product FL6. 1 H NMR(400MHz,DMSO-d 6 )δ12.74(s,1H),8.41(s,1H),7.92(d,J＝9.2Hz,1H),7.85(dd,J＝8.6,1.7Hz,1H),7.75(d,J＝8.7Hz,1H),7.62(td,J＝8.3,6.2Hz,1H),7.51(d,J＝8.1Hz,1H),7.48-7.37(m,2H),7.23(d,J＝2.0Hz,1H),4.37(s,2H),3.66(d,J＝13.0Hz,2H),3.39(d,J＝20.4Hz,2H),3.05(t,J＝10.5Hz,2H),2.37-2.32(m,1H),1.78-1.42(m,4H),1.19-1.00(m,4H)；LRMS(EI)m/z 554(M + )。
EXAMPLE 7 preparation of 5- (4- (((5-cyclopropyl-3- (2, 6-dichlorophenyl) isoxazol-4-yl) methoxy) methyl) -4-fluoropiperidin-1-yl) picolinic acid (FL 7)
The 4-iodo-ethyl benzoate was replaced with 5-bromopyridine-2-carboxylic acid methyl ester, and the remaining required raw materials, reagents and preparation methods were the same as in example 1, to obtain product FL7. 1 H NMR(400MHz,DMSO-d 6 )δ8.13-7.81(m,2H),7.80-7.40(m,4H),4.33(s,2H),3.69-3.25(m,4H),3.20-2.85(m,2H),2.42-2.24(m,1H),1.76-1.37(m,4H),1.19-0.93(m,4H)；LRMS(EI)m/z 521(M + )。
EXAMPLE 8 preparation of 5- (4- (((3- (2-chloro-6-fluorophenyl) -5-cyclopropylisoxazol-4-yl) methoxy) methyl) -4-fluoropiperidin-1-yl) picolinic acid (FL 8)
2, 6-dichlorobenzaldehyde is replaced by 2-chloro-6-fluorobenzaldehyde and 4-iodo-ethyl benzoate is replaced by 5-bromopyridine-2-carboxylic acid methyl ester, and the rest of the required raw materials, reagents and preparation method are the same as those in example 1, so as to obtain a product FL8. 1 H NMR(400MHz,DMSO-d 6 )δ8.09-7.90(m,2H),7.73-7.62(m,2H),7.56-7.26(m,2H),4.36(s,2H),3.64-3.34(m,4H),3.15-2.90(m,2H),2.41-2.20(m,1H),1.75-1.37(m,4H),1.18-0.95(m,4H)；LRMS(EI)m/z 505(M + )。
EXAMPLE 9 preparation of methyl 6- (4- (((5-cyclopropyl-3- (2, 6-dichlorophenyl) isoxazol-4-yl) methoxy) methyl) -4-fluoropiperidin-1-yl) -1-methyl-1H-indole-3-carboxylate (FL 9)
The 4-iodo-ethyl benzoate was replaced with 6-bromo-1-methyl-1H-indole-3-carboxylic acid methyl ester, and the remaining required raw materials, reagents and preparation methods were the same as in example 1, to obtain product FL9. 1 H NMR(400MHz,CDCl 3 )δ8.01(d,J＝8.7Hz,1H),7.67(s,1H),7.40(d,J＝7.8Hz,2H),7.34-7.30(m,1H),7.26(s,1H),7.02(d,J＝8.4Hz,1H),6.79(s,1H),4.39(s,3H),3.89(s,3H),3.77(s,3H),3.43-3.37(m,4H),3.05(t,J＝10.6Hz,2H),2.22-2.14(m,1H),1.91-1.71(m,4H),1.32-1.19(m,4H),1.16-1.11(m,4H)；LRMS(EI)m/z 587(M + )。
EXAMPLE 10 preparation of 6- (4- (((5-cyclopropyl-3- (2, 6-dichlorophenyl) isoxazol-4-yl) methoxy) methyl) -4-fluoropiperidin-1-yl) -1-methyl-1H-indole-3-carboxylic acid (FL 10)
The 4-iodo-ethyl benzoate was replaced with 6-bromo-1-methyl-1H-indole-3-carboxylic acid methyl ester, and the remaining required raw materials, reagents and preparation method were the same as in example 1, to give product FL10. 1 H NMR(400MHz,DMSO-d 6 )δ11.81(s,1H),7.89-7.76(m,2H),7.72-7.49(m,3H),6.96(d,J＝8.5Hz,2H),4.35(s,2H),3.78(s,3H),3.43-3.38(m,4H),2.90(t,J＝13.0Hz,2H),2.40-2.33(m,1H),1.79-1.54(m,4H),1.21-1.06(m,4H)；LRMS(EI)m/z 573(M + )。
EXAMPLE 11 preparation of 6- (4- (((3- (2-chloro-6-fluorophenyl) -5-cyclopropylisoxazol-4-yl) methoxy) methyl) -4-fluoropiperidin-1-yl) -1-methyl-1H-indole-3-carboxylic acid (FL 11)
2, 6-dichlorobenzaldehyde is replaced by 2-chloro-6-fluorobenzaldehyde and 4-iodo-ethyl benzoate is replaced by 6-bromo-1-methyl-1H-indole-3-methyl formate, and the rest of the required raw materials, reagents and preparation method are the same as those in example 1, so that the target product FL11 is obtained. 1 H NMR(400MHz,DMSO-d 6 )δ11.79(s,1H),7.88-7.76(m,2H),7.66-7.60(m,1H),7.52(d,J＝8.1Hz,1H),7.47-7.36(m,1H),7.00-6.88(m,2H),4.38(s,2H),3.78(s,3H),3.44-3.35(m,4H),2.90(t,J＝11.4Hz,2H),2.39-2.32(m,1H),1.75-1.38(m,4H),1.21-1.04(m,4H)；LRMS(EI)m/z 557(M + )。
EXAMPLE 12 preparation of 6- (4- (((3- (2-chloro-6-fluorophenyl) -5-cyclopropylisoxazol-4-yl) methoxy) methyl) -4-fluoropiperidin-1-yl) -1-ethyl-1H-indole-3-carboxylic acid (FL 12)
2, 6-dichlorobenzaldehyde is replaced by 2-chloro-6-fluorobenzaldehyde and 4-iodo-ethyl benzoate is replaced by 6-bromo-1-ethyl-1H-indole-3-methyl formate, and the rest of the required raw materials, reagents and preparation method are the same as those in example 1, so that the target product FL12 is obtained. 1 H NMR(400MHz,DMSO-d 6 )δ11.82(s,1H),7.89(s,1H),7.82(d,J＝8.7Hz,1H),7.67-7.58(m,1H),7.52(d,J＝8.1Hz,1H),7.42(t,J＝8.7Hz,1H),7.02-6.86(m,2H),4.38(s,2H),4.21(q,J＝7.2Hz,2H),3.45-3.35(m,3H),2.89(t,J＝10.9Hz,2H),2.89(t,J＝10.9Hz,2H),2.39-2.33(m,1H),1.72-1.47(m,4H),1.37(t,J＝7.2Hz,2H),1.20-1.07(m,4H)；LRMS(EI)m/z 571(M + )。
EXAMPLE 13 preparation of methyl 2- (4- (((5-cyclopropyl-3- (2, 6-dichlorophenyl) isoxazol-4-yl) methoxy) methyl) -4-fluoropiperidin-1-yl) benzo [ d ] thiazole-6-carboxylate (FL 13)
A100 mL eggplant-type bottle was charged with 5-cyclopropyl-3- (2, 6-dichlorophenyl) -4- (((4-fluoropiperidin-4-yl) methoxy) methyl) isoxazole (2.0 g, 5.01mmol), 2-bromobenzo [ d ] m]Methyl thiazole-6-carboxylate (1.09g, 4.01mmol) and N, N-diisopropylethylamine (776.8mg, 6.01mmol) were dissolved in 50mL of N-methylpyrrolidone, and the mixture was stirred at 60 ℃ overnight. The reaction was monitored by Thin Layer Chromatography (TLC) and after completion of the reaction, water (100 mL) and ethyl acetate (3X 100mL) were added. The organic layer was washed with 200mL of saturated brine, dried over anhydrous Na2SO4, and concentrated in vacuo, petroleum Ether (PE): ethyl Acetate (EA) =5:1 to 11, passing through a column, and spin-drying to obtain 2.04 g of a solid, namely 2- (4- (((5-cyclopropyl-3- (2, 6-dichlorophenyl) isoxazol-4-yl) methoxy) methyl) -4-fluoropiperidine-1-yl) benzo [ d]Thiazole-6-carboxylic acid methyl ester, yield 69%, 1 H NMR(500MHz,CDCl 3 )δ8.41(d,J＝2.9Hz,1H),7.91(dd,J＝15.0,2.9Hz,1H),7.64(dd,J＝16.1,13.7Hz,1H),7.57(d,J＝15.0Hz,1H),7.45(dd,J＝14.9,1.0Hz,2H),4.80(s,2H),4.41(dt,J＝24.8,11.4Hz,2H),3.96(dt,J＝24.8,11.4Hz,2H),3.90(s,3H),3.76(d,J＝50.4Hz,2H),2.73(p,J＝20.2Hz,1H),1.96(ddt,J＝50.4,24.9,11.4Hz,2H),1.66(ddt,J＝50.3,24.9,11.4Hz,2H),1.45-0.88(m,4H)；ESI-MS m/z:591(M+1)。
EXAMPLE 14 preparation of 2- (4- (((5-cyclopropyl-3- (2, 6-dichlorophenyl) isoxazol-4-yl) methoxy) methyl) -4-fluoropiperidin-1-yl) benzo [ d ] thiazole-6-carboxylic acid (FL 14)
To a 250mL two-necked flask was added 2- (4- (((5-cyclopropyl-3- (2, 6-dichlorophenyl) isoxazol-4-yl) methoxy) methyl) -4-fluoropiperidin-1-yl) benzo [ d ] under nitrogen]Thiazole-6-carboxylic acid methyl ester (FL 13,2.05g, 3.56mmol), followed by the addition of 30mL of methanol, 30mL of tetrahydrofuran and 10mL of 1, 4-dioxane for dissolution, followed by the addition of 2N KOH (30mL, 61.0mmol). Stirring was carried out overnight at 70 ℃. The reaction was monitored by Thin Layer Chromatography (TLC) and after completion the solution was spun dry in vacuo and 100mL of water and 50mL of methyl tert-butyl ether were added. The aqueous layer was adjusted to pH 5 with 2N HCl, then extracted with dichloromethane CH2Cl2 (3X 100mL), the organic layer was washed with saturated brine 200mL, dried over anhydrous Na2SO4, and the crude product concentrated in vacuo and recrystallized from MeOH/CH3CN to give 1.4g of a white solid in 70% yield. 1 H NMR(400MHz,DMSO-d 6 )δ12.71(s,1H),8.38(d,J＝1.7Hz,1H),7.86(dd,J＝8.4,1.8Hz,1H),7.66-7.52(m,3H),7.48(d,J＝8.5Hz,1H),4.34(s,2H),3.87(d,J＝12.7Hz,2H),3.41(d,J＝19.8Hz,2H),3.32-3.29(m,2H),2.38-2.31(m,1H),1.79-1.44(m,4H),1.20-1.03(m,4H)；LRMS(EI)m/z 577(M + )。
EXAMPLE 15 preparation of methyl 6- (4- (((3- (2-chloro-6-fluorophenyl) -5-cyclopropylisoxazol-4-yl) methoxy) methyl) -4-fluoropiperidin-1-yl) -1-ethyl-1H-indole-3-carboxylate (FL 15)
Replacing 2, 6-dichlorobenzaldehyde by 2-chloro-6-fluorobenzaldehyde and 4-iodo-ethyl benzoate by 6-bromo-1-ethyl-1The target product FL15 is obtained by the same method as the example 1 with the H-indole-3-methyl formate and other needed raw materials, reagents and preparation method. 1 H NMR(400MHz,CDCl 3 )δ8.02(d,J＝8.0Hz,1H),7.74(s,1H),7.41-7.36m,1H),7.31(d,J＝8.0Hz,1H),7.11(t,J＝8.4Hz,1H),7.01(d,J＝6.8Hz,1H),6.82(s,1H),4.42(s,2H),4.14(q,J＝7.3Hz,2H),3.89(s,3H),3.38(d,J＝19.1Hz,4H),3.05(t,J＝12.9Hz,2H),2.20-2.13(m,1H),1.92-1.65(m,4H),1.50(t,J＝7.3Hz,3H),1.30-1.22(m,2H),1.16-1.11(m,2H)；LRMS(EI)m/z 585(M + )。
EXAMPLE 16 preparation of 6- (4- (((5-cyclopropyl-3- (2, 6-dichlorophenyl) isoxazol-4-yl) methoxy) methyl) -4-fluoropiperidin-1-yl) -1- (2-hydroxyethyl) -1H-indole-3-carboxylic acid (FL 16)
The 4-iodo-ethyl benzoate was replaced with 6-bromo-1- (2- ((trimethylsilyl) oxy) ethyl) -1H-indole-3-carboxylic acid, and the remaining required raw materials, reagents and preparation method were the same as in example 1 to give the target product FL16. 1 H NMR(400MHz,DMSO-d 6 )δ8.17-7.96(m,3H),7.69-7.69(m,3H),7.59-7.55(m,1H),4.37(s,2H),4.30(s,2H),3.79-3.70(m,4H),3.50(s,4H),2.41-2.35(m,1H),2.08-1.80(m,4H),1.20-1.06(m,4H)；LRMS(EI)m/z 603(M + )。
EXAMPLE 17 preparation of methyl 5- (4- (((5-cyclopropyl-3- (2, 6-dichlorophenyl) isoxazol-4-yl) methoxy) methyl) -4-fluoropiperidin-1-yl) picolinate (FL 17)
The 4-iodo-ethyl benzoate was replaced with 5-bromopyridine-2-carboxylic acid methyl ester, and the remaining required raw materials, reagents and preparation methods were the same as in example 1, to obtain the target product FL17. 1 H NMR(400MHz,CDCl 3 )δ8.35(d,J＝2.9Hz,1H),8.01(d,J＝8.8Hz,1H),7.40(d,J＝1.9Hz,1H),7.38(s,1H),7.33(dd,J＝9.3,6.6Hz,1H),7.17(dd,J＝8.9,2.9Hz,1H),4.37(s,2H),3.97(s,3H),3.67(d,J＝13.1Hz,2H),3.36(d,J＝17.9Hz,2H),3.22(td,J＝12.8,2.6Hz,2H),2.17-2.10(m,1H),1.83(t,J＝11.6Hz,2H),1.75-1.50(m,2H),1.33-1.26(m,2H),1.16-1.11(m,2H)；LRMS(EI)m/z 535(M + )。
EXAMPLE 18 preparation of methyl 6- (4- (((5-cyclopropyl-3- (2, 6-dichlorophenyl) isoxazol-4-yl) methoxy) methyl) -4-fluoropiperidin-1-yl) -2-naphthoate (FL 18)
2, 6-twoThe chlorobenzaldehyde is replaced by 2-chloro-6-fluorobenzaldehyde and 4-iodo-ethyl benzoate is replaced by 6-bromo-2-methyl naphthoate, and the other required raw materials, reagents and preparation methods are the same as those in example 1, so as to obtain a target product FL18. 1 H NMR(400MHz,CDCl 3 )δ8.48(s,1H),7.99(d,J＝7.7Hz,1H),7.82(d,J＝8.1Hz,1H),7.68(d,J＝5.3Hz,1H),7.37(td,J＝8.2,5.9Hz,1H),7.31(s,1H),7.29(s,1H),7.09(t,J＝8.4Hz,2H),4.41(s,2H),3.96(s,3H),3.63(d,J＝12.6Hz,2H),3.37(d,J＝19.1Hz,2H),3.20(s,2H),2.20-2.10(m,1H),1.95-1.80(m,2H),1.57(s,2H),1.31-1.24(m,2H),1.15-1.10(m,2H)；LRMS(EI)m/z 568(M + )。
EXAMPLE 19 preparation of 2- (4- (((3- (2-chloro-6-fluorophenyl) -5-cyclopropylisoxazol-4-yl) methoxy) methyl) -4-fluoropiperidin-1-yl) benzo [ d ] thiazole-6-carboxylic acid (FL 19)
2, 6-dichlorobenzaldehyde is replaced by 2-chloro-6-fluorobenzaldehyde, and the rest of the required raw materials, reagents and preparation method are the same as those in example 14, so that the target product FL19 is obtained. 1 H NMR(400MHz,DMSO-d 6 )δ12.69(s,1H),8.38(d,J＝1.7Hz,1H),7.86(dd,J＝8.5,1.8Hz,1H),7.61(td,J＝8.3,6.2Hz,1H),7.49(t,J＝8.0Hz,2H),7.44-7.36(m,1H),4.36(s,2H),3.86(d,J＝12.7Hz,2H),3.53-3.17(m,4H),2.37-2.30(m,1H),1.88-1.43(m,4H),1.20-1.04(m,4H)；LRMS(EI)m/z 561(M + )。
EXAMPLE 20 preparation of 3- ((4- (((5-cyclopropyl-3- (2, 6-dichlorophenyl) isoxazol-4-yl) methoxy) methyl) -4-fluoropiperidin-1-yl) -1H-indazole-6-carboxylic acid (FL 20)
