CN108602811B - FXR receptor agonists - Google Patents

FXR receptor agonists Download PDF

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CN108602811B
CN108602811B CN201780007789.5A CN201780007789A CN108602811B CN 108602811 B CN108602811 B CN 108602811B CN 201780007789 A CN201780007789 A CN 201780007789A CN 108602811 B CN108602811 B CN 108602811B
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CN108602811A (en
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吴永谦
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Xuanzhu Biopharmaceutical Co Ltd
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    • A—HUMAN NECESSITIES
    • A61—MEDICAL OR VETERINARY SCIENCE; HYGIENE
    • A61K—PREPARATIONS FOR MEDICAL, DENTAL OR TOILETRY PURPOSES
    • A61K31/00—Medicinal preparations containing organic active ingredients
    • A61K31/33—Heterocyclic compounds
    • A61K31/395—Heterocyclic compounds having nitrogen as a ring hetero atom, e.g. guanethidine or rifamycins
    • A61K31/41—Heterocyclic compounds having nitrogen as a ring hetero atom, e.g. guanethidine or rifamycins having five-membered rings with two or more ring hetero atoms, at least one of which being nitrogen, e.g. tetrazole
    • A61K31/42—Oxazoles
    • A—HUMAN NECESSITIES
    • A61—MEDICAL OR VETERINARY SCIENCE; HYGIENE
    • A61K—PREPARATIONS FOR MEDICAL, DENTAL OR TOILETRY PURPOSES
    • A61K31/00—Medicinal preparations containing organic active ingredients
    • A61K31/33—Heterocyclic compounds
    • A61K31/395—Heterocyclic compounds having nitrogen as a ring hetero atom, e.g. guanethidine or rifamycins
    • A61K31/41—Heterocyclic compounds having nitrogen as a ring hetero atom, e.g. guanethidine or rifamycins having five-membered rings with two or more ring hetero atoms, at least one of which being nitrogen, e.g. tetrazole
    • A61K31/42—Oxazoles
    • A61K31/422—Oxazoles not condensed and containing further heterocyclic rings
    • A—HUMAN NECESSITIES
    • A61—MEDICAL OR VETERINARY SCIENCE; HYGIENE
    • A61K—PREPARATIONS FOR MEDICAL, DENTAL OR TOILETRY PURPOSES
    • A61K31/00—Medicinal preparations containing organic active ingredients
    • A61K31/33—Heterocyclic compounds
    • A61K31/395—Heterocyclic compounds having nitrogen as a ring hetero atom, e.g. guanethidine or rifamycins
    • A61K31/435—Heterocyclic compounds having nitrogen as a ring hetero atom, e.g. guanethidine or rifamycins having six-membered rings with one nitrogen as the only ring hetero atom
    • A61K31/47—Quinolines; Isoquinolines
    • A61K31/472—Non-condensed isoquinolines, e.g. papaverine
    • A61K31/4725—Non-condensed isoquinolines, e.g. papaverine containing further heterocyclic rings
    • C—CHEMISTRY; METALLURGY
    • C07—ORGANIC CHEMISTRY
    • C07D—HETEROCYCLIC COMPOUNDS
    • C07D261/00—Heterocyclic compounds containing 1,2-oxazole or hydrogenated 1,2-oxazole rings
    • C07D261/02—Heterocyclic compounds containing 1,2-oxazole or hydrogenated 1,2-oxazole rings not condensed with other rings
    • C07D261/06—Heterocyclic compounds containing 1,2-oxazole or hydrogenated 1,2-oxazole rings not condensed with other rings having two or more double bonds between ring members or between ring members and non-ring members
    • C07D261/08—Heterocyclic compounds containing 1,2-oxazole or hydrogenated 1,2-oxazole rings not condensed with other rings having two or more double bonds between ring members or between ring members and non-ring members with only hydrogen atoms, hydrocarbon or substituted hydrocarbon radicals, directly attached to ring carbon atoms
    • C—CHEMISTRY; METALLURGY
    • C07—ORGANIC CHEMISTRY
    • C07D—HETEROCYCLIC COMPOUNDS
    • C07D413/00—Heterocyclic compounds containing two or more hetero rings, at least one ring having nitrogen and oxygen atoms as the only ring hetero atoms
    • C07D413/02—Heterocyclic compounds containing two or more hetero rings, at least one ring having nitrogen and oxygen atoms as the only ring hetero atoms containing two hetero rings
    • C07D413/12—Heterocyclic compounds containing two or more hetero rings, at least one ring having nitrogen and oxygen atoms as the only ring hetero atoms containing two hetero rings linked by a chain containing hetero atoms as chain links

Abstract

The invention provides a compound shown in the following general formula (1), pharmaceutically acceptable salt, ester or stereoisomer thereof, a preparation method of the compound and application of the compound in preparing medicines for preventing and/or treating non-alcoholic fatty liver, primary biliary cirrhosis, lipid metabolism disorder, diabetic complication and malignant tumor.
Figure DDA0001739294160000011
Wherein R is1、R2、R3、R4M, n, W, A, Z, E, F, X, Y are as defined in the specification.

Description

FXR receptor agonists
Technical Field
The invention relates to FXR receptor agonists, pharmaceutically acceptable salts, esters and stereoisomers thereof, pharmaceutical preparations containing the compounds, and applications of the compounds, the pharmaceutically acceptable salts, esters and stereoisomers thereof in preparing medicaments for treating and/or preventing related diseases such as non-alcoholic fatty liver, primary biliary cirrhosis, lipid metabolism disorder, diabetic complications, malignant tumors and the like mediated by FXR receptors.
Background
FXR receptors (farnesoid X receptors) belong to members of the nuclear receptor family of ligand-activated transcription factors and have the typical nuclear receptor structure, i.e., an amino-terminal highly conserved DNA Binding Domain (DBD), a carboxy-terminal Ligand Binding Domain (LBD), an amino-terminal ligand-independent transcriptional activation domain (AF1), a carboxy-terminal ligand-dependent transcriptional activation domain (AF2), and a foot chain domain. FXR forms heterodimers with Retinoid X Receptor (RXR), and when ligand binds to the LBD region of FXR, conformation of FXR is changed, and a binding domain of DNA binds to FXR-responsive element (IR-1) of a target gene promoter, releasing a co-repressor (e.g., NCOR), recruiting a co-activator, and thereby exerting a transcription regulatory effect.
FXR is expressed in various organ tissues including adipose tissues, liver, gastrointestinal tract, kidney and the like, wherein the expression level in the liver is most abundant. The FXR signaling pathway can directly or indirectly regulate the expression of a plurality of downstream genes, such as BSEP, SHP, CYP7A1, FGFR4, OST alpha/beta, SREBP-1C and other genes, and further regulate a plurality of metabolic pathways, such as: triglyceride, cholesterol, blood sugar and energy stability metabolism cholic acid, and can be used for treating cancer, non-alcoholic fatty liver, metabolic disorder, and inflammation. The regulation of cholic acid metabolism by inhibiting the synthesis, binding and transport of cholic acid is the main regulator of cholic acid balance in vivo.
Some natural cholic acid compounds can stimulate FXR receptors, such as chenodeoxycholic acid (CDCA), deoxycholic acid (DCA), lithocholic acid (LCA) and taurine and glycine conjugates of these cholic acids. Except natural compounds, FXR agonists developed internationally at present can be mainly divided into two major classes, one class is steroids, represented by Obeticholic acid (OCA) of Intercept company, and the FXR agonists are in clinical stage III aiming at nonalcoholic fatty liver indications; the other is a new molecular entity, an early developed compound such as GW4604(WO2000/037077), which although having strong agonistic activity is photolabile and less bioavailable, and in addition PX-104(WO2011020615a1) developed by pherex corporation, assigned to Gilead corporation, is currently in phase II clinical studies.
Figure BDA0001739294150000021
However, it is still desired to develop a novel FXR receptor agonist having high efficacy, low toxicity, and good stability.
Disclosure of Invention
The present invention addresses the problem of providing a compound having a novel molecular structure which is capable of effectively agonizing the FXR receptor, increasing the expression levels of BSEP and SHP genes, and effectively inhibiting the expression of the CYP7A1 gene. In addition, in order to achieve a better therapeutic effect and to better satisfy market demand, it is also desired to provide an FXR receptor agonist that is highly effective, low-toxic, and has good stability.
Specifically, the present invention aims to provide an FXR receptor agonist with a novel structure, which has a good drug effect and provides a possibility for the FXR receptor agonist to be used for treating nonalcoholic fatty liver, primary biliary cirrhosis, lipid metabolism disorder, diabetic complications and malignant tumors.
It is another object of the present invention to provide a method for producing the above FXR receptor agonist.
It is another object of the present invention to provide pharmaceutical compositions and formulations containing the above FXR receptor agonists.
Still another object of the present invention is to provide the use of the above FXR receptor agonist for the preparation of a medicament for the prevention and/or treatment of non-alcoholic fatty liver disease, primary biliary cirrhosis, lipid metabolism disorders, diabetic complications, and malignant tumors.
The present inventors have made intensive studies to achieve the above object and as a result, have found that a compound represented by the following general formula (I), a pharmaceutically acceptable salt thereof, an ester thereof and a stereoisomer thereof can effectively stimulate the FXR receptor, thereby completing the present invention.
Specifically, the invention relates to the following technical scheme:
a compound of scheme 1, general formula (i), pharmaceutically acceptable salts thereof, esters thereof, or stereoisomers thereof:
Figure BDA0001739294150000031
wherein the content of the first and second substances,
R1、R2each independently selected from hydrogen atom, cyano group, halogen atom, nitro group, amino group, hydroxyl group, carboxyl group, C1-6Alkyl radical, C1-6Alkoxy radical, C1-6Alkylamino, di-C1-6 alkylamino, C1-6Alkylthio radical, C1-6Alkylcarbonyl, halo C1-6Alkyl, halo C1-6Alkoxy radical, C1-6Alkoxy radical C1-6Alkyl radical, C1-6Alkylcarbonyloxy, C1-6Alkylsulfonyl radical, C1-6Alkylaminosulfonyl, di-C1-6Alkylaminosulfonyl radical, C1-6Alkylsulfonylamino group, C1-6Alkylsulfonyloxy, C2-8Alkenyl or C2-8An alkynyl group;
R3selected from hydrogen atoms, cyano groups, halogen atoms, nitro groups, amino groups, hydroxyl groups, carboxyl groups, or C optionally substituted by one or more substituents P1-6Alkyl, halo C1-6Alkyl, hydroxy C1-6Alkyl, carboxyl C1-6Alkyl radical, C1-6Alkoxy, halo C1-6Alkoxy, hydroxy C1-6Alkoxy, carboxyl C1-6Alkoxy radical, C1-6Alkylsulfonyl radical, C1-6Alkylaminosulfonyl, di-C1-6Alkylaminosulfonyl radical, C1-6Alkylsulfonylamino group, C1-6Alkylsulfonyloxy, 3-8 membered cycloalkyl C1-6Alkyl, 3-8 membered heterocyclic group C1-6An alkyl group;
p is selected from hydroxyl, amino, carboxyl, cyano, nitro, halogen atom, C1-6Alkyl radical, C1-6Alkoxy radical, C1-6Alkylamino, di-C1-6Alkylamino, halogeno C1-6Alkyl, halo C1-6Alkoxy radical, C1-6Alkoxy radical C1-6Alkyl radical, C1-6Alkylcarbonyl group, C1-6Alkylcarbonyloxy, C1-6Alkylsulfonyl radical, C2-8Alkenyl or C2-8An alkynyl group;
R4selected from hydrogen atom, halogen atom, cyano group, nitro group, amino group, hydroxyl group, carboxyl group, C1-6Alkoxy radical, C1-6Alkyl, hydroxy C1-6Alkyl, halo C1-6Alkyl, carboxyl C1-6Alkyl, carboxyl oxy C1-6Alkyl, carboxy amino C1-6Alkyl, amino C1-6Alkyl, aminocarbonyl C1-6Alkyl, hydroxy C1-6Alkoxy, halo C1-6Alkoxy, carboxyl C1-6Alkoxy radical, C2-8Alkenyl radical, C2-8Alkynyl, C1-6Alkylamino radical, C1-6Alkylcarbonyl group, C1-6Alkylcarbonylamino, C1-6Alkylsulfonyl radical, C1-6Alkylsulfonylaminocarbonyl group, C1-6Alkylaminosulfonyl, di-C1-6Alkylamino, 5-8 membered heteroaryl or 3-8 membered heterocyclyl;
w is selected from CH2、NH、O、S、SO、SO2Or CO;
a is selected from NH, O or S;
z is selected from aryl, 5-8 membered heteroaryl, 3-8 membered cycloalkyl or 3-8 membered heterocyclyl, substituted or unsubstituted with one or more substituents Q;
q is selected from cyano, amino, hydroxyl, carboxyl, nitro, halogen atom, C1-6Alkyl radical, C1-6Alkoxy radical, C1-6Alkylamino, di-C1-6Alkylamino, halogeno C1-6Alkyl, halo C1-6Alkoxy radical, C1-6Alkoxy radical C1-6Alkyl radical, C2-8Alkenyl or C2-8An alkynyl group;
e is selected from CH2、NH、O、S、SO、SO2Or CO;
f is selected from absent, CH2、NH、O、S、SO、SO2Or CO;
x is selected from CH or N;
y is selected from CH2、NH、O、S、SO、SO2Or CO;
E. x, Y, F are each independently selected from single bonds;
m is an integer from 0 to 3;
n is an integer from 0 to 4.
A compound of scheme 2, scheme 1, a pharmaceutically acceptable salt thereof, an ester thereof, or a stereoisomer thereof:
wherein the content of the first and second substances,
R1、R2each independently selected from hydrogen atom, cyano group, halogen atom, nitro group, amino group, hydroxyl group, carboxyl group, C1-4Alkyl radical, C1-4Alkoxy radical, C1-4Alkylamino, di-C1-4Alkylamino radical, C1-4Alkylthio radical, C1-4Alkylcarbonyl, halo C1-4Alkyl, halo C1-4Alkoxy radical, C1-4Alkoxy radical C1-4Alkyl radical, C1-4Alkylcarbonyloxy, C1-4Alkylsulfonyl radical, C1-4Alkylaminosulfonyl, di-C1-4Alkylaminosulfonyl radical, C2-6Alkenyl or C2-6An alkynyl group;
R3selected from hydrogen atoms, cyano groups, halogen atoms, nitro groups, amino groups, hydroxyl groups, carboxyl groups, or C optionally substituted by one or more substituents P1-4Alkyl, halo C1-4Alkyl radical, C1-4Alkoxy, halo C1-4Alkoxy, 3-6 membered cycloalkyl C1-4Alkyl, 3-6 membered heterocyclic group C1-4An alkyl group;
p is selected from hydroxyl, amino, carboxyl, cyano, nitro, halogen atom, C1-4Alkyl radical, C1-4Alkoxy radical, C1-4Alkylamino, di-C1-4Alkylamino, halogeno C1-4Alkyl, halo C1-4Alkoxy radical, C1-4Alkoxy radical C1-4Alkyl radical, C1-4Alkylcarbonyl group, C1-4Alkylcarbonyloxy, C1-4Alkylsulfonyl radical, C2-6Alkenyl or C2-6An alkynyl group;
R4selected from hydrogen atom, halogen atom, cyano group, nitro group, amino group, hydroxyl group, carboxyl group, C1-4Alkoxy radical, C1-4Alkyl, hydroxy C1-4Alkyl, halo C1-4Alkyl, carboxyl C1-4Alkyl, carboxyl oxy C1-4Alkyl, carboxy amino C1-4Alkyl, amino C1-4Alkyl, aminocarbonyl C1-4Alkyl, hydroxy C1-4Alkoxy, halo C1-4Alkoxy, carboxyl C1-4Alkoxy radical, C2-6Alkenyl radical, C2-6Alkynyl, C1-4Alkylamino radical, C1-4Alkylcarbonyl group, C1-4Alkylcarbonylamino, C1-4Alkylsulfonyl radical, C1-4Alkylsulfonylaminocarbonyl group, C1-4Alkylaminosulfonyl, di-C1-4Alkylamino, 5-6 membered heteroaryl or 4-7 membered heterocyclyl;
w is selected from CH2、NH、O、S、SO、SO2Or CO;
a is selected from NH, O or S;
z is selected from aryl, 5-6 membered heteroaryl, 3-6 membered cycloalkyl or 4-7 membered heterocyclyl, substituted or unsubstituted with one or more substituents Q;
q is selected from cyano, amino, hydroxyl, carboxyl, nitro, halogen atom, C1-4Alkyl radical, C1-4Alkoxy radical, C1-4Alkylamino, di-C1-4Alkylamino, halogeno C1-4Alkyl, halo C1-4Alkoxy radical, C1-4Alkoxy radical C1-4Alkyl radical, C2-6Alkenyl or C2-6An alkynyl group;
e is selected from CH2NH, O, S or CO;
f is selected from absent, CH2NH, O or S;
x is selected from CH or N;
y is selected from CH2NH, O orS；
E. X, Y, F are each independently selected from single bonds;
m is an integer of 0 to 2;
n is an integer from 0 to 3.
A compound of scheme 3, scheme 2, a pharmaceutically acceptable salt thereof, an ester thereof, or a stereoisomer thereof:
wherein the content of the first and second substances,
R1、R2each independently selected from hydrogen atom, cyano group, halogen atom, nitro group, amino group, hydroxyl group, carboxyl group, C1-4Alkyl radical, C1-4Alkoxy radical, C1-4Alkylamino, di-C1-4Alkylamino radical, C1-4Alkylthio radical, C1-4Alkylcarbonyl, halo C1-4Alkyl, halo C1-4Alkoxy or C1-4Alkoxy radical C1-4An alkyl group;
R3selected from cyano, C optionally substituted by one or more substituents P1-4Alkyl, halo C1-4Alkyl, 3-6 membered cycloalkyl C1-4Alkyl, 3-6 membered heterocyclyl or 3-6 membered heterocyclyl C1-4An alkyl group;
p is selected from hydroxyl, amino, carboxyl, cyano, nitro, halogen atom, C1-4Alkyl radical, C1-4Alkoxy radical, C1-4Alkylamino, di-C1-4Alkylamino, halogeno C1-4Alkyl or halo C1-4An alkoxy group;
R4selected from hydrogen atom, halogen atom, cyano group, nitro group, amino group, hydroxyl group, carboxyl group, C1-4Alkoxy radical, C1-4Alkyl, hydroxy C1-4Alkyl, halo C1-4Alkyl, carboxyl C1-4Alkyl, carboxyl oxy C1-4Alkyl, carboxy amino C1-4Alkyl, amino C1-4Alkyl, aminocarbonyl C1-4Alkyl, hydroxy C1-4Alkoxy, halo C1-4Alkoxy, carboxyl C1-4Alkoxy radical, C2-6Alkenyl radical, C2-6Alkynyl, C1-4Alkylamino radical, C1-4Alkylcarbonyl group, C1-4An alkylcarbonylamino group,C1-4Alkylsulfonyl radical, C1-4Alkylsulfonylaminocarbonyl group, C1-4Alkylaminosulfonyl, di-C1-4Alkylamino, 5-6 membered heteroaryl or 5-6 membered heterocyclyl;
w is selected from CH2、NH、O、S、SO、SO2Or CO;
a is selected from NH, O or S;
z is selected from phenyl, optionally substituted with one or more substituents Q, 5-6 membered heteroaryl containing 1-2N, O and/or S atoms, 5-6 membered cycloalkyl, or 5-6 membered heterocyclyl containing 1-2N, O and/or S atoms;
q is selected from cyano, amino, hydroxyl, carboxyl, nitro, halogen atom, C1-4Alkyl radical, C1-4Alkoxy radical, C1-4Alkylamino, di-C1-4Alkylamino, halogeno C1-4Alkyl or halo C1-4An alkoxy group;
e is selected from CH2NH, O or CO;
f is selected from absent, CH2NH or O;
x is selected from CH or N;
y is selected from CH2NH or O;
E. x, Y, F are each independently selected from single bonds;
m is an integer of 0 to 2;
n is an integer from 0 to 3.
A compound of scheme 4, scheme 3, a pharmaceutically acceptable salt thereof, an ester thereof, or a stereoisomer thereof:
wherein the content of the first and second substances,
R3selected from cyano, C optionally substituted by one or more substituents P1-4Alkyl, halo C1-4Alkyl or C3-6Cycloalkyl radical C1-4An alkyl group;
p is selected from hydroxyl, amino, carboxyl, cyano, nitro, halogen atom, C1-4Alkyl radical, C1-4Alkoxy radical, C1-4Alkylamino, di-C1-4Alkylamino, halogeno C1-4Alkyl or halo C1-4An alkoxy group;
w is selected from NH, O or S;
a is selected from NH, O or S;
z is selected from phenyl, optionally substituted with one or more substituents Q, or a 5-6 membered heteroaryl group containing 1-2N, O and/or S atoms;
q is selected from cyano, amino, hydroxyl, carboxyl, nitro, halogen atom, C1-4Alkyl radical, C1-4Alkoxy radical, C1-4Alkylamino, di-C1-4Alkylamino, halogeno C1-4Alkyl or halo C1-4An alkoxy group;
E. x, Y, F together with the benzene ring form the following structure:
chromanyl, benzo 1, 3-dioxolyl, benzo 1, 4-dioxanyl, benzo 1, 3-dioxanyl, benzotetrahydropyridinyl, chromazinyl, benzotetrahydropyrazinyl, 1,2,3, 4-tetrahydroquinazolinyl, 1,2,3, 4-tetrahydrocinnolinyl, indanyl, tetrahydronaphthyl, tetralone;
n is an integer from 0 to 3.
A compound of scheme 5, scheme 4, a pharmaceutically acceptable salt thereof, an ester thereof, or a stereoisomer thereof:
R1、R2each independently selected from a hydrogen atom, a cyano group, a fluorine atom, a chlorine atom, a bromine atom, a nitro group, an amino group, a hydroxyl group, a carboxyl group, a methyl group, an ethyl group, a propyl group, a butyl group, a methoxy group, a methylamino group, an acetyl group, a trifluoromethyl group, a trifluoroethyl group or a trifluoromethoxy group;
R3selected from the group consisting of methyl, ethyl, n-propyl, isopropyl, n-butyl, isobutyl, tert-butyl, trifluoromethyl, trifluoroethyl, trifluoropropyl, cyano, cyclopropyl, cyclobutyl, cyclopentyl, cyclohexyl, cyclopropylmethyl, cyclobutylmethyl, cyclopentylmethyl, cyclohexylmethyl;
R4selected from the group consisting of a hydrogen atom, a halogen atom, a cyano group, a nitro group, an amino group, a hydroxyl group, a carboxyl group, a methyl group, an ethyl group, a propyl group, a hydroxymethyl group, a hydroxyethyl group, a methoxy group, an ethoxy group, a trifluoromethyl group, a trifluoromethoxy group, a hydroxyl group, a carboxyl group, a methyl group, an ethyl group, a propyl group, a hydroxymethyl group, a hydroxyethyl group, a methoxy group, an ethoxy group, a trifluoromethyl group, a,Ethynyl, methylamino, ethylamino, acetyl, acetamido, methanesulfonyl, dimethylamino, methylsulphonamidocarbonyl, ethylsulphonamidocarbonyl, oxadiazole, thiazole, isothiazole, thiadiazole, triazole or tetrazole;
z is selected from phenyl, pyrrolyl, pyrazolyl, imidazolyl, thiazolyl, isothiazolyl, oxazolyl, isoxazolyl, furanyl, pyridyl, pyrimidinyl, pyrazinyl or pyridazinyl, substituted or unsubstituted with one or more substituents Q;
q is selected from cyano, amino, hydroxyl, carboxyl, nitro, halogen atom, C1-4Alkyl radical, C1-4Alkoxy radical, C1-4Alkylamino, di-C1-4Alkylamino, halogeno C1-4Alkyl or halo C1-4An alkoxy group;
E. x, Y, F together with the benzene ring form the following structure:
Figure BDA0001739294150000081
n is an integer from 0 to 2.
A compound of scheme 6, scheme 5, a pharmaceutically acceptable salt thereof, an ester thereof, or a stereoisomer thereof:
E. x, Y, F together with the benzene ring form the following structure:
Figure BDA0001739294150000082
Figure BDA0001739294150000091
n is selected from 1 or 2.
A compound of scheme 7, as depicted in scheme 6, a pharmaceutically acceptable salt thereof, an ester thereof, or a stereoisomer thereof:
R1、R2each independently selected from hydrogen atom, cyano group, and fluoro groupA chlorine atom, a methyl group, an ethyl group, a propyl group, a butyl group, a methoxy group, a methylamino group, an acetyl group, a trifluoromethyl group, or a trifluoromethoxy group;
R3selected from methyl, ethyl, n-propyl, isopropyl, n-butyl, isobutyl, tert-butyl, trifluoromethyl, trifluoroethyl, cyano, cyclopropyl, cyclobutyl, cyclopentyl, cyclopropylmethyl or cyclobutylmethyl;
R4selected from hydrogen atom, halogen atom, cyano-group, nitryl, amino, hydroxyl, carboxyl, methyl, ethyl, propyl, hydroxymethyl, hydroxyethyl, methoxy, ethoxy, trifluoromethyl, trifluoromethoxy, acetyl, acetamido, methylsulfonaminocarbonyl, ethylsulfonylaminocarbonyl, oxadiazole, thiazole, isothiazole, thiadiazole, triazole or tetrazole;
w is selected from NH, O or S; a is selected from NH, O or S;
z is selected from phenyl, pyrrolyl, pyrazolyl, imidazolyl, thiazolyl, isothiazolyl, oxazolyl, isoxazolyl, furanyl, pyridyl, pyrimidinyl, pyrazinyl or pyridazinyl, substituted or unsubstituted with one or more substituents Q;
q is selected from cyano, amino, hydroxyl, carboxyl, nitro, fluorine atom, chlorine atom, bromine atom, methyl, ethyl, propyl, butyl, methoxy, ethylamino, dimethylamino, trifluoromethyl or trifluoromethoxy;
n is selected from 1.
The compound of scheme 8, scheme 1, pharmaceutically acceptable salts thereof, esters thereof or stereoisomers thereof, having the structure of formula (I-1) below:
Figure BDA0001739294150000101
wherein the content of the first and second substances,
R1、R2each independently selected from hydrogen atom, cyano group, halogen atom, nitro group, amino group, hydroxyl group, carboxyl group, C1-4Alkyl radical, C1-4Alkoxy radical, C1-4Alkylamino, di-C1-4Alkylamino radical, C1-4Alkylthio radical, C1-4Alkylcarbonyl, halo C1-4Alkyl, halo C1-4Alkoxy or C1-4Alkoxy radical C1-4An alkyl group;
R3selected from cyano, C optionally substituted by one or more substituents P1-4Alkyl, halo C1-4Alkyl, 3-6 membered cycloalkyl or 3-6 membered cycloalkyl C1-4An alkyl group;
p is selected from hydroxyl, amino, carboxyl, cyano, nitro, halogen atom, C1-4Alkyl radical, C1-4Alkoxy radical, C1-4Alkylamino, di-C1-4Alkylamino, halogeno C1-4Alkyl or halo C1-4An alkoxy group;
R4selected from hydrogen atom, halogen atom, cyano group, nitro group, amino group, hydroxyl group, carboxyl group, C1-4Alkoxy radical, C1-4Alkyl, hydroxy C1-4Alkyl, halo C1-4Alkyl, carboxyl C1-4Alkyl, amino C1-4Alkyl, aminocarbonyl C1-4Alkyl, hydroxy C1-4Alkoxy, halo C1-4Alkoxy, carboxyl C1-4Alkoxy radical, C2-6Alkenyl radical, C2-6Alkynyl, C1-4Alkylamino radical, C1-4Alkylcarbonyl group, C1-4Alkylcarbonylamino, C1-4Alkylsulfonyl radical, C1-4Alkylsulfonylaminocarbonyl group, C1-4Alkylaminosulfonyl, di-C1-4Alkylamino or 5-6 membered heteroaryl;
w is selected from CH2NH, O, S, SO or SO2；
A is selected from NH, O or S;
z is selected from phenyl or 5-6 membered heteroaryl, substituted or unsubstituted with one or more substituents Q;
q is selected from cyano, amino, hydroxyl, carboxyl, nitro, halogen atom, C1-4Alkyl radical, C1-4Alkoxy radical, C1-4Alkylamino, di-C1-4Alkylamino, halogeno C1-4Alkyl or halo C1-4An alkoxy group;
e is selected from CH2NH, O or CO;
f is selected from absent, CH2NH or O;
x is selected from CH or N;
y is selected from CH2NH or O;
E. x, Y, F are each independently selected from single bonds;
n is an integer from 0 to 3.
The compound of scheme 9, scheme 8, a pharmaceutically acceptable salt thereof, an ester thereof or a stereoisomer thereof,
wherein the content of the first and second substances,
R1、R2each independently selected from a hydrogen atom, a cyano group, a fluorine atom, a chlorine atom, a bromine atom, a nitro group, an amino group, a hydroxyl group, a carboxyl group, a methyl group, an ethyl group, a propyl group, an isopropyl group, a butyl group, an isobutyl group, a tert-butyl group, a methoxy group, a methylamino group, an acetyl group, a trifluoromethyl group, a trifluoroethyl group or a trifluoromethoxy group;
R3selected from methyl, ethyl, n-propyl, isopropyl, n-butyl, isobutyl, tert-butyl, trifluoromethyl, trifluoroethyl, trifluoropropyl, cyano, cyclopropyl, cyclobutyl, cyclopentyl, cyclohexyl, cyclopropylmethyl, cyclopropylethyl, cyclobutylmethyl, cyclopentylmethyl or cyclohexylmethyl;
R4selected from hydrogen atom, halogen atom, cyano group, nitro group, amino group, hydroxyl group, carboxyl group, methyl group, ethyl group, propyl group, isopropyl group, hydroxymethyl group, hydroxyethyl group, methoxy group, ethoxy group, trifluoromethyl group, trifluoromethoxy group, ethynyl group, methylamino group, ethylamino group, acetyl group, acetamido group, methanesulfonyl group, methanesulfonamido carbonyl group, ethanesulfonylaminocarbonyl group, dimethylamino group, pyrazole, imidazole, oxazole, isoxazole, oxadiazole, thiazole, isothiazole, thiadiazole, triazole or tetrazole;
w is selected from CH2NH, O or S; a is selected from NH, O or S;
z is selected from phenyl or 5-6 membered heteroaryl, substituted or unsubstituted with one or more substituents Q;
q is selected from cyano, amino, hydroxyl, carboxyl, nitro, halogen atom, C1-4Alkyl radical, C1-4Alkoxy radical, C1-4Alkylamino, di-C1-4Alkylamino, halogeno C1-4Alkyl or halo C1-4An alkoxy group;
e is selected from CH2NH, O or CO; f is selected from absent, CH2NH or O;
x is selected from CH or N; y is selected from CH2NH or O;
E. x, Y, F are each independently selected from single bonds;
n is selected from 1 or 2.
