CN108264506B

CN108264506B - Isoflavone derivative, preparation method and medical application thereof

Info

Publication number: CN108264506B
Application number: CN201810061682.5A
Authority: CN
Inventors: 向华; 丘荣茂; 陶睿; 郑凡; 李昊霖; 杨启桢; 胡伟康; 张瑾; 刘曼; 陈明琪; 陈德英; 尤启冬
Original assignee: China Pharmaceutical University
Current assignee: China Pharmaceutical University
Priority date: 2018-01-17
Filing date: 2018-01-17
Publication date: 2021-01-26
Anticipated expiration: 2038-01-17
Also published as: CN108264506A

Abstract

The invention relates to the field of medicinal chemistry, in particular to isoflavone derivatives with a general formula (I), a preparation method thereof, a pharmaceutical composition containing the compounds and medical application thereof, and particularly relates to application of the isoflavone derivatives as medicaments for preventing or treating hyperlipidemia, II-type diabetes, atherosclerosis and nonalcoholic steatohepatitis.

Description

Isoflavone derivative, preparation method and medical application thereof

The technical field is as follows:

the invention relates to the field of pharmaceutical chemistry, in particular to isoflavone derivatives. The invention also discloses a preparation method and pharmacological activity thereof, a medicinal composition containing the compounds and medicinal application thereof, in particular application of the compounds as medicaments for preventing or treating hyperlipidemia, type II diabetes, atherosclerosis and non-alcoholic steatohepatitis.

Background art:

dyslipidemia is a systemic disease in which plasma lipid or lipids is/are higher than normal due to abnormal fat metabolism or movement, and is manifested by elevated Total Cholesterol (TC) and/or Triglycerides (TG) or low high density lipoprotein cholesterol (HDL-C) in the blood. Dyslipidemia can lead to metabolic diseases such as atherosclerosis, non-alcoholic fatty liver disease, type II diabetes and the like. The incidence of cardiovascular disease is closely related to the level of lipoproteins in the body, and increased TG, increased LDL-C and decreased HDL-C are major risk factors for cardiovascular disease.

Farnesoid X Receptor (FXR), a member of the nuclear receptor superfamily, is a ligand-activated transcription factor that is highly expressed mainly in the liver, intestinal tract, kidney, adrenal gland. FXR is not only involved in regulating and controlling the metabolism of bile acid and glycolipid of the body, but also related to various metabolic diseases. After FXR is activated, intestinal cells are induced to synthesize and secrete human Fibroblast Growth Factor (FGF)19, and FGF19 activates FGF receptor 4 on the surface of liver cells after entering liver, so that the expression of cholesterol 7 hydroxylase 1 serving as a rate-limiting enzyme for bile acid synthesis is inhibited, and the synthesis of bile acid is reduced. Activating FXR can promote expression of B type I scavenger receptor, inhibit expression of sterol regulatory element binding protein 1c, induce peroxisome proliferator activated receptor alpha expression, promote reverse transport of cholesterol into liver, reduce lipid synthesis, and promote fatty acid beta oxidation.

Isoflavone compounds are widely present in plants and are one of effective components of many medicinal plants. Researches find that isoflavone compounds have obvious influence on cardiovascular system and endocrine system, and have anticancer activity, antioxidant effect, antibacterial effect, antiinflammatory effect, and hepatoprotective effect. The Food and Drug Administration (FDA) of the united states has formally recommended soy protein containing soy isoflavones as a health food for lowering blood cholesterol levels to reduce the risk of coronary heart disease in 10 months 1999.

The invention content is as follows:

the invention discloses an isoflavone derivative with a general formula I, and in vitro experiments prove that the compound can obviously inhibit the lipid accumulation of fat cells. Therefore, the compound of the invention has potential blood fat regulating activity.