The target product FL20 was obtained by substituting 4-iodo-benzoic acid ethyl ester with 3-bromo-1- (tetrahydro-2H-pyran-2-yl) -1H-indazole-6-carboxylic acid methyl ester and the remaining required starting materials, reagents and preparation methods were the same as in example 1. 1 H NMR(400MHz,DMSO-d 6 )δ13.00(s,1H),12.31(s,1H),7.95(s,1H),7.82(d,J＝8.6Hz,1H),7.65-7.60(m,2H),7.56-7.52(m,2H),4.35(s,2H),3.58(d,J＝12.7Hz,2H),3.40(d,J＝20.4Hz,2H),3.09-2.96(m,2H),2.39-2.33(m,1H),1.80-1.57(m,4H),1.20-1.06(m,4H)；LRMS(EI)m/z 560(M + )。
EXAMPLE 21 preparation of 3- (4- (((3- (2-chloro-6-fluorophenyl) -5-cyclopropylisoxazol-4-yl) methoxy) methyl) -4-fluoropiperidin-1-yl) -1H-indazole-6-carboxylic acid (FL 21)
2, 6-dichlorobenzaldehyde is replaced by 2-chloro-6-fluorobenzaldehyde and 4-iodo-ethyl benzoate is replaced by 3-bromo-1- (tetrahydro-2H-pyran-2-yl) -1H-indazole-6-methyl formate, and the rest of the required raw materials, reagents and preparation methods are the same as those in example 1, so that the target product FL21 is obtained. 1 H NMR(400MHz,DMSO-d 6 )δ12.97(s,1H),12.34(s,1H),7.95(s,1H),7.82(d,J＝8.6Hz,1H),7.63-7.57(m,1H),7.54(dd,J＝8.6,1.3Hz,1H),7.50(d,J＝8.6Hz,1H),7.44-7.36(m,1H),4.38(s,2H),3.61-3.54(m,2H),3.39(d,J＝20.6Hz,2H),3.08-2.99(m,2H),2.39-2.32(m,1H),1.78-1.56(m,4H),1.21-1.07(m,4H)；LRMS(EI)m/z 544(M + )。
Example preparation of 3- ((4- (((5-cyclopropyl-3- (2, 6-dichlorophenyl) isoxazol-4-yl) methoxy) methyl) -4-fluoropiperidin-1-yl) -1H-indazole-5-carboxylic acid (FL 22)
The target product FL22 was obtained by substituting 4-iodo-benzoic acid ethyl ester with 3-bromo-1- (tetrahydro-2H-pyran-2-yl) -1H-indazole-5-carboxylic acid methyl ester, and the remaining required starting materials, reagents and preparation methods were the same as in example 1. 1 H NMR(400MHz,DMSO-d 6 )δ12.71(s,1H),12.36(s,1H),8.35(s,1H),7.86(dd,J＝8.8,1.2Hz,1H),7.65-7.60(m,2H),7.55(dd,J＝9.2,6.8Hz,1H),7.41(d,J＝8.8Hz,1H),4.35(s,2H),3.59(d,J＝15.5Hz,2H),3.41(d,J＝20.1Hz,2H),3.17-2.88(m,2H),2.39-2.33(m,1H),1.82-1.54(m,4H),1.21-1.00(m,4H)；LRMS(EI)m/z 560(M + )。
EXAMPLE 23 preparation of 3- (4- (((3- (2-chloro-6-fluorophenyl) -5-cyclopropylisoxazol-4-yl) methoxy) methyl) -4-fluoropiperidin-1-yl) -1H-indazole-5-carboxylic acid (FL 23)
2, 6-dichlorobenzaldehyde is replaced by 2-chloro-6-fluorobenzaldehyde and 4-iodo-ethyl benzoate is replaced by 3-bromo-1- (tetrahydro-2H-pyran-2-yl) -1H-indazole-6-methyl formate, and the rest of the required raw materials, reagents and preparation method are the same as those in example 1, so that the target product FL23 is obtained. 1 H NMR(400MHz,DMSO-d 6 )δ12.71(s,1H),12.36(s,1H),8.34(s,1H),7.86(dd,J＝8.8,1.3Hz,1H),7.61(td,J＝8.3,6.2Hz,1H),7.51(d,J＝8.1Hz,1H),7.43-7.43(m,2H),4.38(s,2H),3.59(d,J＝14.2Hz,2H),3.40(d,J＝20.3Hz,2H),3.16-3.03(m,2H),2.39-2.32(m,1H),1.88-1.49(m,4H),1.20-0.98(m,4H)；LRMS(EI)m/z 544(M + )。
EXAMPLE 24 preparation of 5-cyclopropyl-3- (2, 6-dichlorophenyl) -4- (((4-fluoro-1- (4- (trifluoromethyl) phenyl) piperidin-4-yl) methoxy) methyl) isoxazole (FL 24)
The 4-iodo-ethyl benzoate was replaced with 1-bromo-4- (trifluoromethyl) benzene, and the remaining required raw materials, reagents and preparation methods were the same as in example 1, to obtain the target product FL24. 1 H NMR(400MHz,CD3OD)δ7.63-7.59(m,2H),7.58-7.52(m,1H),7.49(d,J＝8.8Hz,2H),7.04(d,J＝8.8Hz,2H),4.32(s,2H),3.58(d,J＝13.1Hz,2H),3.39(s,2H),3.10-2.94(m,2H),2.37-2.30(m,1H),1.73-1.36(m,4H),1.22-1.00(m,4H)；LRMS(EI)m/z 544(M + )。
EXAMPLE 25 preparation of 3- (2-chloro-6-fluorophenyl) -5-cyclopropyl-4- (((4-fluoro-1- (4- (trifluoromethyl) phenyl) piperidin-4-yl) methoxy) methyl) isoxazole (FL 25)
The target product FL25 was obtained by substituting 2, 6-dichlorobenzaldehyde with 2-chloro-6-fluorobenzaldehyde and 4-iodo-ethyl benzoate with 1-bromo-4- (trifluoromethyl) benzene, and the other required raw materials, reagents and preparation methods were the same as in example 1. 1 H NMR(400MHz,DMSO-d 6 )δ7.62(td,J＝8.3,6.2Hz,1H),7.51-7.48(m,3H),7.46-7.37(m,1H),7.05(d,J＝8.8Hz,2H),4.35(s,2H),3.86-3.50(m,2H),3.36(d,J＝20.2Hz,2H),3.10-2.89(m,2H),2.37-2.30(m,1H),1.67-1.38(m,4H),1.21-1.06(m,4H)；LRMS(EI)m/z 527(M + )。
EXAMPLE 26 preparation of 6- (4- (((5-cyclopropyl-3- (2, 6-difluorophenyl) isoxazol-4-yl) methoxy) methyl) -4-fluoropiperidin-1-yl) -1-methyl-1H-indole-3-carboxylic acid (FL 26)
The target product FL26 was obtained by substituting 2, 6-dichlorobenzaldehyde with 2, 6-difluorobenzaldehyde and ethyl 4-iodo-benzoate with methyl 6-bromo-1-methyl-1H-indole-3-carboxylate, and the other required raw materials, reagents and preparation methods were the same as in example 1. 1 H NMR(400MHz,DMSO-d 6 )δ11.76(s,1H),7.84(s,1H),7.81(d,J＝9.5Hz,1H),7.74-7.59(m,1H),7.40-7.21(m,2H),7.01-6.92(m,2H),4.41(s,2H),3.77(s,3H),3.42-3.36(m,4H),3.03-2.77(m,2H),2.38-2.31(m,1H),1.82-1.42(m,4H),1.19-1.03(m,4H)；LRMS(EI)m/z 540(M + ).
EXAMPLE 27 preparation of 6- (4- (((5-cyclopropyl-3- (2, 6-difluorophenyl) isoxazol-4-yl) methoxy) methyl) -4-fluoropiperidin-1-yl) -1-ethyl-1H-indole-3-carboxylic acid (FL 27)
2, 6-dichlorobenzaldehyde is replaced by 2, 6-difluorobenzaldehyde and 4-iodo-ethyl benzoate is replaced by 6-bromo-1-ethyl-1H-indole-3-methyl formate, and the rest of the required raw materials, reagents and preparation method are the same as those in example 1, so as to obtain the target product FL27. 1 H NMR(400MHz,DMSO-d 6 )δ11.81(s,1H),7.89(s,1H),7.81(d,J＝8.7Hz,1H),7.69-7.58(m,1H),7.33-7.28(m,2H),6.98(d,J＝1.5Hz,1H),6.96-6.92(m,1H),4.41(s,2H),4.20(q,J＝7.2Hz,2H),3.39-3.36(m,4H),3.06-2.77(m,2H),2.38-2.31(m,1H),1.75-1.50(m,4H),1.36(t,J＝7.2Hz,3H),1.18-1.03(m,4H)；LRMS(EI)m/z 554(M + ).
EXAMPLE 28 preparation of 2- (3- (((3- (2-chloro-6-fluorophenyl) -5-cyclopropylisoxazol-4-yl) methoxy) methyl) -3-fluoro-8-azabicyclo [3.2.1] oct-8-yl) benzo [ d ] thiazole-6-carboxylic acid (FL 28)
Replacement of 2, 6-Dichlorobenzaldehyde by 2-chloro-6-fluorobenzaldehyde and (4-fluoro-1-tert-butoxycarbonyl substituted piperidin-4-yl) methanol by tert-butyl 3-fluoro-3- (hydroxymethyl) -8-azabicyclo [3.2.1]The octa-8-carboxylate, the other required raw materials, reagents and preparation method are the same as those of examples 13 and 14, and the target product FL28 is obtained. 1 H NMR(400MHz,DMSO-d 6 )δ12.66(s,1H),8.40(s,1H),7.87(d,J＝8.4Hz,1H),7.65(dd,J＝14.4,7.8Hz,1H),7.59-7.43(m,3H),4.39(s,2H),4.31(s,2H),3.51(d,J＝30.0Hz,2H),2.42-2.35(m,1H),2.06-1.84(m,8H),1.33-1.04(m,4H)；LRMS(EI)m/z 587(M + ).
EXAMPLE 29 preparation of 2- (3- (((3- (2-chloro-6-fluorophenyl) -5-cyclopropylisoxazol-4-yl) methoxy) methyl) -3-fluoro-8-azabicyclo [3.2.1] oct-8-yl) -4-fluorobenzo [ d ] thiazole-6-carboxylic acid (FL 29)
Replacing 2, 6-dichlorobenzaldehyde by 2-chloro-6-fluorobenzaldehyde and 2-bromobenzo [ d]Replacement of thiazole-6-carboxylic acid methyl ester with 2-bromo-4-fluorobenzo [ d]Replacement of methyl thiazole-6-carboxylate and (4-fluoro-1-tert-butoxycarbonyl-substituted-piperidin-4-yl) methanol with tert-butyl 3-fluoro-3- (hydroxymethyl) -8-azabicyclo [3.2.1]The octane-8-carboxylate, the other required raw materials, reagents and preparation method are the same as those of examples 13 and 14, and the target product FL29 is obtained. 1 H NMR(400MHz,DMSO-d 6 )δ13.02(s,1H),8.27(d,J＝1.5Hz,1H),7.68-7.61(m,2H),7.56(d,J＝8.1Hz,1H),7.51-7.43(m,1H),4.39(s,2H),4.34(s,2H),3.51(d,J＝29.8Hz,2H),2.42-2.35(m,1H),2.02-1.91(m,8H),1.20-1.07(m,4H)；LRMS(EI)m/z 605(M + )。
EXAMPLE 30 preparation of 2- (4- (((3- (2-chloro-6-fluorophenyl) -5-cyclopropylisoxazol-4-yl) methoxy) methyl) -4-fluoropiperidin-1-yl) -4-fluorobenzo [ d ] thiazole-6-carboxylic acid (FL 30)
Replacement of 2, 6-Dichlorobenzaldehyde by 2-chloro-6-fluorobenzaldehyde and 2-bromobenzo [ d]Replacement of thiazole-6-carboxylic acid methyl ester with 2-bromo-4-fluorobenzo [ d]Thiazole-6-carboxylic acid methyl ester, the other required raw materials, reagents and preparation method are the same as example 14, and the target product FL30 is obtained. 1 H NMR(400MHz,DMSO-d 6 )δ12.97(s,1H),8.21(d,J＝1.5Hz,1H),7.64(d,J＝1.1Hz,1H),7.62(s,1H),7.59(dd,J＝11.5,1.5Hz,1H),7.54(dd,J＝9.0,7.2Hz,1H),4.34(s,2H),3.79-3.37(m,6H),2.40-2.33(m,1H),1.82-1.69(m,2H),1.54-1.37(m,2H),1.21-1.06(m,4H)；LRMS(EI)m/z 579(M + ).
EXAMPLE 31 preparation of 2- (4- (((5-cyclopropyl-3- (2, 6-dichlorophenyl) isoxazol-4-yl) methoxy) methyl) -4-fluoropiperidin-1-yl) -4-fluorobenzo [ d ] thiazole-6-carboxylic acid (FL 31)
Reacting 2-bromobenzo [ d]Replacement of thiazole-6-carboxylic acid methyl ester with 2-bromo-4-fluorobenzo [ d]Thiazole-6-carboxylic acid methyl ester, the other required raw materials, reagents and preparation method were the same as example 14, to obtain the target product FL31. 1 H NMR(400MHz,DMSO-d 6 )δ12.94(s,1H),8.25(d,J＝1.5Hz,1H),7.63(d,J＝1.9Hz,1H),7.61-7.59(m,2H),7.55(dd,J＝9.3,6.6Hz,1H),4.33(s,2H),3.88(d,J＝11.6Hz,2H),3.46-3.28(m,4H),2.38-2.31(m,1H),1.84-1.50(m,4H),1.19-1.06(m,4H)；LRMS(EI)m/z 595(M + ).
EXAMPLE 32 preparation of methyl 2- (4- (((3- (2-chloro-6-fluorophenyl) -5-cyclopropylisoxazol-4-yl) methoxy) methyl) -4-fluoropiperidin-1-yl) -4-fluorobenzo [ d ] thiazole-6-carboxylate (FL 32)
By replacing 2, 6-dichlorobenzaldehyde by 2-chloro-6-fluorobenzaldehyde and 2-bromobenzo [ d ]]Replacement of thiazole-6-carboxylic acid methyl ester with 2-bromo-4-fluorobenzo [ d]Thiazole-6-carboxylic acid methyl ester, the other required raw materials, reagents and preparation method were the same as example 14, to obtain the target product FL32. 1 H NMR(400MHz,CDCl 3 )δ8.08(d,J＝0.8Hz,1H),7.70(dd,J＝11.3,0.7Hz,1H),7.42(s,1H),7.40(s,1H),7.32(dd,J＝8.7,7.4Hz,1H),4.36(s,2H),3.91(s,3H),3.71-3.43(m,6H),2.17-2.11(m,1H),1.84-1.68(m,2H),1.70-1.57(m,2H),1.31-1.25(m,2H),1.17-1.12(m,2H)；LRMS(EI)m/z 593(M + ).
Example 33- (4- (((5-cyclopropyl-3- (2, 6-dichlorophenyl) isoxazol-4-yl) methoxy) methyl) -3-fluoro-8-azabicyclo [3.2.1] oct-8-yl) -4-fluorobenzo [ d ] thiazole-6-carboxylic acid (FL 33)
By reacting 2-bromobenzo [ d ]]Replacement of thiazole-6-carboxylic acid methyl ester with 2-bromo-4-fluorobenzo [ d]Replacement of methyl thiazole-6-carboxylate and (4-fluoro-1-tert-butoxycarbonyl-substituted-piperidin-4-yl) methanol with tert-butyl 3-fluoro-3- (hydroxymethyl) -8-azabicyclo [3.2.1]The octa-8-carboxylate, the other required raw materials, reagents and preparation method are the same as example 14, and the target product FL33 is obtained. 1 H NMR(400MHz,DMSO-d 6 )δ12.96(s,1H),8.27(d,J＝1.5Hz,1H),7.70(d,J＝1.2Hz,1H),7.68(s,1H),7.64-7.58(m,2H),4.36(s,4H),3.56-3.29(m,4H),2.46-2.32(m,1H),2.05-1.83(m,6H),1.21-1.05(m,4H)；LRMS(EI)m/z 621(M + ).
EXAMPLE 34 preparation of 2- (4- (((5-cyclopropyl-3- (2, 6-dichlorophenyl) isoxazol-4-yl) methoxy) methyl) -3-fluoro-8-azabicyclo [3.2.1] oct-8-yl) benzo [ d ] thiazole-6-carboxylic acid (FL 34)
(4-fluoro-1-tert-butoxycarbonyl-substituted piperidin-4-yl) methanol was replaced with tert-butyl 3-fluoro-3- (hydroxymethyl) -8-azabicyclo [3.2.1]The octa-8-carboxylate, the remaining required starting materials, reagents and preparation were as in example 14. 1 H NMR(400MHz,DMSO-d 6 )δ8.40(d,J＝1.7Hz,1H),7.87(dd,J＝8.5,1.8Hz,1H),7.69(d,J＝1.3Hz,1H),7.67(s,1H),7.60(dd,J＝9.1,7.1Hz,1H),7.53(d,J＝8.5Hz,1H),4.35(s,2H),4.30(s,2H),3.51(d,J＝30.2Hz,2H),2.42-2.31(m,1H),2.01-1.87(m,6H),1.47-1.40(m,2H),1.21-1.07(m,4H)；LRMS(EI)m/z 603(M + ).