A compound of scheme 10, scheme 9, a pharmaceutically acceptable salt thereof, an ester thereof, or a stereoisomer thereof:
wherein the content of the first and second substances,
R1、R2each independently selected from a hydrogen atom, a cyano group, a fluorine atom, a chlorine atom, a methyl group, an ethyl group, a propyl group, a butyl group, a methoxy group, a methylamino group, an acetyl group, a trifluoromethyl group or a trifluoromethoxy group;
R3selected from methyl, ethyl, n-propyl, isopropyl, n-butyl, isobutyl, tert-butyl, trifluoromethyl, trifluoroethyl, cyano, cyclopropyl, cyclobutyl, cyclopentyl, cyclopropylmethyl or cyclobutylmethyl;
R4selected from hydrogen atom, halogen atom, cyano-group, nitryl, amino, hydroxyl, carboxyl, methyl, ethyl, propyl, hydroxymethyl, hydroxyethyl, methoxy, ethoxy, trifluoromethyl, trifluoromethoxy, acetyl, acetamido, methylsulfonylaminocarbonyl, ethylsulfonylaminocarbonyl, oxadiazole, thiazole, isothiazole, thiadiazole, triazole or tetrazole;
w is selected from NH, O or S; a is selected from NH, O or S;
z is selected from phenyl or pyridyl which is substituted or unsubstituted by one or more substituents Q selected from cyano, amino, hydroxyl, carboxyl, nitro, a halogen atom, C1-4Alkyl radical, C1-4Alkoxy radical, C1-4Alkylamino, di-C1-4Alkylamino, halogeno C1-4Alkyl or halo C1-4An alkoxy group;
e is selected from CH2NH, O or CO; f is selected from CH2NH or O;
x is selected from CH or N; y is selected from CH2NH or O;
E. x, Y, F are each independently selected from single bonds;
n is selected from 1 or 2.
A compound of scheme 11, as depicted in scheme 10, a pharmaceutically acceptable salt thereof, an ester thereof, or a stereoisomer thereof:
wherein E, X, Y, F together form a cyclic group that together with the benzene ring forms the following structure:
chromanyl, benzo 1, 4-dioxinyl, benzo 1, 3-dioxinyl, benzotetrahydropyridinyl, chromazinyl, benzotetrahydropyrazinyl, 1,2,3, 4-tetrahydroquinazolinyl, 1,2,3, 4-tetrahydrocinnolinyl, tetrahydronaphthyl, tetralone.
A compound of scheme 12, scheme 11, a pharmaceutically acceptable salt thereof, an ester thereof, or a stereoisomer thereof:
wherein Z is selected from phenyl which is substituted or unsubstituted by one or more substituents Q, and the substituents Q are selected from cyano, amino, hydroxyl, carboxyl, nitro, halogen atoms and C1-4Alkyl radical, C1-4Alkoxy radical, C1-4Alkylamino, di-C1-4Alkylamino, halogeno C1-4Alkyl or halo C1-4An alkoxy group;
E. x, Y, F together with the benzene ring form the following structure:
Figure BDA0001739294150000131
a compound of scheme 13, scheme 12, a pharmaceutically acceptable salt thereof, an ester thereof, or a stereoisomer thereof:
wherein W is selected from O;
a is selected from O;
z is selected from phenyl which is substituted or unsubstituted by 1 to 2 substituents Q selected from cyano, amino, hydroxyl, carboxyl, nitro, fluorine atom, chlorine atom, bromine atom, methyl, ethyl, methoxy, ethoxy, methylamino, dimethylamino, trifluoromethyl or trifluoromethoxy;
E. x, Y, F together with the benzene ring form the following structure:
Figure BDA0001739294150000132
n is selected from 1.
A compound of scheme 14, as depicted in scheme 9, a pharmaceutically acceptable salt thereof, an ester thereof, or a stereoisomer thereof:
wherein the content of the first and second substances,
R1、R2each independently selected from a hydrogen atom, a cyano group, a fluorine atom, a chlorine atom, a methyl group, an ethyl group, a propyl group, a butyl group, a methoxy group, a methylamino group, an acetyl group, a trifluoromethyl group or a trifluoromethoxy group;
R3selected from methyl, ethyl, n-propyl, isopropyl, n-butyl, isobutyl, tert-butyl, trifluoromethyl, trifluoroethyl, cyano, cyclopropyl, cyclobutyl, cyclopentyl, cyclopropylmethyl or cyclobutylmethyl;
R4selected from hydrogen atom, halogen atom, cyano-group, nitryl, amino, hydroxyl, carboxyl, methyl, ethyl, propyl, hydroxymethyl, hydroxyethyl, methoxy, ethoxy, trifluoromethyl, trifluoromethoxy, acetyl, acetamido, methylsulfonaminocarbonyl, ethylsulfonylaminocarbonyl, oxadiazole, thiazole, isothiazole, thiadiazole, triazole or tetrazole;
w is selected from NH, O or S; a is selected from NH, O or S;
z is selected from phenyl or pyridyl which is substituted or unsubstituted by one or more substituents Q selected from cyano, amino, hydroxyl, carboxyl, nitro, a halogen atom, C1-4Alkyl radical, C1-4Alkoxy radical, C1-4Alkylamino, di-C1-4Alkylamino, halogeno C1-4Alkyl or halo C1-4An alkoxy group;
e is selected from CH2NH or O; f is selected from absent;
x is selected from CH or N; y is selected from CH2Or O;
E. x, Y, F are each independently selected from single bonds;
n is selected from 1 or 2.
A compound of scheme 15, scheme 14, a pharmaceutically acceptable salt thereof, an ester thereof, or a stereoisomer thereof:
wherein E, X, Y together form a cyclic group that together with the benzene ring forms the following structure:
a benzodihydropyrrolyl group, a benzodihydrofuranyl group, a benzo 1, 3-dioxolyl group or a indanyl group.
A compound of scheme 16, scheme 15, a pharmaceutically acceptable salt thereof, an ester thereof, or a stereoisomer thereof:
wherein W is selected from O; a is selected from O;
z is selected from phenyl which is substituted or unsubstituted by one or more substituents Q, wherein the substituents Q are selected from cyano, amino, hydroxyl, carboxyl, nitro, halogen atoms and C1-4Alkyl radical, C1-4Alkoxy radical, C1-4Alkylamino, di-C1-4Alkylamino, halogeno C1-4Alkyl or halo C1-4An alkoxy group;
E. x, Y, F together with the benzene ring form the following structure:
Figure BDA0001739294150000151
a compound of scheme 17, scheme 9, a pharmaceutically acceptable salt thereof, an ester thereof, or a stereoisomer thereof:
wherein the content of the first and second substances,
z is selected from phenyl substituted or unsubstituted with one or more substituents Q;
q is selected from cyano, amino, hydroxyl, carboxyl, nitro, halogen atom, methyl, ethyl, propyl, methoxy, ethoxy, methylamino, ethylamino, dimethylamino, trifluoromethyl or trifluoromethoxy;
e is selected from CH2O or CO; f is selected from absent, CH2Or O;
x is selected from CH or N; y is selected from CH2；
E. X, Y, F are each independently selected from single bonds;
n is selected from 1 or 2.
A compound of scheme 18, scheme 17, a pharmaceutically acceptable salt thereof, an ester thereof, or a stereoisomer thereof:
wherein E is selected from O or CH2(ii) a F is selected from absent, CH2Or O;
x is selected from CH; y is selected from CH2；
E. X, Y, F are each independently selected from single bonds;
n is selected from 1.
A compound of scheme 19, scheme 18, a pharmaceutically acceptable salt thereof, an ester thereof, or a stereoisomer thereof:
wherein E is selected from O; f is selected from absent or CH2；
X is selected from CH; y is selected from CH2；
E. X, Y, F are each independently selected from single bonds;
n is selected from 1.
A compound of scheme 20, scheme 19, a pharmaceutically acceptable salt thereof, an ester thereof, or a stereoisomer thereof:
wherein E, X, Y, F together form a cyclic group that together with the benzene ring forms the following structure:
Figure BDA0001739294150000161
a compound of scheme 21, scheme 17, a pharmaceutically acceptable salt thereof, an ester thereof, or a stereoisomer thereof:
wherein E is selected from CH2(ii) a F is selected from absent or CH2；
X is selected from N; y isIs selected from CH2；
E. X, Y, F are each independently selected from single bonds;
n is selected from 1.
A compound of scheme 22, scheme 17, a pharmaceutically acceptable salt thereof, an ester thereof, or a stereoisomer thereof:
wherein E, X, Y, F are each independently selected from CH2；
E. X, Y, F are each independently selected from single bonds;
n is selected from 1.
Any combination of the above groups can be used, and all the resulting embodiments are described herein.
Part of the Compounds of the invention
Figure BDA0001739294150000171
Figure BDA0001739294150000181
Figure BDA0001739294150000191
Detailed Description
The "halogen atom" in the present invention includes fluorine atom, chlorine atom, bromine atom, iodine atom and the like.
"C" according to the invention1-6Alkyl "denotes straight or branched alkyl having 1 to 6 carbon atoms, including for example" C1-4Alkyl group "," C1-3Alkyl "and the like, specific examples include, but are not limited to: methyl, ethyl, n-propyl, isopropyl, n-butyl, 2-methylpropyl, 1-dimethylethyl, n-pentyl, 3-methylbutyl, 2-methylbutyl, 1-ethylpropyl, n-hexyl, 4-methylpentyl, 3-methylpentyl, 2-methylpentylA methyl group, 1-methylpentyl group, 3-dimethylbutyl group, 2-dimethylbutyl group, 1-dimethylbutyl group, 1, 2-dimethylbutyl group, 1, 3-dimethylbutyl group, 2-ethylbutyl group, or 1, 2-dimethylpropyl group, etc.
"C" according to the invention1-4Alkyl "denotes straight or branched alkyl having 1 to 4 carbon atoms, including for example" C1-4Alkyl group "," C1-3Alkyl "and the like, specific examples include, but are not limited to: methyl, ethyl, n-propyl, isopropyl, n-butyl, 2-methylpropyl, 1-methylpropyl, or 1, 1-dimethylethyl, and the like.
"C" according to the invention2-8The "alkenyl group" means a straight chain or branched alkenyl group having 2 to 8 carbon atoms containing at least one double bond, and includes, for example, "C2-6Alkenyl group "," C2-4Alkenyl group "," C2-3Alkenyl groups "and the like, specific examples include, but are not limited to: vinyl group, 1-propenyl group, 2-butenyl group, 3-butenyl group, 2-methyl-1-propenyl group, 1-methyl-2-propenyl group, 1-pentenyl group, 2-pentenyl group, 3-pentenyl group, 2-methyl-1-butenyl group, 3-methyl-1-butenyl group, 2-methyl-3-butenyl group, 1-dimethyl-2-propenyl group, 1-ethyl-2-propenyl group, 2-hexenyl group, 3-hexenyl group, 2-methyl-1-pentenyl group, 3-methyl-1-pentenyl group, 1-methyl-2-pentenyl group, 3-methyl-2-pentenyl group, 2-methyl-3-pentenyl, 1-methyl-4-pentenyl, 3-methyl-4-pentenyl, 1-dimethyl-3-butenyl, 1, 2-dimethyl-3-butenyl, 1, 3-dimethyl-2-butenyl, 2-dimethyl-3-butenyl, 2, 3-dimethyl-2-butenyl, 2, 3-dimethyl-1-butenyl, 2-ethyl-3-butenyl, 2-heptenyl, 3-heptenyl, 4-heptenyl, 1-octenyl, 3-octenyl, 4-octenyl, 1, 3-butadienyl, 1-pentenyl, 3-pentenyl, 2-dimethyl-3-butenyl, 2, 3-heptenyl, 4-octenyl, 2-octenyl, 4-octenyl, 1, 3-butadienyl, 2-pentenyl, 3-pentenyl, 2-dimethyl-3-butenyl, 2-dimethyl-3-butenyl, 2-heptenyl, 3-heptenyl, 4-octenyl, 2-butenenyl, 2, 3-butenyl, 2, 3-butenyl, 2, 3-butenyl, 3, n, 2, 4-pentadienyl, 1, 4-hexadienyl, 2, 4-hexadienyl, 1, 5-heptadienyl, 2, 6-octadienyl, or the like.
"C" according to the invention2-8Alkynyl refers to a straight or branched chain alkynyl group of 2-8 carbon atoms containing a triple bond, including, for example, "C2-6Alkynyl group "," C2-4Alkynyl group "," C2-3Alkynyl "and the like, specific examples include but are not limited toLimited to: ethynyl, 1-propynyl, 2-butynyl, 1-methyl-2-propynyl, 2-pentynyl, 3-pentynyl, 1-methyl-2-butynyl, 2-methyl-3-butynyl, 1-dimethyl-2-propynyl, 1-ethyl-2-propynyl, 2-hexynyl, 3-hexynyl, 1-methyl-2-pentynyl, 1-methyl-3-pentynyl, 2-methyl-3-pentynyl, 1-dimethyl-3-butynyl, 2-ethyl-3-butynyl, 2-heptynyl, 3-heptynyl, 4-methyl-2-hexynyl, 2-ethyl-2-propynyl, 3-pentynyl, 1-methyl-2-propynyl, 2-ethyl-2-propynyl, 2-hexynyl, 3-hexynyl, 1-methyl-2-propynyl, 2-methyl-3-hexynyl, 2-methyl-propynyl, 1-methyl-3-propynyl, 2 hexynyl, 3 hexynyl, 2, p, 5-methyl-2-hexynyl, 2-methyl-3-hexynyl, 5-methyl-3-hexynyl, 2-methyl-4-hexynyl, 4-methyl-5-hexynyl, 2-octynyl, 3-octynyl, 4-methyl-2-heptynyl, 5-methyl-3-heptynyl, 6-methyl-3-heptynyl, 2-methyl-4-heptynyl, 2-methyl-5-heptynyl, 3-methyl-6-heptynyl or the like.
"C" according to the invention1-6Alkoxy radical, C1-6Alkylamino, di-C1-6Alkylamino radical, C1-6Alkylthio radical, C1-6Alkylcarbonyl group, C1-6Alkylcarbonyloxy, C1-6Alkylsulfonyl radical, C1-6Alkylaminosulfonyl, di-C1-6Alkylaminosulfonyl radical, C1-6Alkylsulfonylamino group, C1-6Alkylsulfonyloxy, C1-6Alkylsulfonylaminocarbonyl "means with C1-6alkyl-O-, C1-6alkyl-NH-, (C)1-6Alkyl radical)2-N-、C1-6alkyl-S-, C1-6alkyl-C (O) -, C1-6alkyl-C (O) -O-, C1-6alkyl-SO2-、C1-6alkyl-NH-SO2-、(C1-6Alkyl radical)2-N-SO2-、C1-6alkyl-SO2-NH-、C1-6alkyl-SO2-O-、C1-6alkyl-SO2A group formed by the formula-NH-C (O) -wherein "C" is1-6Alkyl "is as defined above.
"C" according to the invention1-4Alkoxy radical, C1-4Alkylamino, di-C1-4Alkylamino radical, C1-4Alkylthio radical, C1-4Alkylcarbonyl group, C1-4Alkylcarbonyloxy, C1-4Alkylsulfonyl radical, C1-4Alkylaminosulfonyl, di-C1-4Alkylaminosulfonyl radical, C1-4Alkylsulfonylamino group, C1-4Alkylsulfonyloxy, C1-4Alkylsulfonylaminocarbonyl "means with C1-4alkyl-O-, C1-4alkyl-NH-, (C)1-4Alkyl radical)2-N-、C1-4alkyl-S-, C1-4alkyl-C (O) -, C1-4alkyl-C (O) -O-, C1-4alkyl-SO2-、C1-4alkyl-NH-SO2-、(C1-4Alkyl radical)2-N-SO2-、C1-4alkyl-SO2-NH-、C1-4alkyl-SO2-O-、C1-4alkyl-SO2A group formed by the formula-NH-C (O) -wherein "C" is1-4Alkyl "is as defined above.
The "halo C" of the present invention1-6Alkyl, hydroxy C1-6Alkyl, carboxyl C1-6Alkyl, amino C1-6Alkyl radical, C1-6Alkoxy radical C1-6Alkyl, halo C1-6Alkoxy, hydroxy C1-6Alkoxy, carboxyl C1-6Alkoxy, carboxyloxy C1-6Alkyl, carboxy amino C1-6Alkyl, aminocarbonyl C1-6Alkyl "means one or more, for example, 1 to 4, 1 to 3, 1 to 2 halogen atoms, hydroxyl group, amino group, C1-6Alkoxy, carboxyl oxy, carboxyl amino and aminocarbonyl are respectively substituted for C1-6Alkyl radical, C1-6A group formed by a hydrogen atom in an alkoxy group.
The "halo C" of the present invention1-4Alkyl, hydroxy C1-4Alkyl, carboxyl C1-4Alkyl, amino C1-4Alkyl radical, C1-4Alkoxy radical C1-4Alkyl, halo C1-4Alkoxy, hydroxy C1-4Alkoxy, carboxyl C1-4Alkoxy, carboxyloxy C1-4Alkyl, carboxy amino C1-4Alkyl, aminocarbonyl C1-4Alkyl "means one or more, for example, 1 to 4, 1 to 3, 1 to 2 halogen atoms, hydroxyl group, amino group, C1-4Alkoxy, carboxyl oxy, carboxyl amino and aminocarbonyl are respectively substituted for C1-4Alkyl radical, C1-4A group formed by a hydrogen atom in an alkoxy group.
The "3-to 8-membered cycloalkyl group C" of the present invention1-6Alkyl, 3-8 membered heterocyclyl C1-6Alkyl "refers to 3-8 membered cycloalkyl, 3-8 membered heterocyclyl" substituted C1-6Hydrogen atoms in the alkyl group.
The "3-6 membered cycloalkyl group C" of the present invention1-4Alkyl, 3-6 membered heterocyclyl C1-4Alkyl "refers to 3-6 membered cycloalkyl, 3-6 membered heterocyclyl" substituted C1-4Hydrogen atoms in the alkyl group.
The "aryl" as used herein refers to an aromatic ring, such as phenyl, naphthyl, anthracenyl and the like.
The "5-8 membered heteroaryl" in the present invention refers to an unsaturated cyclic group having 5 to 8 ring atoms and containing at least one heteroatom, such as nitrogen, oxygen, and sulfur, including the case where carbon, nitrogen, and sulfur atoms are oxidized. Including, for example, "5-to 7-membered heteroaryl", "5-to 6-membered heteroaryl", "7-to 8-membered heteroaryl", specifically "5-to 8-membered heteroaryl containing 1 to 3O, S and/or N", "5-to 8-membered heteroaryl containing 1 to 2O, S and/or N", "5-to 8-membered heteroaryl containing 2 to 3O, S and/or N", "5-to 6-membered heteroaryl containing 1 to 2N, O and/or S atoms", "6-membered heteroaryl containing 1 to 2N, O and/or S atoms". Specific examples include, but are not limited to, furyl, thienyl, pyrrolyl, thiazolyl, isothiazolyl, thiadiazolyl, oxazolyl, isoxazolyl, oxadiazolyl, imidazolyl, pyrazolyl, 1,2, 3-triazolyl, 1,2, 4-triazolyl, tetrazolyl, 1,2, 3-oxadiazolyl, 1,2, 4-oxadiazolyl, 1,2, 5-oxadiazolyl, 1,3, 4-oxadiazolyl, pyridyl, 2-pyridone, 4-pyridone, pyrimidinyl, pyridazinyl, pyrazinyl, 1,2, 3-triazinyl, 1,3, 5-triazinyl, or 1,2,4, 5-tetrazinyl, and the like.
"3-8 membered cycloalkyl" means a partially saturated or saturated monocyclic cycloalkyl group derived from an alkane moiety of 3-8 carbon atoms with one hydrogen atom removed, and includes, for example, "3-6 membered cycloalkyl", "4-7 membered cycloalkyl", "4-6 membered cycloalkyl", "5-6 membered cycloalkyl", and the like. Specifically, the cycloalkyl group may be a "3-to 8-membered saturated cycloalkyl group", a "3-to 8-membered partially saturated cycloalkyl group", a "5-to 6-membered saturated cycloalkyl group", or a "5-to 6-membered partially saturated cycloalkyl group". 3-8 membered saturated cycloalkyl groups include, but are not limited to: cyclopropane, cyclobutane, cyclopentane, cyclohexane, cycloheptane, cyclooctane, methylcyclopropane, dimethylcyclopropane, methylcyclobutane, dimethylcyclobutane, methylcyclopentane, dimethylcyclopentane, methylcyclohexane, dimethylcyclohexane, etc.; 3-8 membered partially saturated cycloalkyl groups include, but are not limited to: cyclopentenyl, 1, 3-cyclopentadienyl, cyclohexenyl, 1, 4-cyclohexadienyl, cycloheptenyl, 1, 4-cycloheptadienyl, or cyclooctenyl.
"3-8 membered heterocyclyl" refers to a group derived from a saturated or partially saturated monocyclic heterocyclic compound containing 3 to 8 ring atoms and at least one heteroatom (e.g., 1,2,3,4, or 5 heteroatoms) by removal of one hydrogen atom. Including, for example, "3-7 membered heterocyclic group", "3-6 membered heterocyclic group", "3-5 membered heterocyclic group", "4-7 membered heterocyclic group", "4-6 membered heterocyclic group", "5-6 membered heterocyclic group", 6-7 membered heterocyclic group "," 6-8 membered heterocyclic group "and the like. The method specifically comprises the following steps: "a 3-8 membered heterocyclic group containing 1 to 2N, O and/or S atoms", "a 3-8 membered saturated heterocyclic group containing 1 to 2N, O and/or S atoms", "a 5-6 membered saturated heterocyclic group", "a 5-6 membered heterocyclic group containing 1 to 2N, O and/or S atoms", "a 5-6 membered saturated heterocyclic group containing 1 to 2N, O and/or S atoms". The 3-8 membered partially saturated heteromonocyclic group refers to a cyclic group containing a double bond and a hetero atom. The 3-to 8-membered saturated heteromonocyclic group means a heteroatom-containing cyclic group having all saturated bonds. Examples include, but are not limited to: aziridinyl, 2H-aziridinyl, diazacyclopropenyl, 3H-diazacyclopropenyl, azetidinyl, 1, 4-dioxanyl, 1, 3-dioxolanyl, 1, 4-dioxadienyl, tetrahydrofuryl, dihydropyrrolyl, pyrrolidinyl, imidazolidinyl, 4, 5-dihydroimidazolyl, pyrazolidinyl, 4, 5-dihydropyrazolyl, 2, 5-dihydrothienyl, tetrahydrothienyl, 4, 5-dihydrothiazolyl, piperidinyl, piperazinyl, morpholinyl, 4, 5-dihydrooxazolyl, 4, 5-dihydroisoxazolyl, 2, 3-dihydroisoxazolyl, 2H-1, 2-oxazinyl, 6H-1, 3-oxazinyl, 4H-1, 3-thiazinyl, 6H-1, 3-thiazinyl, 2H-pyranyl, 2H-pyran-2-onyl, 3, 4-dihydro-2H-pyranyl, 2, 5-dihydrothienyl, 3, 4-dihydro-2H-pyranyl, 5, 6-dihydro-4H-1, 3-oxazinyl, 1,2,3, 6-tetrahydropyridinyl, 1,2,3, 4-tetrahydropyridinyl, or 2,3,4, 5-tetrahydropyridinyl, and the like.
The term "heteroatom" as used herein means N, O, S, SO and/or SO2Etc., preferably N, O, S.
"partially saturated" as used herein means that the cyclic moiety includes at least one double or triple bond.
In the present invention, "F is selected from absent" and means Y is directly bonded to a phenyl group.
E. X, Y, F together with the benzene ring may form the following structure:
Figure BDA0001739294150000231
Figure BDA0001739294150000232
further preferred is
Figure BDA0001739294150000233
Figure BDA0001739294150000241
In addition, the invention also provides a preparation method of the compound represented by the general formula (I), pharmaceutically acceptable salts, esters and stereoisomers of the compound.
In particular, the preparation method includes, but is not limited to, the following process schemes (wherein each abbreviation is defined as follows: DCM: dichloromethane; DMF: N, N-dimethylformamide; DMSO: dimethyl sulfoxide; EA: ethyl acetate; MeOH: methanol; NBS: N-bromosuccinimide; NCS: N-chlorosuccinimide; PE: petroleum ether; THF: tetrahydrofuran; DIBAL-H: diisobutylaluminum hydride; Pd (dppf) Cl)2: [1,1' -bis (diphenylphosphino) ferrocene]Dichloroation ofPalladium); PPTS: pyridinium p-toluenesulfonate; and (3) DHP: 3, 4-2H-dihydropyran; TFAA: trifluoroacetic anhydride; LiHMDS: lithium bis (trimethylsilyl) amide; TBSCl: tert-butyldimethylsilyl chloride; TBAF: tetrabutylammonium fluoride trihydrate; DEAD: diethyl azodicarboxylate):
Figure BDA0001739294150000242
R1、R2、R3、R4m, n, W, A, Z, E, F, X and Y are as described above, and A' represents a fluorine atom, a chlorine atom, a bromine atom or an iodine atom.
Specific exemplary steps are as follows:
1. preparation of intermediate 1
Dissolving the starting material 1 in organic solvent (such as lower alcohol such as ethanol), slowly adding the starting material 2 in batches, adding alkaline solution (such as NaOH solution) after adding, and heating to 60-90 deg.C for 5-48 hr. After the reaction, the solvent was removed from the reaction mixture under reduced pressure, and the solid was washed with water and dried to obtain intermediate 1.
2. Preparation of intermediate 2
The intermediate 1 is dissolved in an organic solvent (e.g., DMF, etc.), an electrophilic substitution reagent (e.g., N-chlorosuccinimide, etc.) is slowly added in portions, and after the addition is completed, the reaction solution is stirred for 0.2 to 5 hours, and then poured into water. Extraction with an organic solvent (e.g., ethyl acetate, etc.), washing of the organic phase with water and saturated sodium chloride solution, drying, and removal of the solvent afforded intermediate 2.
3. Preparation of intermediate 3
The intermediate 2 is dissolved in an organic solvent (e.g., triethylamine, etc.), and the starting material 3 is added and reacted for 5 to 20 hours. After completion of the reaction, the solvent was removed under reduced pressure and column chromatography (e.g., PE: EA ═ 10:1) gave intermediate 3.
4. Preparation of intermediate 4
Dissolving the intermediate 3 in an organic solvent (such as tetrahydrofuran and the like), cooling in an ice bath, adding a toluene solution of diisobutylaluminum hydride (DIBAL-H), heating to 20-30 ℃ after the addition is finished, reacting for 5-20 hours, cooling in an ice bath, adding a saturated halogenating agent (such as an ammonium chloride solution and the like) to quench the reaction after the reaction is finished, extracting with an organic solvent (such as ethyl acetate and the like), washing an organic phase with a saturated halogenating agent (such as an ammonium chloride solution, a sodium chloride solution and the like), drying, and removing the solvent to obtain an intermediate 4.
5. Preparation of intermediate 5
Dissolving the intermediate 4 in an organic solvent (such as dichloromethane and the like), adding triethylamine, then adding a halide (such as phosphorus trichloride, phosphorus tribromide and the like), and reacting at 20-40 ℃ for 0.5-5 hours. After completion of the reaction, the solvent was removed and column chromatography (e.g., PE: EA ═ 10:1) gave intermediate 5.
6. Preparation of intermediate 6
Intermediate 6 was prepared or purchased.
7. Preparation of intermediate 7
The intermediate 6 is dissolved in an organic solvent (e.g., N-dimethylformamide, acetonitrile, toluene, etc.), an alkaline reagent (e.g., potassium carbonate, cesium carbonate, sodium iodide, etc.) and the intermediate 5 are added, and stirred (0.5 to 24 hours) at 0 to 100 ℃. The intermediate 7 is obtained by diluting the mixture with an organic solvent (e.g., ethyl acetate, etc.), washing the diluted mixture with a saturated halogenating agent (e.g., sodium chloride solution, etc.) or with water, drying, concentrating, and purifying by silica gel column chromatography (e.g., ethyl acetate: petroleum ether: 1-10, dichloromethane: methanol: 15: 1).
8. Preparation of Compounds of formula (I)
Dissolving intermediate 7 in organic solvent (such as methanol/water, tetrahydrofuran, methanol, tetrahydrofuran/methanol, methanol/tetrahydrofuran/water, etc.), adding alkaline compound (such as lithium hydroxide monohydrate, sodium hydroxide, etc.), and stirring at 15-60 deg.C for 8-72 hr. Diluting the reaction solution with water, adjusting pH to 2-7 with acidic solution (such as citric acid, hydrochloric acid, etc.), adding organic solvent (such as ethyl acetate, etc.), separating, washing the organic phase with saturated halogenating agent (such as sodium chloride solution, etc.), drying, concentrating, and purifying (preferably by preparative high performance liquid chromatography, silica gel column chromatography, etc.) to obtain the compound of formula (I).
"pharmaceutically acceptable salts" of the compounds of formula (I) according to the invention are taken to mean the acidic functions present in the compounds of formula (I)Salts of the groups with suitable inorganic or organic cations (bases), including salts with alkali or alkaline earth metals, ammonium salts, and salts with nitrogen-containing organic bases; and basic functional groups present in the compounds of formula (I) (e.g. -NH)2Etc.) with suitable inorganic or organic anions (acids), including with inorganic acids, with organic carboxylic acids.
The "ester" of the compound represented by the formula (I) of the present invention means an ester which can be formed by esterification with an alcohol when a carboxyl group is present in the compound of the formula (I), or an ester which can be formed by esterification with an organic acid, an inorganic acid, an organic acid salt or the like when a hydroxyl group is present in the compound of the formula (I). The ester can be hydrolyzed in the presence of acid or alkali to generate corresponding acid or alcohol.
"stereoisomers" of the compounds of the invention are classified as conformational and configurational isomers, and configurational isomers are also classified as cis-trans isomers and optical isomers. Conformational isomerism is a stereoisomerism phenomenon in which organic molecules having a certain configuration are rotated or twisted due to carbon and carbon single bonds, so that atoms or atom groups of the molecules generate different arrangement modes in space, and the common structures include structures of alkanes and cycloalkanes, such as chair conformation and ship conformation which appear in cyclohexane structure. "stereoisomers" refers to compounds of the invention when they contain one or more asymmetric centers and thus can be present as racemates and racemic mixtures, single enantiomers, diastereomeric mixtures and individual diastereomers. The compounds of the present invention have asymmetric centers that each independently produce two optical isomers, and the scope of the present invention includes all possible optical isomers and diastereomeric mixtures and pure or partially pure compounds. The compounds of the present invention, if they contain an olefinic double bond, include both cis-and trans-isomers, unless otherwise specified. The compounds of the present invention may exist in tautomeric forms having different points of attachment of hydrogen through one or more double bond shifts.