Wherein R is₁、R₂Each independently is hydrogen, halogen, C_1-3Alkyl radical, C_1-3Alkoxy, halo C_1-3Alkyl, halo C_1-3An alkoxy group;

x is O, S, NH;

R₃is hydrogen, halogen, C_1-6Alkyl radical, C_3-6Cycloalkyl radical, C_1-6Alkoxy, halo C_1-6Alkyl, halo C_1-6An alkoxy group;

R₄is hydrogen, halogen, hydroxyl, nitro, cyano, methoxy, tetrazolyl or COOR substituted at any position of a benzene ring₅、CONR₆R₇、 OCH₂COOR₈；

R₅、R₆、R₇、R₈Independently of each other are hydrogen and C_1-6Alkyl radical, C_3-6A cycloalkyl group.

Wherein R is₁、R₂Independently of one another, preferably hydrogen, chlorine, bromine, fluorine, methoxy, monofluoromethoxy, difluoromethoxy, trifluoromethoxy;

x is preferably O;

R₃preferably isopropyl, cyclopropyl;

R₄preferably hydrogen, halogen, hydroxy, nitro, cyano, methoxy, tetrazolyl, COOR, substituted meta or para to the phenyl ring₅、CONR₆R₇、 OCH₂COOR₈；

R₅、R₆、R₇、R₈Independently of one another, preferably hydrogen, methyl, ethylN-propyl, isopropyl, cyclopropyl.

The structures of some of the compounds of the present invention are as follows:

compound numbering	R₁	R₂	X	R₃	R₄
						A1	-Cl	-Cl	O	-CH(CH₃)₂	4-F
A2	-Cl	-Cl	O	-CH(CH₃)₂	4-NO₂
						A3	-Cl	-Cl	O	-CH(CH₃)₂	4-OCH₃
A4	-Cl	-Cl	O	-CH(CH₃)₂	4-OH
						A5	-Cl	-Cl	O	-CH(CH₃)₂	4-OCH₂CO₂CH₂CH₃
A6	-Cl	-Cl	O	-CH(CH₃)₂	4-OCH₂COOH
						A7	-Cl	-Cl	O	-CH(CH₃)₂	4-COOCH₃
A8	-Cl	-Cl	O	-CH(CH₃)₂	4-COOH
						A9	-Cl	-Cl	O	-CH(CH₃)₂	4-CONHCH₃
A10	-Cl	-Cl	O	-CH(CH₃)₂	4-CONHCH₂HC₃
						A11	-Cl	-Cl	O	-CH(CH₃)₂	4-CONHCH(CH₂)₂
A12	-Cl	-Cl	O	-CH(CH₃)₂	4-CON(CH₃)₂

The codes of the compounds in the following pharmacological experiments and examples are equivalent to the structures of the compounds corresponding to the codes in the description.

Compounds of the general formula A1-A12 can be prepared by:

Reagents and conditions：a)NH₂OH·HCl，NaOH，EtOH；b)NCS，DMF；c)NaOCH₃，CH₃OH，THF；d)DIBAL-H，THF；e)SOCl₂，DCM；f)BF₃·Et₂O，Δ；g)BF₃·Et₂O，DMF，CH₃SO₂Cl；h)K₂CO₃，DMF，Δ；i)BBr₃，DCM；j)K₂CO₃， DMF，Δ；k)con HCl，1，4-Dioxane，Δ；l)con HCl，1，4-Dioxane，Δ；m)EDCI，HOBT，HNR₁R₂，DMF.

13T 3-L1 cell induced differentiation experiment

1.1 Experimental materials

DMEM (Dulbecco's Modified Eagle's Medium) Medium was purchased from Kyowa Kai-based Biotechnology, Inc.; fetal Bovine Serum (FBS) was purchased from Gibco BRL of Invitrogen Corporation (Carlsbad, Calif.); insulin injection, qianpang pharmaceutical ltd, Jiangsu; trypsin, Dexamethasone (DEX), 3-isobutyl-1-methylxanthine (IBMX) and other reagents are all made in the domestic analytical pure.