EXAMPLE 35 preparation of methyl 2- (3- (((5-cyclopropyl-3- (2- (trifluoromethyl) phenyl) isoxazol-4-yl) methoxy) methyl) -3-fluoro-8-azabicyclo [3.2.1] oct-8-yl) -4-fluorobenzo [ d ] thiazole-6-carboxylate (FL 35)
2, 6-dichlorobenzeneReplacement of Formaldehyde by 2- (trifluoromethyl) benzaldehyde, 2-bromobenzo [ d ]]Replacement of thiazole-6-carboxylic acid methyl ester with 2-bromo-4-fluorobenzo [ d]Replacement of methyl thiazole-6-carboxylate and (4-fluoro-1-tert-butoxycarbonyl-substituted-piperidin-4-yl) methanol with tert-butyl 3-fluoro-3- (hydroxymethyl) -8-azabicyclo [3.2.1]The octane-8-carboxylate, the other required raw materials, reagents and preparation method are the same as those in example 14, and the target product FL35 is obtained. 1 H NMR(400MHz,CDCl 3 )δ8.31(d,J＝1.6Hz,1H),8.01(dd,J＝8.5,1.5Hz,1H),7.60-7.50(m,2H),7.43-7.38(m,2H),4.47(s,2H),4.44(s,2H),3.92(s,3H),3.51(d,J＝28.0Hz,2H),2.33-2.05(m,5H),1.65-1.49(m,4H),1.28-1.19(m,2H),1.15-1.08(m,2H)；LRMS(EI)m/z 650(M + ).
EXAMPLE 36 preparation of 2- (3- (((5-cyclopropyl-3- (2- (trifluoromethyl) phenyl) isoxazol-4-yl) methoxy) methyl) -3-fluoro-8-azabicyclo [3.2.1] oct-8-yl) -4-fluorobenzo [ d ] thiazole-6-carboxylic acid (FL 36)
By replacing 2, 6-dichlorobenzaldehyde by 2- (trifluoromethyl) benzaldehyde or 2-bromobenzo [ d ]]Replacement of thiazole-6-carboxylic acid methyl ester with 2-bromo-4-fluorobenzo [ d]Replacement of methyl thiazole-6-carboxylate and (4-fluoro-1-tert-butoxycarbonyl-substituted-piperidin-4-yl) methanol with tert-butyl 3-fluoro-3- (hydroxymethyl) -8-azabicyclo [3.2.1]The octa-8-carboxylate, other required raw materials, reagents and preparation method are the same as example 14, and the target product FL36 is obtained. 1 H NMR(400MHz,DMSO-d 6 )δ13.00(s,1H),8.27(d,J＝1.5Hz,1H),7.73-7.69(m,1H),7.67(dd,J＝6.5,1.7Hz,1H),7.64(dd,J＝4.1,1.6Hz,1H),7.61(d,J＝1.5Hz,1H),7.58-7.55(m,1H),4.44(s,2H),4.35(s,2H),3.54(d,J＝29.6Hz,2H),2.41-2.34(m,1H),2.05-1.85(m,6H),1.51(q,J＝6.5Hz,2H),1.21-1.05(m,4H)；LRMS(EI)m/z636(M + ).
EXAMPLE 37 preparation of 2- (3- (((5-cyclopropyl-3- (2- (trifluoromethyl) phenyl) isoxazol-4-yl) methoxy) methyl) -3-fluoro-8-azabicyclo [3.2.1] oct-8-yl) benzo [ d ] thiazole-6-carboxylic acid (FL 37)
Replacement of 2, 6-Dichlorobenzaldehyde by 2- (trifluoromethyl) benzaldehyde and (4-fluoro-1-tert-butoxycarbonyl substituted piperidin-4-yl) methanol by tert-butyl 3-fluoro-3- (hydroxymethyl) -8-azabicyclo [3.2.1]The octa-8-carboxylate, the other required raw materials, reagents and preparation method are the same as example 14, and the target product FL37 is obtained. 1 H NMR(500MHz,CDCl 3 )δ8.58(d,J＝3.1Hz,1H),8.43(dd,J＝14.4,3.7Hz,1H),8.07(dd,J＝14.9,3.0Hz,1H),7.68(d,J＝15.0Hz,1H),7.57-7.29(m,2H),7.12(dd,J＝14.2,3.7Hz,1H),4.80(s,2H),4.77(ddd,J＝12.9,7.4,4.2Hz,2H),4.11(d,J＝50.6Hz,2H),2.72(p,J＝20.1Hz,1H),2.36(ddd,J＝50.6,24.9,6.4Hz,2H),2.11(ddd,J＝50.4,24.9,6.4Hz,2H),2.01-1.76(m,4H),1.34-1.11(m,2H),1.11-0.89(m,2H).LRMS(EI)m/z 618(M + ).
EXAMPLE 38 preparation of 2- (4- (((5-cyclopropyl-3- (2- (trifluoromethyl) phenyl) isoxazol-4-yl) methoxy) methyl) -4-fluoropiperidin-1-yl) -4-fluorobenzo [ d ] thiazole-6-carboxylic acid (FL 38)
Replacement of 2, 6-Dichlorobenzaldehyde by 2- (trifluoromethyl) benzaldehyde and (4-fluoro-1-tert-butoxycarbonyl substituted piperidin-4-yl) methanol by tert-butyl 3-fluoro-3- (hydroxymethyl) -8-azabicyclo [3.2.1]The octane-8-carboxylate, the other required raw materials, reagents and preparation method are the same as those in example 14, and the target product FL38 is obtained. 1 H NMR(500MHz,CDCl 3 )δ8.00(d,J＝8.0,2H),7.53(d,J＝8.4,2H),7.17(s,1H),6.86(s,1H),3.96(s,3H),3.91(s,6H),3.74(s,2H),3.32-3.41(m,3H),2.85-2.92(m,2H),2.52-2.71(m,1H),2.28-2.48(m,1H),2.19-2.30(m,1H),2.07-2.14(m,4H),1.70-1.82(m,4H)；LRMS(EI)m/z 610(M + ).
EXAMPLE 39 preparation of 2- (4- (((5-cyclopropyl-3- (2- (trifluoromethyl) phenyl) isoxazol-4-yl) methoxy) methyl) -4-fluoropiperidin-1-yl) benzo [ d ] thiazole-6-carboxylic acid (FL 39)
2, 6-dichlorobenzaldehyde is replaced by 2- (trifluoromethyl) benzaldehyde, and the other required raw materials, reagents and preparation methods are the same as those in example 14, so that the target product FL39 is obtained. 1 H NMR(500MHz,CDCl 3 )δ8.58(d,J＝3.1Hz,1H),8.43(dd,J＝14.3,3.7Hz,1H),8.07(dd,J＝15.0,2.9Hz,1H),7.68(d,J＝15.0Hz,1H),7.55-7.32(m,2H),7.12(dd,J＝14.2,3.7Hz,1H),4.80(s,2H),4.52(dt,J＝24.9,11.0Hz,2H),4.24(dt,J＝24.9,11.0Hz,2H),3.77(d,J＝50.4Hz,2H),2.89(p,J＝20.2Hz,1H),2.05(ddt,J＝50.4,24.9,11.0Hz,2H),1.74(ddt,J＝50.4,24.7,11.0Hz,2H),1.23(m,2H),1.11-0.87(m,2H).LRMS(EI)m/z 592(M + ).
EXAMPLE 40 preparation of methyl 6- (3- (((5-cyclopropyl-3- (2, 6-dichlorophenyl) isoxazol-4-yl) methoxy) methyl) -3-fluoro-8-azabicyclo [3.2.1] oct-8-yl) -1-methyl-1H-indole-3-carboxylate (FL 40)
(4-fluoro-1-tert-butoxycarbonyl-substituted piperidin-4-yl) methanol was replaced with tert-butyl 3-fluoro-3- (hydroxymethyl) -8-azabicyclo [3.2.1]The octyl-8-carboxylate and 4-iodo-ethyl benzoate were replaced with 6-bromo-1-methyl-1H-indole-3-carboxylic acid methyl ester, and the remaining required raw materials, reagents and preparation method were the same as in example 1, to obtain the target product FL40. 1 H NMR(500MHz,CDCl 3 )δ9.00(s,1H),8.10(d,J＝7.5Hz,1H),7.64(t,J＝7.5Hz,1H),7.46(d,J＝7.4Hz,2H),6.62(dd,J＝7.5,1.4Hz,1H),6.48(d,J＝1.4Hz,1H),4.80(s,2H),4.24-4.15(m,2H),3.93(d,J＝25.3Hz,2H),3.82(d,J＝80.0Hz,6H),2.71(p,J＝10.2Hz,1H),2.22-1.96(m,4H),1.79(ddd,J＝25.3,12.5,3.2Hz,2H),1.54(tdd,J＝5.8,3.8,1.9Hz,2H),1.35-1.13(m,2H),1.10-0.90(m,2H).LRMS(EI)m/z 613(M + ).
EXAMPLE 41 preparation of methyl 6- (3- (((5-cyclopropyl-3- (2- (trifluoromethyl) phenyl) isoxazol-4-yl) methoxy) methyl) -3-fluoro-8-azabicyclo [3.2.1] oct-8-yl) -1-methyl-1H-indole-3-carboxylate (FL 41)
The 2, 6-dichlorobenzaldehyde is replaced by 2- (trifluoromethyl) benzaldehyde, and the (4-fluoro-1-tert-butyloxycarbonyl substituted piperidin-4-yl) methanol is replaced by tert-butyl 3-fluoro-3- (hydroxymethyl) -8-azabicyclo [3.2.1]The octyl-8-carboxylate and 4-iodo-ethyl benzoate were replaced with 6-bromo-1-methyl-1H-indole-3-carboxylic acid methyl ester, and the remaining required raw materials, reagents and preparation method were the same as in example 1 to obtain the target product FL14. 1 H NMR(500MHz,CDCl 3 )δ9.00(s,1H),8.43(dd,J＝14.4,3.7Hz,1H),8.10(d,J＝15.0Hz,1H),7.65-7.35(m,2H),7.12(dd,J＝14.2,3.7Hz,1H),6.62(dd,J＝15.0,2.9Hz,1H),6.48(d,J＝2.9Hz,1H),4.80(s,2H),4.12(tt,J＝8.6,6.4Hz,2H),4.00(d,J＝50.4Hz,2H),3.82(d,J＝80.0Hz,6H),2.80(dd,J＝40.3,20.2Hz,1H),2.11(m,4H),1.86-1.39(m,4H),1.36-0.78(m,4H).LRMS(EI)m/z 628(M + ).
EXAMPLE 42 preparation of 6- (3- (((5-cyclopropyl-3- (2, 6-dichlorophenyl) isoxazol-4-yl) methoxy) methyl) -3-fluoro-8-azabicyclo [3.2.1] oct-8-yl) -1-methyl-1H-indole-3-carboxylic acid (FL 42)
Reacting (4-fluoro-1-tert-butoxycarbonyl)Replacement of the radical-substituted piperidin-4-yl) methanol by tert-butyl 3-fluoro-3- (hydroxymethyl) -8-azabicyclo [3.2.1]The octyl-8-carboxylate and 4-iodo-ethyl benzoate were replaced with 6-bromo-1-methyl-1H-indole-3-carboxylic acid methyl ester, and the remaining required raw materials, reagents and preparation method were the same as in example 1, to obtain the target product FL42. 1 H NMR(500MHz,CDCl 3 )δ9.26(s,1H),7.64(dd,J＝16.1,13.7Hz,1H),7.60-7.28(m,3H),6.72(d,J＝2.9Hz,1H),6.12(dd,J＝15.0,3.1Hz,1H),4.80(s,2H),4.26(tt,J＝8.5,6.4Hz,2H),3.96(d,J＝50.4Hz,2H),3.74(s,3H),3.03(p,J＝20.2Hz,1H),2.34(ddd,J＝50.4,24.8,6.4Hz,2H),2.22-1.84(m,4H),1.81-1.40(m,2H),1.37-0.82(m,4H).LRMS(EI)m/z 599(M + ).
EXAMPLE 43 preparation of 6- (3- (((5-cyclopropyl-3- (2- (trifluoromethyl) phenyl) isoxazol-4-yl) methoxy) methyl) -3-fluoro-8-azabicyclo [3.2.1] oct-8-yl) -1-methyl-1H-indole-3-carboxylic acid (FL 43)
Replacing 2, 6-dichlorobenzaldehyde with 2- (trifluoromethyl) benzaldehyde, replacing (4-fluoro-1-tert-butyloxycarbonyl substituted piperidine-4-yl) methanol with tert-butyl 3-fluoro-3- (hydroxymethyl) -8-azabicyclo [ 3.2.1%]The octyl-8-carboxylate and 4-iodo-ethyl benzoate were replaced with 6-bromo-1-methyl-1H-indole-3-carboxylic acid methyl ester, and the remaining required raw materials, reagents and preparation method were the same as in example 1, to obtain the target product FL43. 1 H NMR(500MHz,CDCl 3 )δ9.26(s,1H),8.43(dd,J＝14.4,3.7Hz,1H),7.78-7.29(m,3H),7.12(dd,J＝14.2,3.7Hz,1H),6.68(d,J＝2.9Hz,1H),6.12(dd,J＝15.0,3.1Hz,1H),4.80(s,2H),4.25(tt,J＝8.6,6.4Hz,2H),4.03(d,J＝50.4Hz,2H),3.74(s,3H),3.13-2.77(m,1H),2.52-1.96(m,6H),1.81(s,1H),1.41-0.73(m,4H).LRMS(EI)m/z 614(M + ).
EXAMPLE 44 preparation of 6- (3- (((3- (2, 6-dichlorophenyl) -5- ((2, 2-difluorocyclopropyl) methyl) isoxazol-4-yl) methoxy) methyl) -4-fluoropiperidin-1-yl) -1-methyl-1H-indole-3-carboxylic acid (FL 44)
Methyl 3-cyclopropyl-3-oxo-propionate was replaced with methyl 4- (2, 2-difluorocyclopropane) -3-oxobutanoate and ethyl 4-iodo-benzoate with methyl 6-bromo-1-methyl-1H-indole-3-carboxylate, and the remaining required raw materials, reagents and preparation methods were the same as in example 1, 1 H NMR(500MHz,CDCl 3 )δ9.26(s,1H),7.89-7.58(m,1H),7.52-7.21(m,3H),6.79(d,J＝2.9Hz,1H),6.12(dd,J＝15.0,3.1Hz,1H),4.80(s,2H),3.76(t,J＝25.2Hz,5H),3.64-3.40(m,3H),3.39-3.18(m,2H),2.94(dd,J＝24.8,14.6Hz,1H),2.35-2.02(m,2H),1.98-1.50(m,3H),0.80(m,2H).LRMS(EI)m/z 623(M + ).
EXAMPLE 45 preparation of 2- (4- (((3- (2, 6-dichlorophenyl) -5- ((2, 2-difluorocyclopropyl) methyl) isoxazol-4-yl) methoxy) methyl) -4-fluoropiperidin-1-yl) -4-fluorobenzo [ d ] thiazole-6-carboxylic acid (FL 45)
Exchange of methyl 3-cyclopropyl-3-oxo-propionate for methyl 4- (2, 2-difluorocyclopropane) -3-oxobutanoate and 2-bromobenzo [ d ]]Replacement of thiazole-6-carboxylic acid methyl ester with 2-bromo-4-fluorobenzo [ d]Thiazole-6-methyl formate, and the other required raw materials, reagents and preparation methods were the same as example 14 to obtain the target product FL45. 1 H NMR(500MHz,CDCl 3 )δ8.35(d,J＝2.9Hz,1H),7.96(dd,J＝15.9,2.9Hz,1H),7.64(dd,J＝16.2,13.7Hz,1H),7.46(dd,J＝14.8,1.1Hz,2H),4.80(s,2H),4.37(dt,J＝24.9,10.9Hz,2H),4.03(dt,J＝24.9,10.9Hz,2H),3.63(d,J＝50.4Hz,2H),3.23(dd,J＝24.7,17.0Hz,1H),2.94(dd,J＝24.8,17.0Hz,1H),2.09(ddt,J＝50.4,24.7,10.9Hz,2H),1.90-1.37(m,3H),1.33-0.54(m,2H).LRMS(EI)m/z 645(M + ).
EXAMPLE 46 preparation of 2- (4- (((5-cyclopropyl-3- (2, 6-difluorophenyl) isoxazol-4-yl) methoxy) methyl) -4-fluoropiperidin-1-yl) benzo [ d ] thiazole-6-carboxylic acid (FL 46)
2, 6-dichlorobenzaldehyde is replaced by 2, 6-difluorobenzaldehyde, and the other required raw materials, reagents and preparation method are the same as those in example 14, so that the target product FL46 is obtained. 1 H NMR(400MHz,DMSO-d 6 )δ13.02(s,1H),8.25(d,J＝1.3Hz,1H),7.63(ddd,J＝11.6,7.7,1.5Hz,2H),7.30(t,J＝8.1Hz,2H),4.40(s,2H),3.88(d,J＝10.3Hz,2H),3.62-3.26(m,6H),2.34(ddd,J＝13.4,8.3,5.3Hz,1H),1.87-1.31(m,4H),1.36-0.86(m,4H).LRMS(EI)m/z 544(M + ).