The invention also provides a pharmaceutical composition which contains the compound shown in the formula (I), pharmaceutically acceptable salts, esters and stereoisomers thereof. In particular, the present invention provides a pharmaceutical composition for the treatment and/or prevention of FXR mediated diseases, which contains the compound represented by the above formula (I), a pharmaceutically acceptable salt thereof, an ester thereof, and a stereoisomer thereof.
The invention further provides a pharmaceutical preparation which comprises the compound shown in the formula (I), pharmaceutically acceptable salts, esters and stereoisomers thereof and one or more pharmaceutically acceptable carriers and/or diluents, and the pharmaceutical preparation can be prepared into any pharmaceutically acceptable dosage form. Administered to a patient in need of such treatment by oral, parenteral, rectal, or pulmonary administration. For oral administration, it can be made into conventional solid preparations such as tablet, capsule, pill, granule, etc.; it can also be made into oral liquid, such as oral solution, oral suspension, syrup, etc. When the composition is formulated into oral preparations, appropriate filler, binder, disintegrating agent, lubricant, etc. can be added. For parenteral administration, it can be made into injection, including injection solution, sterile powder for injection and concentrated solution for injection. The injection can be prepared by conventional method in the existing pharmaceutical field, and can be prepared without adding additives or adding appropriate additives according to the properties of the medicine. For rectal administration, it can be made into suppository, etc. For pulmonary administration, it can be made into inhalant or spray.
The invention also provides application of the compound shown in the formula (I), pharmaceutically acceptable salts, esters and stereoisomers of the compound in preparation of medicines for treating and/or preventing FXR mediated diseases and related diseases. The diseases comprise: (1) chronic intrahepatic or some form of extrahepatic cholestatic disorder, or liver fibrosis resulting from chronic cholestatic disorder or acute intrahepatic cholestatic disorder, cirrhosis of the liver, obstructive or chronic inflammatory disorders of the liver, fatty liver and its complications, alcohol-related fatty liver and its complications, acute liver failure, cholelithiasis, and/or inflammatory bowel disease, primary biliary cirrhosis; conditions and diseases caused by chronic fatty and fibrotic conditions resulting from forced accumulation of lipids, particularly triglycerides, and then activation of liver fibrosis, such as non-alcoholic fatty liver disease or non-alcoholic steatohepatitis; lipid or lipoprotein disorders such as atherosclerosis, dyslipidemia, thrombosis. (2) Clinical complications of type I or type II diabetes mellitus include diabetic nephropathy, diabetic neuropathy, diabetic retinopathy, and other observed outcomes of their clinically overt long-term diabetes. (3) A non-malignant hyperproliferative disease or hyperproliferative disease selected from: hepatocellular carcinoma, colon adenoma and polyposis, colon adenocarcinoma, breast cancer, pancreatic cancer, esophageal cancer, and other forms of gastrointestinal and hepatic neoplastic disease. (4) Dyslipidemia includes atherosclerosis, bile acid disorders, benign intrahepatic cholestasis, progressive familial intrahepatic cholestasis, primary biliary cirrhosis, primary sclerosing cholangitis, cholesterol gallstones, dyslipidemia, fibrosis-related diseases, chronic hepatitis, non-viral hepatitis, inflammatory bowel disease, intestinal flora disorders, liver transplantation, fatty liver, cirrhosis, hepatitis, liver failure, cholestasis, cholelithiasis, non-alcoholic fatty liver disease, diabetes, myocardial infarction, stroke, thrombosis, cancer, and the like.
The present invention also provides a method for treating and/or preventing FXR mediated diseases, comprising the step of administering to a mammal in need of such treatment a compound represented by formula (I) above, a pharmaceutically acceptable salt thereof, an ester thereof, and a stereoisomer thereof.
The compound of the invention has the following advantages:
(1) the compound of formula (I), pharmaceutically acceptable salts, esters and stereoisomers thereof have excellent FXR receptor agonistic activity, and can be safely used for treating and/or preventing related diseases such as non-alcoholic fatty liver, primary biliary cirrhosis, lipid metabolism disorder, diabetic complications, malignant tumors and the like;
(2) the compound of formula (I), pharmaceutically acceptable salts, esters and stereoisomers thereof have good biological stability, longer action and high bioavailability;
(3) the compound of formula (I), pharmaceutically acceptable salts, esters and stereoisomers thereof show lower toxicity, good drug resistance and high safety.
The beneficial effects of the compounds of the present invention are further illustrated below by biological experiments, but this should not be understood as the only beneficial effects of the compounds of the present invention.
Experimental example 1: in vitro biochemical analysis of Compounds of the invention
(1) Test article: the chemical names and preparation methods of the compounds of the present invention are shown in the preparation examples of the respective compounds.
(2) The experimental method comprises the following steps:
dissolving a detection compound in 100% DMSO, diluting by 1000 times, taking 160nL, and then adding 3.84 mu L of detection buffer solution; diluting the Target/Antibody mixture by 2 times, and then adding 8 mu L of solution; adding 4.0. mu.L of coactivated peptide diluted 4 times; incubation at room temperature for 60 minutes; after incubation, the data were detected and analyzed in a fluorescent microplate reader.
(3) Experimental results and conclusions:
TABLE 1 Biochemical analysis of the Compounds of the invention
Figure BDA0001739294150000281
As shown in table 1, the compounds of the present invention have various degrees of agonism on FXR, and are of great significance for treating non-alcoholic fatty liver disease, primary biliary cirrhosis, lipid metabolism disorder, diabetic complications, malignant tumors, and other related diseases, especially compound 1, compound 2, compound 4, compound 5, compound 6, compound 7, and compound 9.
Experimental example 2: in vitro cell level assay for Compounds of the invention
(1) Test article: the chemical names and preparation methods of the compounds of the present invention are shown in the preparation examples of the respective compounds.
(2) The experimental method comprises the following steps:
a polystyrene-TC treated microwell reaction plate (Corning Cat. #3712) was used in this experiment.
1000 × Compound 40nL and 4 μ L of assay medium were added together to the assay plate; adding 32 μ L of cells to the analysis medium, diluting to a suitable cell density, and adding to the assay plate; add 4. mu.L of assay medium to all wells and the final assay volume is 40. mu.L; the test plate was placed at 37 deg.C/5% CO2Incubating for 16-24h in a humidified incubator; then add 8 μ L substrate to the assay plate; incubating the experimental plate for 2h at room temperature in a dark place; detection after incubation in a fluorescent microplate reader (Tecan Safire)2) And the data was analyzed.
(3) Experimental results and conclusions:
TABLE 2 FXR agonistic Activity of Compounds of the invention
Figure BDA0001739294150000291
As shown in Table 2, the compounds of the invention have different degrees of agonism on UAS-bla HEK 293T cells, and have important significance for treating related diseases such as non-alcoholic fatty liver, primary biliary cirrhosis, lipid metabolism disorder, diabetic complications, malignant tumors and the like, in particular to a compound 2, a compound 4 and a compound 9.
Experimental example 3: the compounds of the invention are relatively effective on HepG2 cells and human hepatocytes BSEP, CYP7A1 and SHP mRNA Influence of expression amount
(1) Test article: the chemical names and preparation methods of the compounds of the present invention are shown in the preparation examples of the respective compounds.
PBS stands for phosphate buffer.
Control drug: PX-104, the concrete structure is seen in background technology; PX-102, which is a racemate of PX-104.
(2) The experimental method comprises the following steps:
firstly, laying cells, adding compound and collecting cells
Digesting and collecting cells by using pancreatin, and measuring the cell concentration; resuspending the cells to a density of 7.5e5 cells/mL according to the counting results; 6-well cell culture plates, each well inoculated with 2mL of cells; culturing plateIn an incubator, 5% CO at 37 ℃2The culture was conditioned for 24 hours.
Test compounds were diluted with DMSO to 12150, 4050, 1350, 450, 150, 50, 16.67, 5.56 and 1.85 μ M; mu.L of the stock solution obtained by the previous dilution was added to 5mL of the medium, respectively. The resulting working solution concentrations were 12150, 4050, 1350, 450, 150, 50, 16.67, 5.56, 1.85nM, respectively. A control group culture medium is prepared by using DMSO with the same volume instead of a stock solution; taking out the cell culture plate from the incubator, removing the culture medium, and adding the working solution and the control culture medium; the plates were returned to the incubator at 37 ℃ with 5% CO2The culture was conditioned for 24 hours.
After 24 hours of treatment, the cell culture plate was removed from the incubator, the medium was removed, and the cells were rinsed 3 times with pre-cooled (4 ℃) PBS; add 200. mu.L of pancreatin per well (preheat to 37 ℃) and gently shake to allow the pancreatin to cover the plate bottom evenly. The plate was returned to the incubator and incubated until the cells were detached from the plate bottom. Digestion was stopped by adding 1mL of medium. After gently blowing and beating for several times by using a pipette, sucking all substances in the holes into a centrifugal tube of 1.5mL RNase-free, and centrifuging for 5 minutes at 200 Xg; the supernatant was removed and a cell sample was collected.
② extraction and purification of RNA from cell samples
Cell lysis: fresh RNA lysate (1mL lysate plus 10. mu.L 2-mercaptoethanol); add 600. mu.L of lysate to the cell sample; vortexing vigorously for 1-2 min to lyse the cells completely; the cell lysate was centrifuged at 12,000 Xg for 5 minutes; the supernatant was transferred to a 1.5mL RNase-free centrifuge tube.
RNA extraction and purification: adding an equal amount of 70% ethanol to the cell lysate; violently shaking the centrifugal tube, fully mixing, and dispersing particle precipitates possibly formed after adding ethanol as much as possible; the adsorption column was placed on a collection tube and the mixture was transferred to the adsorption column. Transferring at most 700 μ L each time; centrifuge at 12,000 Xg for 15 seconds at room temperature. Discarding the solution in the collecting tube, and putting the adsorption column on the collecting tube again; the remaining mixture was transferred in its entirety to an adsorption column. Adding 700 μ L of eluent I into the adsorption column; centrifuge at 12,000 Xg for 15 seconds at room temperature. Placing the adsorption column on a new collection tube; adding 500 μ L of eluent II into the adsorption column; centrifuge at 12,000 Xg for 15 seconds at room temperature. Discarding the solution in the collecting tube, and putting the adsorption column on the collecting tube again; adding 500 μ L of eluent II into the adsorption column; centrifuging at room temperature of 12,000 Xg for 1-2 min, and placing the adsorption column on an RNA collection tube; adding 50 μ L RNase-free water to the central position of the adsorption column, and incubating for 1 min at room temperature; the RNA was eluted into collection tubes by centrifugation at 14,000 Xg for 2 minutes at room temperature.
The concentration and mass of the extracted RNA were measured. RNA was stored at-80 ℃.
③ reverse transcription of RNA into cDNA
The RNA extracted in the second step was incubated at 70 ℃ for 5 minutes to denature the RNA. After treatment, the samples were placed on ice;
RNA samples were diluted to 200 ng/. mu.L with RNAse-free water; mu.L of the reverse transcription solution was prepared according to the following table and mixed with 10. mu.L of denatured RNA. The total amount of RNA in the reverse transcription reaction was 2. mu.g. During the experiment, all reagents were placed on ice.
Figure BDA0001739294150000311
Reverse transcription was performed on a G-Storm GS1thermal cycler PCR thermal cycler. The reverse transcription process was set up as follows: 10 minutes at 25 ℃ → 120 minutes at 37 ℃ → 5 minutes at 85 ℃ → 4 ℃. infinity. The reverse transcription product (cDNA) was stored at-20 ℃.
Sample qPCR experiment
And selecting proper cDNA concentration to perform qPCR experiment of the sample according to the qPCR amplification efficiency. And a third step of taking 10 mu L of cDNA sample obtained by reverse transcription and adding 60 mu L of RNase-free water to dilute the cDNA sample by 7 times.
mu.L of the reaction mixture was prepared according to the following table, and 20. mu.L was pipetted into a 96-well PCR reaction plate, and 3 replicates (7. mu.L of 100ng in each reaction well) of the cDNA sample were added.
Figure BDA0001739294150000312
Figure BDA0001739294150000321
qPCR was performed on ABI7500 real-time quantitative PCR instrument programmed as follows: 50 ℃ for 2 min → 95 ℃ for 10 min → 95 ℃ for 15 sec → 60 ℃ for 60 sec, with 40 cycles set between 95 ℃ for 15 sec and 60 ℃ for 60 sec.
(3) Experimental results and conclusions:
TABLE 3 results of the measurement of BSEP, SHP, CYP7A1mRNA at various concentrations
Figure BDA0001739294150000322
TABLE 4 results of measurement of the relative expression amount of BSEP mRNA in HepG2 cells treated with the compound of the present invention
Figure BDA0001739294150000323
TABLE 5 results of measurement of the relative expression amount of BSEP mRNA in HepG2 cells treated with the compound of the present invention
Figure BDA0001739294150000324
TABLE 6 results of measurement of the relative expression amount of BSEP mRNA in HepG2 cells treated with the compound of the present invention
Figure BDA0001739294150000325
TABLE 7 results of measurement of the relative expression amount of BSEP mRNA in HepG2 cells treated with the compound of the present invention
Figure BDA0001739294150000326
TABLE 8 results of measurement of the relative expression amount of BSEP mRNA in HepG2 cells treated with the compound of the present invention
Figure BDA0001739294150000327
TABLE 9 results of measurement of the relative expression amount of BSEP mRNA in HepG2 cells treated with the compound of the present invention
Figure BDA0001739294150000331
Remarking: relative expression of mRNA (% 100 × [ mRNA (test substance)/mRNA (PX-102 or PX-104) ]
As shown in Table 3, the compound 1 of the present invention has good agonistic effect on BSEP (EC50nM) and SHP (EC50nM), and good inhibitory effect on CYP7A 1; as shown in tables 4-9, the compounds of the present invention have a good expression effect on BSEP mRNA in HepG2 cells, and have important significance in the treatment of non-alcoholic fatty liver disease.
Experimental example 4: beagle canine pharmacokinetic experiments with the compounds of the invention
And (3) testing the sample: the compounds of the examples of the invention, self-made, the chemical names and the preparation methods thereof are shown in the preparation examples of the compounds.
Preparation of a test solution of the compound of the embodiment of the invention:
the compound of the present example was administered intravenously (iv) in the formulation of 5% DMSO + 10% PEG400+ 85% (28% HP- β -CD).
Preparation of 28% HP-beta-CD: weighing 2.8g of HP-beta-CD (hydroxypropyl beta cyclodextrin), adding a small amount of sterile water for injection, dissolving by ultrasonic, adding sterile water for injection to constant volume of 10ml, and mixing by vortex.
Weighing 18.10mg of the compound of the embodiment of the invention, adding 1.65mL of DMSO, carrying out vortex dissolution, adding 3.3mL of PEG400, carrying out vortex mixing, adding 28.05mL of 28% HP-beta-CD, carrying out vortex mixing, and carrying out heat preservation at 50 ℃ for 20 minutes to obtain an intravenous injection administration solution with the concentration of 0.5 mg/mL.
Oral administration (po) of the compounds of the examples of the invention was formulated with 0.1% tween 80+ 2% HPC.
Preparation of 0.1% tween 80+ 2% HPC: weighing 2g of HPC (hydroxypropyl cellulose), adding purified water to 100ml, continuously stirring to dissolve, adding 0.1ml of Tween 80, stirring to dissolve, and shaking uniformly to obtain a blank solvent of 0.1% Tween 80+ 2% of HPC.
Precisely weighing 35.04mg of the compound of the embodiment of the invention, adding 64mL of the solvent, grinding by a tissue grinder for 5 minutes to uniformly disperse, and grinding into fine and uniform suspension to obtain the intragastric administration suspension solution with the concentration of 0.5 mg/mL.
Experimental methods
The test liquid medicine is administrated according to the following method:
Figure BDA0001739294150000341
blood sampling time points:
iv: 0.083h,0.25h,0.5h,1h,2h,4h,6h,8h,24h and 48h after the medicine is taken.
po: 0.167h,0.5h,1h,2h,4h,6h,8h,24h,48h and 72h after the medicine is taken.
Plasma collection: the animals were fixed, and about 400. mu.l of blood was collected through the forelimb vein at each time point and placed in the presence of EDTA-K2In an anticoagulation tube, the blood sample is evenly mixed, centrifuged for 6 minutes at 8000 rpm at 4 ℃ to separate plasma, and frozen in a refrigerator at-80 ℃.
Plasma sample analysis:
plasma sample analysis protein precipitation method: 20 mul of plasma was aspirated, 200 mul of acetonitrile solution containing 50ng/ml of Tolbutamide (Tolbutamide) as an internal standard was added, 10 minutes of vortexing was performed at 1000 rpm, 20 minutes of centrifugation was performed at 4000 rpm, 100 mul of supernatant was aspirated, 100 mul of water was added, vortexing was performed, and LC-MS/MS analysis was performed.
The experimental results are as follows:
the compound of the embodiment of the invention has high exposure, longer half-life and bioavailability of more than 50%.
Detailed Description
The present invention will be described in further detail with reference to the following examples. It should not be understood that the scope of the above-described subject matter of the present invention is limited to the following examples. All the technologies realized based on the above contents of the present invention belong to the scope of the present invention.
Preparation example 1: (E) preparation of (E) -2, 6-dichlorobenzaldehyde oxime
Figure BDA0001739294150000342
2, 6-Dichlorobenzaldehyde (25g,0.14mol) was dissolved in ethanol (200mL), hydroxylamine hydrochloride (11g,0.16mol) was added slowly in portions, and after the addition, NaOH solution (6.4g,0.16mol, dissolved in 100mL of water) was added and heated to 90 ℃ for reaction for 24 hours. After the reaction was completed, the solvent was removed from the reaction solution under reduced pressure, and the solid was washed with water (200mL) and dried to obtain the title compound (25.9 g) in 97% yield.
Preparation example 2: preparation of (Z) -2, 6-dichloro-N-hydroxybenzylidene chloride
Figure BDA0001739294150000351
(E) -2, 6-Dichlorobenzaldehyde oxime (25.9g,0.136mol) was dissolved in DMF (300mL), N-chlorosuccinimide (18.2g,0.136mol) was added gradually in portions, and after the addition was completed, the reaction mixture was stirred at 25 ℃ for one hour, and poured into water (500 mL). Extraction was performed with ethyl acetate (500mL), and the organic phase was washed with water (200mL) and saturated sodium chloride solution (200mL), dried over anhydrous sodium sulfate, and the solvent was removed to give the title compound 28g, which was carried on to the next step without purification.
Preparation example 3: preparation of 5-cyclopropyl-3- (2, 6-dichlorophenyl) isoxazole-4-carboxylic acid ethyl ester
Figure BDA0001739294150000352
(Z) -2, 6-dichloro-N-hydroxybenzylidene chloride (28g,0.125mol) was dissolved in triethylamine (100mL), and ethyl 3-cyclopropyl-3-oxopropionate (19.5g,0.125mol) was added to the solution to react at 25 ℃ for 12 hours. After the reaction was completed, the solvent was removed under reduced pressure, and column chromatography (PE: EA ═ 10:1) was performed to obtain 24.4g of the title compound in 55.1% yield in two steps.
Preparation example 4: preparation of (5-cyclopropyl-3- (2, 6-dichlorophenyl) isoxazol-4-yl) methanol
Figure BDA0001739294150000353
Dissolving 5-cyclopropyl-3- (2, 6-dichlorophenyl) isoxazole-4-carboxylic acid ethyl ester (20g,61.3mmol) in tetrahydrofuran (300mL), cooling in an ice bath, adding a toluene solution (1.5mol/L,123mL,0.184mol) of diisobutylaluminum hydride (DIBAL-H), heating to 25 ℃ to react for 12 hours after the addition is finished, cooling in an ice bath, adding a saturated ammonium chloride solution (200mL) to quench the reaction, extracting with ethyl acetate (500mL), washing the organic phase with a saturated ammonium chloride solution (200mL) and a saturated sodium chloride solution (200mL), drying over anhydrous sodium sulfate, and removing the solvent to obtain the title compound 16g with a yield of 92.0%.
Preparation example 5-1: preparation of 4- (bromomethyl) -5-cyclopropyl-3- (2, 6-dichlorophenyl) isoxazole
Figure BDA0001739294150000354
(5-cyclopropyl-3- (2, 6-dichlorophenyl) isoxazol-4-yl) methanol (5.0g,17.6mmol) was added to dichloromethane (30mL), triethylamine (1.78g,17.6mmol) was added, then phosphorus tribromide (4.77g,17.6mmol) was added, and the reaction was carried out at 30 ℃ for 2 hours. After completion of the reaction, the solvent was removed, and column chromatography (PE: EA ═ 10:1) was performed to obtain 4.3g of the title compound in 70.4% yield.
Preparation examples 5 to 2: preparation of 4- (chloromethyl) -5-cyclopropyl-3- (2, 6-dichlorophenyl) isoxazole
Figure BDA0001739294150000361
(5-cyclopropyl-3- (2, 6-dichlorophenyl) isoxazol-4-yl) methanol (5.0g,17.6mmol) was added to dichloromethane (30mL), triethylamine (1.78g,17.6mmol) was added, then phosphorus trichloride (2.42g,17.6mmol) was added, and the reaction was carried out at 30 ℃ for 2 hours. After completion of the reaction, the solvent was removed, and column chromatography (PE: EA ═ 10:1) was performed to obtain the title compound 3.60g in 67.7% yield.
Example 12- (2-chloro-4- ((5-cyclopropyl-3- (2, 6-dichlorophenyl) isoxazol-4-yl) methoxy) phenyl) Preparation of chroman-6-carboxylic acid (Compound 1)
Figure BDA0001739294150000362
(1) Preparation of (E) -1- (5-bromo-2-hydroxyphenyl) -3- (2-chloro-4-hydroxyphenyl) propan-2-en-1-one
Figure BDA0001739294150000363
1- (5-bromo-2-hydroxyphenyl) ethanone (40.7g,189.26mmol) and 2-chloro-4-hydroxybenzaldehyde (30g,191.61mmol) were dissolved in ethanol (1000mL) and potassium hydroxide (42.56g,760mmol) was added. The reaction solution was stirred at room temperature for 24h, quenched (5L) with ice water, adjusted to pH 2 with 2mol/L HCl, and the solid precipitated and filtered to give the title compound (42g crude).
(2) Preparation of 6-bromo-2- (2-chloro-4-hydroxyphenyl) chroman-4-one
Figure BDA0001739294150000371
(E) -1- (5-bromo-2-hydroxyphenyl) -3- (2-chloro-4-hydroxyphenyl) propan-2-en-1-one (42g crude) was dissolved in acetic acid (500mL), concentrated hydrochloric acid (100mL) was added, and the reaction was heated to reflux for 4 days. Cooled to room temperature, diluted with ethyl acetate (2L), washed with saturated sodium chloride solution (500 mL. times.4), dried over anhydrous sodium sulfate and concentrated in vacuo to give the title compound (34g crude).
(3) Preparation of 4- (6-bromo-chroman-2-yl) -3-chlorophenol
Figure BDA0001739294150000372
Mercury dichloride (39.4g,0.1449mol) is dissolved in 5mol/L hydrochloric acid (500mL), zinc powder (92.7g,1.449mol) is added in portions, the temperature is controlled to be less than 20 ℃, after the addition, the mixture is stirred at room temperature for 30 minutes, liquid is poured off, 5mol/L hydrochloric acid (500mL) is added to the solid, the mixture is stirred at room temperature for 5 minutes, and the liquid is poured off. To the residual solid were added 5mol/L hydrochloric acid (1000mL) and a toluene solution (500mL) of 6-bromo-2- (2-chloro-4-hydroxyphenyl) chroman-4-one (34g of crude product), and after completion of the addition, the mixture was heated to 80 ℃ to react for 4 hours, cooled to room temperature, extracted with ethyl acetate (400mL × 2), combined with the organic phase, dried over anhydrous sodium sulfate, concentrated, and the crude product was purified by silica gel column chromatography (ethyl acetate: petroleum ether ═ 1:5) to give the title compound 11g, which was 16.9% in 3 steps.
(4) Preparation of 2- (2-chloro-4-hydroxyphenyl) chroman-6-carbonitrile
Figure BDA0001739294150000373
4- (6-bromo-chroman-2-yl) -3-chlorophenol (11g,32.5mmol) was dissolved in N-methylpyrrolidone (200mL), zinc cyanide (2g,17mmol) and tetrakis (triphenylphosphine) palladium (1.87g,1.62mmol) were added, and the mixture was heated to 110 ℃ under nitrogen for reaction overnight. Cooled to room temperature, diluted with ethyl acetate (500mL), washed with saturated sodium chloride solution (200mL × 3), dried over anhydrous sodium sulfate, concentrated and the crude product purified by silica gel column chromatography (ethyl acetate: petroleum ether ═ 1:5) to give the title compound 3.1g, 33% yield.
(5) Preparation of methyl 2- (2-chloro-4-hydroxyphenyl) chroman-6-carboxylate
Figure BDA0001739294150000381
2- (2-chloro-4-hydroxyphenyl) -chroman-6-carbonitrile (3.1g,10.85mmol) was dissolved in methanol (100mL), concentrated sulfuric acid (10mL) was added dropwise with stirring at room temperature, and after completion of addition, the mixture was heated to reflux for 4 days. After cooling to room temperature, ethyl acetate (500mL) was added for dilution, washed with saturated sodium chloride solution (200mL × 3), dried over anhydrous sodium sulfate, concentrated in vacuo, and the crude product was purified by silica gel column chromatography (EA: PE ═ 1:5) to give the title compound (2g crude product).
(6) Preparation of methyl 2- (2-chloro-4- ((5-cyclopropyl-3- (2, 6-dichlorophenyl) isoxazol-4-yl) methoxy) phenyl) chroman-6-carboxylate
Figure BDA0001739294150000382
Methyl 2- (2-chloro-4-hydroxyphenyl) chroman-6-carboxylate (2g crude) was dissolved in N, N-dimethylformamide (30mL), and a solution of sodium iodide (1.89g,12.60mmol), potassium carbonate (2.61g,18.90mmol) and 4- (chloromethyl) -5-cyclopropyl-3- (2, 6-dichlorophenyl) isoxazole (2.27g,7.50mmol) in N, N-dimethylformamide (20mL) was added, and the mixture was heated to 60 ℃ and stirred overnight. Ethyl acetate (200mL) was added for dilution, washed with a saturated sodium chloride solution (50mL × 3), dried over anhydrous sodium sulfate, concentrated in vacuo, and the crude product was purified by silica gel column chromatography (ethyl acetate: petroleum ether ═ 1:5) to give the product (1.2g crude product).
(7) Preparation of 2- (2-chloro-4- ((5-cyclopropyl-3- (2, 6-dichlorophenyl) isoxazol-4-yl) methoxy) phenyl) chroman-6-carboxylic acid
Figure BDA0001739294150000383
Methyl 2- (2-chloro-4- ((5-cyclopropyl-3- (2, 6-dichlorophenyl) isoxazol-4-yl) methoxy) phenyl) chroman-6-carboxylate (1.2g,2.00mmol) was dissolved in methanol/water (100mL/20mL), lithium hydroxide monohydrate (252mg,6.00mmol) was added, and stirring was carried out overnight at room temperature. The reaction solution was diluted with water (100mL), the pH was adjusted to 3 with aqueous citric acid, ethyl acetate (500mL) was added, the layers were separated, the organic phase was washed with saturated sodium chloride solution (100 mL. times.2), dried over anhydrous sodium sulfate, concentrated in vacuo, and the crude product was purified by preparative HPLC to give the title compound 45mg, 0.73% yield over 3 steps.
Molecular formula C29H22Cl3NO5Molecular weight 570.85LC-MS (negative, m/z):570(M + H)+
1H-NMR(300MHz,CD3OD)δ:7.82-7.77(m,2H),7.54-7.38(m,4H),6.90-6.79(m,3H),4.94-4.85(m,2H),3.09-2.98(m,1H),2.86-2.81(m,2H),2.37-2.25(m,2H),1.97-1.90(m,1H),1.30-1.19(m,4H).
Example 22- (2-chloro-4- ((5-cyclopropyl-3- (2, 6-dichlorophenyl) isoxazol-4-yl) methoxy) benzene 2, 3-dihydrobenzofuran-5-carboxylic acid (Compound 2)
Figure BDA0001739294150000391
(1) Preparation of 2-chloro-4- ((tetrahydro-2H-pyran-2-yl) oxy) benzaldehyde
Figure BDA0001739294150000392
2-chloro-4-hydroxybenzaldehyde (1.0g,6.38mmol), 3, 4-dihydropyran (1.1g,13.1mmol) and pyridinium p-toluenesulfonate (0.8g,3.18mmol) were added in this order to methylene chloride (100mL) and reacted at 25 ℃ for 4 hours, after completion of the reaction, concentrated, and the residue was subjected to silica gel column chromatography (petroleum ether: ethyl acetate ═ 10:1) to give the title compound 1.2g in 78.1% yield.
(2) Preparation of methyl 2- (2-chloro-4- ((tetrahydro-2H-pyran-2-yl) oxy) phenyl) -2, 3-dihydrobenzofuran-5-carboxylate
Figure BDA0001739294150000393
2-chloro-4- ((tetrahydro-2H-pyran-2-yl) oxy) benzaldehyde (1.1g,4.57mmol), methyl 3-methyl-4-nitrobenzoate (1.07g,5.48mmol), N, N-diisopropylethylamine (1.24g,9.6mmol) and tetrabutylammonium fluoride (8.2mL,8.2mmol,1M tetrahydrofuran solution) were sequentially added to tetrahydrofuran (10mL) to react at 80 ℃ for 16 hours, the solvent was removed after completion of the reaction, and the residue was subjected to silica gel column chromatography (petroleum ether: ethyl acetate ═ 20:1) to give the title compound 1.1g, yield 61.9%.
(3) Preparation of methyl 2- (2-chloro-4-hydroxyphenyl) -2, 3-dihydrobenzofuran-5-carboxylate
Figure BDA0001739294150000401
Methyl 2- (2-chloro-4- ((tetrahydro-2H-pyran-2-yl) oxy) phenyl) -2, 3-dihydrobenzofuran-5-carboxylate (1.1g,2.83mmol) and p-toluenesulfonic acid (0.2g,1.16mmol) were added to methanol (20mL) and reacted at 25 ℃ for 2 hours, after completion of the reaction, the solvent was removed and the residue was subjected to silica gel column chromatography (petroleum ether: ethyl acetate ═ 5:1) to give the title compound 0.54g in 62.6% yield.
(4) Preparation of methyl 2- (2-chloro-4- ((5-cyclopropyl-3- (2, 6-dichlorophenyl) isoxazol-4-yl) methoxy) phenyl) -2, 3-dihydrobenzofuran-5-carboxylate
Figure BDA0001739294150000402
The procedure was as in example 1, reaction step (6).