1.2 oil Red O staining and semi-quantitative detection of intracellular lipid content

3T3-L1 cells were cultured in a conventional medium, and 2 days after the cells were completely confluent (in this case, day 0 of differentiation), differentiation medium I (DMEM medium containing 10% FBS, 0.5mmol/L IBMX, 10mg/L insulin, 1. mu. mol/L DEX) was changed for 2 days. On day 2 of differentiation, cells were cultured in differentiation medium II (DMEM medium containing 10% FBS and 10mg/L insulin). On day 4 of differentiation and thereafter, the corresponding medium was DMEM medium containing 10. mu. mol/L of the target compound and 10. mu. mol/L of the positive drug GW4064, and the control was non-dosed complete medium. And (4) differentiating the cells, discarding culture solution in the holes at the 6 th day, washing the cells twice by PBS, and adding the cells fixed by 10% formaldehyde for 2 hours. PBS was washed twice and stained with 0.5% oil red for 2 hours. PBS was washed 3 times. Drying at 37 deg.C, adding isopropanol, extracting for 10min, and measuring absorbance at 492 nm. The inhibition rate was calculated as follows:

as a result: the oil red staining method shows that the lipid content in fat cells after administration is obviously lower than that of a control group, which indicates that the new compound can obviously inhibit lipid accumulation.

Results of the experiment

Compound numbering	Inhibition ratio (%)
		GW4064	30.90
A1	33.30
		A2	23.45
A3	28.25
		A4	24.17
A5	29.04
		A6	28.73
A7	32.95
		A8	39.46
A9	35.3
		A10	19.20
A11	30.65
		A12	27.50

The invention further relates to a pharmaceutical composition consisting of the compound of the general formula and a pharmaceutically acceptable carrier.

The compounds of the present invention may be formulated for administration either alone or in combination with one or more pharmaceutically acceptable carriers. Can be administered in oral dosage forms, such as common tablet and capsule, sustained release tablet and capsule, controlled release tablet and capsule, dripping pill, dispersible powder, granule, etc.; can also be prepared into injection. These pharmaceutical formulations may contain the active ingredient in combination with a carrier, for example, in a total amount of 0.05% to 90%, more usually between about 15% to 60% by weight of the active ingredient. The dosage of the compound can be 0.001-100 mg/kg/day, and the dosage range can also be deviated according to different disease degrees or different dosage forms.

Detailed Description

The preparation of the partial compounds was carried out as follows:

¹H-NMR was measured by means of a Bruker model AV300 (300MHz) NMR spectrometer (TMS as internal standard) and a Mass spectrometer Agilent1100LC-MSD-Trap/SL type mass spectrometer (ESI-MS).

The silica gel for column chromatography is 100-200 mesh, 200-300 mesh or 300-400 mesh silica gel (Qingdao ocean chemical plant), and the eluent is petroleum ether-ethyl acetate system or dichloromethane-methanol system. Thin Layer Chromatography (TLC) using GF254 thin layer chromatography plate (tai jiang friend silica gel development ltd); the TLC development system is a petroleum ether-ethyl acetate system or a dichloromethane-methanol system, and a small amount of acetic acid is added when necessary; TLC was illuminated under model ZF7 three-way uv analyzer (henan consortium seoul instruments ltd).

Example 1

Preparation of 2, 6-Dichlorobenzaldehyde oxime (2)

Hydroxylamine hydrochloride (10.9g) and sodium hydroxide (6.27g) were dissolved in water at room temperature, added dropwise to a solution of 2, 6-dichlorobenzaldehyde (25g) in ethanol (200mL) and stirred at 90 ℃ for 24 hours, cooled to room temperature, the ethanol was spin-dried, filtered, the paint cake was washed with water and dried in the infrared to give 26.5g of product with a yield of 96.5%. Ms (esi): 189.9[ M + H ] M/z]⁺。

Example 2

Preparation of 2, 6-dichloro-N-hydroxy-chlorobenzaldehyde oxime (3)

2, 6-Dichlorobenzaldehyde oxime (26g) was dissolved in DMF (160mL), NCS (18.5g) was added portionwise at room temperature, and after stirring for 1 hour, the solution was poured into water (800mL), extracted with ethyl acetate, and the ethyl acetate layers were combined, washed with water and saturated brine, respectively, dried over anhydrous sodium sulfate, and the solvent was dried to give 30g of a colorless transparent oily product in 97% yield. Ms (esi): m/z 223.9[ M + H ]]⁺。