EXAMPLE 47 preparation of 2- (4- (((5-cyclopropyl-3- (2, 6-difluorophenyl) isoxazol-4-yl) methoxy) methyl) -4-fluoropiperidin-1-yl) -4-fluorobenzo [ d ] thiazole-6-carboxylic acid (FL 47)
Replacement of 2, 6-Dichlorobenzaldehyde by 2, 6-Difluorobenzaldehyde and 2-Bromobenzo [ d]Thiazole-6-carboxylic acid methyl ester replacementTo 2-bromo-4-fluorobenzo [ d]The thiazole-6-methyl formate, the other required raw materials, reagents and preparation method are the same as those of example 14, and the target product DC69 is obtained with the yield of 80%. 1 H NMR(500MHz,CDCl 3 )δ8.32(d,J＝2.9Hz,1H),7.64(dd,J＝16.0,3.0Hz,1H),7.53-7.19(m,1H),7.21-6.83(m,2H),4.78(s,2H),4.56(dt,J＝24.9,11.4Hz,2H),4.00-3.78(m,4H),2.91(p,J＝20.1Hz,1H),2.12-1.68(m,4H),1.41-0.71(m,4H).；LRMS(EI)m/z 562(M + ).
EXAMPLE 48 2- (3- (((3- (2, 6-dichlorophenyl) -5- ((2, 2-difluorocyclopropyl) methyl) isoxazol-4-yl) methoxymethyl) -3-fluoro-8-azabicyclo [3.2.1] oct-8-yl) -4-fluorobenzo [ d ] thiazole-6-carboxylic acid (FL 48)
Replacement of methyl 3-cyclopropyl-3-oxo-propionate with methyl 4- (2, 2-difluorocyclopropane) -3-oxobutanoate, and (4-fluoro-1-tert-butoxycarbonyl-substituted piperidin-4-yl) methanol with tert-butyl 3-fluoro-3- (hydroxymethyl) -8-azabicyclo [3.2.1]Octa-8-carboxylate and 2-bromobenzo [ d]Replacement of thiazole-6-carboxylic acid methyl ester with 2-bromo-4-fluorobenzo [ d]Thiazole-6-carboxylic acid methyl ester, the other required raw materials, reagents and preparation method were the same as example 14, to obtain the target product FL48. 1 H NMR(500MHz,CDCl 3 )δ9.00-8.07(m,1H),8.05(dd,J＝14.9,3.0Hz,1H),7.63(t,J＝7.5Hz,1H),7.47(d,J＝7.4Hz,2H),4.82(s,2H),4.66(tt,J＝8.5,6.4Hz,2H),4.27(d,J＝50.4Hz,1H),3.29-2.91(m,2H),2.60-1.90(m,4H),1.89-1.31(m,5H),1.26-0.66(m,2H)；LRMS(EI)m/z 671(M + ).
EXAMPLE 49 preparation of 2- (3- (((3- (2, 6-dichlorophenyl) -5- ((2, 2-difluorocyclopropyl) methyl) isoxazol-4-yl) methoxymethyl) -3-fluoro-8-azabicyclo [3.2.1] oct-8-yl) benzo [ d ] thiazole-6-carboxylic acid (FL 49)
Replacement of methyl 3-cyclopropyl-3-oxo-propionate with methyl 4- (2, 2-difluorocyclopropane) -3-oxobutanoate and (4-fluoro-1-tert-butoxycarbonyl-piperidin-4-yl) methanol with tert-butyl 3-fluoro-3- (hydroxymethyl) -8-azabicyclo [3.2.1]The octa-8-carboxylate, the other required raw materials, reagents and preparation method are the same as example 32, and the target product FL49 is obtained. 1 H NMR(500MHz,CDCl 3 )δ9.01-8.08(m,1H),8.07(dd,J＝14.9,3.0Hz,1H),7.90-7.53(m,2H),7.46(dd,J＝14.9,1.0Hz,2H),4.80(s,2H),4.64(tt,J＝8.5,6.4Hz,2H),4.25(d,J＝50.4Hz,1H),3.29-2.90(m,2H),2.60-1.93(m,4H),1.89-1.32(m,5H),1.26-0.67(m,2H)；LRMS(EI)m/z653(M + ).
EXAMPLE 50 2- (3- (((3- (2, 6-dichlorophenyl) -5- ((2, 2-difluorocyclopropyl) methyl) isoxazol-4-yl) methoxymethyl) -3-fluoro-8-azabicyclo [3.2.1] oct-8-yl) -1-methyl-1H-indole-3-carboxylic acid (FL 50)
Replacement of methyl 3-cyclopropyl-3-oxo-propionate with methyl 4- (2, 2-difluorocyclopropane) -3-oxobutanoate, and (4-fluoro-1-tert-butoxycarbonyl-substituted piperidin-4-yl) methanol with tert-butyl 3-fluoro-3- (hydroxymethyl) -8-azabicyclo [3.2.1]The octyl-8-carboxylate and 4-iodo-ethyl benzoate were replaced with 6-bromo-1-methyl-1H-indole-3-carboxylic acid methyl ester, and the remaining required raw materials, reagents and preparation method were the same as in example 1 to obtain the target product FL50. 1 H NMR(500MHz,CDCl 3 )δ9.26(s,1H),7.64(dd,J＝16.1,13.7Hz,1H),7.49-7.32(m,3H),6.73(d,J＝2.9Hz,1H),6.12(dd,J＝15.0,3.1Hz,1H),4.80(s,2H),4.56-4.24(m,2H),4.19(dd,J＝72.1,28.4Hz,2H),3.98-3.77(m,2H),3.74(s,3H),3.52(dd,J＝24.8,16.4Hz,1H),3.19-2.48(m,3H),2.02-1.24(m,3H),1.27-0.29(m,4H).LRMS(EI)m/z 649(M + ).
EXAMPLE 51 preparation of 2- (3- (((3- (2, 6-dichlorophenyl) -5- ((2, 2-difluorocyclopropyl) methyl) isoxazol-4-yl) methoxymethyl) -4-fluoropiperidin-1-yl) -1-ethyl-1H-indole-3-carboxylic acid (FL 51)
The methyl 3-cyclopropyl-3-oxo-propionate was replaced with methyl 4- (2, 2-difluorocyclopropane) -3-oxobutanoate and ethyl 4-iodo-benzoate was replaced with methyl 6-bromo-1-ethyl-1H-indole-3-carboxylate, and the remaining required raw materials, reagents and preparation methods were the same as in example 1 to obtain the target product FL51. 1 H NMR(500MHz，CDCl 3 )δ9.26(s,1H),7.64(dd,J＝16.1,13.7Hz,1H),7.56-7.23(m,3H),6.79(d,J＝3.1Hz,1H),6.12(dd,J＝15.0,3.1Hz,1H),4.80(s,2H),4.24(q,J＝12.6Hz,2H),3.77(d,J＝50.4Hz,2H),3.52(dt,J＝24.9,12.9Hz,2H),3.23(ddd,J＝25.0,13.5,6.6Hz,3H),2.85(dd,J＝24.8,15.2Hz,1H),2.32-2.00(m,2H),1.94-1.48(m,3H),1.06(t,J＝12.6Hz,3H),1.24-0.39(m,2H)；LRMS(EI)m/z 637(M + ).
EXAMPLE 52 preparation of 2- (3- (((3- (2, 6-dichlorophenyl) -5- ((2, 2-difluorocyclopropyl) methyl) isoxazol-4-yl) methoxymethyl) -3-fluoro-8-azabicyclo [3.2.1] oct-8-yl) -1-ethyl-1H-indole-3-carboxylic acid (FL 52)
Replacement of methyl 3-cyclopropyl-3-oxo-propionate with methyl 4- (2, 2-difluorocyclopropane) -3-oxobutanoate, and (4-fluoro-1-tert-butoxycarbonyl-substituted piperidin-4-yl) methanol with tert-butyl 3-fluoro-3- (hydroxymethyl) -8-azabicyclo [3.2.1]The octyl-8-carboxylate and 4-iodo-ethyl benzoate were replaced with 6-bromo-1-ethyl-1H-indole-3-carboxylic acid methyl ester, and the remaining required raw materials, reagents and preparation method were the same as in example 1, to obtain the target product FL52. 1 H NMR(500MHz,CDCl 3 )δ9.26(s,1H),7.64(dd,J＝16.1,13.7Hz,1H),7.57-7.33(m,3H),6.71(d,J＝3.1Hz,1H),6.12(dd,J＝15.0,3.1Hz,1H),4.80(s,2H),4.47-4.08(m,6H),4.07-3.75(m,2H),3.46(dd,J＝24.8,12.4Hz,1H),3.16-2.78(m,3H),2.15-1.47(m,3H),1.40-0.16(m,7H)；LRMS(EI)m/z 663(M + ).
EXAMPLE 53 preparation of 2- (3- (((3- (2, 6-dichlorophenyl) -5- ((2, 2-difluorocyclopropyl) methyl) isoxazol-4-yl) methoxymethyl) -4-fluoropiperidin-1-yl) benzo [ d ] thiazole-6-carboxylic acid (FL 53)
Methyl 3-cyclopropyl-3-oxo-propionate was replaced with methyl 4- (2, 2-difluorocyclopropane) -3-oxobutanoate, and the remaining required raw materials, reagents and preparation methods were the same as in example 14, to obtain the target product FL53. 1 H NMR(500MHz,CDCl 3 )δ8.58(d,J＝1.6Hz,1H),8.07(dd,J＝7.5,1.4Hz,1H),7.83-7.57(m,2H),7.46(d,J＝7.4Hz,2H),4.80(s,2H),4.50(dt,J＝12.3,5.5Hz,2H),4.19(dt,J＝12.4,5.5Hz,2H),3.88(dd,J＝12.4,7.6Hz,1H),3.81(d,J＝25.3Hz,2H),2.84(dd,J＝12.4,7.6Hz,1H),2.02(ddt,J＝25.2,12.3,5.4Hz,2H),1.74(ddt,J＝25.1,12.4,5.5Hz,2H),1.69-1.51(m,1H),1.42-1.20(m,1H),0.78-0.64(m,1H).LRMS(EI)m/z 627(M + ).
EXAMPLE 54 preparation of 2- (3- (((3- (2, 6-dichlorophenyl) -5- (2, 2-difluorocyclopropyl) isoxazol-4-yl) methoxymethyl) -3-fluoro-8-azabicyclo [3.2.1] oct-8-yl) -4-fluorobenzo [ d ] thiazole-6-carboxylic acid (FL 54)
Replacement of methyl 3-cyclopropyl-3-oxo-propionate with methyl 4- (2, 2-difluorocyclopropane) -3-oxobutanoate with methyl 3- (2, 2-difluorocyclopropyl-3-oxopropanoate, (4-fluoro-1-tert-butoxycarbonyl)Replacement of substituted piperidin-4-yl) methanol with tert-butyl 3-fluoro-3- (hydroxymethyl) -8-azabicyclo [3.2.1]Octa-8-carboxylate and 2-bromobenzo [ d]Replacement of thiazole-6-carboxylic acid methyl ester with 2-bromo-4-fluorobenzo [ d]Thiazole-6-carboxylic acid methyl ester, the other required raw materials, reagents and preparation method were the same as example 14, to obtain the target product FL54. 1 H NMR(500MHz,CDCl 3 )δ8.35(d,J＝3.1Hz,1H),7.74(dd,J＝15.9,3.1Hz,1H),7.64(dd,J＝16.1,13.7Hz,1H),7.46(dd,J＝14.9,1.0Hz,2H),4.77-4.71(m,2H),4.70(s,2H),4.31(d,J＝50.6Hz,2H),3.33(tt,J＝41.8,23.1Hz,1H),2.84(ddd,J＝50.4,24.8,6.7Hz,2H),2.17-1.95(m,2H),1.83(ddd,J＝50.5,24.8,6.7Hz,2H),1.71-1.48(m,2H),1.39-0.98(m,2H)；LRMS(EI)m/z 657(M + ).
EXAMPLE 55 preparation of 3- (((3- (2, 6-dichlorophenyl) -5- (2, 2-difluorocyclopropyl) isoxazol-4-yl) methoxymethyl) -3-fluoro-8-azabicyclo [3.2.1] oct-8-yl) benzo [ d ] thiazole-6-carboxylic acid (FL 55)
Exchange of methyl 3-cyclopropyl-3-oxo-propionate for methyl 4- (2, 2-difluorocyclopropane) -3-oxobutanoate for 3- (2, 2-difluorocyclopropyl-3-oxopropanoic acid and (4-fluoro-1-tert-butoxycarbonyl-piperidin-4-yl) methanol for tert-butyl 3-fluoro-3- (hydroxymethyl) -8-azabicyclo [3.2.1]The octa-8-carboxylate, the other required raw materials, reagents and preparation method are the same as example 14, and the target product FL55 is obtained. 1 H NMR(500MHz,CDCl 3 )δ8.58(d,J＝3.1Hz,1H),8.07(dd,J＝14.9,3.0Hz,1H),7.83-7.53(m,2H),7.46(dd,J＝14.9,1.0Hz,2H),4.89-4.59(m,4H),4.29(d,J＝50.4Hz,2H),3.26(tt,J＝41.8,23.1Hz,1H),2.76(ddd,J＝50.5,24.8,6.8Hz,2H),2.18-1.97(m,2H),1.84(ddd,J＝50.4,24.8,6.7Hz,2H),1.66-1.42(m,2H),1.37-0.99(m,2H).LRMS(EI)m/z 639(M + )。
EXAMPLE 56 preparation of 2- (3- (((3- (2, 6-dichlorophenyl) -5- (2, 2-difluorocyclopropyl) isoxazol-4-yl) methoxymethyl) -3-fluoro-8-azabicyclo [3.2.1] oct-8-yl) -1-methyl-1H-indole-3-carboxylic acid (FL 56)
Replacement of methyl 3-cyclopropyl-3-oxo-propionate with methyl 4- (2, 2-difluorocyclopropane) -3-oxobutanoate with methyl 3- (2, 2-difluorocyclopropyl-3-oxopropanoate and (4-fluoro-1-tert-butoxycarbonyl-substituted piperidin-4-yl) -methanol with tert-butyl 3-fluoro-3- (hydroxymethyl)-8-azabicyclo [3.2.1]The octyl-8-carboxylate and 4-iodo-ethyl benzoate were replaced with 6-bromo-1-methyl-1H-indole-3-carboxylic acid methyl ester, and the remaining required raw materials, reagents and preparation method were the same as in example 1, to obtain the target product FL56. 1 H NMR(500MHz,CDCl 3 )δ9.26(s,1H),7.64(dd,J＝16.1,13.7Hz,1H),7.56-7.26(m,3H),6.62(d,J＝2.9Hz,1H),6.12(dd,J＝15.0,3.1Hz,1H),4.76(s,2H),4.26(s,2H),3.81(d,J＝50.4Hz,2H),3.74(s,3H),2.88(tt,J＝41.9,23.0Hz,1H),2.34-2.02(m,6H),1.72-0.90(m,4H)；LRMS(EI)m/z635(M + )。
EXAMPLE preparation of 2- (3- (((5- (2, 2-difluorocyclopropyl) -3- (2- (trifluoromethoxy) phenyl) isoxazol-4-yl) methoxy) methyl) -3-fluoro-8-azabicyclo [3.2.1] oct-8-yl) -4-fluorobenzo [ d ] thiazole-6-carboxylic acid (FL 57)
2, 6-dichlorobenzaldehyde is replaced by 2- (trifluoromethyl) benzaldehyde, and the rest of the required raw materials, reagents and preparation method are the same as example 54, so that the target product FL57 is obtained. 1 H NMR(500MHz,CDCl 3 )δ8.43(dd,J＝14.4,3.7Hz,1H),8.35(d,J＝3.1Hz,1H),7.74(dd,J＝16.0,3.0Hz,1H),7.43(ddd,J＝18.1,14.0,3.3Hz,2H),7.12(dd,J＝14.2,3.7Hz,1H),4.81(s,2H),4.73(ddd,J＝8.4,6.5,2.0Hz,2H),4.19(d,J＝50.4Hz,2H),3.01(tt,J＝41.8,22.9Hz,1H),2.52-1.65(m,8H),1.55-0.76(m,2H)；LRMS(EI)m/z672(M + ).