(5) Preparation of 2- (2-chloro-4- ((5-cyclopropyl-3- (2, 6-dichlorophenyl) isoxazol-4-yl) methoxy) phenyl) -2, 3-dihydrobenzofuran-5-carboxylic acid
Figure BDA0001739294150000403
The procedure was as in example 1, reaction step (7).
Molecular formula C28H20Cl3NO5Molecular weight 556.8LC-MS (M/z):556.2(M + H)+
1H-NMR(400MHz,MeOD)δ:7.90(d,J＝4.4Hz,1H),7.86(s,1H),7.42-7.51(m,3H),7.27(d,J＝8.8Hz,1H),6.89(d,J＝4.4Hz,1H),6.86(s,1H),6.73(d,J＝8.8Hz,1H),6.06(t,J＝8.8Hz,1H),4.90(s,2H),3.74-3.81(m,1H),3.01-3.07(m,1H),2.32(t,J＝6.0Hz,1H),1.18-1.21(m,4H).
Example 32- (2-chloro-4- ((5-cyclopropyl-3- (2, 6-dichlorophenyl) isoxazol-4-yl) methoxy) benzene Radical) -2, 3-dihydrobenzo [ b][1,4]Preparation of Dioxin-6-carboxylic acid (Compound 3)
Figure BDA0001739294150000411
(1) Preparation of 2-chloro-4- ((tetrahydro-2H-pyran-2-yl) oxy) benzaldehyde
Figure BDA0001739294150000412
2-chloro-4-hydroxybenzaldehyde (2g,12.77mmol) and 3, 4-2H-dihydropyran (1.5g,17.83mmol) were added to dichloromethane (50mL), pyridinium p-toluenesulfonate (0.3g,1.19mmol) was added, and the reaction was stirred at 25 ℃ for 6 hours. The reaction solution was concentrated, and the resulting crude product was purified by silica gel column chromatography (petroleum ether: ethyl acetate: 50:1) to obtain 2.6g of the title compound in 83.9% yield.
(2) Preparation of 2- (3-chloro-4-vinylphenoxy) tetrahydro-2H-pyran
Figure BDA0001739294150000413
Methyltriphenylphosphine bromide (4g,11.17mmol) and potassium tert-butoxide (1.5g,13.37mmol) were added to tetrahydrofuran (80mL), cooled to 0 ℃ under nitrogen, a solution of 2-chloro-4- ((tetrahydro-2H-pyran-2-yl) oxy) benzaldehyde (2.5g,10.4mmol) in tetrahydrofuran (20mL) was added and the reaction stirred at 25 ℃ for 3 hours. Water (100mL) and ethyl acetate (100mL) were added, liquid separation was performed, the aqueous phase was extracted with ethyl acetate (100mL × 3), the organic phases were combined, dried over anhydrous sodium sulfate, filtered, concentrated, and the resulting crude product was purified by silica gel column chromatography (petroleum ether: ethyl acetate ═ 50:1) to obtain 2g of the title compound in 80.6% yield.
(3) Preparation of 1- (2-chloro-4- ((tetrahydro-2H-pyran-2-yl) oxy) phenyl) ethane-1, 2-diol
Figure BDA0001739294150000421
2- (3-chloro-4-vinylphenoxy) tetrahydro-2H-pyran (1.6g,6.7mmol) was dissolved in a mixed solvent of tert-butanol (20mL) and water (20mL), cooled to 0 ℃ and AD-mix-beta (2g) was added, and the reaction was stirred at 25 ℃ for 6 hours. Water (50mL) and ethyl acetate (50mL) were added, liquid separation was performed, the aqueous phase was extracted with ethyl acetate (50mL × 3), the organic phases were combined, dried over anhydrous sodium sulfate, filtered, concentrated, and the resulting crude product was purified by silica gel column chromatography (petroleum ether: ethyl acetate ═ 3:1) to obtain the title compound 1.2g, yield 65.7%.
(4) Preparation of 2- ((tert-butyldimethylsilyl) oxy) -1- (2-chloro-4- ((tetrahydro-2H-pyran-2-yl) oxy) phenyl) ethanol
Figure BDA0001739294150000422
1- (2-chloro-4- ((tetrahydro-2H-pyran-2-yl) oxy) phenyl) ethane-1, 2-diol (1g,3.67mmol) and imidazole (0.5g,7.34mmol) were dissolved in dichloromethane (20mL), tert-butyldimethylchlorosilane (0.7g,4.64mmol) was added, and the reaction was stirred at 25 ℃ for 16 hours. The reaction solution was concentrated, and the resulting crude product was purified by silica gel column chromatography (petroleum ether: ethyl acetate: 10:1) to obtain 1.2g of the title compound in 84.7% yield.
(5) Preparation of methyl 4- (2- ((tert-butyldimethylsilyl) oxy) -1- (2-chloro-4- ((tetrahydro-2H-pyran-2-yl) oxy) phenyl) ethoxy) -3-iodobenzoate
Figure BDA0001739294150000423
2- ((tert-butyldimethylsilyl) oxy) -1- (2-chloro-4- ((tetrahydro-2H-pyran-2-yl) oxy) phenyl) ethanol (1g,2.58mmol), methyl 4-hydroxy-3-iodobenzoate (0.7g,2.52mmol) and triphenylphosphine (0.8g,3.05mmol) were dissolved in tetrahydrofuran (20mL), cooled to 0 deg.C, diethyl azodicarboxylate (0.6g,3.44mmol) was added under nitrogen, and the reaction was stirred at 25 deg.C for 16 hours. The reaction solution was concentrated, and the resulting crude product was purified by silica gel column chromatography (petroleum ether: ethyl acetate: 5:1) to obtain 0.9g of the title compound in 55.2% yield.
(6) Preparation of methyl 4- (1- (2-chloro-4- ((tetrahydro-2H-pyran-2-yl) oxy) phenyl) -2-hydroxyethoxy) -3-iodobenzoate
Figure BDA0001739294150000431
4- (2- ((tert-butyldimethylsilyl) oxy) -1- (2-chloro-4- ((tetrahydro-2H-pyran-2-yl) oxy) phenyl) ethoxy) -3-iodobenzoate (0.9g,1.39mmol) was dissolved in tetrahydrofuran (20mL), tetrabutylammonium fluoride trihydrate (0.44g,1.39mmol) was added, and the reaction was stirred at 25 ℃ for 2 hours. The reaction solution was concentrated, and the resulting crude product was purified by silica gel column chromatography (petroleum ether: ethyl acetate: 5:1) to obtain 0.7g of the title compound in 94.6% yield.
(7) Preparation of methyl 2- (2-chloro-4- ((tetrahydro-2H-pyran-2-yl) oxy) phenyl) -2, 3-dihydrobenzo [ b ] [1,4] dioxine-6-carboxylate
Figure BDA0001739294150000432
Dissolving methyl 4- (1- (2-chloro-4- ((tetrahydro-2H-pyran-2-yl) oxy) phenyl) -2-hydroxyethoxy) -3-iodobenzoate (0.6g,1.13mmol) in 1, 4-dioxane (10mL), adding sodium tert-butoxide (0.11g,1.14mmol), 1, 10-phenanthroline (25mg,0.13mmol), cuprous iodide (24mg,0.13mmol), heating to 100 ℃ under the protection of nitrogen, and stirring for reaction for 16 hours. The reaction solution was concentrated, and the resulting crude product was purified by silica gel column chromatography (petroleum ether: ethyl acetate: 5:1) to obtain the title compound 0.25g in 54.4% yield.
(8) Preparation of methyl 2- (2-chloro-4-hydroxyphenyl) -2, 3-dihydrobenzo [ b ] [1,4] dioxine-6-carboxylate
Figure BDA0001739294150000441
2- (2-chloro-4- ((tetrahydro-2H-pyran-2-yl) oxy) phenyl) -2, 3-dihydrobenzo [ b ] [1,4] dioxin-6-carboxylate (0.25g,0.62mmol) was dissolved in anhydrous ethanol (5mL), pyridinium p-toluenesulfonate (20mg,0.08mmol) was added, and the reaction was stirred by heating to 80 ℃ for 2 hours. The reaction solution was concentrated, and the resulting crude product was purified by silica gel column chromatography (petroleum ether: ethyl acetate ═ 2:1) to obtain 0.16g of the title compound in 80.0% yield.
(9) Preparation of 4- (bromomethyl) -5-cyclopropyl-3- (2, 6-dichlorophenyl) isoxazole
Figure BDA0001739294150000442
(5-cyclopropyl-3- (2, 6-dichlorophenyl) isoxazol-4-yl) methanol (0.5g,1.76mmol) and triphenylphosphine (0.6g,2.29mmol) were dissolved in tetrahydrofuran (5mL), cooled to 0 deg.C, carbon tetrabromide (0.76g,2.29mmol) was added, and the reaction was stirred at 25 deg.C for 2 hours. The reaction solution was concentrated, and the resulting crude product was purified by silica gel column chromatography (petroleum ether: ethyl acetate: 5:1) to obtain the title compound (0.5 g) in 81.9% yield.
(10) Preparation of methyl 2- (2-chloro-4- ((5-cyclopropyl-3- (2, 6-dichlorophenyl) isoxazol-4-yl) methoxy) phenyl) -2, 3-dihydrobenzo [ b ] [1,4] dioxane-6-carboxylate
Figure BDA0001739294150000443
Methyl 2- (2-chloro-4-hydroxyphenyl) -2, 3-dihydrobenzo [ b ] [1,4] dioxine-6-carboxylate (0.12g,0.37mmol) and 4- (bromomethyl) -5-cyclopropyl-3- (2, 6-dichlorophenyl) isoxazole (0.13g,0.37mmol) were dissolved in N, N-dimethylformamide (5mL), potassium carbonate (0.1g,0.72mmol) was added, and the reaction was stirred at 80 ℃ for 16 hours. The reaction solution was concentrated, and the resulting crude product was purified by silica gel column chromatography (petroleum ether: ethyl acetate ═ 2:1) to obtain 0.15g of the title compound in 68.2% yield.
(11) Preparation of 2- (2-chloro-4- ((5-cyclopropyl-3- (2, 6-dichlorophenyl) isoxazol-4-yl) methoxy) phenyl) -2, 3-dihydrobenzo [ b ] [1,4] dioxine-6-carboxylic acid
Figure BDA0001739294150000451
Methyl 2- (2-chloro-4- ((5-cyclopropyl-3- (2, 6-dichlorophenyl) isoxazol-4-yl) methoxy) phenyl) -2, 3-dihydrobenzo [ b ] [1,4] dioxine-6-carboxylate (0.12g,0.2mmol) was dissolved in tetrahydrofuran (3mL) and methanol (3mL), and a solution of lithium hydroxide monohydrate (12mg,0.28mmol) in water (1mL) was added and the reaction was stirred at 25 ℃ for 16 hours. The reaction solution was concentrated, and the resulting crude product was purified by silica gel column chromatography (dichloromethane: methanol 20:1) and Combiflash (acetonitrile/water 10% -35% -80%) to give the title compound 20mg in 17.1% yield.
The molecular formula is as follows: c28H20Cl3NO6Molecular weight: 572.8 LC-MS (M/z):572.1,574.1(M + H)+
1H-NMR(400MHz,MeOD)δ:7.41-7.61(m,6H),6.95(d,J＝8.4Hz,1H),6.89(s,1H),6.85(dd,J1＝8.8Hz,J＝2.6Hz,1H),5.42(dd,J1＝8.4Hz,J＝2.0Hz,1H),4.96(s,2H),4.45(dd,J1＝11.6Hz,J＝2.4Hz,1H),3.90-3.95(m,1H),2.32-2.42(m,1H),1.22-1.24(m,4H).
Example 42- (2-chloro-4- ((5-cyclopropyl-3- (2, 6-dichlorophenyl) isoxazol-4-yl) methoxy) benzene Preparation of 1,2,3, 4-tetrahydroisoquinoline-6-carboxylic acid (Compound 4)
Figure BDA0001739294150000452
(1) Preparation of 4- ((4-bromo-3-chlorophenoxy) methyl) -5-cyclopropyl-3- (2, 6-dichlorophenyl) isoxazole
Figure BDA0001739294150000453
4-bromo-3-chlorophenol (11g,53.02mmol) was dissolved in N, N-dimethylformamide (150mL), and a solution of sodium iodide (16g,106.74mmol), potassium carbonate (44g,318.36mmol) and 4- (chloromethyl) -5-cyclopropyl-3- (2, 6-dichlorophenyl) isoxazole (17.6g,58.17mmol) in N, N-dimethylformamide (50mL) was added, and the mixture was warmed to 60 ℃ and stirred overnight. Cooled to room temperature, diluted with ethyl acetate (1L), washed with saturated sodium chloride solution (200mL × 3), dried over anhydrous sodium sulfate, concentrated in vacuo, and the crude product purified by silica gel column chromatography (ethyl acetate: petroleum ether ═ 1:10) to give the title compound (19.6g, 78.5% yield over 2 steps).
(2) Preparation of methyl 2- (2-chloro-4- ((5-cyclopropyl-3- (2, 6-dichlorophenyl) isoxazol-4-yl) methoxy) phenyl) -1,2,3, 4-tetrahydroisoquinoline-6-carboxylate
Figure BDA0001739294150000461
4- ((4-bromo-3-chlorophenoxy) methyl) -5-cyclopropyl-3- (2, 6-dichlorophenyl) isoxazole (2.1g,4.43mmol) was dissolved in 1, 4-dioxane (60mL), 1,2,3, 4-tetrahydroisoquinoline-6-carboxylic acid methyl ester hydrochloride (1g,4.39mmol), cesium carbonate (4.3g,13.20mmol) and Brettphos-Pd (100mg,0.11mmol) were added, and the mixture was heated to reflux for 3 days. The reaction mixture was added with water (20mL) and ethyl acetate (300mL), washed with saturated sodium chloride solution (50mL × 3), the organic phase was dried over anhydrous sodium sulfate, concentrated in vacuo, and the residue was purified by silica gel column chromatography (ethyl acetate: petroleum ether ═ 1:5) to give the title compound (180mg crude).
(3) Preparation of 2- (2-chloro-4- ((5-cyclopropyl-3- (2, 6-dichlorophenyl) isoxazol-4-yl) methoxy) phenyl) -1,2,3, 4-tetrahydroisoquinoline-6-carboxylic acid
Figure BDA0001739294150000462
Referring to the preparation method of step (7) of example 1, methyl 2- (2-chloro-4- ((5-cyclopropyl-3- (2, 6-dichlorophenyl) isoxazol-4-yl) methoxy) phenyl) -1,2,3, 4-tetrahydroisoquinoline-6-carboxylate (180mg crude product) was added to give the title compound (8mg, 0.3% yield over 2 steps).
Molecular formula C29H23Cl3N2O4Molecular weight 569.86LC-MS (positive, m/z):571(M+H)+
1H-NMR(300MHz,CD3OCD3)δ:7.86-7.81(m,2H),7.60-7.52(m,3H),7.28-7.25(m,1H),7.16(d,J＝8.7Hz,1H),6.95(d,J＝2.7Hz,1H),6.82(dd,J1＝3Hz,J2＝8.7Hz,1H),4.96(s,2H),4.21(s,2H),3.30-3.26(m,2H),3.08-2.95(m,2H),2.47-2.38(m,1H),1.23-1.17(m,4H).
example 52- (2-chloro-4- ((5-cyclopropyl-3- (2, 6-dichlorophenyl) isoxazol-4-yl) methoxy) benzene Preparation of 1,2,3, 4-tetrahydroisoquinoline-7-carboxylic acid (Compound 5)
Figure BDA0001739294150000471
(1) Preparation of methyl 4- (2-aminoethyl) benzoate
Figure BDA0001739294150000472
Methyl 4- (cyanomethyl) benzoate (6.0g,34.2mmol) was dissolved in saturated hydrogen chloride solution (10mL) of dioxane, methanol (10mL), palladium on carbon (10%) (0.6g) were added, hydrogen gas was replaced, the mixture was reacted at 25 ℃ for 24 hours, filtered, the cake was washed with methanol (20mL), and the filtrate was concentrated and subjected to column chromatography (dichloromethane: methanol 10:1) to obtain the title compound (1.5g, yield 24.6%).
(2) Preparation of methyl 4- (2- (2,2, 2-trifluoroacetylamino) ethyl) benzoate
Figure BDA0001739294150000473
Methyl 4- (2-aminoethyl) benzoate (1.5g,8.4mmol) was added to trifluoroacetic anhydride (10mL) and the reaction stirred at 25 ℃ for 3 hours, then the reaction solution was poured into 20mL of ice water and stirred for 30 minutes, the precipitated solid was filtered, washed with n-hexane (10mL) and dried to give the title compound (1.5g, 65.2% yield).
(3) Preparation of methyl 2- (2,2, 2-trifluoroacetyl) -1,2,3, 4-tetrahydroisoquinoline-7-carboxylate
Figure BDA0001739294150000474
To a mixed solution of glacial acetic acid (6mL) and concentrated sulfuric acid (9mL), methyl 4- (2- (2,2, 2-trifluoroacetamido) ethyl) benzoate (1.5g,5.5mmol), paraformaldehyde (250mg,8.3mmol) were added under ice bath, and after stirring at 25 ℃ for 16 hours, the reaction solution was poured into 150mL of ice water, extracted with ethyl acetate (100mL × 3), and the organic phases were combined and subjected to column chromatography (petroleum ether: ethyl acetate ═ 5:1) to give the title compound (1.3g, yield 82.3%).
(4) Preparation of methyl 1,2,3, 4-tetrahydroisoquinoline-7-carboxylate
Figure BDA0001739294150000481
Methyl 2- (2,2, 2-trifluoroacetyl) -1,2,3, 4-tetrahydroisoquinoline-7-carboxylate (1.2g,4.2mmol) was dissolved in a mixed solution of methanol (10mL) and water (4mL), potassium carbonate (0.9g,6.5mmol) was added, the reaction was stirred at 25 ℃ for 3 hours, after methanol was evaporated under reduced pressure, water (50mL) was added, ethyl acetate was extracted (30mL × 3), the organic phases were combined, and column chromatography was performed (petroleum ether: ethyl acetate ═ 5:1) to give the title compound (0.6g, yield 75%).
(5) Preparation of 1-bromo-2-chloro-4-methoxybenzene
Figure BDA0001739294150000482
4-bromo-3-chlorophenol (0.3g,1.4mol) was dissolved in N, N-dimethylformamide (20mL), potassium carbonate (0.3g,2.2mol), iodomethane (0.3g,2.1mmol) were added, the reaction was stirred at 25 ℃ for 2 hours, ethyl acetate (50mL) was added, water (50mL) was added for washing, a saturated sodium chloride solution (50mL) was added for washing, the organic phase was dried over anhydrous sodium sulfate, and column chromatography was performed (petroleum ether: ethyl acetate ═ 10:1) to give the title compound (0.3g, 93.7% yield).
(6) Preparation of 2- (2-chloro-4-methoxyphenyl) -1,2,3, 4-tetrahydroisoquinoline-7-carboxylic acid
Figure BDA0001739294150000483
1-bromo-2-chloro-4-methoxybenzene (0.3g,1.4mmol), methyl 1,2,3, 4-tetrahydroisoquinoline-7-carboxylate (0.3g,1.6mmol) were dissolved in toluene (10mL), 1 '-binaphthyl-2, 2' -bis-diphenylphosphine (87mg,0.14mmol), tris (dibenzylideneacetone) dipalladium (130mg,0.14mmol), sodium tert-butoxide (270mg,2.8mmol) were added, the reaction mixture was heated to 100 ℃ for 8 hours, after completion of the reaction, the reaction mixture was concentrated, and column chromatography was performed on the residue (petroleum ether: ethyl acetate ═ 2:1) to give the title compound (80mg, 18% yield).
(7) Preparation of 2- (2-chloro-4-hydroxyphenyl) -1,2,3, 4-tetrahydroisoquinoline-7-carboxylic acid
Figure BDA0001739294150000491
In ice bath, 2- (2-chloro-4-methoxyphenyl) -1,2,3, 4-tetrahydroisoquinoline-7-carboxylic acid (80mg,0.26mmol) was dissolved in dichloromethane (5mL), boron tribromide (125mg,0.5mmol) was added, the reaction was stirred at 25 ℃ for 6 hours, cooled to 0 ℃, 2mL of a saturated sodium bicarbonate solution was added to the reaction solution, the organic phase was separated, and the solution was dried over anhydrous sodium sulfate to dryness to obtain the title compound (75mg, yield 98.8%).
(8) Preparation of methyl 2- (2-chloro-4-hydroxyphenyl) -1,2,3, 4-tetrahydroisoquinoline-7-carboxylate
Figure BDA0001739294150000492
2- (2-chloro-4-hydroxyphenyl) -1,2,3, 4-tetrahydroisoquinoline-7-carboxylic acid (75mg,0.24mmol) was dissolved in methanol (5mL), thionyl chloride (60mg,0.50mmol) was added dropwise, the mixture was heated to 90 ℃ to react for 2 hours, and the reaction mixture was concentrated and subjected to column chromatography (dichloromethane: methanol ═ 20:1) to give the title compound (60mg, yield 78.6%).
(9) Preparation of methyl 2- (2-chloro-4- ((5-cyclopropyl-3- (2, 6-dichlorophenyl) isoxazol-4-yl) methoxy) phenyl) -1,2,3, 4-tetrahydroisoquinoline-7-carboxylate
Figure BDA0001739294150000493
4- (bromomethyl) -5-cyclopropyl-3- (2, 6-dichlorophenyl) isoxazole (66mg,0.19mmol), methyl 2- (2-chloro-4-hydroxyphenyl) -1,2,3, 4-tetrahydroisoquinoline-7-carboxylate (60mg,0.19mmol) were dissolved in N, N-dimethylformamide (5mL), cesium carbonate (130mg,0.4mmol) was added, the reaction was stirred at 40 ℃ for 3 hours, water (20mL) was added, ethyl acetate was extracted (15mL × 3), the organic phases were combined, and column chromatography was performed (dichloromethane: methanol ═ 15:1) to give the title compound (70mg, yield 63.1%).
(10) Preparation of 2- (2-chloro-4- ((5-cyclopropyl-3- (2, 6-dichlorophenyl) isoxazol-4-yl) methoxy) phenyl) -1,2,3, 4-tetrahydroisoquinoline-7-carboxylic acid
Figure BDA0001739294150000501
Methyl 2- (2-chloro-4- ((5-cyclopropyl-3- (2, 6-dichlorophenyl) isoxazol-4-yl) methoxy) phenyl) -1,2,3, 4-tetrahydroisoquinoline-7-carboxylate (70mg,0.12mmol) was dissolved in tetrahydrofuran (5mL), sodium hydroxide (10mg,0.25mmol) was added, after stirring at 25 ℃ for 12 hours, 1N HCl solution was added to adjust pH to 6, the solution was evaporated to dryness, and the residue was subjected to column chromatography (dichloromethane: methanol ═ 10:1) to give the title compound (40mg, yield 58.5%).
The molecular formula is as follows: c29H23Cl3N2O4Molecular weight: 569.9 LC-MS (m/z): 571.1(M + H)+
1H-NMR(400MHz,DMSO-d6)δ:7.90(d,J＝8.8Hz,1H),7.85(s,1H),7.42(d,J＝7.2Hz,1H),7.32-7.36(m,1H),7.28(s,1H),7.00(d,J＝8.8Hz,1H),6.90(d,J＝2.4Hz,1H),6.71(dd,J1＝8.8Hz,J2＝6.0Hz,1H),4.79(s,2H),4.23(s,2H),3.30-3.33(m,2H),3.02-3.09(m,2H),2.11-2.19(m,1H),1.27-1.32(m,2H),1.14-1.16(m,2H).
Example 62- (2-chloro-4- ((5-cyclopropyl-3- (2, 6-dichlorophenyl) isoxazol-4-yl) methoxy) benzene Preparation of 1,2,3, 4-tetrahydroisoquinoline-5-carboxylic acid (Compound 6)
Figure BDA0001739294150000502
(1) Preparation of 5-cyanoisoquinoline
Figure BDA0001739294150000503
Adding 5-bromoisoquinoline (2.0g,9.6mmol) and zinc cyanide (0.68g,5.8mmol) into N, N-dimethylformamide (100mL), adding tetrakis (triphenylphosphine) palladium (0.45g,0.39mmol) under the protection of nitrogen, heating to 100 ℃ for reaction for 16 hours, detecting the reaction completion by TLC, cooling to 25 ℃, adding water (200mL), suction filtering, washing a filter cake with water (100mL), and vacuum drying to obtain the title compound (0.9g, yield 60.8%).
(2) Preparation of isoquinoline-5-carboxylic acid
Figure BDA0001739294150000511
5-cyanoisoquinoline (0.9g,5.8mmol) was added to 50% sulfuric acid solution (30mL), heated to 100 ℃ for reaction for 16 h, cooled to 25 ℃, poured into ice water (100mL), stirred to precipitate a white precipitate, filtered with suction, and the filter cake was dried under vacuum to give the title compound (0.8g, 80% yield).
(3) Preparation of isoquinoline-5-carboxylic acid methyl ester
Figure BDA0001739294150000512
Isoquinoline-5-carboxylic acid (0.8g,4.62mmol) was added to methanol (30mL), concentrated sulfuric acid (0.5mL) was added with stirring, the temperature was raised to 83 ℃ for 24 hours, the reaction was detected by TLC to be complete, the reaction was concentrated under reduced pressure, and the crude product was used directly in the next step.
(4) Preparation of methyl 1,2,3, 4-tetrahydroisoquinoline-5-carboxylate
Figure BDA0001739294150000513
Methyl isoquinoline-5-carboxylate (crude product) was added to glacial acetic acid (30mL), and platinum dioxide (86mg) was added with stirring to displace hydrogen and reacted at 25 ℃ for 4 hours. TLC detection reaction was complete, suction filtration was performed, the filtrate was concentrated, saturated aqueous sodium bicarbonate (50mL) and ethyl acetate (100mL) were added, the mixture was separated, the organic phase was dried over anhydrous sodium sulfate, suction filtration was performed, and the filtrate was concentrated to give the title compound (0.87g, 98.9% yield).
(5) Preparation of methyl 2- (2-chloro-4-nitrophenyl) -1,2,3, 4-tetrahydroisoquinoline-5-carboxylate
Figure BDA0001739294150000514
Methyl 1,2,3, 4-tetrahydroisoquinoline-5-carboxylate (0.84g,4.4mmol), 2-chloro-1-fluoro-4-nitrobenzene (0.85g,4.8mmol) and cesium carbonate (2.87g,8.8mmol) were added to N, N-dimethylacetamide (50mL), warmed to 70 ℃ for 16 hours, TLC detected complete, water (150mL) and ethyl acetate (150mL) were added, the organic phase was separated, dried over anhydrous sodium sulfate, filtered, concentrated in vacuo, and the crude product was subjected to silica gel column chromatography (petroleum ether: ethyl acetate ═ 2:1) to afford the title compound (860mg, 56.2% yield).
(6) Preparation of methyl 2- (4-amino-2-chlorophenyl) -1,2,3, 4-tetrahydroisoquinoline-5-carboxylate
Figure BDA0001739294150000521
Methyl 2- (2-chloro-4-nitrophenyl) -1,2,3, 4-tetrahydroisoquinoline-5-carboxylate (0.86g,2.5mmol) was added to tetrahydrofuran (20mL), and Raney nickel (86mg, 10% by mass) was added under nitrogen protection, and the mixture was reacted with hydrogen at 25 ℃ for 1.5 hours. The reaction was completed by TLC, suction filtered, and the filtrate was concentrated to give the title compound (730mg, 92.2% yield).
(7) Preparation of methyl 2- (2-chloro-4-hydroxyphenyl) -1,2,3, 4-tetrahydroisoquinoline-5-carboxylate
Figure BDA0001739294150000522
2- (4-amino-2-chlorophenyl) -1,2,3, 4-tetrahydroisoquinoline-5-carboxylic acid methyl ester (700mg,2.2mmol) was added to a mixed solvent of 1mol/L sulfuric acid (10mL) and methanol (2mL), the temperature was reduced to 0 deg.C, sodium nitrite (167mg,2.42mmol) was added, 50% sulfuric acid (10mL) was added after 1 hour of reaction, the temperature was increased to 100 deg.C for 2 hours, the pH was adjusted to 7 with 1mol/L aqueous sodium hydroxide solution, ethyl acetate (200mL) was added for extraction, the organic phase was concentrated in vacuo, and the crude product was subjected to silica gel column chromatography (petroleum ether: ethyl acetate ═ 1:3) to give the title compound (130mg, 18.6% yield).
(8) Preparation of methyl 2- (2-chloro-4- ((5-cyclopropyl-3- (2, 6-dichlorophenyl) isoxazol-4-yl) methoxy) phenyl) -1,2,3, 4-tetrahydroisoquinoline-5-carboxylate
Figure BDA0001739294150000523
The procedure was as in example 5, reaction step (9).
(9) Preparation of 2- (2-chloro-4- ((5-cyclopropyl-3- (2, 6-dichlorophenyl) isoxazol-4-yl) methoxy) phenyl) -1,2,3, 4-tetrahydroisoquinoline-5-carboxylic acid
Figure BDA0001739294150000531
The procedure was as in example 5, reaction step (10).
Molecular formula C29H23Cl3N2O4Molecular weight 569.9 LC-MS (m/z): 569.1(M + H)+
1H-NMR(400MHz,DMSO-d6)δ:7.99-7.96(m,1H),7.43-7.41(m,2H),7.36-7.29(m,3H),7.00(d,J＝8.8Hz,1H),6.90(d,J＝2.8Hz,1H),6.72-6.69(m,1H),4.78(s,2H),4.22(s,2H),3.43-3.41(m,2H),3.30-3.27(m,2H),2.19-2.15(m,1H),1.19-1.14(m,4H).
Example 76- (2-chloro-4- ((5-cyclopropyl-3- (2, 6-dichlorophenyl) isoxazol-4-yl) methoxy) benzene Preparation of yl) -5,6,7, 8-tetrahydronaphthalene-2-carboxylic acid (Compound 7)
Figure BDA0001739294150000532
(1) 6-bromo-3, 4-dihydronaphthalen-2-yl trifluoromethanesulfonate
Figure BDA0001739294150000533
6-bromo-3, 4-dihydronaphthalen-2 (1H) -one (10g,44.43mmol) and tetrahydrofuran (250mL) were added under nitrogen in a three-necked flask, cooled to-78 deg.C, and lithium bis (trimethylsilyl) amide (89.2mL,89.2mmol) was added dropwise. After stirring at-78 ℃ for 1 hour, a solution of 1,1, 1-trifluoro-N-phenyl-N- (trifluoromethyl) sulfonyl) methanesulfonamide (19.1g,53.46mmol) in tetrahydrofuran (50mL) was added dropwise, the mixture was stirred for 30 minutes, and the mixture was returned to room temperature and stirred for 30 minutes. The reaction was quenched with 30mL of water, extracted with ethyl acetate (3 × 200mL), the organic phases were combined, washed with water (3 × 50mL), dried over anhydrous magnesium sulfate and concentrated, and the residue was purified by silica gel column (ethyl acetate: petroleum ether ═ 1:100-1:50) to give 4.6g of the title compound in 29% yield.