Example 3

Preparation of 3- (2, 6-dichlorophenyl) -5-methyl-isoxazole-4-carboxylic acid methyl ester (4)

Sodium (0.21g) was added to anhydrous methanol (20mL) under ice-bath, a tetrahydrofuran (3mL) solution of methyl isobutyrylacetate (1.3mL) was added dropwise after stirring for half an hour, a tetrahydrofuran (5mL) solution of 2, 6-dichloro-N-hydroxy-chlorobenzaldehyde oxime (1g) was added dropwise after reacting for half an hour, the solvent was dried by spinning after stirring for 15 hours at room temperature, extracted with ethyl acetate, washed with saturated brine, dried over anhydrous magnesium sulfate, and the solvent was dried by spinning to obtain an oil. And (5) performing column chromatography separation and purification to obtain 0.35g of a white solid product with the yield of 58%. Ms (esi): 314.1[ M + H ] M/z]⁺。

Example 4

Preparation of 3- (2, 6-dichlorophenyl) -4-hydroxymethyl-5-methyl-isoxazole (5)

A toluene solution of diisobutylaluminum hydride (16.4mL, 1.5M) was added dropwise to a solution of methyl 3- (2, 6-dichlorophenyl) -5-methyl-isoxazole-4-carboxylate (7g) in tetrahydrofuran (50mL) under nitrogen atmosphere, and the mixture was allowed to return to room temperature and stirred for 15 hours. Under ice bath, methanol and water are slowly and sequentially added dropwise to generate a gel-like suspension. After filtration, the mixture was extracted with ethyl acetate, washed with water and saturated brine in this order, dried over anhydrous magnesium sulfate, and the solvent was dried by spin-drying to obtain 5.8g of a white solid product with a yield of 90%. Ms (esi): 286.0[ M + H ] M/z]⁺。

Example 5

Preparation of 3- (2, 6-dichlorophenyl) -4-chloromethyl 5-methyl-isoxazole (6)

3- (2, 6-dichlorophenyl) -4-hydroxymethyl-5-methyl-isoxazole (2.9g) was dissolved in DCM (50mL), thionyl chloride (1mL) was slowly added dropwise in an ice bath, and after stirring at room temperature for 4 hours, the solution was poured into a cold saturated aqueous sodium bicarbonate solution, extracted with ethyl acetate, washed with water and saturated brine in this order, dried over anhydrous magnesium sulfate, and the solvent was spin-dried to give 3.4g of a colorless oil with a yield of 98%.¹H-NMR(300MHz，CDCl₃) δ：7.46-7.27(m，3H)，4.15(s，2H)，3.35(m，1H)，1.39(d，6H).

Example 6

Preparation of 1- (2, 4-dihydroxyphenyl) -2- (4-fluorophenyl) ethanone (7a)

To the reaction flaskP-fluorophenylacetic acid (7g, 42.1mmol) and resorcinol (6.9g, 63.15mmol) were added thereto, 150ml of boron trifluoride ether solution was added thereto under nitrogen protection. The reaction is carried out for 7h at 80 ℃. After cooling to room temperature, 400ml of saturated sodium carbonate solution was added, extraction was performed with ethyl acetate, washing was performed with saturated sodium bicarbonate solution and saturated brine, drying was performed with anhydrous sodium sulfate, filtration and spin-drying were performed to obtain a deep red liquid, which was directly dropped. Ms (esi): m/z 245[ M-H ]]⁺。

Example 7

Preparation of 1- (2, 4-dihydroxyphenyl) -2- (4-nitrophenyl) ethanone (7b)

The same synthesis method as that of the compound (7a) is adopted to obtain red liquid which is directly added downwards. Ms (esi): 272[ M-H ] M/z]^-。

Example 8

Preparation of 1- (2, 4-dihydroxyphenyl) -2- (4-methoxyphenyl) ethanone (7c)

The same synthesis method as that of the compound (7a) is adopted to obtain red liquid which is directly added downwards. Ms (esi): 257[ M-H ] M/z]^-。

Example 9

Preparation of methyl 4- (2- (2, 4-dihydroxyphenyl) -2-oxoethyl) benzoate (7d)