Example 58 preparation of 2- (3- (((5- ((2, 2-difluorocyclopropyl) methyl) -3- (2- (trifluoromethoxy) phenyl) isoxazol-4-yl) methoxy) methyl) -3-fluoro-8-azabicyclo [3.2.1] oct-8-yl) -1-methyl-1H-indole-3-carboxylic acid (FL 58)
2, 6-dichlorobenzaldehyde is replaced by 2- (trifluoromethyl) benzaldehyde, and the rest of the required raw materials, reagents and preparation methods are the same as those of example 50, so that the target product FL58 is obtained. 1 H NMR(500MHz,CDCl 3 )δ9.25(s,1H),8.42(dd,J＝14.2,3.7Hz,1H),7.59-7.32(m,3H),7.11(dd,J＝14.2,3.7Hz,1H),6.69(d,J＝3.0Hz,1H),6.11(dd,J＝15.0,2.9Hz,1H),4.79(s,2H),4.55-4.10(m,4H),3.73(s,3H),3.34(dd,J＝24.7,16.3Hz,1H),2.64(dd,J＝24.7,16.3Hz,1H),2.50-2.08(m,6H),1.77-1.24(m,3H),1.12-0.83(m,2H)；LRMS(EI)m/z 664(M + ).
EXAMPLE 59 preparation of 2- (3- (((5- ((2, 2-difluorocyclopropyl) methyl) -3- (2- (trifluoromethoxy) phenyl) isoxazol-4-yl) methoxy) methyl) -3-fluoro-8-azabicyclo [3.2.1] oct-8-yl) -4-fluorobenzo [ d ] thiazole-6-carboxylic acid (FL 59)
2, 6-dichlorobenzaldehyde is replaced by 2- (trifluoromethyl) benzaldehyde, and the other required raw materials, reagents and preparation method are the same as example 48 to obtain the target product FL59. 1 H NMR(500MHz,CDCl 3 )δ8.43(dd,J＝14.4,3.7Hz,1H),8.35(d,J＝3.1Hz,1H),7.94(dd,J＝16.0,3.0Hz,1H),7.59-7.30(m,2H),7.12(dd,J＝14.2,3.7Hz,1H),4.80(s,2H),4.67-4.56(m,2H),4.41(d,J＝50.4Hz,2H),3.21(dd,J＝24.7,14.3Hz,1H),2.98(dd,J＝24.9,14.2Hz,1H),2.45(ddd,J＝50.5,24.8,6.7Hz,2H),2.08-1.84(m,2H),1.85-1.46(m,5H),1.15-0.51(m,2H).LRMS(EI)m/z 686(M + ).
EXAMPLE 60 preparation of 2- (4- (((5-cyclopropyl-3- (2- (trifluoromethoxy) phenyl) isoxazol-4-yl) oxy) methyl) -4-fluoropiperidin-1-yl) -4-fluorobenzo [ d ] thiazole-6-carboxylic acid (FL 60)
The preparation method is the same as that of example 38, and the target product FL60 is obtained. 1 H NMR(500MHz,CDCl 3 )δ8.00(d,J＝8.0,2H),7.53(d,J＝8.4,2H),7.17(s,1H),6.86(s,1H),3.96(s,3H),3.91(s,6H),3.32-3.41(m,3H),2.85-2.92(m,2H),2.52-2.71(m,1H),2.28-2.48(m,1H),2.19-2.30(m,1H),2.07-2.14(m,4H),1.70-1.82(m,4H)；LRMS(EI)m/z 610(M+).；LRMS(EI)m/z 596(M + ).
EXAMPLE 61 preparation of 2- (4- (((5-cyclopropyl-3- (2- (trifluoromethoxy) phenyl) isoxazol-4-yl) oxy) methyl) -3-fluoro-8-azabicyclo [3.2.1] oct-8-yl) -4-fluorobenzo [ d ] thiazole-6-carboxylic acid (FL 61)
The preparation method is the same as example 36, and the target product FL61 is prepared. 1 H NMR(CDCl 3 ,500MHz)8.27(d,J＝1.5Hz,1H),7.73-7.69(m,1H),7.67(dd,J＝6.5,1.7Hz,1H),7.64(dd,J＝4.1,1.6Hz,1H),7.61(d,J＝1.5Hz,1H),7.58-7.55(m,1H),4.35(s,2H),3.54(d,J＝29.6Hz,2H),2.41-2.34(m,1H),2.05-1.85(m,6H),1.51(q,J＝6.5Hz,2H),1.21-1.05(m,4H)；LRMS(EI)m/z 636(M+).LRMS(EI)m/z 622(M + ).
EXAMPLE 62 preparation of 2- (4- (((5-cyclopropyl-3- (2, 6-dichlorophenyl) isoxazol-4-yl) oxy) methyl) -4-fluoropiperidin-1-yl) -4-fluorobenzo [ d ] thiazole-6-carboxylic acid (FL 62)
The preparation method is the same as example 31, and the target product FL62 is prepared. 1 H NMR(CDCl 3 ,500MHz)8.25(d,J＝1.5Hz,1H),7.63(d,J＝1.9Hz,1H),7.61-7.59(m,2H),7.55(dd,J＝9.3,6.6Hz,1H),3.88(d,J＝11.6Hz,2H),3.46-3.28(m,4H),2.38-2.31(m,1H),1.84-1.50(m,4H),1.19-1.06(m,4H)；LRMS(EI)m/z581(M + ).
EXAMPLE 63 preparation of 2- (4- (((5-cyclopropyl-3- (2, 6-dichlorophenyl) isoxazol-4-yl) oxy) methyl) -3-fluoro-8-azabicyclo [3.2.1] oct-8-yl) -4-fluorobenzo [ d ] thiazole-6-carboxylic acid (FL 63)
The preparation method is the same as example 33, and the target product FL63 is prepared. 1 H NMR(CDCl 3 ,500MHz)8.27(d,J＝1.5Hz,1H),7.70(d,J＝1.2Hz,1H),7.68(s,1H),7.64-7.58(m,2H),4.36(s,2H),3.56-3.29(m,4H),2.46-2.32(m,1H),2.05-1.83(m,6H),1.21-1.05(m,4H)；LRMS(EI)m/z 607(M + ).
EXAMPLE 64 preparation of 2- (4- (((5-cyclopropyl-3- (2, 6-dichlorophenyl) isoxazol-4-yl) oxy) methyl) -3-fluoro-8-azabicyclo [3.2.1] oct-8-yl) -1-methyl-1H-indole-3-carboxylic acid (FL 64)
The preparation method is the same as example 42, and the target product FL64 is prepared. 1 H NMR(CDCl 3 ,500MHz)δ9.00(s,1H),8.43(dd,J＝14.4,3.7Hz,1H),8.10(d,J＝15.0Hz,1H),7.65-7.35(m,2H),7.12(dd,J＝14.2,3.7Hz,1H),6.62(dd,J＝15.0,2.9Hz,1H),6.48(d,J＝2.9Hz,1H),4.12(tt,J＝8.6,6.4Hz,2H),4.00(d,J＝50.4Hz,2H),3.82(d,J＝80.0Hz,6H),2.80(dd,J＝40.3,20.2Hz,1H),2.11(m,4H),1.86-1.39(m,4H),1.36-0.78(m,4H).LRMS(EI)m/z 585(M + ).
EXAMPLE 65 preparation of 2- (4- (((5-cyclopropyl-3- (2, 6-dichlorophenyl) isoxazol-4-yl) oxy) methyl) -4-fluoropiperidin-1-yl) -1-methyl-1H-indole-3-carboxylic acid (FL 65)
The preparation method is the same as that of example 10, and the target product FL65 is prepared. 1 H NMR(CDCl 3 ,500MHz)δ9.18(s,1H),7.58(dd,J＝16.1,13.6Hz,1H),7.48-7.25(m,3H),7.01(d,J＝2.9Hz,1H),6.07(dd,J＝14.8,2.9Hz,1H),4.47(d,J＝50.2Hz,2H),3.71(s,3H),3.34(ddt,J＝117.5,24.7,13.1Hz,4H),2.75(p,J＝20.1Hz,1H),2.18-1.61(m,4H),1.19-0.76(m,4H).LRMS(EI)m/z 559(M + ).
EXAMPLE 66 preparation of 7- (3- (((5- ((2, 2-difluorocyclopropyl) methyl) -3- (2- (trifluoromethoxy) phenyl) isoxazol-4-yl) methoxy) methyl) -3-fluoro-8-azabicyclo [3.2.1] oct-8-yl) imidazo [1,2-a ] pyridine-3-carboxylic acid (FL 66)
Exchange of 6-bromo-1-methyl-1H-indole-3-carboxylic acid methyl ester for 7-bromoimidazo [1,2-a ]]Pyridine-3-methyl formate, and the other required raw materials, reagents and preparation methods were the same as in example 58, to obtain the target product FL66. 1 H NMR(500MHz,CDCl 3 )δ8.43(dd,J＝14.4,3.7Hz,1H),8.33(d,J＝15.0Hz,1H),7.66(s,1H),7.56-7.30(m,2H),7.12(dd,J＝14.2,3.7Hz,1H),6.65(d,J＝3.1Hz,1H),6.56(dd,J＝14.9,3.0Hz,1H),4.80(s,2H),4.29-4.16(m,3H),4.13(s,1H),3.57(dd,J＝24.8,14.7Hz,1H),2.80-2.39(m,3H),2.12-1.83(m,4H),1.79-1.36(m,3H),1.14-0.60(m,2H)；LRMS(EI)m/z 651(M + ).
EXAMPLE 67 preparation of 2- (4- (((5-cyclopropyl-3- (2- (trifluoromethoxy) phenyl) isoxazol-4-yl) methoxy) methyl) -3-fluoro-8-azabicyclo [3.2.1] oct-8-yl) imidazo [1,2-a ] pyridine-3-carboxylic acid (FL 67)
The 2, 6-dichlorobenzaldehyde is replaced by 2- (trifluoromethyl) benzaldehyde, and the (4-fluoro-1-tert-butyloxycarbonyl substituted piperidin-4-yl) methanol is replaced by tert-butyl 3-fluoro-3- (hydroxymethyl) -8-azabicyclo [3.2.1]Conversion of octyl-8-carboxylate and ethyl 4-iodo-benzoate to 7-bromoimidazo [1,2-a ]]Pyridine-3-methyl formate, and the other required raw materials, reagents and preparation methods are the same as example 1, and the target product FL67 is prepared. 1 H NMR(500MHz,CDCl 3 )δ8.43(dd,J＝7.5,1.6Hz,1H),8.33(d,J＝7.5Hz,1H),7.66(s,1H),7.43(dtd,J＝30.9,7.4,1.5Hz,1H),7.12(dd,J＝7.4,1.5Hz,1H),6.70(d,J＝1.4Hz,1H),6.57(dd,J＝7.5,1.6Hz,1H),4.80(s,1H),4.35-4.05(m,1H),3.94(d,J＝25.1Hz,1H),3.15(ddd,J＝25.3,12.4,3.3Hz,1H),2.56(p,J＝10.2Hz,1H),2.21-1.95(m,2H),1.90-1.51(m,4H),1.40-1.13(m,2H),1.13-0.84(m,2H).LRMS(EI)m/z 601(M + ).
EXAMPLE 68 preparation of 2- (4- (((5-cyclopropyl-3- (2- (trifluoromethoxy) phenyl) isoxazol-4-yl) methoxy) methyl) -4-fluoropiperidin-1-yl) imidazo [1,2-a ] pyridine-3-carboxylic acid (FL 68)
Replacement of 2, 6-Dichlorobenzaldehyde by 2- (trifluoromethyl) benzaldehyde and ethyl 4-iodo-benzoate by 7-bromoimidazo [1,2-a ]]Pyridine-3-methyl formate, other needed raw materials, reagents and preparation methods are the same as the examples, and the target product FL68 is prepared. 1 H NMR(500MHz,CDCl 3 )δ8.39(dd,J＝14.2,3.7Hz,1H),8.29(d,J＝14.8Hz,1H),7.62(s,1H),7.52-7.30(m,2H),7.09(dd,J＝14.2,3.7Hz,1H),6.75-6.50(m,2H),4.78(s,2H),3.79(d,J＝50.2Hz,2H),3.43(dt,J＝25.0,12.9Hz,2H),3.12(dt,J＝24.8,12.9Hz,2H),2.86-2.55(m,1H),2.09(ddt,J＝50.5,25.1,12.9Hz,2H),1.71(ddt,J＝50.6,24.8,13.0Hz,2H),1.41-0.82(m,4H).LRMS(EI)m/z 575(M + ).
EXAMPLE 69 preparation of 6- (3- (((5-cyclopropyl-3- (2, 6-dichlorophenyl) isoxazol-4-yl) methoxy) methyl) -4-fluoropiperidin-1-yl) imidazo [1,2-a ] pyridine-3-carboxylic acid (FL 69)
Replacement of 4-iodo-benzoic acid ethyl ester by 7-bromoimidazo [1,2-a ]]Pyridine-3-methyl formate, and the other required raw materials, reagents and preparation methods are the same as example 1, and the target product FL69 is prepared. 1 H NMR(500MHz,CDCl 3 )δ8.33(d,J＝15.0Hz,1H),7.98-7.55(m,2H),7.46(dd,J＝14.9,1.0Hz,2H),6.93(d,J＝3.1Hz,1H),6.80(dd,J＝14.9,3.0Hz,1H),4.80(s,2H),3.83(d,J＝50.6Hz,2H),3.65-3.34(m,3H),3.14(dt,J＝24.9,13.2Hz,2H),2.26-1.57(m,4H),1.45-0.73(m,4H).LRMS(EI)m/z 560(M + ).
EXAMPLE 70 preparation of 6- (3- (((5-cyclopropyl-3- (2, 6-dichlorophenyl) isoxazol-4-yl) methoxy) methyl) -3-fluoro-8-azabicyclo [3.2.1] oct-8-yl) imidazo [1,2-a ] pyridine-3-carboxylic acid (FL 70)
(4-fluoro-1-tert-butoxycarbonyl-substituted-piperidin-4-yl) -methanol was replaced with tert-butyl-3-fluoro-3- (hydroxymethyl) -8-azabicyclo [3.2.1] methyl]Conversion of octyl-8-carboxylate and ethyl 4-iodo-benzoate to 7-bromoimidazo [1,2-a ]]Pyridine-3-methyl formate, and the other required raw materials, reagents and preparation methods are the same as example 1, and the target product FL70 is prepared. 1 H NMR(500MHz,CDCl 3 )δ8.33(d,J＝15.0Hz,1H),7.74-7.53(m,2H),7.46(dd,J＝14.9,1.0Hz,2H),6.66(d,J＝3.0Hz,1H),6.58(dd,J＝14.9,3.0Hz,1H),4.80(s,2H),4.46(d,J＝50.4Hz,2H),4.34-4.15(m,2H),3.04(p,J＝20.0Hz,1H),2.72(ddd,J＝50.4,24.8,6.7Hz,2H),2.51-2.05(m,4H),1.84-1.49(m,2H),1.45-0.58(m,4H).LRMS(EI)m/z 586(M + ).
EXAMPLE 71 preparation of 2- (4- (((3- (2-chloro-6-fluorophenyl) -5-cyclopropylisoxazol-4-yl) methoxy) methyl) -4-fluoropiperidin-1-yl) imidazo [1,2-a ] pyridine-3-carboxylic acid (FL 71)
Replacement of 2, 6-Dichlorobenzaldehyde by 2-chloro-6-fluorobenzaldehyde and ethyl 4-iodo-benzoate by 7-bromoimidazo [1,2-a ]]Pyridine-3-methyl formate, and the other required raw materials, reagents and preparation methods are the same as example 1, and the target product FL71 is prepared. 1 H NMR(500MHz,CDCl 3 )δ8.33(d,J＝14.9Hz,1H),7.66(s,1H),7.51-7.11(m,3H),7.00-6.71(m,2H),4.80(s,2H),3.87(d,J＝50.4Hz,2H),3.30(dtd,J＝37.6,25.0,12.9Hz,4H),2.86(p,J＝20.0Hz,1H),2.32-1.51(m,4H),1.46-0.80(m,4H).LRMS(EI)m/z 544(M + ).
EXAMPLE 72 preparation of 2- (4- (((3- (2-chloro-6-fluorophenyl) -5-cyclopropylisoxazol-4-yl) methoxy) methyl) -3-fluoro-8-azabicyclo [3.2.1] oct-8-yl) -imidazo [1,2-a ] pyridine-3-carboxylic acid (FL 72)
Replacing 2, 6-dichlorobenzaldehyde with 2-chloro-6-fluorobenzaldehyde, replacing (4-fluoro-1-tert-butyloxycarbonyl substituted piperidine-4-yl) methanol with tert-butyl 3-fluoro-3- (hydroxymethyl) -8-azabicyclo [3.2.1]Replacement of octa-8-carboxylate and ethyl 4-iodo-benzoate by 7-bromoimidazo [1,2-a]Pyridine-3-methyl formate, other needed raw materials, reagents and preparation methods are the same as the examples, and the target product FL72 is prepared. 1 H NMR(500MHz,CDCl 3 )δ8.33(d,J＝7.5Hz,1H),7.66(s,1H),7.50-7.11(m,3H),6.78-6.51(m,2H),4.80(s,2H),4.42(d,J＝25.1Hz,2H),4.26-3.97(m,2H),2.66(p,J＝10.1Hz,1H),2.55-2.04(m,6H),1.40-0.81(m,6H).LRMS(EI)m/z 570(M + ).