(2) Preparation of 2- (2-chloro-4-methoxyphenyl) -4,4,5, 5-tetramethyl-1, 3, 2-dioxaborolan
Figure BDA0001739294150000541
1-bromo-2-chloro-4-methoxybenzene (10g,45.15mmol), 1, 4-dioxane (200mL), pinacol diboron (12g,47.24mmol), potassium acetate (13.4g,136.54mmol), Pd (dppf) Cl were added to a three-necked flask2Dichloromethane complex (1.86g,2.28 mmol). Nitrogen gasThe displacement was carried out three times, followed by heating to 90 ℃ for reaction overnight. The reaction mixture was cooled, diluted with ethyl acetate (50mL) and washed with water (3X 100 mL). The residue was dried over anhydrous magnesium sulfate and concentrated, and purified by silica gel column (ethyl acetate: petroleum ether: 1:100-1:20) to obtain 7g of the title compound in 58% yield.
(3) Synthesis of 7-bromo-3- (2-chloro-4-methoxyphenyl) -1, 2-dihydronaphthalene
Figure BDA0001739294150000542
A three-necked flask was charged with an aqueous solution (5mL) of 2- (2-chloro-4-methoxyphenyl) -4,4,5, 5-tetramethyl-1, 3, 2-dioxaborolan (3g,11.17mmol), 1, 4-dioxane (40mL), 6-bromo-3, 4-dihydronaphthalen-2-yl trifluoromethanesulfonate (4.39g,12.29mmol) and sodium carbonate (2.97g,27.76mmol), Pd (dppf) Cl2DCM (89.9mg) was purged with nitrogen three times and heated to 80 ℃ for reaction overnight. Cooled and diluted with ethyl acetate (50 mL). The mixture was washed with water (3X 20 mL). The organic phase was dried over anhydrous sodium sulfate, concentrated and the residue was purified over a silica gel column (ethyl acetate: petroleum ether: 1:100-1:30) to give 1.5g of the title compound in 38% yield.
(4) Preparation of 6- (2-chloro-4-methoxyphenyl) -7, 8-dihydronaphthalene-2-carbonitrile
Figure BDA0001739294150000543
In a single-necked flask, 7-bromo-3- (2-chloro-4-methoxyphenyl) -1, 2-dihydronaphthalene (1.5g,4.29mmol), DMF (30mL), dicyano zinc (748mg,6.37mmol), and tetrakis (triphenylphosphine) palladium (497mg,0.43mmol) were added, and the reaction was allowed to proceed overnight at 125 ℃ with nitrogen purging three times. After cooling, insoluble solids were filtered off and diluted with ethyl acetate (50 mL). The mixture was washed with water (3X 20 mL). The organic phase was dried over anhydrous sodium sulfate, concentrated, and the residue was purified over a silica gel column (ethyl acetate: petroleum ether: 1:50-1: 20). 0.9g of the title compound is obtained in 71% yield.
(5) Preparation of methyl 6- (2-chloro-4-methoxyphenyl) -7, 8-dihydronaphthalene-2-carboxylate
Figure BDA0001739294150000551
6- (2-chloro-4-methoxyphenyl) -7, 8-dihydronaphthalene-2-carbonitrile (900mg,3.04mmol) and methanol (15mL) were added in a single-necked flask, followed by dropwise addition of H at 0 deg.C2SO4(5mL) after the addition, the reaction was allowed to proceed overnight at 80 deg.C, cooled and diluted with ethyl acetate (50 mL). The mixture was washed with water (3X 20 mL). The organic phase was dried over anhydrous sodium sulfate, concentrated, and the residue was purified on a silica gel column (ethyl acetate: petroleum ether: 1:50-1:30) to obtain 0.7g of the title compound in 70% yield.
(6) Preparation of methyl 6- (2-chloro-4-methoxyphenyl) -5,6,7, 8-tetrahydronaphthalene-2-carboxylate
Figure BDA0001739294150000552
In a 50mL single-necked flask were added methyl 6- (2-chloro-4-methoxyphenyl) -7, 8-dihydronaphthalene-2-carboxylate (700mg,2.13mmol), ethyl acetate (20mL), and PtO2(96mg) was hydrogenated at 0 ℃ for 1 hour. Insoluble solids were filtered off and the filtrate was concentrated to give 690mg of product in 98% yield.
(7) Preparation of methyl 6- (2-chloro-4-hydroxyphenyl) -5,6,7, 8-tetrahydronaphthalene-2-carboxylate
Figure BDA0001739294150000553
In a 50mL single-necked flask, methyl 6- (2-chloro-4-methoxyphenyl) -5,6,7, 8-tetrahydronaphthalene-2-carboxylate (680mg,2.06mmol) and methylene chloride (30mL) were added, followed by cooling to 0 ℃ and dropwise addition of BF3(Me2S) (2.68 g). The reaction mixture was stirred at 0 ℃ for 5 hours. Then, dichloromethane (50mL) was added for dilution and washing with water (3X 30 mL). The organic layer was separated, dried, concentrated and the residue was purified by column (ethyl acetate: petroleum ether: 1:20-1:5) to give 390mg of the product in 60% yield.
(8) Preparation of methyl (2-chloro-4- ((5-cyclopropyl-3- (2, 6-dichlorophenyl) isoxazol-4-yl) methoxy) phenyl) -5,6,7, 8-tetrahydronaphthalene-2-carboxylate
Figure BDA0001739294150000561
In a 50mL single-necked flask were added (5-cyclopropyl-3- (2, 6-dichlorophenyl) isoxazol-4-yl) methanol (524mg,1.84mmol) and dichloromethane (10mL), followed by dropwise addition of SOCl at 0 deg.C2(871mg,7.32mmol) and N, N-dimethylformamide (2 drops), stirred at 0 ℃ for 30 minutes, concentrated and added with methyl 6- (2-chloro-4-hydroxyphenyl) -5,6,7, 8-tetrahydronaphthalene-2-carboxylate (390mg,1.23mmol), N, N-dimethylformamide (10mL), sodium iodide (55mg,0.369mmol) and K2CO3(1.02g,7.33 mmol). The mixture was stirred at 60 ℃ overnight. The reaction mixture was cooled, diluted with ethyl acetate (30mL) and washed with water (3X 30 mL). The organic layer was separated, dried and concentrated to yield 750mg of the title compound.
(9) Preparation of 6- (2-chloro-4- ((5-cyclopropyl-3- (2, 6-dichlorophenyl) isoxazol-4-yl) methoxy) phenyl) -5,6,7, 8-tetrahydronaphthalene-2-carboxylic acid
Figure BDA0001739294150000562
A50 mL single vial was charged with an aqueous solution (1mL) of methyl 6- (2-chloro-4- ((5-cyclopropyl-3- (2, 6-dichlorophenyl) isoxazol-4-yl) methoxy) phenyl) -5,6,7, 8-tetrahydronaphthalene-2-carboxylate (750mg,1.29mmol), methanol (20mL), and sodium hydroxide (103mg,2.58mmol), heated to 60 deg.C, stirred overnight, cooled to concentrate, and the residue was purified by prep-HPLC to give 100mg of the title compound.
Molecular formula C30H24Cl3NO4Molecular weight 568.87 LC-MS (ES, M/z):566(M + H)+
1H-NMR(DMSO,ppm):δ:7.5-7.7(m,5H),7.25-7.3(m,3H),6.91(s,1H),6.7-6.8(m,1H),4.9(s,2H),2.8-2.99(m,5H),1.8-1.9(m,2H),1.11-1.29(m,5H).
Example 82- (2-chloro-4- ((5-cyclopropyl-3- (2, 6-dichlorophenyl)) Isoxazol-4-yl) methoxy) phenyl) Preparation of isoindoline-5-carboxylic acid (Compound 8)
Figure BDA0001739294150000571
(1) Preparation of methyl 3, 4-dimethylbenzoate
Figure BDA0001739294150000572
3, 4-Dimethylbenzoic acid (5.0g, 33.3mmol) was dissolved in methanol (50mL), thionyl chloride (7.9g,66.4mmol) was added dropwise under an ice-water bath, the temperature was raised to 25 ℃ and stirring was continued for 6 hours, the solvent was removed and the residue was used for the next step.
(2) Preparation of methyl 3, 4-bis (bromomethyl) benzoate
Figure BDA0001739294150000573
Methyl 3, 4-dimethylbenzoate (5.4g,32.9mmol), N-bromosuccinimide (12.8g,71.9mmol) and a catalytic amount of benzoyl peroxide (100mg,0.4mmol) were dissolved in carbon tetrachloride (50mL), heated to 80 ℃ and reacted for 12 hours. Cooled to 25 ℃, filtered, and the filtrate was washed with saturated sodium bicarbonate solution (20mL), dried over anhydrous sodium sulfate, filtered, and the filtrate was concentrated to give the title compound 10.6g, 100.0% yield.
(3) Preparation of methyl 2- (4-methoxybenzyl) isoindole-5-carboxylate
Figure BDA0001739294150000574
Methyl 3, 4-bis (bromomethyl) benzoate (10.6g,32.9mmol) was dissolved in tetrahydrofuran (50mL), p-methoxybenzylamine (4.5g,32.8mmol) and triethylamine (6.6g,65.3mmol) were added, stirring was carried out at 25 ℃ for 16 hours, the solvent was removed, ethyl acetate (100mL) and water (30mL) were added, liquid separation was carried out, the organic phase was removed, and the residue was separated by column chromatography (petroleum ether: ethyl acetate ═ 2:1) to give the title compound 4.5g, yield 46%.
(4) Preparation of dihydroisoindoline-5-carboxylic acid methyl ester
Figure BDA0001739294150000575
Methyl 2- (4-methoxybenzyl) isoindole-5-carboxylate (1.7g,5.7mmol) was dissolved in dichloromethane (50mL), 1-chloroethyl chloroformate (5.7g,39.9mmol) was added, the reaction was stirred at 25 ℃ for 30 hours, methanol (5mL) was added, and stirring was continued for 1 hour. The solvent was removed, ethyl acetate (100mL) and a saturated sodium bicarbonate solution (20mL) were added, the layers were separated, concentrated, and the residue was separated by column chromatography (dichloromethane: methanol ═ 20:1) to give the title compound 0.7g in 70% yield.
(5) Preparation of 1-bromo-2-chloro-4-methoxybenzene
Figure BDA0001739294150000581
4-bromo-3-chlorophenol (2.0g,9.6mmol) was dissolved in N, N-dimethylformamide (20mL), potassium carbonate (2.0g,14.5mol) and iodomethane (1.6g,11.3mmol) were added, the reaction was stirred at 25 ℃ for 2 hours, ethyl acetate (50mL) and water (50mL) were added, the layers were separated, the organic layer was dried over anhydrous sodium sulfate, and filtered. Concentration and column chromatography of the residue (petroleum ether: ethyl acetate: 10:1) gave the title compound 1.7g, 80% yield.
(6) Preparation of methyl 2- (2-chloro-4-methoxyphenyl) isoindoline-5-carboxylate
Figure BDA0001739294150000582
1-bromo-2-chloro-4-methoxybenzene (0.62g,2.8mmol), methyl isoindoline-5-carboxylate (0.5g,2.8mmol) were dissolved in toluene (10mL), 1 '-binaphthyl-2, 2' -bis-diphenylphosphine (0.35g,0.56mmol), tris (dibenzylideneacetone) dipalladium (0.26g,0.28mmol), sodium tert-butoxide (0.82g,8.5mmol) were added, the reaction mixture was heated to 100 ℃ for reaction for 8 hours, the reaction solution was concentrated, and the residue was subjected to column chromatography (petroleum ether: ethyl acetate ═ 2:1) to give 0.15g of a product in 16.9% yield.
(7) Preparation of 2- (2-chloro-4-hydroxyphenyl) isoindoline-5-carboxylic acid
Figure BDA0001739294150000583
Methyl 2- (2-chloro-4-methoxyphenyl) isoindoline-5-carboxylate (0.15g,0.47mmol) was dissolved in dichloromethane (5mL) under ice bath, boron tribromide (1.9mL,1.9mmol) was added, the reaction was stirred at 25 ℃ for 6 hours, cooled to 0 ℃, a saturated sodium bicarbonate solution (2mL) was added to the reaction solution, concentrated, and the residue was subjected to column chromatography (dichloromethane: methanol ═ 30:1) to give the title compound 0.07g, 50% yield.
(8) Preparation of methyl 2- (2-chloro-4-hydroxyphenyl) isoindoline-5-carboxylate
Figure BDA0001739294150000591
2- (2-chloro-4-hydroxyphenyl) isoindoline-5-carboxylic acid (0.07g,0.24mmol) was dissolved in methanol (5mL), thionyl chloride (0.06g,0.50mmol) was added dropwise, the reaction solution was heated to 90 ℃ for 2 hours, the reaction solution was concentrated, and the residue was subjected to column chromatography (dichloromethane: methanol ═ 30:1) to give the title compound 0.05g, in 68.5% yield.
(9) Preparation of methyl 2- (2-chloro-4- ((5-cyclopropyl-3- (2, 6-dichlorophenyl) isoxazol-4-yl) methoxy) phenyl) isoindoline-5-carboxylate
Figure BDA0001739294150000592
Referring to the preparation method in step (9) of example 5, 4- (bromomethyl) -5-cyclopropyl-3- (2, 6-dichlorophenyl) isoxazole (0.055g,0.16mmol), methyl 2- (2-chloro-4-hydroxyphenyl) isoindoline-5-carboxylate (0.05g,0.16mmol) were added to give 0.03g of a product in 33.0% yield.
(10) Preparation of 2- (2-chloro-4- ((5-cyclopropyl-3- (2, 6-dichlorophenyl) isoxazol-4-yl) methoxy) phenyl) isoindoline-5-carboxylic acid
Figure BDA0001739294150000593
Methyl 2- (2-chloro-4- ((5-cyclopropyl-3- (2, 6-dichlorophenyl) isoxazol-4-yl) methoxy) phenyl) isoindoline-5-carboxylate (0.03g,0.05mmol) was dissolved in methanol (5mL), lithium hydroxide monohydrate (21mg,0.5mmol) was added, the reaction was stirred at 25 ℃ for 48 hours, concentrated, water (5mL) was added, 1N HCl solution was added to adjust pH to 6, concentrated, and the residue was subjected to column chromatography (dichloromethane: methanol ═ 15:1) to give the title compound 15mg in 53.9% yield.
Molecular formula C28H21Cl3N2O4Molecular weight 555.8 LC-MS (M/z):555.1(M + H)+
1H-NMR(400MHz,MeOD)δ:7.94(s,2H),7.48-7.53(m,3H),7.37(d,J＝8.4Hz,1H),7.12(d,J＝9.2Hz,1H),6.81(d,J＝2.8Hz,1H),6.72(dd,J＝9.2Hz,2.8Hz,1H),4.67(s,2H),4.58(s,4H),2.31-2.37(m,1H),0.88-0.92(m,4H).
Example 9(R) -2- (2-chloro-4- ((5-cyclopropyl-3- (2, 6-dichlorophenyl) isoxazol-4-yl) methoxy) benzene Preparation of Yl) chroman-6-carboxylic acid (Compound 9)
Figure BDA0001739294150000601
(1) Preparation of methyl 2- (2-chloro-4-hydroxyphenyl) chroman-6-carboxylate
Figure BDA0001739294150000602
Specific preparation method reference is made to the preparation of example 1, steps (1) to (5).
(2) Resolution of (R) -2- (2-chloro-4-hydroxyphenyl) chroman-6-carboxylic acid methyl ester and (S) -2- (2-chloro-4-hydroxyphenyl) chroman-6-carboxylic acid methyl ester
Figure BDA0001739294150000603
The crude methyl 2- (2-chloro-4-hydroxyphenyl) chroman-6-carboxylate (4.5g) was separated by Chiral preparative column (Chiral-Prep-HPLC): chiral column, DAICEL CHIRALPAK AD-H; mobile phase, mobile phase a: n-hexane (0.1% TFA), mobile phase B: isopropanol, a: B ═ 80: 20. To give methyl (R) -2- (2-chloro-4-hydroxyphenyl) chroman-6-carboxylate (2.3g, crude product) and methyl (S) -2- (2-chloro-4-hydroxyphenyl) chroman-6-carboxylate (1.2 g).
(3) Preparation of (R) -methyl 2- (2-chloro-4- ((5-cyclopropyl-3- (2, 6-dichlorophenyl) isoxazol-4-yl) methoxy) phenyl) chroman-6-carboxylate
Figure BDA0001739294150000611
A solution of (5-cyclopropane-3- (2, 6-dichlorophenyl) isoxazol-4-yl) methanol (1.066g,3.75mmol) in methylene chloride (10mL) was added to a 50mL round-bottomed flask, sulfinyl chloride (10mL) was slowly added dropwise to the solution while stirring at room temperature, and after completion of the addition, stirring was carried out at room temperature for 2.5 hours, and concentration under reduced pressure was carried out to give a crude product. A100 mL three-necked flask was further charged with a DMF solution (40mL) of methyl (R) -2- (2-chloro-4-hydroxyphenyl) chroman-6-carboxylate (1g,3.14mmol), potassium carbonate (2.167g), and sodium iodide (1.4g) in this order. While stirring at room temperature, the crude DMF solution obtained above was slowly added dropwise thereto, and after completion of the addition, the mixture was stirred at 60 ℃ for 18 hours. Ethyl acetate (200mL) was added to the reaction mixture, which was then washed with saturated brine (50 mL. times.3), and the organic phase was separated, dried over anhydrous sodium sulfate, and concentrated under reduced pressure to give the product (crude product, 1.6 g).
(4) Preparation of (R) -2- (2-chloro-4- ((5-cyclopropyl-3- (2, 6-dichlorophenyl) isoxazol-4-yl) methoxy) phenyl) chroman-6-carboxylic acid
Figure BDA0001739294150000612
A solution of (R) -methyl 2- (2-chloro-4- ((5-cyclopropyl-3- (2, 6-dichlorophenyl) isoxazol-4-yl) methoxy) phenyl) chroman-6-carboxylate (1.6g, crude) in methanol/water (20/5mL) was added to a 50mL round-bottomed flask, and lithium hydroxide (328.8mg) was added thereto in portions at 0 ℃ and, after completion of the addition, stirred at room temperature for 2 days. The reaction mixture was adjusted to pH 4-5 with citric acid, ethyl acetate (100mL) was added, the mixture was washed with saturated brine (50 mL. times.2), and the organic phase was dried over anhydrous magnesium sulfate and concentrated under reduced pressure. The residue was subjected to flash separation (acetonitrile, water, 1% aqueous ammonia) to give the title compound 270mg in 15.1% yield in two steps.
Molecular formula C29H22Cl3NO5Molecular weight 570.85LC-MS (ES, M/z):568.1(M-1)-
1H-NMR(300MHz,DMSO,ppm):δ12.5(brs,1H),7.52-7.75(m,5H),7.37-7.40(d,J＝8.7Hz,1H),6.97-6.97(d,J＝2.4Hz,1H),6.82-6.89(m,2H),5.32-5.36(m,1H),4.94(s,2H),2.98-3.02(m,1H),2.79-2.84(m,1H),2.44(m,1H),2.14-2.19(m,1H),1.95-2.07(m,1H),1.10-1.23(m,4H).
Example 10(S) -2- (2-chloro-4- ((5-cyclopropyl-3- (2, 6-dichlorophenyl) isoxazol-4-yl) methoxy) Preparation of phenyl) chroman-6-carboxylic acid (Compound 10)
Figure BDA0001739294150000621
(1) Preparation of methyl (S) -2- (2-chloro-4- ((5-cyclopropyl-3- (2, 6-dichlorophenyl) isoxazol-4-yl) methoxy) phenyl) chroman-6-carboxylate
Figure BDA0001739294150000622
Preparation method referring to example 9, step (3), the title compound was obtained as crude product.
(2) Preparation of (S) -2- (2-chloro-4- ((5-cyclopropyl-3- (2, 6-dichlorophenyl) isoxazol-4-yl) methoxy) phenyl) chroman-6-carboxylic acid
Figure BDA0001739294150000623
Preparation method referring to example 9, step (4), the title compound (150mg) was obtained in 16.7% yield over two steps.
Molecular formula C29H22Cl3NO5Molecular weight 570.85LC-MS 568.1(M-1)-
1H NMR(300MHz,DMSO-d6,ppm):δ7.52-7.73(m,5H),7.37-7.40(d,J＝8.4Hz,1H),6.96-6.97(d,J＝2.4Hz,1H),6.82-6.87(m,2H),5.31-5.35(m,1H),4.94(s,2H),2.97-3.01(m,1H),2.73-2.83(m,1H),2.43(m,1H),2.18-2.27(m,1H),1.94-1.99(m,1H),1.12-1.21(m,4H).
Example 11(S) -2- (2-chloro-4- ((5-cyclopropyl-3- (2, 6-dichlorophenyl) isoxazol-4-yl) methoxy) Preparation of phenyl) -2, 3-dihydrobenzofuran-5-carboxylic acid (Compound 11)
Figure BDA0001739294150000631
2- (2-chloro-4- ((5-cyclopropyl-3- (2, 6-dichlorophenyl) isoxazol-4-yl) methoxy) phenyl) -2, 3-dihydrobenzofuran-5-carboxylic acid (291mg,0.52mmol, prepared as described in example 2, steps 1 to 5) was resolved under conditions: column DAICEL CHIRALPAK AD-3(4.6 mm. times.50 mm,3.0 μm), column temp:25 ℃, phase A: n-Hexane (0.1% TFA), phase B: Ethanlol, total flow:1.0mL/min, Wavelength: from 190nm to 500nm, 60mg of product is obtained, the yield is 20.6%, and the ee value is 99.66%.
The molecular formula is as follows: c28H20Cl3NO5Molecular weight 556.8LC-MS (M/z):554 (M-H)+)
1H-NMR(300MHz,DMSO)δ:7.80(d,J＝5.1Hz,2H),7.59-7.63(m,2H),7.52-7.56(m,1H),7.27(d,J＝9.0Hz,1H),7.01(d,J＝2.7Hz,1H),6.96(d,J＝3.9Hz,1H),6.78(dd,J1＝2.7Hz,J2＝5.7Hz,1H),6.03-6.09(m,1H),4.93(s,2H),3.71-3.80(m,1H),3.04-3.13(m,1H),2.43-2.45(m,1H),1.10-1.22(m,4H).
Example 12(R) -2- (2-chloro-4- ((5-cyclopropyl-3- (2, 6-dichlorophenyl) isoxazol-4-yl) methoxy) Preparation of phenyl) -2, 3-dihydrobenzofuran-5-carboxylic acid (Compound 12)
Figure BDA0001739294150000632
2- (2-chloro-4- ((5-cyclopropyl-3- (2, 6-dichlorophenyl) isoxazol-4-yl) methoxy) phenyl) -2, 3-dihydrobenzofuran-5-carboxylic acid (291mg,0.52mmol, prepared according to the method described in example 2, steps 1 to 5) was resolved under the same conditions as in example 11 to give the product 90mg in 30.9% yield and 90.48% ee.
The molecular formula is as follows: c28H20Cl3NO5Molecular weight 556.8LC-MS (M/z):556(M + H)+)
1H-NMR(300MHz,DMSO)δ:7.80(d,J＝5.7Hz,2H),7.59-7.63(m,2H),7.52-7.56(m,1H),7.27(d,J＝9.0Hz,1H),7.01(d,J＝2.7Hz,1H),6.96(d,J＝3.6Hz,1H),6.78(dd,J1＝2.7Hz,J2＝5.7Hz,1H),6.03-6.09(m,1H),4.93(s,2H),3.71-3.80(m,1H),3.04-3.13(m,1H),2.43-2.45(m,1H),1.10-1.22(m,4H).
Example 132- (2-chloro-4- ((5-cyclopropyl-3- (2, 6-dichlorophenyl) isoxazol-4-yl) methoxy) benzene Preparation of Yl) chroman-5-carboxylic acid (Compound 13)
Figure BDA0001739294150000641
(1) Preparation of methyl 2-formyl-3-hydroxybenzoate
Figure BDA0001739294150000642
Methyl 3-hydroxybenzoate (30.00g,197.2mmol) was added to trifluoroacetic acid (500mL), HMTA (33.20g,236.8mmol) was slowly added thereto, the temperature was raised to 80 ℃ for reaction for 6 hours, the reaction mixture was poured into 1.5L of ice water, 1L of ethyl acetate was added for extraction, the organic phase was washed successively with 800mL each of water and saturated brine, dried over anhydrous sodium sulfate, and concentrated to obtain 20.30g of the title compound in 57.1% yield by column chromatography (PE: EA ═ 8: 1).
(2) Preparation of 1- (2-chloro-4-methoxyphenyl) ethan-1-one
Figure BDA0001739294150000643
M-chloroanisole (30.00g,210.4mmol) was added dropwise to a solution of acetyl chloride (19.80g,252.2mmol) and anhydrous aluminum chloride (36.45g,273.4mmol) in DCM (200mL) at 0 deg.C, and the reaction was continued for 2 hours after completion of the dropwise addition. The reaction solution was poured into ice water (500mL), 1M HCl (300mL) and DCM (800mL) were added for extraction, the organic phase was dried over anhydrous sodium sulfate, concentrated, and the residue was subjected to column chromatography (PE: EA ═ 10:1) to give the title compound 13.7g in 35.3% yield.
(3) Preparation of 2-bromo-1- (2-chloro-4-methoxyphenyl) ethan-1-one
Figure BDA0001739294150000651
1- (2-chloro-4-methoxyphenyl) ethan-1-one (8.01g,43.4mmol) was dissolved in dichloromethane (200mL), and bromine (6.94g,43.4mmol) was added dropwise in an ice-water bath, and the reaction was continued for 2 hours after completion of the addition. The reaction solution was directly concentrated and subjected to column chromatography (PE: EA ═ 8:1) to obtain 8.34g of a product, with a yield of 73.0%.
(4) Preparation of diethyl (2- (2-chloro-4-methoxyphenyl) -2-oxoethyl) phosphonate
Figure BDA0001739294150000652
2-bromo-1- (2-chloro-4-methoxyphenyl) ethan-1-one (8.34g,31.7mmol) and triethyl phosphite (7.89g,47.6mmol) were added to toluene (100mL) and the temperature was raised to 110 ℃ for reaction for 24 hours. The reaction solution was directly concentrated and subjected to column chromatography (PE: EA ═ 5:1) to obtain 4.40g of a product, with a yield of 43.3%.
(5) Preparation of methyl 2- (3- (2-chloro-4-methoxyphenyl) -3-oxoprop-1-en-1-yl) -3-hydroxybenzoate
Figure BDA0001739294150000653
Diethyl (2- (2-chloro-4-methoxyphenyl) -2-oxoethyl) phosphonate (4.40g,13.7mmol), methyl 2-formyl-3-hydroxybenzoate (4.90g,27.4mmol) and DBU (4.20g,27.4mmol) were added to THF (100mL) and reacted for 3 hours. Water and ethyl acetate were added and extracted 200mL each, the organic phase was dried over anhydrous sodium sulfate, concentrated, and the residue was subjected to column chromatography (PE: EA ═ 5:1) to give the title compound 2.08g, yield 43.7%.
(6) Preparation of methyl 2- (3- (2-chloro-4-methoxyphenyl) -3-oxopropyl) -3-hydroxybenzoate
Figure BDA0001739294150000661
Methyl 2- (3- (2-chloro-4-methoxyphenyl) -3-oxoprop-1-en-1-yl) -3-hydroxybenzoate (1.50g,4.3mmol) was added to THF (50mL), and Pt/C (150mg) was added and the reaction was carried out under a hydrogen atmosphere for 2 hours. Suction was applied, the filter cake was washed with THF (10mL), and the filtrate was used directly for the next reaction.
(7) Preparation of methyl 2- (3- (2-chloro-4-methoxyphenyl) -3-hydroxypropyl) -3-hydroxybenzoate
Figure BDA0001739294150000662
The filtrate (60mL) from the above step was placed in a 250mL flask, and sodium borohydride (197mg,5.2mmol) was added and stirred for 1 hour. Water and ethyl acetate were added and extracted 100mL each, and the organic phase was dried over anhydrous sodium sulfate, concentrated, and subjected to column chromatography (PE: EA ═ 1:1) to give the title compound 1.45g, in 95.6% yield in two steps.
(8) Preparation of methyl 2- (2-chloro-4-methoxyphenyl) chroman-5-carboxylate
Figure BDA0001739294150000663
Methyl 2- (3- (2-chloro-4-methoxyphenyl) -3-hydroxypropyl) -3-hydroxybenzoate (1.45g,4.1mmol), triphenylphosphine (5.37g,20.5mmol) and DEAD (3.57g,20.5mmol) were added to THF (50mL) and reacted for 2h. The reaction mixture was directly spin-dried, and column chromatography (PE: EA ═ 25:1) was performed on the residue to give the title compound 800mg, in 58.1% yield.
(9) Preparation of methyl 2- (2-chloro-4-hydroxyphenyl) chroman-5-carboxylate
Figure BDA0001739294150000664
Methyl 2- (2-chloro-4-methoxyphenyl) chroman-5-carboxylate (800mg,2.4mmol) was dissolved in DCM (20mL) and 1M boron tribromide in DCM solution (7.2mL) was added dropwise at-10 ℃ and the reaction was continued for 2 hours after completion of the dropwise addition. Quench with 5mL of water. Water (50mL) and DCM (80mL) were added for extraction, the organic phase was washed with saturated brine (60mL), dried over anhydrous sodium sulfate, concentrated and the residue was subjected to column chromatography (PE: EA ═ 10:1) to give the title compound 90mg, yield 11.7%.
(10) Preparation of methyl 2- (2-chloro-4- ((5-cyclopropyl-3- (2, 6-dichlorophenyl) isoxazol-4-yl) methoxy) phenyl) chroman-5-carboxylate
Figure BDA0001739294150000671
Methyl 2- (2-chloro-4-hydroxyphenyl) chroman-5-carboxylate (90mg,0.28mmol), 4- (bromomethyl) -5-cyclopropyl-3- (2, 6-dichlorophenyl) isoxazole (145mg,0.42mmol) and potassium carbonate (77mg,0.56mmol) were added to DMF (5mL) and reacted for 2 hours. Water and ethyl acetate were added thereto, and the mixture was extracted 80mL each, and the organic phase was washed with saturated brine (60mL), dried over anhydrous sodium sulfate, and concentrated, and the residue was subjected to column chromatography (PE: EA ═ 3:1) to give 120mg of the title compound in a yield of 72.7%.