The same synthesis method as that of the compound (7a) is adopted to obtain red liquid which is directly added downwards. Ms (esi): 285[ M-H ] M/z]^-。

Example 10

Preparation of 7-hydroxy-3- (4-fluorophenyl) -4H-benzopyran-4-one (8a)

1- (2, 4-dihydroxyphenyl) -2- (4-fluorophenyl) ethanone (5.14g, 19.9mmol) was dissolved in DMF (11.1ml, 59.7mmol), boron trifluoride etherate (8.7ml, 99.5mmol) was added under ice-bath, and the temperature was raised to 60 ℃ with stirring. And dropwise adding the DMF solution of the uniformly mixed methanesulfonyl chloride into the raw materials, stirring for 5 minutes, heating to 95 ℃, and reacting for 2 hours under the protection of nitrogen. After the reaction is finished, the reaction liquid is poured into a cold saturated sodium carbonate solution, yellow solid is separated out, and the reaction liquid is filtered and dried under infrared. Silica gel column chromatography is carried out to obtain 3.2g of white solid with yield of 60.0%.¹H-NMR (300MHz，d₆-DMSO)δ：10.86(s，1H)，8.38(s，1H)，7.96(d，1H)，7.59(m，2H)，7.24(m，2H)，6.94(dd，1H)，6.85(d， 1H).

Example 11

Preparation of 7-hydroxy-3- (4-nitrophenyl) -4H-benzopyran-4-one (8b)

The same synthesis as for compound (8a) was used to give a yellow solid with a yield of 50%.¹H-NMR(300MHz，d₆-DMSO)δ：10.91(s， 1H)，8.60(s，1H)，8.29(d，2H)，8.01(d，1H)，7.91(d，2H)，6.99(dd，1H)，6.90(d，1H).

Example 12

Preparation of 7-hydroxy-3- (4-methoxyphenyl) -4H-benzopyran-4-one (8c)

The same synthesis as for compound (8a) was carried out to give a white solid in 58.2% yield. Ms (esi): m/z 269[ M + H ═]¹。

Example 13

Preparation of methyl 4- (7-hydroxy-4-oxo-4H-benzopyran-3-enyl) benzoate (8d)

The same synthesis as that of the compound (8a) was carried out to obtain a pale yellow solid with a yield of 54.2%.¹H-NMR(300MHz，d₆-DMSO)δ：10.88(s， 1H)，8.52(s，1H)，8.01(d，2H)，8.01(dd，3H)，7.77(d，2H)，6.99(dd，1H)，6.90(d，1H)，3.88(s，3H).

Example 14

Preparation of 7- ((3- (2, 6-dichlorophenyl) -5-isopropylisoxazol-4-yl) oxymethyl) -3- (4-fluorophenyl) -4H-benzopyran (A1)

3- (2, 6-dichlorophenyl) -4-chloromethyl 5-methyl-isoxazole and 1- (2, 4-dihydroxyphenyl) -2- (4-fluorophenyl) ethanone were dissolved in DMF, and potassium carbonate and potassium iodide were added to react at 60 ℃ for 8 hours. After completion of the reaction, it was cooled to room temperature, poured into water, extracted with ethyl acetate, and dried over anhydrous sodium sulfate. Spin-drying and column chromatography to obtain white solid with 85.6% yield.¹H NMR(300MHz，CDCl₃)δ8.06(d，J＝8.9Hz，1H)， 7.83(s，1H)，7.44(dd，J＝8.6，5.5Hz，2H)，7.35(d，J＝1.9Hz，1H)，7.32(s，1H)，7.25(dd，J＝9.3，6.6Hz，1H)， 7.04(t，J＝8.7Hz，2H)，6.77(dd，J＝8.9，2.2Hz，1H)，6.67(d，J＝2.2Hz，1H)，4.76(s，2H)，3.28(dt，J＝13.8， 6.9Hz，1H)，1.37(d，J＝6.9Hz，6H).