EXAMPLE 73 preparation of 2- (4- (((3- (2-chloro-6-fluorophenyl) -5-cyclopropylisoxazol-4-yl) oxy) methyl) -3-fluoro-8-azabicyclo [3.2.1] oct-8-yl) -imidazo [1,2-a ] pyridine-3-carboxylic acid (FL 73)
The preparation method is the same as example 72, and the target product FL73 is prepared. 1 H NMR(500MHz,CDCl 3 )δ8.33(d,J＝15.0Hz,1H),7.66(s,1H),7.46-7.14(m,3H),6.86-6.49(m,2H),5.23(d,J＝50.4Hz,2H),4.39-4.05(m,2H),2.94-2.14(m,7H),1.86-0.73(m,6H).LRMS(EI)m/z 556(M + ).
Figure BDA0001653521430000511
EXAMPLE 74 preparation of methyl 2- (6- (3- (((5-cyclopropyl-3- (2, 6-dichlorophenyl) isoxazol-4-yl) methoxy) methyl) -4-fluoropiperidin-1-yl) -1-methyl-1H-indole-3-amide) acetate (FL 74)
2- (6- (3- (((5-cyclopropyl-3- (2, 6-dichlorophenyl) isoxazol-4-yl) methoxy) methyl) -4-fluoropiperidin-1-yl) -1-methyl-1H-indole-3-carboxylic acid (0.06mmo 1), glycine methyl ester (0.06mmo 1), 2- (7-azobenzotriazol) -N, N, N ', N' -tetramethyluronium hexafluorophosphate (0.065mmo 1), N, N-diisopropylethylamine (0.05 mI) was added to dichloromethane (2 mL), the reaction was monitored by Thin Layer Chromatography (TLC), after completion of the reaction, the solution was vacuum-dried, saturated sodium bicarbonate was then added, ethyl acetate was extracted, and the organic layer was dried over anhydrous Na 2 SO 4 Dry, petroleum Ether (PE): ethyl Acetate (EA) = (0-100%) through column to obtain FL74. 1 H NMR(500MHz,CDCl 3 )δ8.37(s,1H),7.64(dd,J＝16.2,13.7Hz,1H),7.52-7.35(m,3H),7.04(s,1H),6.83(d,J＝2.9Hz,1H),6.12(dd,J＝15.0,2.9Hz,1H),4.80(s,2H),3.87(s,2H),3.78(d,J＝36.3Hz,4H),3.67(d,J＝40.8Hz,4H),3.34(ddt,J＝111.0,24.9,12.5Hz,4H),2.87(dd,J＝40.2,20.1Hz,1H),1.86(ddtd,J＝62.8,37.3,24.8,12.5Hz,4H),1.42-0.74(m,4H).LRMS(EI)m/z 644(M + ).
EXAMPLE 75 preparation of 2- (6- (3- (((5-cyclopropyl-3- (2, 6-dichlorophenyl) isoxazol-4-yl) methoxy) methyl) -4-fluoropiperidin-1-yl) -1-methyl-1H-indole-3-carboxamide) acetic acid (FL 75)
To a 250mL two-necked flask was added methyl 2- (6- (3- (((5-cyclopropyl-3- (2, 6-dichlorophenyl) isoxazol-4-yl) methoxy) methyl) -4-fluoropiperidin-1-yl) -1-methyl-1H-indole-3-amide) acetate (0.05 mmol) under nitrogen, followed by 2mL of methanol and 2mL of 1, 4-dioxane for dissolution, followed by 4N LiOH (4 mL). Stirring was carried out overnight at 70 ℃. The reaction was monitored by Thin Layer Chromatography (TLC) and after completion the solution was spun dry in vacuo and water and methyl tert-butyl ether were added. The aqueous layer was adjusted to 2N HClpH 5, then dichloromethane CH2Cl2 extraction was added, organic layer washed with brine, dried over anhydrous Na2SO4, crude product concentrated in vacuo, dichloromethane (PE): passing methanol (EA) through the column to obtain FL75. 1 H NMR(500MHz,CDCl 3 )δ8.30(s,1H),7.77(s,1H),7.58(dd,J＝16.1,13.6Hz,1H),7.53-7.32(m,3H),7.09(d,J＝3.1Hz,1H),6.07(dd,J＝14.9,3.0Hz,1H),4.76(s,2H),3.73(d,J＝20.1Hz,4H),3.61(d,J＝39.4Hz,3H),3.34(dt,J＝24.6,11.3Hz,2H),3.12(dt,J＝24.7,11.2Hz,2H),2.89(p,J＝19.9Hz,1H),2.09-1.58(m,4H),1.40-0.80(m,4H).LRMS(EI)m/z 630(M + ).
EXAMPLE 76 preparation of 2- (6- (3- (((5-cyclopropyl-3- (2- (trifluoromethoxy) phenyl) isoxazol-4-yl) methoxy) methyl) -3-fluoro-8-azabicyclo [3.2.1] oct-8-yl) -1-methyl-1H-indole-3-carboxamide) acetic acid (FL 76)
The preparation method is the same as that of example 75, and the target product FL76 is prepared. 1 H NMR(500MHz,CDCl 3 )δ8.43(dd,J＝14.2,3.7Hz,1H),8.37(s,1H),7.57-7.35(m,3H),7.12(dd,J＝14.3,3.8Hz,1H),6.59(d,J＝2.9Hz,1H),6.53(s,1H),6.12(dd,J＝15.0,3.1Hz,1H),4.80(s,2H),4.25(tt,J＝8.6,6.4Hz,2H),3.98(d,J＝50.4Hz,2H),3.74(s,3H),3.60(s,2H),3.04(p,J＝20.1Hz,1H),2.39(ddd,J＝50.4,24.7,6.4Hz,2H),2.26-1.93(m,4H),1.75-1.39(m,2H),1.33-0.68(m,4H).LRMS(EI)m/z 671(M + ).
EXAMPLE 77 preparation of 2- (6- (3- (((5- ((2, 2-difluorocyclopropyl) methyl) -3- (2- (trifluoromethoxy) phenyl) isoxazol-4-yl) methoxy) methyl) -3-fluoroazabicyclo [3.2.1] oct-8-yl) -4-fluorobenzo [ d ] thiazole-6-amide) acetic acid (FL 77)
The preparation method is the same as example 75, and the target product FL77 is prepared. 1 H NMR(500MHz,CDCl 3 )δ8.43(dd,J＝14.4,3.7Hz,1H),8.17(d,J＝2.9Hz,1H),7.48-7.33(m,2H),7.30-7.06(m,2H),5.33(s,1H),4.80(s,2H),4.51-4.14(m,2H),4.04(d,J＝50.6Hz,2H),3.84(s,2H),3.12(ddd,J＝169.2,24.8,13.8Hz,2H),2.24(ddd,J＝50.4,24.7,6.6Hz,2H),1.99-1.20(m,7H),1.24-0.30(m,2H).LRMS(EI)m/z 743(M + ).
EXAMPLE 78 preparation of 2- (6- (3- (((5- (2, 2-difluorocyclopropyl) -3- (2- (trifluoromethoxy) phenyl) isoxazol-4-yl) methoxy) methyl) -3-fluoroazabicyclo [3.2.1] oct-8-yl) -1-methyl-1H-indole-3-carboxamide) acetic acid (FL 78)
The preparation method is the same as example 75, and the target product FL78 is prepared. 1 H NMR(500MHz,CDCl 3 )δ8.43(dd,J＝14.4,3.7Hz,1H),8.37(s,1H),7.74(s,1H),7.53-7.29(m,3H),7.12(dd,J＝14.2,3.7Hz,1H),6.73(d,J＝2.9Hz,1H),6.12(dd,J＝15.0,3.1Hz,1H),4.88(s,2H),4.47-4.27(m,2H),4.21(d,J＝50.4Hz,2H),3.99-3.80(m,2H),3.74(s,3H),3.60(s,2H),3.14-2.66(m,3H),1.63(ddd,J＝50.4,24.8,13.3Hz,2H),1.41-0.94(m,4H).LRMS(EI)m/z 707(M + ).
EXAMPLE 79 preparation of 2- (6- (3- (((5- (2, 2-difluorocyclopropyl) -3- (2- (trifluoromethoxy) phenyl) isoxazol-4-yl) methoxy) methyl) -3-fluoroazabicyclo [3.2.1] oct-8-yl) -4-fluorobenzo [ d ] thiazole-6-carboxamide) acetic acid (FL 79)
The preparation method is the same as that of example 75, and the target product FL79 is prepared. 1 H NMR(500MHz,CDCl 3 )δ8.43(dd,J＝14.4,3.7Hz,1H),8.17(d,J＝2.9Hz,1H),7.46(dddd,J＝33.1,29.1,15.5,3.2Hz,3H),7.12(dd,J＝14.2,3.7Hz,1H),6.14(s,1H),4.73(s,2H),4.66(tt,J＝8.6,6.4Hz,2H),4.14(d,J＝50.4Hz,2H),3.84(s,2H),3.08(tt,J＝41.8,23.0Hz,1H),2.46(ddd,J＝50.4,24.9,6.4Hz,2H),2.30-1.67(m,6H),1.41-0.90(m,2H).LRMS(EI)m/z 729(M + ).
EXAMPLE 80 preparation of 1H NMR (500MHz, CDCl3) LRMS (EI) M/z (M +). 2- (6- (3- (((5- ((2, 2-difluorocyclopropyl) methyl) -3- (2- (trifluoromethoxy) phenyl) isoxazol-4-yl) methoxy) methyl) -3-fluoroazabicyclo [3.2.1] oct-8-yl) -1-methyl-1H-indole-3-amide) acetic acid (FL 80)
The preparation method is the same as example 75, and the target product FL80 is prepared. 1 H NMR(500MHz,CDCl 3 )δ8.43(dd,J＝14.3,3.7Hz,1H),8.37(s,1H),7.61-7.35(m,3H),7.12(dd,J＝14.2,3.7Hz,1H),6.62(d,J＝3.1Hz,1H),6.30(s,1H),6.12(dd,J＝15.0,3.1Hz,1H),4.80(s,2H),4.31(tt,J＝8.6,6.4Hz,2H),4.08(d,J＝50.4Hz,2H),3.93(dd,J＝24.7,15.8Hz,1H),3.74(s,3H),3.60(s,2H),2.61-2.32(m,3H),2.28-2.02(m,4H),1.86-1.44(m,3H),1.17-0.67(m,2H).LRMS(EI)m/z 721(M + ).
EXAMPLE 81 preparation of 2- (6- (3- (((5-cyclopropyl-3- (2- (trifluoromethoxy) phenyl) isoxazol-4-yl) methoxy) methyl) -3-fluoro-8-azabicyclo [3.2.1] oct-8-yl) -4-fluorobenzo [ d ] thiazole-6-carboxamide) acetic acid (FL 81)
The preparation method is the same as example 75, and the target product FL81 is prepared. 1 H NMR(500MHz,CDCl 3 )δ8.43(dd,J＝14.4,3.7Hz,1H),8.17(d,J＝2.9Hz,1H),7.54(dd,J＝16.0,3.0Hz,1H),7.51-7.32(m,2H),7.12(dd,J＝14.2,3.7Hz,1H),6.21(s,1H),4.80(s,2H),4.55(tt,J＝8.6,6.4Hz,2H),4.01(d,J＝50.4Hz,2H),3.84(s,2H),2.76(p,J＝20.1Hz,1H),2.25(ddd,J＝50.4,24.9,6.4Hz,2H),2.04-1.47(m,6H),1.42-0.48(m,4H).LRMS(EI)m/z 693(M + ).
EXAMPLE 82 preparation of 2- (6- (3- (((5-cyclopropyl-3- (2, 6-dichlorophenyl) isoxazol-4-yl) methoxy) methyl) -3-fluoro-8-azabicyclo [3.2.1] oct-8-yl) -1-methyl-1H-indole-3-carboxamide) acetic acid (FL 82)
The preparation method is the same as example 75, and the target product FL82 is prepared. 1 H NMR(500MHz,CDCl 3 )δ8.36(s,1H),7.63(dd,J＝16.1,13.6Hz,1H),7.52-7.35(m,3H),6.58(d,J＝3.1Hz,1H),6.29-6.01(m,2H),4.79(s,2H),4.18(tt,J＝8.6,6.4Hz,2H),3.93(d,J＝50.4Hz,2H),3.73(s,3H),3.59(s,2H),2.71(p,J＝20.2Hz,1H),2.34-1.90(m,4H),1.82(ddd,J＝50.4,24.9,6.4Hz,2H),1.66-1.38(m,2H),1.28-0.99(m,4H).LRMS(EI)m/z 656(M + ).
EXAMPLE 83 preparation of 2- (2- (3- (((3- (2, 6-dichlorophenyl) -5- ((2, 2-difluorocyclopropyl) methyl) isoxazol-4-yl) methoxy) methyl) -3-fluoro-8-azabicyclo [3.2.1] oct-8-yl) -4-fluorobenzo [ d ] thiazole-6-amide) acetic acid (FL 83)
The preparation method is the same as example 75, and the target product FL83 is prepared. 1 H NMR(500MHz,CDCl 3 )δ8.15(d,J＝2.9Hz,1H),7.63(dd,J＝16.1,13.7Hz,1H),7.51(dd,J＝16.0,3.0Hz,1H),7.44(dd,J＝14.8,1.1Hz,2H),6.12(s,1H),4.79(s,2H),4.76-4.60(m,2H),4.46(d,J＝50.2Hz,2H),3.83(s,2H),3.06(ddd,J＝80.4,24.8,14.2Hz,2H),2.72(ddd,J＝50.4,24.8,6.7Hz,2H),2.21-1.42(m,7H),1.07-0.47(m,2H).LRMS(EI)m/z 728(M + ).
EXAMPLE 84 preparation of 2- (6- (3- (((5-cyclopropyl-3- (2, 6-dichlorophenyl) isoxazol-4-yl) methoxy) methyl) -3-fluoro-8-azabicyclo [3.2.1] oct-8-yl) -4-fluorobenzo [ d ] thiazole-6-carboxamide) acetic acid (FL 84)
The preparation method is the same as example 75, and the target product FL84 is prepared. 1 H NMR(500MHz,CDCl 3 )δ8.15(d,J＝2.9Hz,1H),7.63(dd,J＝16.1,13.7Hz,1H),7.52(dd,J＝15.9,2.9Hz,1H),7.44(dd,J＝14.9,1.0Hz,2H),6.20(s,1H),4.79(s,2H),4.64(tt,J＝8.4,6.4Hz,2H),4.08(d,J＝50.4Hz,2H),3.83(s,2H),2.73(dd,J＝40.3,20.1Hz,1H),2.16-1.49(m,8H),1.54-0.62(m,4H).LRMS(EI)m/z 678(M + ).
EXAMPLE 85 preparation of 2- (2- (3- (((3- (2, 6-dichlorophenyl) -5- ((2, 2-difluorocyclopropyl) methyl) isoxazol-4-yl) oxy) methyl) -3-fluoro-8-azabicyclo [3.2.1] oct-8-yl) -4-fluorobenzo [ d ] thiazole-6-amide) acetic acid (FL 85)
The preparation method is the same as that of example 75, and the target product FL85 is prepared. 1 H NMR(500MHz,CDCl 3 )δ8.17(d,J＝2.9Hz,1H),7.64(dd,J＝16.1,13.7Hz,1H),7.54(dd,J＝16.0,3.0Hz,1H),7.46(dd,J＝14.9,1.0Hz,2H),6.20(s,1H),4.94(d,J＝50.4Hz,2H),4.58(tt,J＝8.6,6.4Hz,2H),3.84(s,2H),3.33(ddd,J＝92.3,24.7,15.6Hz,2H),2.32(ddd,J＝50.6,24.9,6.4Hz,2H),1.90(ddd,J＝50.4,24.7,6.3Hz,2H),1.79-1.47(m,5H),1.14-0.64(m,2H).LRMS(EI)m/z 714(M + ).
EXAMPLE 86 preparation of 2- (6- (3- (((5-cyclopropyl-3- (2, 6-dichlorophenyl) isoxazol-4-yl) oxy) methyl) -3-fluoro-8-azabicyclo [3.2.1] oct-8-yl) -4-fluorobenzo [ d ] thiazole-6-carboxamide) acetic acid (FL 86)
The preparation method is the same as example 75, and the target product FL86 is prepared. 1 H NMR(500MHz,CDCl 3 )δ8.17(d,J＝2.9Hz,1H),7.73(s,1H),7.60(ddd,J＝19.0,16.0,8.3Hz,2H),7.46(dd,J＝14.9,1.0Hz,2H),5.24(d,J＝50.4Hz,2H),4.98-4.62(m,2H),3.84(s,2H),2.74(p,J＝20.2Hz,1H),2.42(ddd,J＝50.4,24.8,6.7Hz,2H),2.24-1.83(m,4H),1.79-1.51(m,2H),1.45-0.64(m,4H).LRMS(EI)m/z 634(M + ).