(11) Preparation of 2- (2-chloro-4- ((5-cyclopropyl-3- (2, 6-dichlorophenyl) isoxazol-4-yl) methoxy) phenyl) chroman-5-carboxylic acid
Figure BDA0001739294150000672
Methyl 2- (2-chloro-4- ((5-cyclopropyl-3- (2, 6-dichlorophenyl) isoxazol-4-yl) methoxy) phenyl) chroman-5-carboxylate (120mg,0.2mmol) was added to THF (2mL), methanol (2mL), water (2mL) and lithium hydroxide monohydrate (51mg,1.2mmol) were added and the reaction was allowed to react for 6 hours. The pH was adjusted to 2 with 1M HCl, water (30mL) and ethyl acetate (60mL) were added for extraction, the organic phase was washed with saturated brine (30mL), dried over anhydrous sodium sulfate, concentrated, and the residue was subjected to column chromatography (DCM: MeOH ═ 20:1) to give the title compound 28mg, 23.9% yield.
Molecular formula C29H22Cl3NO5Molecular weight 570.85LC-MS (M/e):570.1(M + H)+)
1H-NMR(400MHz,DMSO)δ:7.65-7.60(m,2H),7.59-7.51(m,1H),7.48-7.39(m,2H),7.25-7.17(m,1H),7.03(d,J＝1.2Hz,1H),7.01(d,J＝1.2Hz,1H),6.90-6.80(m,1H),5.30-5.25(m,1H),4.94(s,2H),3.20-3.10(m,2H),2.49-2.40(m,1H),2.22-2.10(m,1H),1.89-1.80(m,1H),1.23-1.17(m,2H),1.17-1.11(m,2H).
Example 142- (2-chloro-4- ((5-cyclopropyl-3- (2, 6-dichlorophenyl) isoxazol-4-yl) methoxy) benzene Preparation of Yl) chroman-7-carboxylic acid (Compound 14)
Figure BDA0001739294150000681
(1) Preparation of (E) -1- (4-bromo-2-hydroxyphenyl) -3- (2-chloro-4-hydroxyphenyl) prop-2-en-1-one
Figure BDA0001739294150000682
Prepared by the method of preparation in step (1) of reference example 1,1- (4-bromo-2-hydroxyphenyl) ethan-1-one (27.4g,127.42mmol), 2-chloro-4-hydroxybenzaldehyde (20g,127.74mmol) was added to give 36g of the product (yield 80%).
(2) Preparation of 7-bromo-2- (2-chloro-4-hydroxyphenyl) chroman-4-one
Figure BDA0001739294150000683
In a 3L four-necked flask, (E) -1- (4-bromo-2-hydroxyphenyl) -3- (2-chloro-4-hydroxyphenyl) prop-2-en-1-one (36g,101.81mmol) was dissolved in a mixed solvent of methanol (720mL) and hydrochloric acid (12N,720mL), and the temperature was raised to 73 ℃ to react for 72 hours. The reaction mixture was poured into 3L of ice water and stirred to form a large amount of precipitate, which was filtered off with suction and the filter cake was dried to obtain 21g of the product (58% yield).
(3) Preparation of 4- (7-bromochroman-2-yl) -3-chlorophenol
Figure BDA0001739294150000691
Referring to the preparation of step (3) of example 1, 7-bromo-2- (2-chloro-4-hydroxyphenyl) chroman-4-one (21g,59.39mmol) was added to give 13g of the product (yield 64%).
(4) Preparation of 2- (2-chloro-4-hydroxyphenyl) chroman-7-carbonitrile
Figure BDA0001739294150000692
A250 mL three-necked round-bottomed flask was charged with 4- (7-bromochroman-2-yl) -3-chlorophenol (13g,38.28mmol), according to the preparation method of step (4) of example 1, to obtain 6.0g of a product (yield 55%).
(5) Preparation of methyl 2- (2-chloro-4-hydroxyphenyl) chroman-7-carboxylate
Figure BDA0001739294150000693
Referring to the preparation of step (5) of example 1,2- (2-chloro-4-hydroxyphenyl) chroman-7-carbonitrile (6g,21.00mmol) was added to give 2.5g of a product (yield 37%).
(6) Preparation of methyl 2- (2-chloro-4- ((5-cyclopropyl-3- (2, 6-dichlorophenyl) isoxazol-4-yl) methoxy) phenyl) chroman-7-carboxylate
Figure BDA0001739294150000694
Referring to the preparation method of step (6) of example 1, methyl 2- (2-chloro-4-hydroxyphenyl) chroman-7-carboxylate (2.5g,7.84mmol), 4- (chloromethyl) -5-cyclopropyl-3- (2, 6-dichlorophenyl) isoxazole (2.84g,9.39mmol) were added to give 1.2g of a product (yield 26%).
(7) Preparation of 2- (2-chloro-4- ((5-cyclopropyl-3- (2, 6-dichlorophenyl) isoxazol-4-yl) methoxy) phenyl) chroman-7-carboxylic acid
Figure BDA0001739294150000695
A100 mL three-necked flask was charged with a mixed solvent of methyl 2- (2-chloro-4- ((5-cyclopropyl-3- (2, 6-dichlorophenyl) isoxazol-4-yl) methoxy) phenyl) chroman-7-carboxylate (1.2g,2.05mmol) in methanol and water, cooled to 0 ℃, added portionwise with lithium hydroxide monohydrate (260mg,6.19mmol), and heated to 30 ℃ for 2 days. The pH was adjusted to 4 with hydrochloric acid (1mol/L), ethyl acetate (500mL) and saturated brine (500mL) were added, the organic phase was separated, washed with saturated brine (2 × 500mL), dried over anhydrous sodium sulfate, concentrated, and the residue was subjected to column chromatography (petroleum ether: ethyl acetate 4:1 to 1:1) to give 0.28g of the product (yield 24%).
Molecular formula C29H22Cl3NO5Molecular weight 570.85LC-MS (ES, M/z) 570(M +1)+
1H NMR:(300MHz,DMSO-d6,ppm):δ12.82(brs,1H),7.64-7.39(m,5H),7.32-7.31(d,J＝1.2Hz,1H),7.26-7.23(m,1H),6.70-6.69(d,J＝2.4Hz,1H),6.86-6.85(m,1H),5.31-5.28(d,J＝9.0Hz,1H),4.93(s,2H),3.07-2.73(m,2H),2.47-2.45(m,1H),2.19-2.14(m,1H),1.99-1.90(m,1H),1.23-1.134(m,4H).
Example 152- (2-chloro-4- ((5-cyclopropyl-3- (2, 6-dichlorophenyl) isoxazol-4-yl) methoxy) benzene Preparation of Yl) chroman-8-carboxylic acid (Compound 15)
Figure BDA0001739294150000701
(1) Preparation of methyl salicylate
Figure BDA0001739294150000702
Salicylic acid (20.0g,0.14mol) was added to methanol (100ml), thionyl chloride (20.7g,0.17mol) was added dropwise to the system at 25 ℃, the reaction was stirred up to 50 ℃ for 12 hours, the reaction solution was spun dry, methylene chloride (500ml) was added to the reaction system, the pH was adjusted to 7 with sodium bicarbonate, the organic phase was separated, dried over anhydrous sodium sulfate, and concentrated to give the title compound (20.0g, yield 90.9%).
(2) Preparation of 3-formyl-2-hydroxybenzoic acid methyl ester
Figure BDA0001739294150000711
Methyl salicylate (20.0g,0.13mol) was added to trifluoroacetic acid (200mL), urotropin (28.0g,0.2mol), cuprous oxide (14.8g,0.13mol) were added to the system, stirred to 60 ℃ for reaction for 24 hours, the reaction solution was poured into water (1000mL), dichloromethane (500mL) was added, the organic phase was separated, the solvent was removed by rotary evaporation, and the residue was purified by silica gel column chromatography (ethyl acetate: petroleum ether: 1:10) to give the title compound (7.0g, yield 29.5%).
(3) Preparation of diethyl (2- (2-chloro-4-methoxyphenyl) -2-oxyethyl) phosphonate
Figure BDA0001739294150000712
M-chloroanisole (15.0g,0.11mol) was added to trifluoroacetic anhydride (115.5g,0.55mol), diethylphosphonoacetic acid (25.5g,0.13mol) was added, phosphoric acid (15.0g) was added dropwise at 25 ℃, the reaction was stirred at 25 ℃ for 24 hours, the reaction solution was poured into ice water (500ml), extraction was performed with dichloromethane (500ml), the organic phase was separated, the reaction solution was concentrated, and the residue was purified by column chromatography (dichloromethane: methanol ═ 50:1) to give the title compound (18.0g, crude product).
(4) Preparation of methyl (E, Z) -3- (3- (2-chloro-4-methoxyphenyl) -3-oxoprop-1-en-1-yl) -2-hydroxybenzoate
Figure BDA0001739294150000713
Diethyl (2- (2-chloro-4-methoxyphenyl) -2-oxyethyl) phosphonate (5.9g,18.5mmol), methyl 3-aldehyde-2-hydroxybenzoate (3.0g,16.6mmol), 1, 5-diazabicyclo [5.4.0] undec-5-ene (3.4g,22.2mmol) were added to tetrahydrofuran (50ml), the reaction was stirred at 25 ℃ for 24 hours, the reaction solution was concentrated, and the residue was purified by silica gel column chromatography (ethyl acetate: petroleum ether ═ 1:10) to give the title compound (2.0g, yield 34.7%).
(5) Preparation of methyl 3- (3- (2-chloro-4-methoxyphenyl) -3-oxopropyl) -2-hydroxybenzoate
Figure BDA0001739294150000721
Methyl (Z) -3- (3- (2-chloro-4-methoxyphenyl) -3-oxoprop-1-en-1-yl) -2-hydroxybenzoate (1.0g,2.9mmol) was added to tetrahydrofuran (20ml), and then, hydrogen gas was used for substitution 3 times, hydrogenation was performed at 25 ℃ for 12 hours, the reaction mixture was filtered, and the filtrate was concentrated to obtain the title compound (900mg, yield 90%).
(6) Preparation of methyl 3- (3- (2-chloro-4-methoxyphenyl) -3-hydroxypropyl) -2-hydroxybenzoate
Figure BDA0001739294150000722
Methyl 3- (3- (2-chloro-4-methoxyphenyl) -3-oxopropyl) -2-hydroxybenzoate (900mg,2.6mmol) was added to tetrahydrofuran (20ml), sodium borohydride (97.5mg,2.6mmol) was added, the reaction was stirred at 25 ℃ for 2 hours, water (50ml) and ethyl acetate (30ml) were added to the reaction mixture, the organic phase was separated, dried over anhydrous sodium sulfate, and concentrated to give the product (900mg, crude).
(7) Preparation of methyl 2- (2-chloro-4-methoxyphenyl) chroman-8-carboxylate
Figure BDA0001739294150000723
Methyl 3- (3- (2-chloro-4-methoxyphenyl) -3-hydroxypropyl) -2-hydroxybenzoate (900mg,2.6mmol) was added to tetrahydrofuran (20ml), triphenylphosphine (1.3g,5.0mmol) and diethyl azodicarboxylate (0.89g,5.13mmol) were added, the reaction was stirred at 25 ℃ for 12 hours, the reaction solution was concentrated, and column chromatography (petroleum ether: ethyl acetate ═ 20:1) was performed to obtain a product (300mg, yield 35.0%).
(8) Preparation of methyl 2- (2-chloro-4-hydroxyphenyl) chroman-8-carboxylate
Figure BDA0001739294150000731
Methyl 2- (2-chloro-4-methoxyphenyl) chroman-8-carboxylate (300.0mg,0.9mmol) was added to dichloromethane (20ml), the temperature was reduced to-25 ℃, boron tribromide (1.1g,4.5mmol) was added dropwise, reaction was carried out for 1 hour, methanol (10ml) was added dropwise to the reaction solution, pH was adjusted to 7 with an aqueous sodium bicarbonate solution, and the organic phase was separated and subjected to column chromatography (petroleum ether: ethyl acetate ═ 20:1) to give the title compound (120mg, yield 41.8%).
(9) Preparation of methyl 2- (2-chloro-4- (5-cyclopropyl-3- (2, 6-dichlorophenyl) isoxazole-4-methoxy) phenyl) chroman-8-carboxylate
Figure BDA0001739294150000732
Methyl 2- (2-chloro-4-hydroxyphenyl) chroman-8-carboxylate (120.0mg,0.38mmol), 4-bromomethyl-5-cyclopropyl-3- (2, 6-dichlorophenyl) isoxazole (131.9mg,0.38mmol) were added to N, N-dimethylformamide (10ml) and cesium carbonate (245.3mg,0.75mmol), heated to 50 ℃ for 1 hour, the reaction solution was poured into water (50ml), extracted with ethyl acetate (100ml), the organic phase was separated, dried over anhydrous sodium sulfate, and the solvent was removed by rotary evaporation to give the title compound (100mg, yield 45.5%).
(10) Preparation of 2- (2-chloro-4- ((5-cyclopropyl-3- (2, 6-dichlorophenyl) isoxazol-4-yl) methoxy) phenyl) chroman-8-carboxylic acid
Figure BDA0001739294150000733
Methyl 2- (2-chloro-4- ((5-cyclopropyl-3- (2, 6-dichlorophenyl) isoxazol-4-yl) methoxy) phenyl) chroman-8-carboxylate (100mg,0.17mmol), lithium hydroxide (20.5mg,0.85mmol), tetrahydrofuran (1ml), water (1ml) were added, stirred at 25 ℃ for 12 hours, ethyl acetate (100ml) was added to separate an organic phase, dried over anhydrous sodium sulfate, and the solvent was removed by rotary evaporation to give the title compound (60.8mg, yield 62.0%).
The molecular formula is as follows: c29H22Cl3NO5Molecular weight: 570.85LC-MS (M/e): 569.8,571.7(M + H)+)
1H NMR(CDCl3)δ:8.05-8.06(d,J＝6.4Hz,1H),7.32-7.43(m,5H),7.05-7.09(m,1H),6.88(s,1H),6.77-6.80(m,1H),5.59-5.62(d,J＝10.4Hz,1H),4.84(s,2H),2.87-3.15(m,2H),2.05-2.17(m,3H),1.16-1.33(m,5H)。
Example 163- (2-chloro-4- ((5-cyclopropyl-3- (2, 6-dichlorophenyl) isoxazol-4-yl) methoxy) benzene Preparation of Yl) chroman-7-carboxylic acid (Compound 16)
Figure BDA0001739294150000741
(1) Preparation of ethyl 3-hydroxy-4-methylbenzoate
Figure BDA0001739294150000742
3-hydroxy-4-methylbenzoic acid (5.0g,32.8mmol) is weighed and added to 100mL of ethanol, 0.5mL of concentrated sulfuric acid is added dropwise, the temperature is raised to 81 ℃ for reaction for 24 hours, concentration is carried out, water and 100mL of ethyl acetate are added respectively, an organic phase is obtained by separation, anhydrous sodium sulfate is dried, filtration is carried out, and the filtrate is concentrated to obtain a product (5.17g, the yield is 87.5%).
(2) Preparation of 1- (bromomethyl) -2-chloro-4-methoxybenzene
Figure BDA0001739294150000743
2-chloro-4-methoxy-1-methylbenzene (4.46g,28.5mmol) was added to 60mL of carbon tetrachloride, NBS (5.06g,28.4mmol) and AIBN (468mg,2.85mmol) were added, the temperature was raised to 80 ℃ to react for 2.0 hours, the temperature was lowered to 25 ℃, suction filtration was performed, the filtrate was concentrated, and the residue was subjected to column chromatography (PE: EA ═ 10:1) to obtain a product (5.52g, yield 82.3%).
(3) Preparation of 2- (2-chloro-4-methoxyphenyl) acetonitrile
Figure BDA0001739294150000751
1- (bromomethyl) -2-chloro-4-methoxybenzene (5.52g,23.4mmol) and potassium carbonate (6.46g,46.8mmol) were added to 100mL acetonitrile, and TMSCN (2.78g,28.0mmol) was added with stirring. The temperature is raised to 60 ℃ for reaction for 8.0 hours, TLC detects that the reaction is complete, concentration is carried out, water (100mL) and ethyl acetate (150mL) are added, an organic phase is obtained by layer separation, concentration is carried out, and the residue is subjected to column chromatography (PE: EA is 5:1) to obtain a product (4.1g, the yield is 96.5%).
(4) Preparation of ethyl 3- (methoxymethoxy) -4-methylbenzoate
Figure BDA0001739294150000752
Dissolving ethyl 3-hydroxy-4-methylbenzoate (5.17g,28.7mmol) in DCM (100mL), cooling to 0 ℃, adding DIPEA (8.14g,63.1mmol) and chloromethyl methyl ether (4.62g,57.4mmol), heating to 25 ℃ for 24 hours, detecting complete reaction by TLC, concentrating, adding water and ethyl acetate each 150mL, separating to obtain an ethyl acetate phase, washing the EA phase with water, washing with saturated sodium chloride aqueous solution, drying with anhydrous sodium sulfate, filtering, and concentrating the filtrate to obtain a product (5.8g, yield 90.1%).
(5) Preparation of 2- (2-chloro-4-methoxyphenyl) acetic acid
Figure BDA0001739294150000753
2- (2-chloro-4-methoxyphenyl) acetonitrile (4.1g,22.6mmol) was added to a mixed solvent of HAc (60mL) and water (60mL), concentrated sulfuric acid (20mL) was added dropwise with stirring, the temperature was raised to 100 ℃ to react for 8.0 hours, the reaction solution was poured into ice water, white precipitate was generated with stirring, suction filtration was performed, and the filter cake was dried to obtain the product (3.84g, yield 84.8%).
(6) Preparation of ethyl 2- (2-chloro-4-methoxyphenyl) acetate
Figure BDA0001739294150000754
2- (2-chloro-4-methoxyphenyl) acetic acid (3.84g,19.1mmol) and ethanol (2.64g,57.4mmol) were added to 60mL of dichloromethane, triethylamine (1.93g,19.1mmol) and EDCi (3.67g,19.1mmol) were added, DMAP (93mg,0.76mmol) was added with stirring, the reaction was allowed to react at 25 ℃ for 4.0 hours, TLC was performed to detect completion of the reaction, concentration was performed, 100mL each of water and ethyl acetate was added, ethyl acetate phase was obtained by separation, ethyl acetate phase was washed with 1N HCl (50mL), saturated aqueous sodium chloride solution was washed, dried over anhydrous sodium sulfate, filtration was performed, and the filtrate was concentrated to obtain a product (4.1g, yield 93.8%).
(7) Preparation of ethyl 4- (bromomethyl) -3- (methoxymethoxy) benzoate
Figure BDA0001739294150000761
Ethyl 3- (methoxymethoxy) -4-methylbenzoate (2.24g,10mmol) was added to 40mL of carbon tetrachloride, NBS (1.78g,10mmol) and AIBN (164mg,1.0mmol) were added, the temperature was raised to 80 ℃ to react for 2.0 hours, the temperature was lowered to 25 ℃, suction filtration was performed, the filtrate was concentrated, and the residue was subjected to column chromatography (PE: EA ═ 5:1) to give the product (2.7g, yield 89.1%).
(8) Preparation of ethyl 4- (2- (2-chloro-4-methylphenyl) -3-ethoxy-3-oxopropyl) -3- (methoxymethoxy) benzoate
Figure BDA0001739294150000762
Dissolving ethyl 2- (2-chloro-4-methoxyphenyl) acetate (1.1g,4.81mmol) in 20mL tetrahydrofuran, cooling to 0 ℃, adding NaHMDS (2.0M,4.8mL,9.6mmol), stirring at 0 ℃ for 1.0 hour, adding ethyl 4- (bromomethyl) -3- (methoxymethoxy) benzoate (1.46g,4.82mmol), heating to 25 ℃ for 18 hours, adding water (50mL) and EA (100mL), separating to obtain an EA phase, concentrating, and subjecting the residue to column chromatography (PE: EA ═ 2:1) to obtain a product (1.5g, yield 69.1%).
(9) Preparation of ethyl 4- (2- (2-chloro-4-methoxyphenyl) -3-ethoxy-3-oxopropyl) -3-hydroxybenzoate
Figure BDA0001739294150000763
Ethyl 4- (2- (2-chloro-4-methylphenyl) -3-ethoxy-3-oxopropyl) -3- (methoxymethoxy) benzoate (1.5g,3.33mmol) was added to a mixed solvent of 10mL TFA and DCM (20mL), reacted at 25 ℃ for 4.0 hours, LC-MS checked for completion, concentrated, and the residue was subjected to column chromatography (PE: DCM ═ 1:3) to give the product (620mg, 45.9% yield).
(10) Preparation of ethyl 4- (2- (2-chloro-4-methoxyphenyl) -3-hydroxypropyl) -3-hydroxybenzoate
Figure BDA0001739294150000771
Ethyl 4- (2- (2-chloro-4-methoxyphenyl) -3-ethoxy-3-oxopropyl) -3-hydroxybenzoate (620mg,1.52mmol) was added to 20mL of tetrahydrofuran, the temperature was reduced to 0 ℃ and lithium aluminum hydride (58mg,1.53mmol) was slowly added. After 40min reaction at 0 ℃, 1ml of water is added for quenching reaction, suction filtration is carried out, filtrate is concentrated, and the residue is subjected to column chromatography (EA: DCM 1:4) to obtain the product (130mg, 23.5 percent of yield).
(11) Preparation of ethyl 3- (2-chloro-4-methoxyphenyl) chroman-7-carboxylate
Figure BDA0001739294150000772
Ethyl 4- (2- (2-chloro-4-methoxyphenyl) -3-hydroxypropyl) -3-hydroxybenzoate (130mg,0.36mmol) was added to 20mL of tetrahydrofuran, the temperature was reduced to 0 ℃, triphenylphosphine (283mg,1.08mmol) and DEAD (188mg,1.08mmol) were added, reaction was carried out at 25 ℃ for 18 hours, LC-MS checked for completion, concentration was carried out, and the residue was subjected to column chromatography (PE: EA ═ 25:1) to give the product (120mg, 96.0% yield).
(12) Preparation of ethyl 3- (2-chloro-4-hydroxyphenyl) chroman-7-carboxylate
Figure BDA0001739294150000773
Reacting 3- (2-chloro-4-methoxyphenyl) chroman-7-carboxylic acidEthyl ester (120mg,0.35mmol) was added to 5mL DCM, cooled to 0 deg.C and 1M BBr added3Then the reaction solution was heated to 10 ℃ for 1.5 hours, and the reaction was completed by LC-MS detection, 1mL of methanol was added, and the mixture was concentrated to obtain a residue, which was subjected to column chromatography (PE: EA: 9:1) to obtain a product (100mg, 85.8% yield).
(13) Preparation of ethyl 3- (2-chloro-4- ((5-cyclopropyl-3- (2, 6-dichlorophenyl) isoxazol-4-yl) methoxy) phenyl) chroman-7-carboxylate
Figure BDA0001739294150000781
Ethyl 3- (2-chloro-4-hydroxyphenyl) chroman-7-carboxylate (100mg,0.30mmol) and 4- (bromomethyl) -5-cyclopropyl-3- (2, 6-dichlorophenyl) isoxazole (114mg,0.33mmol) were added to 10mL DMF, potassium carbonate (83mg,0.60mmol) was added with stirring, the temperature was raised to 50 ℃ for reaction for 2.0 hours, LC-MS detected the formation of the product, the temperature was lowered to 25 ℃,30 mL water was added, precipitation occurred by sonication, filtration was carried out, and the filter cake was dried to give the product (130mg, yield 72.2%).
(14) Preparation of 3- (2-chloro-4- ((5-cyclopropyl-3- (2, 6-dichlorophenyl) isoxazol-4-yl) methoxy) phenyl) chroman-7-carboxylic acid
Figure BDA0001739294150000782
Ethyl 3- (2-chloro-4- ((5-cyclopropyl-3- (2, 6-dichlorophenyl) isoxazol-4-yl) methoxy) phenyl) chroman-7-carboxylate (130mg,0.22mmol) was dissolved in a mixed solvent of THF (4mL), MeOH (2mL) and water (1mL), lithium hydroxide monohydrate (37mg,0.88mmol) was added at 25 ℃ for 18 hours, LC-MS checked for completion of the reaction, ethyl acetate (50mL) and water (50mL) were added, the ethyl acetate phases were separated to obtain an ethyl acetate phase, which was concentrated, and the residue was subjected to silica gel column chromatography (DCM: MeOH ═ 10:1) to obtain a product (80mg, yield 63.7%).
The molecular formula is as follows: c29H22Cl3NO5Molecular weight: 570.85LC-MS (M/e):570.1(M + H)+)
1H-NMR(400MHz,MeOD)δ:7.52-7.42(m,5H),7.20(d,J＝8.0Hz,1H),7.13(d,J＝8.8Hz,1H),6.86(d,J＝2.8Hz,1H),6.72(dd,J1＝2.8Hz,J2＝8.8HZ,1H),4.90(s,2H),4.30(dd,J1＝3.2Hz,J2＝10.4HZ,1H),4.06(t,1H),3.63-3.61(m,1H),3.05(d,J＝7.6Hz,2H),2.34-2.30(m,1H),1.21-1.19(m,4H)。
Example 172- (4- ((5-cyclopropyl-3- (2, 6-dichlorophenyl) isoxazol-4-yl) methoxy) phenyl) benzo Preparation of dihydropyran-6-carboxylic acid (Compound 17)
Figure BDA0001739294150000791
(1) Preparation of methyl 4-hydroxy-3-iodobenzoate
Figure BDA0001739294150000792
Methyl 4-hydroxybenzoate (3.55g,23.3mmol) was dissolved in acetic acid (20mL), a solution of iodine chloride (3.78g,23.3mmol) in acetic acid (5mL) was added dropwise, and the reaction was heated to 65 ℃ and stirred for 16 hours. The reaction was filtered, and the filter cake was washed with water (20mL) and dried to give the title compound (4.0g, 61.7% yield).
(2) Preparation of 1- (4-methoxyphenyl) ethane-1-one
Figure BDA0001739294150000793
1- (4-hydroxyphenyl) ethan-1-one (5g,36.8mmol) was dissolved in acetonitrile (50mL), potassium carbonate (10g,72.3mmol) was added, cooled to 0 deg.C, iodomethane (6.3g,44.4mmol) was added, and the reaction was stirred at 25 deg.C for 3 hours. The reaction mixture was filtered, and the filtrate was concentrated to give the title compound (5.2g, yield 94.5%).
(3) Preparation of 1- (4-methoxyphenyl) -2-propenyl-1-one
Figure BDA0001739294150000794
1- (4-methoxyphenyl) ethan-1-one (5g,33.3mmol), N-methyltrifluoroacetic acid aniline (11g,49.7mmol) and paraformaldehyde (10g,333.3mmol) were added to tetrahydrofuran (100mL), and the reaction was stirred with heating to 80 ℃ for 24 hours. The reaction mixture was filtered, ethyl acetate (100mL) and water (150mL) were added to the filtrate, the mixture was separated, the organic phase was dried over anhydrous sodium sulfate, filtered, and the filtrate was concentrated to give the crude title compound (3.9g) which was used directly in the next reaction.
(4) Preparation of methyl 4-hydroxy-3- (3- (4-methoxyphenyl) -3-oxopropyl-1-en-1-yl) benzoate
Figure BDA0001739294150000801
1- (4-methoxyphenyl) -2-propen-1-one (3.5g, crude), methyl 4-hydroxy-3-iodobenzoate (6g,21.6mmol), triethylamine (7g,69.2mmol), triphenylphosphine (0.3g,1.14mmol), palladium acetate (0.4g,1.78mmol) were added to acetonitrile (100mL), heated to 90 ℃ and the reaction stirred for 16 h. The reaction solution was concentrated, and the resulting crude product was purified by silica gel column chromatography (petroleum ether: ethyl acetate ═ 5:1) to give the title compound (0.9g, two-step yield 8.6%).
(5) Preparation of methyl 4-hydroxy-3- (3- (4-methoxyphenyl) -3-oxopropyl) benzoate
Figure BDA0001739294150000802
Methyl 4-hydroxy-3- (3- (4-methoxyphenyl) -3-oxopropyl-1-en-1-yl) benzoate (0.9g,2.88mmol) was dissolved in methanol (10mL), palladium on carbon (0.1g) was added, and the reaction was stirred at 25 ℃ under a pressure of hydrogen for 16 hours. The reaction mixture was filtered, and the filtrate was concentrated to give the title compound (0.85g, yield 94.4%).
(6) Preparation of methyl 4-hydroxy-3- (3-hydroxy-3- (4-methoxyphenyl) propyl) benzoate
Figure BDA0001739294150000803
Methyl 4-hydroxy-3- (3- (4-methoxyphenyl) -3-oxopropyl) benzoate (0.85g,2.7mmol) was dissolved in anhydrous ethanol (10mL), sodium borohydride (0.2g,5.3mmol) was added, and the reaction was stirred at 25 ℃ for 16 hours. The reaction was quenched by addition of dilute hydrochloric acid (20mL, 1M), extracted with ethyl acetate (30mL × 3), the organic phases were combined, dried over anhydrous sodium sulfate, filtered, the filtrate was concentrated, and the resulting crude product was purified by silica gel column chromatography (petroleum ether: ethyl acetate ═ 2:1) to give the title compound (0.5g, yield 58.1%).
(7) Preparation of methyl 2- (4-methoxyphenyl) chroman-6-carboxylate
Figure BDA0001739294150000811
Methyl 4-hydroxy-3- (3-hydroxy-3- (4-methoxyphenyl) propyl) benzoate (0.5g,1.58mmol) and triphenylphosphine (0.5g,1.91mmol) were dissolved in tetrahydrofuran (10mL), cooled to 0 deg.C, diethyl azodicarboxylate (0.4g,2.3mmol) was added, and the reaction was stirred at 25 deg.C under nitrogen for 16 hours. Water (20mL) and ethyl acetate (20mL) were added for separation, the aqueous phase was extracted with ethyl acetate (30mL × 3), the organic phases were combined, dried over anhydrous sodium sulfate, filtered, the filtrate was concentrated, and the resulting crude product was purified by silica gel column chromatography (petroleum ether: ethyl acetate ═ 20:1) to give the title compound (0.37g, yield 78.7%).
(8) Preparation of 2- (4-hydroxyphenyl) chroman-6-carboxylic acid
Figure BDA0001739294150000812
Methyl 2- (4-methoxyphenyl) chroman-6-carboxylate (0.37g,1.24mmol) was dissolved in dichloromethane (10mL), cooled to-60 deg.C, boron tribromide (0.93g,3.71mmol) was added, and the reaction was stirred at 25 deg.C for 6 hours. The reaction was quenched with water (20mL), extracted with ethyl acetate (30mL × 3), the organic phases combined, dried over anhydrous sodium sulfate, filtered, and the filtrate concentrated to give the title compound (0.3g, 89.5% yield).