Figure BDA0001653521430000551
EXAMPLE 87 preparation of 2- (6- (3- (((5-cyclopropyl-3- (2, 6-dichlorophenyl) isoxazol-4-yl) methoxy) methyl) -3-fluoro-8-azabicyclo [3.2.1] oct-8-yl) -1-methyl-1H-indole-3-carboxamide) ethanesulfonic acid (FL 87)
Dissolving 2- (6- (3- (((5-cyclopropyl-3- (2, 6-dichlorophenyl) isoxazol-4-yl) methoxy) methyl) -4-fluoropiperidin-1-yl) -1-methyl-1H-indole-3-carboxylic acid (0.1 mmol) in tetrahydrofuran (1.0 mL), adding N-methylmaleimide (0.7 mmol), stirring at room temperature until the acid is dissolved, adding 2-chloro-4, 6-dimethoxy-1, 3, 5-triazine (0.15 mmol), reacting at 50 ℃ for 20 minutes to generate precipitates, adding aminoethanesulfonic acid (0.40 mmol, dissolving in DMA 4 mL) under stirring, sealing at 80 ℃ for 2 hours, reacting, cooling to room temperature, adding water, extracting with ethyl acetate, and extracting an organic layer with anhydrous Na 2 SO 4 Drying, spin-drying, and purifying with column to obtain pure FL87. 1 H NMR(500MHz,CDCl 3 )δ8.37(s,1H),7.64(dd,J＝16.1,13.7Hz,1H),7.52-7.38(m,3H),7.19(d,J＝2.9Hz,1H),6.87(s,1H),6.68(s,1H),6.12(dd,J＝15.0,3.1Hz,1H),4.80(s,2H),3.71(t,J＝25.2Hz,5H),3.59-3.05(m,8H),2.73(p,J＝20.1Hz,1H),2.49-1.84(m,4H),1.48-0.65(m,4H).LRMS(EI)m/z 680(M + ).
EXAMPLE 88 preparation of 2- (6- (3- (((5-cyclopropyl-3- (2, 6-dichlorophenyl) isoxazol-4-yl) methoxy) methyl) -4-fluoropiperidin-1-yl) -1-methyl-1H-indole-3-amide) ethanesulfonic acid (FL 88)
The preparation method is the same as example 87, and the target product FL88 is prepared. 1 H NMR(500MHz,CDCl 3 )δ8.37(s,1H),7.61(s,1H),7.54-7.39(m,3H),6.68(s,1H),6.58(d,J＝2.9Hz,1H),6.51(s,1H),6.12(dd,J＝14.9,3.0Hz,1H),4.80(s,2H),4.18(tt,J＝8.6,6.4Hz,2H),3.93(d,J＝50.4Hz,2H),3.74(s,3H),3.56-3.11(m,4H),2.92-2.54(m,1H),2.37-1.92(m,4H),1.79(ddd,J＝50.4,24.7,6.4Hz,2H),1.67-1.41(m,2H),1.38-0.77(m,4H).LRMS(EI)m/z 706(M + ).
EXAMPLE 89 preparation of 2- (2- (3- (((3- (2, 6-dichlorophenyl) -5- ((2, 2-difluorocyclopropyl) methyl) isoxazol-4-yl) methoxy) methyl) -3-fluoro-8-azabicyclo [3.2.1] oct-8-yl) -4-fluorobenzo [ d ] thiazole-6-amide) ethanesulfonic acid (FL 89)
The preparation method is the same as example 87, and the target product FL89 is prepared. 1 H NMR(500MHz,CDCl 3 )δ8.17(d,J＝2.9Hz,1H),7.71-7.58(m,1H),7.52(dd,J＝15.9,2.9Hz,1H),7.47(d,J＝0.6Hz,1H),7.44(d,J＝1.4Hz,1H),6.66(s,1H),6.41(s,1H),4.80(s,2H),4.79-4.70(m,2H),4.15(d,J＝50.4Hz,2H),3.68(td,J＝16.6,1.5Hz,2H),3.33(td,J＝16.8,1.5Hz,2H),3.19(dd,J＝24.7,16.1Hz,1H),3.04-2.70(m,3H),2.21-1.03(m,8H),0.85-0.66(m,1H).LRMS(EI)m/z778(M + ).
EXAMPLE 90 preparation of 2- (6- (3- (((5-cyclopropyl-3- (2, 6-dichlorophenyl) isoxazol-4-yl) methoxy) methyl) -3-fluoro-8-azabicyclo [3.2.1] oct-8-yl) -4-fluorobenzo [ d ] thiazole-6-amide) ethanesulfonic acid (FL 90)
And the other required raw materials, reagents and preparation methods are the same as the embodiment, so that the target product FL90 is prepared. 1 H NMR(500MHz,CDCl 3 )δ8.37(s,1H),7.64(dd,J＝16.1,13.7Hz,1H),7.52-7.38(m,3H),7.19(d,J＝2.9Hz,1H),6.87(s,1H),6.68(s,1H),6.12(dd,J＝15.0,3.1Hz,1H),4.80(s,2H),3.71(t,J＝25.2Hz,5H),3.59-3.05(m,8H),2.73(p,J＝20.1Hz,1H),2.49-1.84(m,2H),1.48-0.65(m,4H).LRMS(EI)m/z 728(M + ).
EXAMPLE 91 preparation of 2- (6- (3- (((5-cyclopropyl-3- (2- (trifluoromethoxy) phenyl) isoxazol-4-yl) methoxy) methyl) -3-fluoro-8-azabicyclo [3.2.1] oct-8-yl) -1-methyl-1H-indole-3-amide) ethanesulfonic acid (FL 91)
The preparation method is the same as example 87, and the target product FL91 is prepared. 1 H NMR(500MHz,CDCl 3 )δ8.43(dd,J＝14.4,3.7Hz,1H),8.37(s,1H),7.55-7.25(m,3H),7.12(dd,J＝14.2,3.7Hz,1H),6.66(s,1H),6.60(d,J＝2.9Hz,1H),6.54(s,1H),6.12(dd,J＝15.0,3.1Hz,1H),4.80(s,2H),4.14(tt,J＝8.6,6.4Hz,2H),3.97(d,J＝50.4Hz,2H),3.74(s,3H),3.54-3.19(m,4H),2.68(p,J＝20.1Hz,1H),2.48-1.71(m,6H),1.68-0.41(m,6H).LRMS(EI)m/z 721(M + ).
EXAMPLE preparation of 2- (6- (3- (((5-cyclopropyl-3- (2- (trifluoromethoxy) phenyl) isoxazol-4-yl) methoxy) methyl) -4-fluoropiperidin-1-yl) -1-methyl-1H-indole-3-amide) ethanesulfonic acid (FL 92)
The preparation method is the same as example 87, and the target product FL92 is prepared. 1 H NMR(500MHz,CDCl 3 )δ8.43(dd,J＝14.4,3.7Hz,1H),8.37(s,1H),7.54-7.27(m,3H),7.20(d,J＝3.1Hz,1H),7.12(dd,J＝14.2,3.7Hz,1H),6.70(s,1H),6.52(s,1H),6.12(dd,J＝15.0,3.1Hz,1H),4.80(s,2H),3.76(t,J＝25.2Hz,5H),3.64-3.04(m,8H),2.78(p,J＝20.1Hz,1H),2.14-1.68(m,4H),1.48-0.77(m,4H).LRMS(EI)m/z 695(M + ).
EXAMPLE 93 preparation of 2- (6- (3- (((5- (2, 2-difluorocyclopropyl) -3- (2- (trifluoromethoxy) phenyl) isoxazol-4-yl) methoxy) methyl) -3-fluoroazabicyclo [3.2.1] oct-8-yl) -4-fluorobenzo [ d ] thiazole-6-amide) ethanesulfonic acid (FL 93)
The preparation method was the same as in example 87, and the target product FL93 was obtained. 1 H NMR(500MHz,CDCl 3 )δ8.43(dd,J＝14.4,3.7Hz,1H),8.17(d,J＝2.9Hz,1H),7.54(dd,J＝16.0,3.0Hz,1H),7.51-7.35(m,2H),7.12(dd,J＝14.2,3.7Hz,1H),6.57(d,J＝72.5Hz,2H),4.80(s,2H),4.79-4.73(m,2H),4.27(d,J＝50.4Hz,2H),3.72-2.89(m,6H),2.52(ddd,J＝50.5,24.8,6.6Hz,2H),2.23-1.41(m,7H),1.32-0.40(m,2H).LRMS(EI)m/z(M + ).
EXAMPLE 94 preparation of 2- (6- (3- (((5-cyclopropyl-3- (2- (trifluoromethoxy) phenyl) isoxazol-4-yl) methoxy) methyl) -3-fluoro-8-azabicyclo [3.2.1] oct-8-yl) -4-fluorobenzo [ d ] thiazole-6-amide) ethanesulfonic acid (FL 94)
The preparation method is the same as example 87, and the target product FL94 is prepared. 1 H NMR(500MHz,CDCl 3 )δ8.37(s,1H),7.64(dd,J＝16.1,13.7Hz,1H),7.52-7.38(m,3H),7.19(d,J＝2.9Hz,1H),6.87(s,1H),6.68(s,1H),6.12(dd,J＝15.0,3.1Hz,1H),4.80(s,2H),3.71(t,J＝25.2Hz,5H),3.59-3.05(m,8H),2.73(p,J＝20.1Hz,1H),2.49-1.84(m,2H),1.48-0.65(m,4H).LRMS(EI)m/z743(M + ).
EXAMPLE 95 preparation of 2- (6- (3- (((5- (2, 2-difluorocyclopropyl) -3- (2- (trifluoromethoxy) phenyl) isoxazol-4-yl)) methyl) -3-fluoroazabicyclo [3.2.1] oct-8-yl) -4-fluorobenzo [ d ] thiazole-6-amide) ethanesulfonic acid (FL 95)
The preparation method is the same as example 87, and the target product FL95 is prepared. 1 H NMR(500MHz,CDCl 3 )δ8.43(dd,J＝14.4,3.7Hz,1H),8.17(d,J＝2.9Hz,1H),7.46(dddd,J＝29.1,18.4,15.6,3.3Hz,3H),7.12(dd,J＝14.2,3.7Hz,1H),6.67(s,1H),6.39(s,1H),4.75-4.47(m,4H),3.89-3.57(m,3H),3.33(td,J＝16.8,1.5Hz,2H),2.82(dd,J＝24.8,16.6Hz,1H),2.52-1.51(m,9H),1.19-0.59(m,2H).LRMS(EI)m/z 779(M + ).
EXAMPLE 96 preparation of 2- (6- (3- (((5-cyclopropyl-3- (2- (trifluoromethoxy) phenyl) isoxazol-4-yl) oxy) methyl) -3-fluoro-8-azabicyclo [3.2.1] oct-8-yl) -4-fluorobenzo [ d ] thiazole-6-amide) ethanesulfonic acid (FL 96)
The preparation method is the same as example 87, and the target product FL96 is prepared. 1 H NMR(500MHz,CDCl 3 )δ8.43(dd,J＝14.4,3.7Hz,1H),8.17(d,J＝2.9Hz,1H),7.45(dddd,J＝33.1,29.1,15.5,3.2Hz,3H),7.12(dd,J＝14.2,3.7Hz,1H),6.65(s,1H),6.50(s,1H),5.00-4.68(m,4H),3.68(td,J＝16.3,1.4Hz,4H),2.94-2.24(m,3H),2.16-1.44(m,6H),1.50-0.74(m,4H).LRMS(EI)m/z 729(M + ).
EXAMPLE preparation of 6- (4- (((3- (2, 6-dichlorophenyl) -5- (2- (trifluoromethyl) cyclopropyl) isoxazol-4-yl) methoxy) methyl) -4-fluoropiperidin-1-yl) -1-methyl-1H-indole-3-carboxylic acid (FL 97)
The preparation method is the same as example 1, and the target product FL97 is prepared. 1 H NMR(500MHz,CDCl 3 )δ9.26(s,1H),7.64(dd,J＝16.1,13.7Hz,1H),7.47(d,J＝0.6Hz,1H),7.44(dd,J＝8.2,6.8Hz,2H),7.21(d,J＝2.9Hz,1H),6.12(dd,J＝15.0,3.1Hz,1H),4.93(s,2H),3.76(s,1H),3.74(s,3H),3.66(s,1H),3.44(dt,J＝24.9,11.3Hz,2H),3.20(dt,J＝24.9,11.3Hz,2H),2.07(ddt,J＝50.4,24.7,11.3Hz,2H),1.91-1.61(m,3H),1.57-1.17(m,2H),0.94(ddd,J＝24.3,21.7,19.5Hz,1H).LRMS(EI)m/z 641(M + ).
EXAMPLE 98 preparation of 2- (4- (((3- (2, 6-dichlorophenyl) -5- (2- (trifluoromethyl) cyclopropyl) isoxazol-4-yl) methoxy) methyl) -4-fluoropiperidin-1-yl) -4-fluorobenzo [ d ] thiazole-6-carboxylic acid (FL 98)
The preparation method is the same as example 13, and the target product FL98 is prepared. 1 H NMR(500MHz,CDCl 3 )δ8.35(d,J＝3.1Hz,1H),7.90(dd,J＝15.9,2.9Hz,1H),7.64(dd,J＝16.1,13.7Hz,1H),7.46(dd,J＝14.9,1.0Hz,2H),4.72(s,2H),4.61(dt,J＝24.9,11.4Hz,2H),4.05(dt,J＝24.7,11.4Hz,2H),3.79(d,J＝50.4Hz,2H),2.04(ddt,J＝50.5,24.9,11.4Hz,2H),1.90-1.32(m,4H),1.18(ddd,J＝24.5,21.2,19.6Hz,1H),0.96(ddd,J＝24.6,21.7,19.9Hz,1H).LRMS(EI)m/z 663(M + ).
EXAMPLE preparation of 99- (3- (((3- (2, 6-dichlorophenyl) -5- (2- (trifluoromethyl) cyclopropyl) isoxazol-4-yl) methoxy) methyl) -4-fluoropiperidin-1-yl) -3-fluoro-8-azabicyclo [3.2.1] oct-8-yl) -4-fluorobenzo [ d ] thiazole-6-carboxylic acid (FL 99)
The preparation method is the same as example 13, and the target product FL98 is prepared. 1 H NMR(500MHz,CDCl 3 )δ8.35(d,J＝3.1Hz,1H),7.95(dd,J＝15.9,2.9Hz,1H),7.64(dd,J＝16.1,13.7Hz,1H),7.46(dd,J＝14.9,1.0Hz,2H),4.91(s,2H),4.72(tt,J＝8.4,6.4Hz,2H),4.21(d,J＝50.4Hz,2H),2.24(ddd,J＝50.4,24.7,6.4Hz,2H),2.09-1.54(m,7H),1.55-1.35(m,1H),1.24(ddd,J＝24.4,21.2,19.7Hz,1H),0.90(ddd,J＝24.5,21.6,19.7Hz,1H).LRMS(EI)m/z 689(M + ).
EXAMPLE 100 preparation of 6- (3-fluoro-3- (((3- (2- (trifluoromethoxy) phenyl) -5- (2- (trifluoromethyl) cyclopropyl) isoxazol-4-yl) methoxy) methyl) -8-azabicyclo [3.2.1] oct-8-yl) -1-methyl-1H-indole-3-carboxylic acid (FL 100)
The preparation method is the same as example 1, and the target product FL100 is prepared. 1 H NMR(500MHz,CDCl 3 )δ9.26(s,1H),8.43(dd,J＝14.4,3.7Hz,1H),7.82-7.25(m,3H),7.12(dd,J＝14.2,3.7Hz,1H),6.64(d,J＝2.9Hz,1H),6.12(dd,J＝15.0,3.1Hz,1H),4.92(s,2H),4.45(s,1H),4.41-4.24(m,3H),3.74(s,3H),2.82(ddd,J＝50.4,24.8,6.7Hz,2H),2.05(dddd,J＝47.4,31.5,17.8,7.0Hz,4H),1.72(td,J＝21.4,17.9Hz,1H),1.59-1.15(m,4H),0.89(ddd,J＝24.4,21.7,19.4Hz,1H).LRMS(EI)m/z 682(M + ).
EXAMPLE 101 preparation of 4-fluoro-2- (3-fluoro-3- (((3- (2- (trifluoromethoxy) phenyl) -5- (2- (trifluoromethoxy) cyclopropyl) isoxazol-4-yl) methoxy) methyl) -8-azabicyclo [3.2.1] oct-8-yl) benzo [ d ] thiazole-6-carboxylic acid (FL 101)
The preparation method is the same as example 13, and the target product FL101 is prepared. 1 H NMR(500MHz,CDCl 3 )δ8.40(dd,J＝14.2,3.7Hz,1H),8.32(d,J＝2.9Hz,1H),7.92(dd,J＝16.0,3.0Hz,1H),7.53-7.31(m,2H),7.09(dd,J＝14.2,3.7Hz,1H),4.75(s,2H),4.61(tt,J＝8.6,6.4Hz,2H),4.29(d,J＝50.4Hz,2H),2.65(ddd,J＝50.4,24.7,6.4Hz,2H),2.37-1.91(m,4H),1.88-1.34(m,4H),1.32-0.80(m,2H)..LRMS(EI)m/z 704M + ).
EXAMPLE 102 preparation of (FL 102) 4- (4-fluoro-4- (((3- (2- (trifluoromethoxy) phenyl) -5- (2- (trifluoromethyl) cyclopropyl) isoxazol-4-yl) methoxy) methyl) piperidin-1-yl) -1-methyl-1H-indole-3-carboxylic acid
The preparation method is the same as that of example 1, and the target product FL102 is prepared. 1 H NMR(500MHz,CDCl 3 )δ9.26(s,1H),8.43(dd,J＝14.4,3.7Hz,1H),7.78-7.36(m,3H),7.24-7.01(m,2H),6.12(dd,J＝15.0,3.1Hz,1H),4.91(s,2H),3.74(t,J＝25.2Hz,5H),3.32(ddt,J＝105.9,24.7,11.3Hz,4H),2.22-1.56(m,5H),1.57-0.83(m,3H).LRMS(EI)m/z 656(M + ).
Pharmacological Activity test examples
Example 1
The screening method comprises the following steps:
number 142
Name FXR-luciferase model agonist screening
The instrument comprises:
microplate detector: enVision (TM) (PerkinElmer)
Materials:
screening plates: viewplate-96 white 96 microwell plate with clear bottom
The detection kit comprises: fluorescent insect fluorescence reporter gene detection kit (Britelite plus type)
The process is as follows:
co-transfecting the FXR expression plasmid and the FXRE luciferase reporter plasmid to 293T cells, culturing the cells in a 96-hole flat-bottom microplate, ensuring the expression of the plasmids, and adding an FXR agonist. The activation degree of the agonist is calculated and quantitatively determined by detecting the fluorescence signal intensity of the luciferase. Wherein during primary screening, 10 μ M of OCA acts on cells, respectively determining the relative activity of the compound to be tested on positive compound, re-screening the compound with relative activity higher than 50% of that of the positive compound, and calculating the dose dependence relationship, i.e. EC 50 The value is obtained.
Sample treatment:
the samples were dissolved in DMSO and stored at low temperature, and the concentration of DMSO in the final system was controlled within a range that did not affect the detection activity.
Data processing and results description:
the activity of the sample is tested under a single concentration condition, e.g., 20. Mu.g/ml, for primary screening. For samples that exhibit activity under certain conditions, e.g., a% inhibition greater than 50, the activity is tested for dose dependence, i.e., IC 50 /EC 50 Values, obtained by nonlinear fitting of sample concentrations by sample activity, were calculated as Graphpad Prism 4, the model used for fitting was sigmoidal-response (variable slope), and for most inhibitor screening models, the bottom and top of the fitted curve were set at 0 and 100. In general, each sample was tested with multiple wells (n.gtoreq.2) and the results were expressed as Standard Deviation (SD) or Standard Error (SE). Generally, each test has a reported compound as a reference.
Figure BDA0001653521430000591
Through activity tests, a plurality of compounds of the invention have excellent pharmacological activity. EC of multiple compounds such as FL3 and FL10 at cellular level 50 Less than 10nM, better than the positive compounds OCA (obeticholic acid) and FL4-13, and stronger pharmacological activity.
FL4-13：
Figure BDA0001653521430000601
Example 2 Thioacetamide (TAA) -induced hepatic fibrosis model experiment in rats
To further evaluate the efficacy of the compounds, we performed TAA-induced rat liver fibrosis model experiments.
The molding method comprises the following steps: SD male rats of 6-8 weeks old are intraperitoneally injected with 150mg/kg TAA physiological saline solution three times a week after 4 consecutive injections, and administration of compounds FL4-14 (FL 10), FL4-15 (FL 11) (20 mg/kg P.O qd) and OCA and FL4-13 (LY-2562175) at the same administration dose as a positive control is started, and TAA molding is continued during administration for four weeks. The model can observe the increase of liver coefficient, the increase of serum alkaline phosphatase (ALP) level, the up-regulation of alpha-smooth muscle actin (alpha-SMA) and type I collagen (Col 1 alpha 1 mRNA) expression in the liver and the increase of the content of liver collagen in pathological sections (Tianlang scarlet staining).
The experimental results show that: FL4-14 (FL 10) in the compound can obviously reduce ALP level in serum (figure 1) and down-regulate Col1 alpha 1mRNA expression (figure 2). The compound FL4-14 (FL 10) reduced the collagen content in the pathological section of liver, showed an anti-hepatic fibrosis effect equivalent to that of positive OCA, but better than the control compound FL4-13 (LY-2562175) (FIG. 3). And FL4-14 has no obvious influence on the contents of cholesterol (TC), high Density Lipoprotein (HDL) and Low Density Lipoprotein (LDL) in the serum of a rat, while the positive drug OCA can obviously increase the contents of TC, HDL and LDL in the serum of the rat, which suggests that FL4-14 (FL 10) can possibly avoid the side effect caused by LDL rise caused by clinical OCA, and is also superior to the positive control compound FL4-13 (figure 4). In addition, the liver coefficients of the FL4-14 (FL 10) group have no obvious change, and the effect of the positive drug OCA on the significant increase of the liver coefficients is avoided (figure 5).
Example 3 pharmacokinetic testing
Preliminary pharmacokinetic tests were performed on select compound FL36 and control compound FL 4-13.
1. Material
1.1 animals and groups
Male SD rats, 12.
1.2 drugs and reagents
FL4-13; FL36; DMSO (dimethyl sulfoxide); etOH; PEG300 (polyethylene glycol); naCl; HPMC (hydroxypropyl methyl cellulose)
2. Method of producing a composite material
2.1 methods of administration
Figure BDA0001653521430000611
2.2 rat pharmacokinetic parameters are given in the table below
Figure BDA0001653521430000612
The analysis data shows that: the exposure (AUC) of the compound FL36 in plasma is much higher than that of the positive control compound FL4-13 under the same administration dose, and the compound shows good pharmacokinetic property.
Example 4 preliminary test for acute toxicity in mice
1. Material
1.1 animals and groups
Female C57 mice, 6-8 weeks old, 6 mice, SPF grade.
1.2 drugs and reagents
FL4-14(FL10)(MW:572.45)
DMSO，Tween80，MC
2. Method of producing a composite material
2.1 Experimental procedures
Taking 6 female ICR mice, after fasting without water inhibition for 12 hours, orally gavage the test substance FL4-14 mg/kg with an administration volume of 10mL/kg, the solvent 10% DMSO +10% Tween80+80% MC 0.5% by weight. Mice were observed for a toxic response immediately after dosing and for 7 consecutive days.
2.2 animal body weights and observations after administration are as follows
Figure BDA0001653521430000621
And (4) conclusion: the primary acute toxicity test is mainly to observe whether the compound has obvious toxic reaction or death at the administration dose of 500mg/kg, and the test result shows that: the compounds of the present application have good safety.
All documents referred to herein are incorporated by reference into this application as if each were individually incorporated by reference. Furthermore, it should be understood that various changes or modifications of the present invention can be made by those skilled in the art after reading the above teachings of the present invention, and these equivalents also fall within the scope of the appended claims of the present application.

Claims (13)

1. A fluoroisoxazole compound having a structure represented by the following formula I, or a pharmaceutically acceptable salt thereof, or a mixture thereof:
Figure FDA0003994654340000011
wherein:
m is 1;
n is 0 or 1;
p is 0 or 1;
Figure FDA0003994654340000012
selected from the group consisting of:
Figure FDA0003994654340000013
wherein the content of the first and second substances,
R 8 selected from the group consisting of: hydrogen, halogen, C 1 -C 6 Alkyl, halo C 1 -C 6 Alkyl radical, C 1 -C 6 Alkoxy radical, C 1 -C 6 Alkoxycarbonyl, halogeno C 1 -C 6 Alkoxy radical, C 2 -C 6 Alkenyl radical, C 2 -C 6 Alkynyl, C 3 -C 8 Cycloalkyl, cyano, nitro, amino, hydroxy, hydroxymethyl, carboxy, mercapto, C 6 -C 10 Aryl and 3-12 membered heterocyclyl;
t is 0, 1 or 2, 3, 4 or 5;
X l and X 2 Each independently is N, CH or CR 8 ；
X 3 Is O or S;
R 9 is-X 4 -CO 2 R 5 、C 1 -C 6 Alkyl, halo C 1 -C 6 Alkyl, -X 4 -CONR 5 R 6 、-X 4 -CONR 5 (CH 2 ) y1 CO 2 R 6 、-X 4 -CONR 5 (CH 2 ) y2 SO 3 R 6 (ii) a y1 and y2 are each independently 1,2, 3 or 4; x 4 Is a bond, C1-C6 alkylene, C 1-2 Alkylene, cyclopropyl or epoxyethyl;
R 5 and R 6 Each independently is hydrogen, C 1-6 Alkyl, halo C 1 -C 6 Alkyl radical, C 2 -C 6 Alkenyl, hydroxymethyl, or 5-7 membered heterocyclyl;
R 10 is hydrogen, C 1 -C 6 Hydroxyalkyl radical, C 1 -C 6 Alkyl, halo C 1 -C 6 Alkyl radical, C 1 -C 6 Alkoxy radical, C 1 -C 6 Alkoxycarbonyl, halogeno C 1 -C 6 Alkoxy radical, C 2 -C 6 Alkenyl radical, C 2 -C 6 Alkynyl or C 3 -C 8 A cycloalkyl group;
R 1 selected from the group consisting of: a substituted or unsubstituted 6-20 membered aromatic ring group, a substituted or unsubstituted 6-20 membered aromatic heterocyclic group; wherein, the substitution refers to that the hydrogen atoms on the group are substituted by 1,2, 3 or 4 substituents selected from the following group: deuterium, tritium, halogen, C 1 -C 6 Alkyl radical, C 1 -C 6 Alkoxy, halo C 1 -C 6 Alkoxy, -O [ (CH) 2 ) q O] r R 5 、C 3 -C 8 Cycloalkoxy, halo C 3 -C 8 Cycloalkoxy, cyano, nitro, hydroxy, hydroxymethyl, carboxy, and C 6 An aryl group; wherein q and r are each 1,2, 3 or 4; wherein the 6-20 membered aromatic ring group is phenyl, and the 6-20 membered aromatic heterocyclic group is pyridyl; r 1 In, R 5 Is hydrogen, C 1-6 Alkyl, or halo C 1 -C 6 An alkyl group;
R 2 is 1 or 2 substituents independently selected from the group consisting of: hydrogen, halogen, substituted or unsubstituted C 1 -C 6 An alkyl group; wherein, the substitution refers to that the hydrogen atoms on the group are substituted by 1,2, 3 or 4 substituents selected from the following group: halogen;
R 3 and R 4 Each independently selected from the group consisting of: hydrogen, C 1 -C 6 An alkyl group; or R 3 And R 4 Are connected together to form C 1 -C 6 An alkylene group.
2. The fluoroisoxazole compound according to claim 1, or a pharmaceutically acceptable salt thereof, or a mixture thereof, wherein R is 8 Selected from the group consisting of: hydrogen, halogen, C 1 -C 6 An alkyl group;
t is 0, 1 or 2;
X l and X 2 Each independently is N, CH or CR 8 ；
X 3 Is O or S;
R 9 is-X 4 -CO 2 R 5 Halogen substituted C 1 -C 6 Alkyl, -X 4 -CONR 5 (CH 2 ) y1 CO 2 R 6 、-X 4 -CONR 5 (CH 2 ) y2 SO 3 R 6 (ii) a y1 and y2 are each independently 1 or 2; x 4 Is a bond;
R 5 and R 6 Each independently of the other is hydrogen, C 1-6 An alkyl group;
R 10 is hydrogen, C 1 -C 6 Hydroxyalkyl, C 1 -C 6 An alkyl group.
3. The fluoroisoxazole compound according to claim 1 or 2, or a pharmaceutically acceptable salt thereof, or a mixture thereof,
Figure FDA0003994654340000021
selected from the group consisting of:
Figure FDA0003994654340000022
wherein R is 8 、R 9 、R 10 And t is as defined in claim 1 or 2.
4. A fluorine-containing isoxazole compound with a structure shown as the following general formula I, or a pharmaceutically acceptable salt thereof, or a mixture thereof,
Figure FDA0003994654340000031
wherein R is 1 Selected from the group consisting of: phenyl, pyridyl; the above groups are substituted or unsubstituted; wherein, the substituted refers to that the hydrogen atom on the group is substituted by 1,2, 3 or 4 substituents selected from the following group: hydrogen, halogen, nitro, cyano, trifluoromethyl, trifluoroethyl, trifluoropropyl, trifluoromethoxy, trifluoroethoxy, phenyl, methoxycarbonyl, ethoxycarbonyl, propoxycarbonyl, C 1-6 Alkyl radical, C l-6 Cycloalkyl radical, C 1-6 Alkoxy radical, C l-6 A cycloalkoxy group;
m、n、p、
Figure FDA0003994654340000032
R 2 、R 3 and R 4 As defined in claim 1.
5. The fluoroisoxazole compound according to claim 4, or a pharmaceutically acceptable salt thereof, or a mixture thereof, wherein C is 3-6 Cycloalkyl is cyclopropyl.
6. A fluoroisoxazole compound, or a pharmaceutically acceptable salt thereof, or a mixture thereof, selected from the group consisting of:
Figure FDA0003994654340000033
Figure FDA0003994654340000041
Figure FDA0003994654340000051
Figure FDA0003994654340000061
7. the fluoroisoxazole compound of claim 6, or a pharmaceutically acceptable salt thereof, or a mixture thereof, wherein the compound is selected from the group consisting of: FL3, FL10, FL11, FL12, FL16, FL30, FL36, and FL37.
8. A pharmaceutical composition comprising a therapeutically effective amount of a fluoroisoxazole compound according to any one of claims 1 to 7, or a pharmaceutically acceptable salt thereof, or a mixture thereof, and a pharmaceutically acceptable carrier.
9. An FXR agonist characterized in that it comprises a fluoroisoxazole compound according to any one of claims 1 to 7, or a pharmaceutically acceptable salt thereof, or a mixture thereof.
10. Use of fluoroisoxazoles according to claims 1 to 7, or pharmaceutically acceptable salts thereof, or mixtures thereof, for the preparation of a medicament for the prophylaxis or treatment of FXR mediated diseases.
11. The use according to claim 10, wherein the disease is selected from the group consisting of: non-alcoholic steatohepatitis, fatty liver, hepatic fibrosis, primary biliary cirrhosis, and hyperlipidemia.
12. The use according to claim 10, wherein the disease is non-alcoholic fatty liver disease.
13. The process for producing a fluorine-containing isoxazole compound according to claim 1,
(a) The method comprises the following steps:
Figure FDA0003994654340000071
wherein the content of the first and second substances,
Figure FDA0003994654340000072
R 1 、R 2 、R 3 、R 4 n, m and p are as defined in formula I in claim 1; x is halogen;
step j: reacting intermediate 12 and intermediate 9 in an organic solvent in the presence of 18-crown-6 and a base to form intermediate 13;
step k: deprotecting intermediate 13 in an organic solvent to form intermediate 14;
step l: under the protection of nitrogen, in an organic solvent, in the presence of alkali, a palladium catalyst and a ligand, reacting an intermediate 14 with an intermediate 15 to form a final product I; or
Step m: intermediate 14 and intermediate 15 are reacted in an organic solvent in the presence of a base under nitrogen to form the final product I.
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Citations (3)

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Publication number Priority date Publication date Assignee Title
CN104513213A (en) * 2013-09-28 2015-04-15 山东亨利医药科技有限责任公司 Fxr agonist
WO2017118294A1 (en) * 2016-01-06 2017-07-13 广州市恒诺康医药科技有限公司 Fxr receptor modulator, preparation method therefor, and uses thereof
WO2017128896A1 (en) * 2016-01-26 2017-08-03 江苏豪森药业集团有限公司 Fxr agonist and preparation method and use thereof

Family Cites Families (4)

* Cited by examiner, † Cited by third party
Publication number Priority date Publication date Assignee Title
TW200906823A (en) * 2007-07-16 2009-02-16 Lilly Co Eli Compounds and methods for modulating FXR
CA2986045A1 (en) * 2015-05-22 2016-12-01 Genentech, Inc. Substituted benzamides and methods of use thereof
WO2018039384A1 (en) * 2016-08-23 2018-03-01 Ardelyx, Inc. Isoxazolyl-carbonyloxy azabicyclo[3.2.1]octanyl compounds as fxr activators
WO2018059314A1 (en) * 2016-09-28 2018-04-05 四川科伦博泰生物医药股份有限公司 Azabicycle derivatives and preparation method and use thereof

Patent Citations (3)

* Cited by examiner, † Cited by third party
Publication number Priority date Publication date Assignee Title
CN104513213A (en) * 2013-09-28 2015-04-15 山东亨利医药科技有限责任公司 Fxr agonist
WO2017118294A1 (en) * 2016-01-06 2017-07-13 广州市恒诺康医药科技有限公司 Fxr receptor modulator, preparation method therefor, and uses thereof
WO2017128896A1 (en) * 2016-01-26 2017-08-03 江苏豪森药业集团有限公司 Fxr agonist and preparation method and use thereof

Non-Patent Citations (2)

* Cited by examiner, † Cited by third party
Title
Discovery and Optimization of Non-bile Acid FXR Agonists as Preclinical Candidates for the Treatment of Nonalcoholic Steatohepatitis;Junyou Li等;《J. Med. Chem.》;20200929;第63卷;第12748-12772页 *
法尼醇X受体激动剂的研究进展;年四昀等;《中国药物化学杂志》;20170228;第27卷(第1期);第57-66页 *

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