(9) Preparation of methyl 2- (4-hydroxyphenyl) chroman-6-carboxylate
Figure BDA0001739294150000813
2- (4-hydroxyphenyl) chroman-6-carboxylic acid (0.3g,1.11mmol) is dissolved in anhydrous methanol (10mL), sulfoxide chloride (0.13g,1.1mmol) is added dropwise, and the reaction is stirred at 60 ℃ for 16 hours. The reaction mixture was concentrated, ethyl acetate (50mL) was added, and the mixture was washed with a saturated aqueous solution of sodium hydrogencarbonate (20mL), dried over anhydrous sodium sulfate, filtered, and the filtrate was concentrated to give a crude product, which was purified by silica gel column chromatography (petroleum ether: ethyl acetate 5:1) to obtain the title compound (0.2g, yield 63.4%).
(10) Preparation of methyl 2- (4- ((5-cyclopropyl-3- (2, 6-dichlorophenyl) isoxazol-4-yl) methoxy) phenyl) chroman-6-carboxylate
Figure BDA0001739294150000821
Methyl 2- (4-hydroxyphenyl) chroman-6-carboxylate (0.15g,0.53mmol) and 4- (bromomethyl) -5-cyclopropyl-3- (2, 6-dichlorophenyl) isoxazole (0.2g,0.58mmol) were dissolved in acetonitrile (10mL), potassium carbonate (0.15g,1.1mmol) was added, and the reaction was heated to 80 ℃ and stirred for 6 hours. The reaction solution was concentrated, and the resulting crude product was purified by silica gel column chromatography (petroleum ether: ethyl acetate ═ 1:1) to obtain the title compound (0.1g, yield 34.5%).
(11) Preparation of 2- (4- ((5-cyclopropyl-3- (2, 6-dichlorophenyl) isoxazol-4-yl) methoxy) phenyl) chroman-6-carboxylic acid
Figure BDA0001739294150000822
Methyl 2- (4- ((5-cyclopropyl-3- (2, 6-dichlorophenyl) isoxazol-4-yl) methoxy) phenyl) chroman-6-carboxylate (0.1g,0.18mmol) was dissolved in tetrahydrofuran (3mL) and methanol (3mL), and a solution of lithium hydroxide monohydrate (15mg,0.36mmol) in water (1mL) was added, followed by stirring at 25 ℃ for 16 hours. The reaction solution was concentrated, and the resulting crude product was purified by silica gel column chromatography (dichloromethane: methanol ═ 20:1) to give the title compound (50mg, yield 51.5%).
The molecular formula is as follows: c29H23Cl2NO5Molecular weight: 536.41 LC-MS (M/e):536.2(M + H)+)
1H-NMR(400MHz,DMSO)δ:7.71(s,1H),7.66(d,J＝8.4Hz,1H),7.61(d,J＝8Hz,2H),7.53-7.58(m,1H),7.27(d,J＝8.8Hz,2H),6.80-6.85(m,3H),5.08(d,J＝10Hz,1H),4.86(s,1H),2.91-3.00(m,1H),2.73-2.80(m,1H),2.42-2.50(m,1H),2.08-2.15(m,1H),1.95-2.02(m,1H),1.10-1.23(m,4H).
Example 182- (6- ((5-cyclopropyl-3- (2, 6-dichlorophenyl) isoxazol-4-yl) methoxy) pyridine-3- Preparation of Yl) chroman-6-carboxylic acid (Compound 18)
Figure BDA0001739294150000831
(1) Preparation of 1- (6- (benzyloxy) pyridin-3-yl) ethanone
Figure BDA0001739294150000832
1- (6-hydroxypyridin-3-yl) ethanone (3.0g,21.9mmol) was dissolved in toluene (30mL), silver carbonate (9.1g,33.3mmol) was added, benzyl bromide (3.8g,22.2mmol) was slowly added dropwise, the dropwise addition was complete, the temperature was raised to 100 ℃ and the reaction was stirred for 12 hours, the reaction was filtered and the filtrate was concentrated to give the title compound (3.9g, 78.3% yield).
(2) Preparation of 1- (6- (benzyloxy) pyridin-3-yl) prop-2-en-1-one
Figure BDA0001739294150000833
1- (6- (benzyloxy) pyridin-3-yl) ethanone (3.8g,16.7mmol), N-methyltrifluoroacetic acid aniline (5.5g,24.9mmol) and paraformaldehyde (5.0g,166.7mmol) were added to tetrahydrofuran (50mL), and the reaction was stirred at 80 ℃ for 24 hours. The reaction mixture was filtered, ethyl acetate (100mL) and water (150mL) were added to the filtrate, the layers were separated, and the organic layer was dried over anhydrous sodium sulfate and concentrated to give the crude title compound (2.0g) which was used directly in the next reaction.
(3) Preparation of methyl (E) -3- (3- (6- (benzyloxy) pyridin-3-yl) -3-oxoprop-1-en-1-yl) -4-hydroxybenzoate
Figure BDA0001739294150000834
Crude 1- (6- (benzyloxy) pyridin-3-yl) prop-2-en-1-one (2.0g), methyl 4-hydroxy-3-iodobenzoate (2.3g,8.4mmol), triethylamine (2.6g,25.7mmol), triphenylphosphine (100mg,0.38mmol) and palladium acetate (128mg,0.57mmol) were added to acetonitrile (30mL) and the reaction was stirred at 90 ℃ for 12 hours. The reaction was concentrated and purified by silica gel column chromatography (petroleum ether: ethyl acetate ═ 5:1) to give the title compound (0.5g, two-step yield 7.7%).
(4) Preparation of methyl 4-hydroxy-3- (3- (6-hydroxypyridin-3-yl) -3-oxopropyl) benzoate
Figure BDA0001739294150000841
(E) -3- (3- (6- (benzyloxy) pyridin-3-yl) -3-oxoprop-1-en-1-yl) -4-hydroxybenzene (0.5g,1.3mmol) was dissolved in methanol (5mL), and 10% palladium on carbon (50mg) was added thereto, followed by stirring at 25 ℃ under hydrogen pressure for 12 hours. Filtration and concentration of the filtrate yielded the product (0.3g, 77.5% yield).
(5) Preparation of methyl 4-hydroxy-3- (3-hydroxy-3- (6-hydroxypyridin-3-yl) propyl) benzoate
Figure BDA0001739294150000842
Methyl 4-hydroxy-3- (3- (6-hydroxypyridin-3-yl) -3-oxopropyl) benzoate (0.3g,1.0mmol) was dissolved in anhydrous methanol (5mL), sodium borohydride (115mg,3.0mmol) was added, the reaction was stirred at 25 ℃ for 1 hour, the temperature was reduced to 0 ℃ and diluted hydrochloric acid (1M) was added to adjust pH 7, the mixture was extracted with ethyl acetate (20mL × 3), the organic phases were combined, dried over anhydrous sodium sulfate, concentrated, and subjected to silica gel column chromatography (dichloromethane: methanol ═ 10:1) to obtain the title compound (290mg, 96.7% yield).
(6) Preparation of methyl 2- (6-hydroxypyridin-3-yl) chroman-6-carboxylate
Figure BDA0001739294150000843
Methyl 4-hydroxy-3- (3-hydroxy-3- (6-hydroxypyridin-3-yl) propyl) benzoate (290mg,0.96mmol) and triphenylphosphine (0.76g,2.9mmol) were dissolved in tetrahydrofuran (10mL), cooled to 0 deg.C, diethyl azodicarboxylate (0.5g,2.9mmol) was added, and the reaction was stirred at 25 deg.C under nitrogen for 12 hours. Water (10mL) and ethyl acetate (20mL) were added for separation, the aqueous phase was extracted with ethyl acetate (20mL × 3), the organic phases were combined, dried over anhydrous sodium sulfate, concentrated, and the crude product was subjected to silica gel column chromatography (dichloromethane: methanol ═ 40:1) to give the title compound (112mg, yield 41.0%).
(7) Preparation of methyl 2- (6- ((5-cyclopropyl-3- (2, 6-dichlorophenyl) isoxazol-4-yl) methoxy) pyridin-3-yl) chroman-6-carboxylate
Figure BDA0001739294150000851
Methyl 2- (6-hydroxypyridin-3-yl) chroman-6-carboxylate (100mg,0.35mmol), 4- (bromomethyl) -5-cyclopropyl-3- (2, 6-dichlorophenyl) isoxazole (122mg,0.35mmol), silver carbonate (193mg,0.7mmol) were dissolved in toluene (5mL), warmed to 100 ℃ and stirred for 12 hours, the reaction was filtered and the crude product was subjected to silica gel column chromatography (petroleum ether: ethyl acetate ═ 3:1) to give the title compound (127mg, 65.8% yield).
(8) Preparation of 2- (6- ((5-cyclopropyl-3- (2, 6-dichlorophenyl) isoxazol-4-yl) methoxy) pyridin-3-yl) chroman-6-carboxylic acid
Figure BDA0001739294150000852
Methyl 2- (6- ((5-cyclopropyl-3- (2, 6-dichlorophenyl) isoxazol-4-yl) methoxy) pyridin-3-yl) chroman-6-carboxylate (120mg,0.22mmol) was dissolved in tetrahydrofuran (3mL) and methanol (3mL), and an aqueous solution (1mL) of lithium hydroxide monohydrate (47mg,1.1mmol) was added, followed by stirring at 25 ℃ for 16 hours. Dilute hydrochloric acid (1M) was added to adjust pH to 7, and the reaction solution was concentrated and subjected to silica gel column chromatography (dichloromethane: methanol to 20:1) to give the title compound (40mg, yield 33.8%).
The molecular formula is as follows: c28H22Cl2N2O5Molecular weight: 537.39 LC-MS (M/e):537.1(M + H)+)
1H-NMR(400MHz,CDCl3)δ:8.08(s,1H),7.89(d,J＝7.2Hz,2H),7.61(dd,J1＝8.8Hz,J2＝6.4Hz,1H),7.38-7.41(m,2H),7.28-7.34(m,1H),6.91(d,J＝8.8Hz,1H),6.67(d,J＝8.8Hz,1H),5.31(s,2H),5.09(dd,J1＝10.0Hz,J2＝2.0Hz,1H),3.01-3.04(m,1H),2.86-2.90(m,1H),2.32-2.36(m,1H),2.20-2.25(m,1H),2.09-2.12(m,1H),1.28-1.31(m,2H),1.13-1.18(m,2H).
Example 19(2- (2-chloro-4- ((5-cyclopropyl-3- (2, 6-dichlorophenyl) isoxazol-4-yl) methoxy) benzene Preparation of Yl) chroman-6-yl) methanol (Compound 19)
Figure BDA0001739294150000861
2- (2-chloro-4- ((5-cyclopropyl-3- (2, 6-dichlorophenyl) isoxazol-4-yl) methoxy) phenyl) chroman-6-carboxylic acid (220mg,0.39mmol) was added to a 50mL three-necked round bottom flask, dissolved in 20mL tetrahydrofuran under nitrogen, cooled to 0 deg.C, borane (1mol/L in THF) (1.94mL) was added dropwise, raised to 25 deg.C and stirred for 1.0 hour. The reaction solution was cooled to 0 ℃ and hydrochloric acid (2N) was added to quench the reaction. Diluting with 100mL of ethyl acetate, washing with saturated brine (2X 30mL), separating the layers to obtain an ethyl acetate phase, drying over anhydrous sodium sulfate, concentrating, and isolating under high pressure to obtain the product (203mg, 93.5% yield).
Molecular formula C29H24Cl3NO4Molecular weight 556.86 LC-MS (ES, M/z):578(M + Na)
1H NMR:(CD3OD,ppm)δ:7.38-7.54(m,4H),7.09-7.11(m,2H),6.77-6.85(m,3H),5.27-5.31(dd,J1＝2.1Hz,J2＝10.2Hz,1H),4.93(s,2H),4.51(s,2H),2.76-3.00(m,2H),2.21-2.37(m,2H),1.86-1.89(m,1H),1.20-1.23(m,4H).
Example 202- (2-chloro-4- ((5-cyclopropyl-3- (2, 6-dichlorophenyl) isoxazol-4-yl) methoxy) benzene Preparation of yl) -N- (methylsulfonyl) chroman-6-carboxamide (Compound 20)
Figure BDA0001739294150000862
A100-mL three-necked flask was charged with 2- (2-chloro-4- ((5-cyclopropyl-3- (2, 6-dichlorophenyl) isoxazol-4-yl) methoxy) phenyl) -chroman-6-carboxylic acid (200mg,0.35mmol), dissolved in dichloromethane (50mL), EDCI (134mg,0.70mmol), 4-dimethylaminopyridine (128mg,1.05mmol), methanesulfonamide (66.5mg,0.70 mmol). Stir at room temperature overnight under nitrogen. The reaction mixture was added to methylene chloride (100mL), followed by washing with saturated brine (3X 50mL), and the organic layer was separated, dried over anhydrous sodium sulfate, concentrated, and purified by column chromatography to give 168mg of the product in 74% yield.
Molecular formula C30H25Cl3N2O6S molecular weight 647.95 LC-MS (ES, M/z):647(M +1)+
1HNMR(DMSO,ppm):δ11.89(brs,1H),7.82(s,1H),7.71-7.74(dd,J1＝2.1Hz,J2＝8.4Hz,1H),7.61-7.64(m,2H),7.52-7.57(m,1H),7.38-7.40(d,J＝8.7Hz,1H),6.98-6.99(d,J＝2.7Hz,1H),6.85-6.92(m,1H),6.82-6.83(m,1H),5.35-5.38(m,1H),4.94(s,2H),3.32-3.34(d,J＝6.6Hz,3H),2.71-3.06(m,2H),1.93-2.49(m,3H),1.13-1.21(m,4H).
Example 214- ((4- (6- (2H-tetrazol-5-yl) chroman-2-yl) -3-chlorophenoxy) methyl) - Preparation of 5-cyclopropyl-3- (2, 6-dichlorophenyl) isoxazole (Compound 21)
Figure BDA0001739294150000871
(1) Preparation of 2- (2-chloro-4- ((5-cyclopropyl-3- (2, 6-dichlorophenyl) isoxazol-4-yl) methoxy) phenyl) chroman-6-carbonitrile
Figure BDA0001739294150000872
Referring to the preparation of step (6) of example 1,2- (2-chloro-4-hydroxyphenyl) chroman-6-carbonitrile (3g,10.50mmol), 4- (chloromethyl) -5-cyclopropyl-3- (2, 6-dichlorophenyl) isoxazole (3.802g,12.57mmol) was added. 2.8g of product are obtained, with a yield of 48.4%.
(2) Preparation of 4- ((4- (6- (2H-tetrazol-5-yl) chroman-2-yl) -3-chlorophenoxy) methyl) -5-cyclopropyl-3- (2, 6-dichlorophenyl) isoxazole
Figure BDA0001739294150000881
A50 mL round-bottomed flask was charged with 2- (2-chloro-4- ((5-cyclopropyl-3- (2, 6-dichlorophenyl) isoxazol-4-yl) methoxy) phenyl) chroman-6-carbonitrile (500mg,0.91mmol), ammonium chloride (975mg,18.23mmol), N, N-dimethylformamide (15mL), NaN3(900mg,13.84 mmol). The reaction mixture is heated to 120 ℃ and reacted for 12 hours, and NaHSO is added into the system3The reaction was quenched (30 mL). Ethyl acetate (3X 50mL) was added for extraction. The combined organic phases are concentratedThe title compound was obtained in 261mg, 48.4% yield.
Molecular formula C29H22Cl3N5O3Molecular weight 594.88 LC-MS (ES, M/z):594(M +1)+
1HNMR(DMSO-d6,ppm):δ7.79(s,1H),7.75(d,J＝8.4Hz,1H),7.61-7.64(m,2H),7.52-7.57(m,1H),7.43(d,J＝8.7Hz,1H),6.97(d,J＝2.4Hz,1H),6.82-6.88(m,2H),5.32(d,J＝8.4Hz,1H),4.94(s,2H),3.03-3.10(m,1H),2.80-2.88(m,1H),2.45(d,J＝3.3Hz,1H),2.14-2.19(m,1H),1.92-2.00(m,1H),1.10-1.22(m,4H).
Example 222- (6- ((5-cyclopropyl-3- (2, 6-dichlorophenyl) isoxazol-4-yl) methoxy) -2- (trifluoro-phenyl) Preparation of methyl) pyridin-3-yl) chroman-6-carboxylic acid (Compound 23)
Figure BDA0001739294150000882
(1) Preparation of 6-oxo-2- (trifluoromethyl) -1, 6-dihydropyridine-3-carboxylic acid ethyl ester
Figure BDA0001739294150000883
A2000 mL three-necked flask was charged with ethyl 6-oxo-2- (trifluoromethyl) -1,4,5, 6-tetrahydropyridine-3-carboxylate (90g,379.5mmol), CCl4(900mL), NBS (81.88g,460.05 mmol). The reaction mixture was warmed to 80 ℃ for 24 hours, cooled to room temperature, filtered to remove solids, and the filtrate was washed with saturated brine (3X 2000 mL). The separated organic layer was dried over anhydrous sodium sulfate, concentrated under reduced pressure, and the residue was purified by column chromatography (ethyl acetate/petroleum ether ═ 1:10) to give the title compound 50g in 56% yield.
(2) Preparation of 6- (benzyloxy) -2- (trifluoromethyl) nicotinic acid ethyl ester
Figure BDA0001739294150000891
A1000 mL three-necked flask was charged with ethyl 6-oxo-2- (trifluoromethyl) -1, 6-dihydropyridine-3-carboxylate (50g,212.6mmol), toluene (500mL), Ag2CO3(76g,276.7mmol) and BnBr (43.7g,255.5 mmol). The reaction mixture was heated to 50 ℃ for 12 hours, cooled to room temperature, diluted with ethyl acetate (1000mL), washed with saturated brine (3 × 1000mL), the organic layer was separated, dried over anhydrous sodium sulfate, concentrated under reduced pressure, and the residue was purified by column chromatography (ethyl acetate/petroleum ether ═ 1:20 to 1:10) to give 37.5g of the title compound in 54% yield.
(3) Preparation of (6- (benzyloxy) -2- (trifluoromethyl) pyridin-3-yl) methanol
Figure BDA0001739294150000892
A2000 mL three-necked flask was charged with a solution of ethyl 6- (benzyloxy) -2- (trifluoromethyl) nicotinate (30g,92.23mmol) in tetrahydrofuran (300mL), the temperature was reduced to-78 deg.C under nitrogen, DIBAL-H (1M in tolumen, 277mL) was added dropwise, and stirring was continued at-78 deg.C for 3 hours. The reaction was quenched by the addition of methanol (140mL) dropwise, allowed to warm to room temperature for 5 minutes, potassium sodium tartrate (280mL aqueous solution) was added dropwise, the solid was filtered off, the filtrate was diluted with ethyl acetate (2000mL) and washed with saturated brine (3X 2000 mL). The separated organic layer was dried over anhydrous sodium sulfate, concentrated under reduced pressure, and the residue was purified by column chromatography (ethyl acetate/petroleum ether ═ 1:6 to 1:4) to give 24g of the title compound in 92% yield.
(4) Preparation of 6- (benzyloxy) -2- (trifluoromethyl) nicotinaldehyde
Figure BDA0001739294150000893
A1000 mL three-necked flask was charged with a solution of (6- (benzyloxy) -2- (trifluoromethyl) pyridin-3-yl) methanol (24g,84.73mmol) in methylene chloride (500 mL). The temperature was lowered to 0 ℃ and Dess-Martin periodinane (43.1g,101.6mmol) was added in portions. The reaction mixture was stirred at room temperature for 18 hours. The solid was filtered off, the filtrate was diluted with dichloromethane (500mL) and washed with saturated brine (3X 1000 mL). The organic layer was separated, dried over anhydrous sodium sulfate, concentrated under reduced pressure, and the residue was purified by column chromatography (ethyl acetate/petroleum ether ═ 1:8 to 1:6) to give 23g of a product with a yield of 97%.
(5) Preparation of (E) -3- (6- (benzyloxy) -2- (trifluoromethyl) pyridin-3-yl) -1- (5-bromo-2-hydroxyphenyl) prop-2-en-1-one
Figure BDA0001739294150000901
A2000 mL three-necked flask was charged with a solution of 6- (benzyloxy) -2- (trifluoromethyl) nicotinaldehyde (23g,81.78mmol) in ethanol (460mL) and 1- (5-bromo-2-hydroxyphenyl) ethan-1-one (17.5g,81.38 mmol). Then, a KOH solution (32.1g,572.2mmol) was added in portions, the mixture was heated at 60 ℃ and stirred for 48 hours, and the reaction was quenched by adding ice/water (1000 mL). The pH of the solution was adjusted to 7 with hydrochloric acid (6 mol/L). The solid was filtered off and dried under reduced pressure to give 36g of product in 92% yield.
(6) Preparation of 6-bromo-2- (6-hydroxy-2- (trifluoromethyl) pyridin-3-yl) chroman-4-one
Figure BDA0001739294150000902
To a 2000mL three-necked flask were added (E) -3- (6- (benzyloxy) -2- (trifluoromethyl) pyridin-3-yl) -1- (5-bromo-2-hydroxyphenyl) prop-2-en-1-one (36g,75.27mmol), acetic acid (360mL), hydrochloric acid (12N,360mL), and sulfuric acid (1 mL). The reaction mixture was reacted at 88 ℃ for 16 hours, cooled to room temperature, poured into ice water (2000mL), filtered to give a solid, and dried under reduced pressure to give 23.6g of the title compound in 81% yield.
(7) Preparation of 5- (6-bromochromen-2-yl) -6- (trifluoromethyl) pyridin-2-ol
Figure BDA0001739294150000903
HgCl was added to a 250-mL three-necked flask2(6.96g) and hydrochloric acid (5N) (150mL), then the temperature was lowered to 0 ℃ and Zn (16.4g) was added in portions. The system was stirred at room temperature for 30 minutes, the liquid phase was partitioned, hydrochloric acid (5N,100mL) was added to the remaining solid, stirring at room temperature for 10 minutes was conducted, the liquid phase was partitioned, and hydrochloric acid (5N,150mL) and a solution of 6-bromo-2- (6-hydroxy-2- (trifluoromethyl) pyridin-3-yl) chroman-4-one (6.6g,17.00mmol) in toluene (100mL) were added to the remaining solid. The reaction mixture was stirred at 80 ℃ for 16 hours, cooled, ethyl acetate (500mL) was added, and the organic layer was washed with saturated brine (3X 500 mL). The organic phase was separated, dried over anhydrous sodium sulfate, concentrated under reduced pressure, and the residue was purified by column chromatography (ethyl acetate/petroleum ether: 1:10 to 1:5) to obtain 3g of product with a yield of 47%.
(8) Preparation of 2- (6-hydroxy-2- (trifluoromethyl) pyridin-3-yl) chroman-6-carbonitrile
Figure BDA0001739294150000911
A100 mL three-necked flask was charged with 5- (6-bromobenzodihydropyran-2-yl) -6- (trifluoromethyl) pyridin-2-ol (3g,8.02mmol), NMP (30mL), ZnCN2(1.12g,9.5mmol)，Pd(PPh3)4(930mg,0.81 mmol). The reaction was carried out at 120 ℃ for 16 hours under nitrogen protection. The reaction solution was cooled to room temperature, ethyl acetate (200mL) was added for dilution, the solid was filtered off, the filtrate was washed with saturated brine (3 × 200mL), the organic phase was separated, anhydrous sodium sulfate was added for drying, the concentration was performed under reduced pressure, and the residue was purified by column chromatography (ethyl acetate/petroleum ether: 1:10 to 1:6) to obtain 1.7g of a product, with a yield of 66%.
(9) Preparation of methyl 2- (6-hydroxy-2- (trifluoromethyl) pyridin-3-yl) chroman-6-carboxylate
Figure BDA0001739294150000912
2- (6-hydroxy-2- (trifluoromethyl) pyridin-3-yl) chroman-6-carbonitrile (1.7g,5.31mmol), methanol (30mL) was added to a 100mL three-necked flask, followed by concentrated sulfuric acid (3mL) dropwise at room temperature. The reaction was refluxed for three days, cooled to room temperature, poured into ice water (200mL), the solid collected, washed with water (3X 50mL), and the solid was dried under reduced pressure to give the title compound 1.3g in 69% yield.
(10) Preparation of methyl 2- (6- ((5-cyclopropyl-3- (2, 6-dichlorophenyl) isoxazol-4-yl) methoxy) -2- (trifluoromethyl) pyridin-3-yl) chroman-6-carboxylate
Figure BDA0001739294150000921
A100 mL three-necked flask was charged with methyl 2- (6-hydroxy-2- (trifluoromethyl) pyridin-3-yl) chroman-6-carboxylate (1.3g,3.68mmol) in N, N-dimethylformamide (30mL), K2CO3(2.55g,18.45mmol), NaI (1.66g,11.07mmol), 4- (chloromethyl) -5-cyclopropyl-3- (2, 6-dichlorophenyl l) -isoxazole (1.33g,4.40 mmol). The reaction mixture was reacted at 60 ℃ for 16 hours, cooled to room temperature, diluted with ethyl acetate (200mL) and washed with saturated brine (3X 200 mL). The organic phase was separated, dried over anhydrous sodium sulfate and concentrated under reduced pressure to give 0.7g of the product in 31% yield.
(11) Preparation of 2- (6- ((5-cyclopropyl-3- (2, 6-dichlorophenyl) isoxazol-4-yl) methoxy) -2- (trifluoromethyl) pyridin-3-yl) chroman-6-carboxylic acid
Figure BDA0001739294150000922
A250 mL three-necked flask was charged with a solution of methyl 2- (6- ((5-cyclopropyl-3- (2, 6-dichlorophenyl) isoxazol-4-yl) methoxy) -2- (trifluoromethyl) pyridin-3-yl) chroman-6-carboxylate (700mg,1.13mmol) in methanol (100mL), water (10mL) and LiOH. H2O (143mg,3.40 mmol). The reaction solution was stirred at room temperature for 3 days. Concentrated under reduced pressure, and the residue was added to ice water (100mL) and adjusted to pH 7 with hydrochloric acid (1 mol/L). Filtration gave a solid which was washed with water (3X 100mL) and n-hexane (3X 100mL) to give 210mg of the title compound in 31% yield.
Molecular formula C29H21Cl2F3N2O5Molecular weight 605.39LC-MS (ES, M/z) 605.1(M +1)+
1HNMR(300MHz,DMSO-d6,ppm):δ12.60(brs,1H),8.08-8.05(d,J＝9.0Hz,1H),7.77(s,1H),7.71-7.68(m,1H),7.57-7.46(m,3H),7.00-6.98(d,J＝8.7Hz,1H),6.90-6.87(d,J＝8.7Hz,1H),5.40-5.27(m,3H),3.11-3.00(m,1H),2.90-2.85(m,1H),2.57-2.55(m,1H),2.09-2.00(m,2H),1.23-1.14(m,4H).
Example 232- (4- ((5-cyclopropyl-3- (2, 6-dichlorophenyl) isoxazol-4-yl) methoxy) -2-fluorobenzene Yl) preparation of chroman-6-carboxylic acid (Compound 24)
Figure BDA0001739294150000931
(1) Preparation of 1- (2-fluoro-4-methoxyphenyl) prop-2-en-1-one
Figure BDA0001739294150000932
Referring to the preparation method in step (3) of example 17, 1- (2-fluoro-4-methoxyphenyl) ethan-1-one (1.7g,10.1mmol) was added to give 2.0g of a product (crude product).
(2) Preparation of methyl (E) -3- (3- (2-fluoro-4-methoxyphenyl) -3-oxoprop-1-en-1-yl) -4-hydroxybenzoate
Figure BDA0001739294150000933
The production method in step (4) of reference example 17 gave a product (0.9g, two-step yield 27.0%).
(3) Preparation of methyl 3- (3- (2-fluoro-4-methoxyphenyl) -3-oxopropyl) -4-hydroxybenzoate
Figure BDA0001739294150000934
Referring to the preparation method of step (5) of example 17, (E) -methyl 3- (3- (2-fluoro-4-methoxyphenyl) -3-oxoprop-1-en-1-yl) -4-hydroxybenzoate (0.83g,2.51mmol) was added to give 0.8g of a product in 95.8% yield.
(4) Preparation of methyl 3- (3- (2-fluoro-4-methoxyphenyl) -3-hydroxypropyl) -4-hydroxybenzoate
Figure BDA0001739294150000935
Methyl 3- (3- (2-fluoro-4-methoxyphenyl) -3-oxopropyl) -4-hydroxybenzoate (800mg,2.41mmol) was dissolved in methanol (20mL), cooled to 0 deg.C, and NaBH was added4(146mg,3.86mmol), and reacted at 0 ℃ for 2 hours. The reaction mixture was poured into water (50mL), extracted with ethyl acetate (100 mL. times.3), and the organic layers were combined, dried over anhydrous sodium sulfate, filtered, and concentrated to give 1.1g of the product (crude).
(5) Preparation of methyl 2- (2-fluoro-4-methoxyphenyl) chroman-6-carboxylate
Figure BDA0001739294150000941
Methyl 3- (3- (2-fluoro-4-methoxyphenyl) -3-hydroxypropyl) -4-hydroxybenzoate (1.1g, crude) was added to phosphoric acid (10mL), and the mixture was heated to 90 ℃ for 30 minutes. Then poured into water (50mL), neutralized with sodium carbonate to neutrality, then extracted with ethyl acetate (100mL × 3), the organic layers were combined, concentrated, and the residue was subjected to silica gel column chromatography (petroleum ether: ethyl acetate ═ 10:1) to give 700mg of the product in 91.9% yield in two steps.
(6) Preparation of methyl 2- (2-fluoro-4-hydroxyphenyl) chroman-6-carboxylate
Figure BDA0001739294150000942
Methyl 2- (2-fluoro-4-methoxyphenyl) chroman-6-carboxylate (700mg,2.21mmol) was dissolved in DCM (30mL) and cooledTo-20 ℃ N2Adding BBr slowly under protection3(2.7g,10.77mmol), and the reaction was carried out for 4 hours. After completion of the reaction, water (20mL) was slowly added, followed by extraction with DCM (50mL × 3), and the organic layers were combined, concentrated, and the residue was subjected to silica gel column chromatography (petroleum ether: ethyl acetate ═ 3:1) to give 350mg of the product in 52.3% yield.
(7) Preparation of methyl 2- (4- ((5-cyclopropyl-3- (2, 6-dichlorophenyl) isoxazol-4-yl) methoxy) -2-fluorophenyl) chroman-6-carboxylate
Figure BDA0001739294150000943
Methyl 2- (2-fluoro-4-hydroxyphenyl) chroman-6-carboxylate (0.3g,0.99mmol) and 4- (bromomethyl) -5-cyclopropyl-3- (2, 6-dichlorophenyl) isoxazole (0.35g,1.0mmol) and potassium carbonate (0.28g,2.02mmol) were added sequentially to DMF (20mL), heated to 60 ℃ for reaction for 4.6 hours, poured into water (100mL), solid precipitated, filtered, and the filter cake was subjected to silica gel column chromatography (petroleum ether: ethyl acetate 10:1) to give 0.43g of the product in 76.2% yield.
(8) Preparation of 2- (4- ((5-cyclopropyl-3- (2, 6-dichlorophenyl) isoxazol-4-yl) methoxy) -2-fluorophenyl) chroman-6-carboxylic acid
Figure BDA0001739294150000951
Methyl 2- (4- ((5-cyclopropyl-3- (2, 6-dichlorophenyl) isoxazol-4-yl) methoxy) -2-fluorophenyl) chroman-6-carboxylate (0.3g,0.528mmol) was dissolved in THF (10mL), water (10mL) and lithium hydroxide monohydrate (110mg,2.62mmol) were added, and the mixture was heated to 50 ℃ and stirred for 24 hours. THF was evaporated, water (10mL) was added, the pH was adjusted to 2-3 with dilute hydrochloric acid (1M) to 0 ℃, a solid was precipitated, filtered, and the filter cake was washed with water (20mL) and acetonitrile (20mL) in that order and dried to give the product (200mg, yield 68.4%).
The molecular formula is as follows: c29H22Cl2FNO5Molecular weight: 554.40 LC-MS (M/e):555.2 (M)+)
1H-NMR(400MHz,DMSO)δ:12.55(s,1H),7.73(s,1H),7.67(d,J＝8.4Hz,1H),7.61-7.64(m,2H),7.52-7.55(m,1H),7.34(t,J＝8.4Hz,1H),6.85(d,J＝8.4Hz,1H),6.76(d,J＝10.8Hz,1H),6.68(d,J＝8.4Hz,1H),5.29(d,J＝10.0Hz,1H),4.86(s,2H),2.96-3.05(m,1H),2.78-2.82(m,1H),2.33-2.42(m 1H),1.95-2.10(m 1H),1.11-1.21(m,4H)。
Example 242- (4- ((5-cyclopropyl-3- (2, 6-dichlorophenyl) isoxazol-4-yl) methoxy) -2- (trifluoro-phenyl) Preparation of methyl) phenyl) chroman-6-carboxylic acid (Compound 26)
Figure BDA0001739294150000952
(1) Preparation of 4-methoxy-2-trifluoromethylbenzaldehyde
Figure BDA0001739294150000953
The compound 1-bromo-4-methoxy-2-trifluorotoluene (12.7g,49.8mmol) was placed in a 250ml three-necked flask, 150ml of THF was added thereto to dissolve it, and after sealing, N was added2Ventilation was performed 3 times. The reaction flask was placed under dry ice conditions at-78 ℃ and stirred. 22ml of n-butyllithium (2.5M) was added to the above reaction flask, and after stirring at-78 ℃ for 30 minutes, 4g of DMF was slowly added thereto, and after continuing the reaction for about 15 minutes, the mixture was allowed to stand at room temperature for reaction, and the progress of the reaction was monitored by TLC. After the reaction was completed, the solvent was concentrated and poured into 150ml of a saturated aqueous NaCl solution, extracted 3 times with ethyl acetate (150ml × 3), the organic phase was dried with anhydrous sodium sulfate, and concentrated and subjected to silica gel column chromatography (EA: PE ═ 1:30) to obtain a product (4.8g, yield 47.1%).
(2) Preparation of 1- (4-methoxy-2- (trifluoromethyl) phenyl) prop-2-en-1-ol
Figure BDA0001739294150000961
The compound 4-methoxy-2-trifluoromethylbenzaldehyde (4.7g,23.0mmol) was placed inAdding 100ml THF into a 250ml three-mouth bottle, dissolving, sealing, and adding N2Ventilation was performed 3 times. 27.6ml of vinyl magnesium bromide (1M) was slowly added to the above reaction flask under ice bath condition, and after about 15min of reaction, the reaction flask was moved to room temperature to continue the reaction for about 2h to complete the reaction. Saturated NH for reaction solution4The Cl solution was quenched, extracted 3 times with ethyl acetate (150 ml. times.3), the organic phase was dried over anhydrous sodium sulfate, and the solvent was concentrated to give the crude product (5.75g) which was used in the next reaction without further purification.
(3) Preparation of 1- (4-methoxy-2- (trifluoromethyl) phenyl) prop-2-en-1-one
Figure BDA0001739294150000962
The crude compound 1- (4-methoxy-2- (trifluoromethyl) phenyl) prop-2-en-1-ol obtained in the above step was dissolved in dichloromethane, and dessimutan oxidant (12.7g,30.0mmol) was added thereto with stirring at room temperature, and stirred at room temperature for 12 hours. After the reaction was completed, the reaction solution was concentrated, and the residue was subjected to column chromatography to obtain the product (2.7g, two-step yield 51.0%).
(4) Preparation of methyl 4-hydroxy-3-iodobenzoate
Figure BDA0001739294150000963
Referring to the preparation of step (1) of example 17, methyl 4-hydroxybenzoate (11.8g,77.6mmol) was added to give a product (7.6g, yield 35.2%).
(5) Preparation of (E) -4-hydroxy-3- (3- (4-methoxy-2- (trifluoromethyl) phenyl) -3-oxoprop-1-en-1-yl) -benzoic acid methyl ester
Figure BDA0001739294150000971
1- (4-methoxy-2- (trifluoromethyl) phenyl) prop-2-en-1-one (2.7g,11.7mmol) and methyl 4-hydroxy-3-iodobenzoate (3.3g,11.7mmol) were reacted according to step (4) of example 17 to give a product (3.8g, yield 85.4%).
(6) Preparation of methyl 4-hydroxy-3- (3- (4-methoxy-2- (trifluoromethyl) phenyl) -3-oxopropyl) -benzoate
Figure BDA0001739294150000972
The crude product obtained by the preparation process of step (5) of reference example 17 was used directly in the next reaction.
(7) Preparation of 4-hydroxy-3- (3-hydroxy-3- (4-methoxy-2- (trifluoromethyl) phenyl) -propyl) -benzoic acid methyl ester
Figure BDA0001739294150000973
Referring to the preparation of step (4) of example 23, the product was obtained (3.50g, two-step yield 91.1%).
(8) Preparation of methyl 2- (4-methoxy-2- (trifluoromethyl) phenyl) chroman-6-carboxylate
Figure BDA0001739294150000974
The compound 4-hydroxy-3- (3-hydroxy-3- (4-methoxy-2- (trifluoromethyl) phenyl) propyl) -benzoic acid methyl ester (1.92g,5mmol), triphenylphosphine (1.57g,6mmol), diethyl azodicarboxylate (1.04g,6mmol) was dissolved in tetrahydrofuran (30mL) and the mixture was stirred at 0 ℃. After the reaction was completed, the solvent was concentrated, 100mL of clear water was added thereto, extraction was performed 3 times with ethyl acetate (100 mL. times.3), the organic layers were combined, dried over anhydrous sodium sulfate, filtered, and concentrated, and the residue was subjected to silica gel column chromatography to obtain a product (1.29g, yield 70.5%).
(9) Preparation of methyl 2- (4-hydroxy-2- (trifluoromethyl) phenyl) chroman-6-carboxylate
Figure BDA0001739294150000981
The compound methyl 2- (4-methoxy-2- (trifluoromethyl) phenyl) chroman-6-carboxylate (550mg,1.5mmol) was dissolved in dichloromethane (10mL), a solution of boron tribromide in dichloromethane (1M,7.5mL) was slowly added dropwise, and the reaction was continued at-78 ℃ for 3 hours after the addition was completed. After the reaction was completed, the reaction solution was allowed to warm to room temperature, poured into ice water, extracted three times with ethyl acetate (50 ml. times.3), the organic phases were washed with saturated aqueous sodium chloride solution, combined, concentrated and subjected to column chromatography to give the product (200mg, yield 37.9%).
(10) Preparation of methyl 2- (4- ((5-cyclopropyl-3- (2, 6-dichlorophenyl) isoxazol-4-yl) methoxy) -2- (trifluoromethyl) phenyl) chroman-6-carboxylate
Figure BDA0001739294150000982
Methyl 2- (4-hydroxy-2- (trifluoromethyl) phenyl) chroman-6-carboxylate (100mg,0.28mmol) is dissolved in DMF (5mL), potassium carbonate (58mg,0.42mmol) is added, stirring is carried out at 25 ℃ for 10 minutes, then 4- (bromomethyl) -5-cyclopropyl-3- (2, 6-dichlorophenyl) isoxazole (97mg,0.28mmol) is added and reaction is carried out at 25 ℃ for 6 hours. After the reaction was completed, the mixture was slowly poured into ice water to precipitate a solid, filtered, and the filter cake was washed with water (20mL), collected and dried to obtain a crude product (154mg), which was used directly in the next reaction.
(11) Preparation of 2- (4- ((5-cyclopropyl-3- (2, 6-dichlorophenyl) isoxazol-4-yl) methoxy) -2- (trifluoromethyl) phenyl) chroman-6-carboxylic acid
Figure BDA0001739294150000983
Methyl 2- (4- ((5-cyclopropyl-3- (2, 6-dichlorophenyl) isoxazol-4-yl) methoxy) -2- (trifluoromethyl) phenyl) chroman-6-carboxylate (154mg) was dissolved in THF (2mL), methanol (2mL) and water (1mL) were added, and lithium hydroxide monohydrate (32mg,0.75mmol) was further added, and the mixture was allowed to react at 30 ℃ for 2 hours. After the reaction was completed, the reaction solution was poured into 50mL of clear water, extracted with ethyl acetate three times (50 mL. times.3), the organic phase was washed with a saturated sodium chloride solution, dried over anhydrous sodium sulfate, concentrated, and subjected to silica gel column chromatography to obtain the final product (50mg, 30% yield in two steps).
The molecular formula is as follows: c30H22Cl2F3NO5Molecular weight: 604.40 LC-MS (M/e):604.1(M + H)+
1H-NMR(400MHz,CDCl3)δ:7.91(s,1H),7.88(d,J＝2.0Hz,1H),7.59(d,J＝8.8,1H),7.41-7.43(m,2H),7.32-7.36(m,1H),7.23(s,1H),7.09(d,J＝2.0Hz,1H),7.04(dd,J1＝2.4Hz,J2＝8.4Hz,1H),5.38(d,J＝10.4Hz,1H),4.89(s,2H),2.91-3.08(m,1H),2.85-2.87(m,1H),2.15-2.19(m,1H),1.90-1.99(m,1H),1.16-1.24(m,4H)。
Example 252- (2, 6-dichloro-4- ((5-cyclopropyl-3- (2, 6-dichlorophenyl) isoxazol-4-yl) methoxy) Preparation of phenyl) chroman-6-carboxylic acid (Compound 27)
Figure BDA0001739294150000991
(1) Preparation of methyl 4-hydroxy-3-iodobenzoate
Figure BDA0001739294150000992
Referring to the preparation method of step (1) of example 17, methyl 4-hydroxybenzoate (10.1g,66.4mol) was added to obtain a product (15.0g, yield 81.3%).
(2) Preparation of (3, 5-dichlorophenoxy) triisopropylsilane
Figure BDA0001739294150000993
3, 5-dichlorophenol (11.5g,70.6mmol), DIPEA (13.7g,106.2mmol), DMAP (861mg,7.06mmol) were added to DCM (200mL), TIPSCl (16.33g,84.7mmol) was added in portions while cooling on ice, and then the reaction was continued for 8 hours while rising to 25 ℃. 1N HCl (126mL) was added to the system, the solution was extracted and separated, and NaHCO was used in order3Washed (120mL) with NaCl solution (150mL), the organic phase was dried over anhydrous sodium sulfate, and the solvent was removed by rotary evaporation to give the product (20.0g, 88.8% yield).
(3) Preparation of 2, 6-dichloro-4-hydroxybenzaldehyde
Figure BDA0001739294150001001
(3, 5-Dichlorophenoxy) triisopropylsilane (20.0g,62.7mmol) was added to anhydrous THF (200mL), and nBuLi (2.4mol/L,28.7mL,68.9mmol) was added dropwise at-78 deg.C, with continued stirring for 1 hour. DMF (6.87g,94.1mmol) was added dropwise thereto, and the reaction was continued at-78 ℃ for 1 hour. The reaction was returned to 25 ℃ for 6 hours. The mixture was extracted with 100mL of 1N HCl and 200mL of EA, and the organic phase was dried over anhydrous sodium sulfate, concentrated, and the solvent was removed by rotary evaporation. DCM (100mL) was added to precipitate a solid, which was filtered off with suction under reduced pressure to give the product (9.5g, 79.3% yield).
(4) Preparation of 2, 6-dichloro-4-methoxybenzaldehyde
Figure BDA0001739294150001002
2, 6-dichloro-4-hydroxybenzaldehyde (9.5g,49.7mmol) was added to DMF (200mL), potassium carbonate (20.6g,149.3mmol) was added to the system, methyl iodide (14.1g,99.3mmol) was added dropwise under ice bath, and the system was returned to 25 ℃ for reaction for 12 hours. The system was poured into 400mL of water, filtered under reduced pressure, the filter cake was washed with water (10mL) and dried to give the product (8.0g, 78.5% yield).
(5) Preparation of 1- (2, 6-dichloro-4-methoxyphenyl) prop-2-en-1-ol
Figure BDA0001739294150001003
2, 6-dichloro-4-methoxybenzaldehyde (8.0g,39.0mmol) was added to 100mL of THF, vinylmagnesium bromide (1M/L,46.8mL,46.8mmol) was added dropwise in ice bath, and the reaction was continued at 25 ℃ for about 6 hours. Then saturated N is addedH4The Cl solution was quenched (30mL), the separated layers were extracted with ethyl acetate (200mL) and water (100mL), the organic phase was dried over anhydrous sodium sulfate, and the solvent was removed by rotary evaporation to give the product (7.5g, 82.5% yield).
(6) Preparation of 1- (2, 6-dichloro-4-methoxyphenyl) prop-2-en-1-one
Figure BDA0001739294150001011
1- (2, 6-dichloro-4-methoxyphenyl) prop-2-en-1-ol (7.5g,32.2mmol) was dissolved in dichloromethane (200mL), dessimutan oxidant (16.4g,38.7mmol) was added portionwise under ice bath and the reaction continued at 25 ℃ for 12 h. The system was suction filtered under reduced pressure, the filtrate was spin-dried, and the residue was subjected to column chromatography (PE: EA ═ 10:1) to give the product (3.6g, yield 48.4%).
(7) Preparation of methyl (E) -3- (3- (2, 6-dichloro-4-methoxyphenyl) -3-oxoprop-1-en-1-yl) -4-hydroxybenzoate
Figure BDA0001739294150001012
1- (2, 6-dichloro-4-methoxyphenyl) prop-2-en-1-one (3.6g,15.6mmol), methyl 4-hydroxy-3-iodobenzoate (4.77g,17.2mmol), triethylamine (3.15g,31.2mmol), triphenylphosphine (409mg,1.56mmol) and palladium acetate (175mg,0.78mmol) were added in this order to acetonitrile (150mL), and the mixture was heated to 90 ℃ for 12 hours under nitrogen protection. After completion of the reaction, concentration was performed, and the residue was subjected to silica gel column chromatography (PE: EA ═ 3:1) to obtain a product (3.0g, yield 50.5%).
(8) Preparation of methyl 3- (3- (2, 6-dichloro-4-methoxyphenyl) -3-oxopropyl) -4-hydroxybenzoate
Figure BDA0001739294150001013
Methyl (E) -3- (3- (2, 6-dichloro-4-methoxyphenyl) -3-oxoprop-1-en-1-yl) -4-hydroxybenzoate (3.0g,7.9mmol), PtO2(300mg) was added to methanol (10)0mL) was added thereto, and the reaction was carried out at 25 ℃ for 4 hours under a hydrogen atmosphere. After the reaction was completed, the solvent was removed by filtration and rotary evaporation, and the residue was subjected to silica gel column chromatography (PE: EA ═ 4:1) to give the product (2.2g, yield 72.7%).
(9) Preparation of methyl 3- (3- (2, 6-dichloro-4-methoxyphenyl) -3-hydroxypropyl) -4-hydroxybenzoate
Figure BDA0001739294150001014
Methyl 3- (3- (2, 6-dichloro-4-methoxyphenyl) -3-oxopropyl) -4-hydroxybenzoate (1.0g,2.6mmol) was dissolved in THF (20mL) and NaBH was added slowly under ice bath4(290mg,7.8mmol), and reacted at 25 ℃ for about 6 hours. The reaction was quenched with 1mL of water and subjected to silica gel column chromatography (PE: EA. RTM. 5:1) to obtain the product (850mg, 84.9% yield).
(10) Preparation of methyl 2- (2, 6-dichloro-4-methoxyphenyl) chroman-6-carboxylate
Figure BDA0001739294150001021
Methyl 3- (3- (2, 6-dichloro-4-methoxyphenyl) -3-hydroxypropyl) -4-hydroxybenzoate (850mg,2.2mmol), diethyl azodicarboxylate (957mg,5.5mmol) was dissolved in tetrahydrofuran (40mL), triphenylphosphine (1.44g,5.5mmol) was added under ice bath, the temperature was raised to 25 ℃ for 12 hours, the solvent was removed by rotary evaporation, and the residue was subjected to silica gel column chromatography (PE: EA ═ 20:1) to give a product (600mg, 74.3% yield).
(11) Preparation of methyl 2- (2, 6-dichloro-4-hydroxyphenyl) chroman-6-carboxylate
Figure BDA0001739294150001022
Methyl 2- (2, 6-dichloro-4-methoxyphenyl) chroman-6-carboxylate (600mg,1.6mmol) is dissolved in dichloromethane (20mL) and a solution of boron tribromide in dichloromethane (1M/L,8.2mL,8.2mmol) in DCM is slowly added dropwise at-78 deg.C and then slowly brought to 25 deg.C for 2h of reaction. The reaction was quenched by adding 1mL of methanol, and subjected to silica gel column chromatography (PE: EA ═ 3:1) to give the product (60mg, yield 10.6%).
(12) Preparation of methyl 2- (2, 6-dichloro-4- ((5-cyclopropyl-3- (2, 6-dichlorophenyl) isoxazol-4-yl) methoxy) phenyl) benzopyran-6-carboxylate
Figure BDA0001739294150001023
Methyl 2- (2, 6-dichloro-4-hydroxyphenyl) chroman-6-carboxylate (60mg,0.17mmol), 4- (bromomethyl) -5-cyclopropyl-3- (2, 6-dichlorophenyl) isoxazole (59mg,0.17mmol), cesium carbonate (111mg,0.34mmol) were added to DMF (10mL) and reacted at 50 ℃ for 2 hours. Ethyl acetate (50mL) and water (30mL) were added to the mixture to extract a liquid, the organic phase was removed by rotary evaporation, and the residue was subjected to column chromatography (PE: EA: 3:1) to give the product (70mg, 66.5% yield).
(13) Preparation of 2- (2, 6-dichloro-4- ((5-cyclopropyl-3- (2, 6-dichlorophenyl) isoxazol-4-yl) methoxy) phenyl) chroman-6-carboxylic acid
Figure BDA0001739294150001031
Methyl 2- (2, 6-dichloro-4- ((5-cyclopropyl-3- (2, 6-dichlorophenyl) isoxazol-4-yl) methoxy) phenyl) chroman-6-carboxylate (70mg,0.11mmol) was added to methanol (4mL) and THF (4mL), and 2mL aqueous solution of lithium hydroxide monohydrate (24mg,0.57mmol) was added, reacted at 50 ℃ for 12 hours, the system was cooled to 25 ℃ and the pH was adjusted to 3-4 with 1M HCl. The system was extracted with ethyl acetate (50mL) and water (30mL), the organic phase was removed by rotary evaporation, and the residue was subjected to column chromatography (DCM: MeOH ═ 40:1) to give the product (15mg, 22.5% yield).
Molecular formula C29H21Cl4NO5Molecular weight 605.29LC-MS (M/e):606.1(M + H)+)
1H-NMR(400MHz,CDCl3)δ:7.95-7.70(m,2H),7.50-7.25(m,3H),6.90-6.62(m,3H),5.85-5.65(m,1H),4.80(s,2H),3.55-3.45(m,1H),3.16-2.45(m,3H),2.20-1.83(m,2H),1.38-1.02(m,4H).
Example 266- (2-chloro-4- ((5-cyclopropyl-3- (2, 6-dichlorophenyl) isoxazol-4-yl) methoxy) benzene Preparation of yl) -5-oxo-5, 6,7, 8-tetrahydronaphthalene-2-carboxylic acid (Compound 28)
Figure BDA0001739294150001032
(1) Preparation of 2- (4-bromo-3-chlorophenoxy) tetrahydro-2H-pyran
Figure BDA0001739294150001033
Referring to the preparation method in step (1) of example 2, 4-bromo-3-chlorophenol (10.0g,48.2mmol), 3, 4-dihydropyran (10.5g,124.8mmol) was added to give 12g of a product in 85.4% yield.
(2) Preparation of 5-oxo-5, 6,7, 8-tetrahydronaphthalen-2-yl trifluoromethanesulfonate
Figure BDA0001739294150001041
6-hydroxy-3, 4-dihydronaphthalen-1 (2H) -one (11g,67.8mmol) was dissolved in dichloromethane (150mL), triethylamine (10.3g,101.8mmol) was added dropwise down to-5 deg.C, trifluoromethanesulfonic acid (23g,153.3mmol) was added slowly and the addition was completed, reaction was carried out at 25 deg.C for 4 hours, after completion of the reaction, concentration was carried out, and the residue was chromatographed on silica gel column (petroleum ether: ethyl acetate ═ 20:1) to give 13g of product in 65.2% yield.
(3) Preparation of methyl 5-oxo-5, 6,7, 8-tetrahydronaphthalene-2-carboxylate
Figure BDA0001739294150001042
5-oxo-5, 6,7, 8-tetrahydronaphthalen-2-yl trifluoromethanesulfonate (10g,34mmol) was dissolved in DMF (70mL) and methanol (20mL), triethylamine (7g,69.2mmol), DPPP (450mg,1.09mmol), palladium acetate (450mg,2.0mmol), carbon monoxide balloon were added in this order to react at 70 ℃ for 16 hours, after completion of the reaction, the reaction mixture was poured into water, filtered, and the filter cake was chromatographed on silica gel column (petroleum ether: ethyl acetate: 5:1) to give 6.2g of the product in 89.3% yield.
(4) Preparation of methyl 6- (2-chloro-4- ((tetrahydro-2H-pyran-2-yl) oxy) phenyl) -5-oxo-5, 6,7, 8-tetrahydronaphthalene-2-carboxylate
Figure BDA0001739294150001043
Methyl 5-oxo-5, 6,7, 8-tetrahydronaphthalene-2-carboxylate (0.6g,2.94mmol), 2- (4-bromo-3-chlorophenoxy) tetrahydro-2H-pyran (1.17g,4.0mmol), Pd2(dba)3(270mg,0.29mmol), Xantphos (340mg,0.58mmol) and cesium carbonate (1.9g,5.8mmol) were added to toluene (30mL) in this order, reacted at 120 ℃ for 20 hours under nitrogen, concentrated, and the residue was chromatographed on silica gel column (petroleum ether: ethyl acetate 10:1) to give 200mg of the product in 16.4% yield.
(5) Preparation of methyl 6- (2-chloro-4-hydroxyphenyl) -5-oxo-5, 6,7, 8-tetrahydronaphthalene-2-carboxylate
Figure BDA0001739294150001051
Methyl 6- (2-chloro-4- ((tetrahydro-2H-pyran-2-yl) oxy) phenyl) -5-oxo-5, 6,7, 8-tetrahydronaphthalene-2-carboxylate (0.2g,0.48mmol) was dissolved in dichloromethane (10mL), trifluoroacetic acid (2mL) was added, and the reaction was carried out at 25 ℃ for 2 hours. Concentration was carried out, and the residue was subjected to silica gel column chromatography (petroleum ether: ethyl acetate: 5:1) to give 100mg of the product, yield: 63.0%.
(6) Preparation of methyl 6- (2-chloro-4- ((5-cyclopropyl-3- (2, 6-dichlorophenyl) isoxazol-4-yl) methoxy) phenyl) -5-oxo-5, 6,7, 8-tetrahydronaphthalene-2-carboxylate
Figure BDA0001739294150001052
Methyl 6- (2-chloro-4-hydroxyphenyl) -5-oxo-5, 6,7, 8-tetrahydronaphthalene-2-carboxylate (0.1g,0.3mmol) and 4- (bromomethyl) -5-cyclopropyl-3- (2, 6-dichlorophenyl) isoxazole (0.125g,0.36mmol) and potassium carbonate (83mg,0.6mmol) were added in this order to DMF (10mL) and reacted at 60 ℃ for 4 hours, after completion of the reaction, poured into water, filtered, and the filter cake was subjected to silica gel column chromatography (petroleum ether: ethyl acetate ═ 5:1-2:1) to give 145mg of the product in 81.0% yield.
(7) Preparation of 6- (2-chloro-4- ((5-cyclopropyl-3- (2, 6-dichlorophenyl) isoxazol-4-yl) methoxy) phenyl) -5-oxo-5, 6,7, 8-tetrahydronaphthalene-2-carboxylic acid
Figure BDA0001739294150001053
Methyl 6- (2-chloro-4- ((5-cyclopropyl-3- (2, 6-dichlorophenyl) isoxazol-4-yl) methoxy) phenyl) -5-oxo-5, 6,7, 8-tetrahydronaphthalene-2-carboxylate (140mg,0.23mmol) was dissolved in methanol (4mL), tetrahydrofuran (8mL), water (8mL), lithium hydroxide monohydrate (50mg,1.19mmol) was added, and the mixture was stirred at 25 ℃ for 16 hours. After completion of the reaction, methanol and tetrahydrofuran were distilled off under reduced pressure, water (10mL) was added to the residue, the pH was adjusted to 2 with dilute hydrochloric acid (1M), extraction was performed with ethyl acetate (20mL × 3), the organic layers were combined, concentrated, and the residue was subjected to silica gel column chromatography (petroleum ether: ethyl acetate: 1) to give a product (40mg, yield: 29.8%).
Molecular formula C30H22Cl3NO5Molecular weight 582.86 LC-MS (M/e):582.2(M + H)+)
1H-NMR(400MHz,CDCl3)δ:8.16(s,1H),8.04(d,J＝8.8Hz,2H),7.42(d,J＝7.6Hz,2H),7.32-7.37(m,1H),7.03(d,J＝8.8Hz,1H),6.90(d,J＝2.0Hz,1H),6.73(dd,J＝2.0Hz,J＝8.8Hz,1H),4.79(s,2H),4.21(dd,J＝4.0Hz,J＝12.8Hz,1H),3.12-3.26(m,2H),2.31-2.42(m,2H),2.10-2.21(m,1H),1.21-1.32(m,2H),1.16-1.19(m,2H).

Claims (9)

1. A compound of formula (I-1) or a pharmaceutically acceptable salt thereof:
Figure FDA0003274062510000011
wherein the content of the first and second substances,
R1、R2each independently selected from hydrogen atom, halogen atom, C1-4Alkyl or C1-4An alkoxy group;
R3selected from 3-6 membered cycloalkyl or 3-6 membered cycloalkyl C1-4An alkyl group;
R4selected from carboxyl;
w is selected from O or S;
a is selected from O or S;
z is selected from phenyl or 5-6 membered heteroaryl, substituted or unsubstituted with one or more substituents Q;
q is selected from cyano, amino, hydroxyl, carboxyl, nitro, halogen atom, C1-4Alkyl radical, C1-4Alkoxy, halo C1-4Alkyl or halo C1-4An alkoxy group;
E. x, Y, F together with the benzene ring form the following structure:
Figure FDA0003274062510000012
Figure FDA0003274062510000021
n is an integer from 0 to 3.
2. The compound of claim 1, or a pharmaceutically acceptable salt thereof:
wherein the content of the first and second substances,
R1、R2each independently selected from a hydrogen atom, a cyano group, a fluorine atom, a chlorine atom, a bromine atom, a methyl group, an ethyl group, a propyl group, an isopropyl group, a butyl group, an isobutyl group, a tert-butyl group or a methoxy group;
R3selected from cyclopropyl, cyclobutyl, cyclopentyl, cyclohexyl, cyclopropylmethyl, cycloPropylethyl, cyclobutylmethyl, cyclopentylmethyl, or cyclohexylmethyl;
n is selected from 1 or 2.
3. The compound of claim 2, or a pharmaceutically acceptable salt thereof:
wherein the content of the first and second substances,
R1、R2each independently selected from hydrogen atom, cyano group, fluorine atom, chlorine atom, methyl group, ethyl group, propyl group, butyl group or methoxy group;
R3selected from cyclopropyl, cyclobutyl, cyclopentyl, cyclopropylmethyl or cyclobutylmethyl;
z is selected from phenyl or pyridyl, substituted or unsubstituted with one or more substituents Q;
n is selected from 1 or 2.
4. The compound of claim 3, or a pharmaceutically acceptable salt thereof:
wherein the content of the first and second substances,
w is selected from O;
a is selected from O;
z is selected from phenyl which is substituted or unsubstituted by 1 to 2 substituents Q selected from cyano, amino, hydroxyl, carboxyl, nitro, fluorine atom, chlorine atom, bromine atom, methyl, ethyl, methoxy, ethoxy, trifluoromethyl or trifluoromethoxy;
n is selected from 1.
5. The compound of claim 1, or a pharmaceutically acceptable salt thereof, wherein the compound is selected from:
Figure FDA0003274062510000031
Figure FDA0003274062510000041
Figure FDA0003274062510000051
6. a pharmaceutical composition comprising a compound according to any one of claims 1 to 5, or a pharmaceutically acceptable salt thereof, together with one or more pharmaceutically acceptable carriers and/or diluents.
7. Use of a compound according to any one of claims 1 to 5 or a pharmaceutically acceptable salt thereof for the manufacture of a medicament for the treatment and/or prevention of FXR mediated diseases and related diseases selected from liver fibrosis, cirrhosis, fatty liver, acute liver failure, cholelithiasis, inflammatory bowel disease, lipid or lipoprotein disorders, clinical complications of type I or type II diabetes, hyperproliferative disorders.
8. The use according to claim 7, wherein the disease or related disorder is selected from the group consisting of non-alcoholic fatty liver disease, atherosclerosis, dyslipidemia, thrombosis, diabetic nephropathy, diabetic neuropathy, diabetic retinopathy, hepatocellular carcinoma, polyposis, colon adenocarcinoma, breast cancer, pancreatic cancer or esophageal cancer.
9. The use according to claim 8, wherein the disease and related disorders is nonalcoholic steatohepatitis.
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