Example 15

Preparation of 7- ((3- (2, 6-dichlorophenyl) -5-isopropylisoxazol-4-yl) oxymethyl) -3- (4-nitrophenyl) -4H-benzopyran (A2)

The same synthesis as that of compound (a1) was carried out to give a yellow solid in 88.7% yield.¹H NMR(300MHz，d₆-DMSO)δ8.64(s，1H)， 8.27(d，J＝8.9Hz，2H)，7.97(d，J＝8.9Hz，1H)，7.89(d，J＝8.8Hz，2H)，7.68-7.58(m，2H)，7.53(dd，J＝9.3， 6.6Hz，1H)，7.13(d，J＝2.1Hz，1H)，6.87(dd，J＝8.9，2.2Hz，1H)，5.02(s，2H)，3.50(dt，J＝14.0，6.9Hz，1H)， 1.34(t，J＝6.7Hz，6H).

Example 16

Preparation of 7- ((3- (2, 6-dichlorophenyl) -5-isopropylisoxazol-4-yl) oxymethyl) -3- (4-methoxyphenyl) -4H-benzopyran (A3)

The same synthesis method as that for compound (a1) was used, resulting in a white solid with a yield of 83.2%.¹H NMR(300MHz，d₆-DMSO)δ8.64(s， 1H)，8.27(d，J＝8.9Hz，2H)，7.97(d，J＝8.9Hz，1H)，7.89(d，J＝8.8Hz，2H)，7.68-7.58(m，2H)，7.53(dd，J＝ 9.3，6.6Hz，1H)，7.13(d，J＝2.1Hz，1H)，6.87(dd，J＝8.9，2.2Hz，1H)，5.02(s，2H)，3.50(dt，J＝14.0，6.9Hz， 1H)，1.34(t，J＝6.7Hz，6H).

Example 17

Preparation of methyl 4- (7- ((3- (2, 6-dichlorophenyl) -5-isopropylisoxazol-4-yl) oxymethyl) -4-oxo-4H-benzopyran-3-enyl) carboxylate (A7)

The same synthesis as that of compound (a1) was carried out to give a yellow solid with a yield of 85.3%.¹H NMR(300MHz，d₆-DMSO)δ8.53(s， 1H)，7.97(t，J＝7.6Hz，3H)，7.74(d，J＝8.1Hz，2H)，7.60(d，J＝7.3Hz，2H)，7.57-7.48(m，1H)，7.10(s，1H)， 6.86(d，J＝8.9Hz，1H)，5.03(s，2H)，3.86(s，3H)，3.51(dt，J＝13.9，6.9Hz，1H)，1.36(d，J＝6.7Hz，6H).

Example 18

Preparation of 7- ((3- (2, 6-dichlorophenyl) -5-isopropylisoxazol-4-yl) oxymethyl) -3- (4-hydroxyphenyl) -4H-benzopyran (A4)

7- ((3- (2, 6-dichlorophenyl) -5-isopropylisoxazol-4-yl) oxymethyl) -3- (4-methoxyphenyl) -4H-benzopyran was dissolved in anhydrous dichloromethane, and a solution of 1N boron tribromide in dichloromethane was added under ice-bath to react at room temperature for 4 hours. After the reaction, a saturated sodium bicarbonate solution was added dropwise in an ice bath, extracted with ethyl acetate, and dried over anhydrous sodium sulfate. The solvent is dried by spinning and the column chromatography is carried out to obtain white solid with the yield of 85.4 percent.¹H NMR(300 MHz，d₆-DMSO)δ9.55(s，1H)，8.36(s，1H)，7.97(d，J＝8.8Hz，1H)，7.60(dd，J＝20.2，7.0Hz，2H)，7.41(d，J ＝8.1Hz，2H)，7.08(s，1H)，6.83(d，J＝7.3Hz，3H)，5.03(s，2H)，3.62-3.43(m，1H)，1.38(d，J＝6.6Hz，6H).

Example 19

Preparation of ethyl 2- (4- (7- ((3- (2, 6-dichlorophenyl) -5-isopropylisoxazol-4-yl) oxymethyl) -4 oxo-4H-benzopyran-3-yl) phenoxyacetate (A5)

Dissolving 7- ((3- (2, 6-dichlorophenyl) -5-isopropyl isoxazol-4-yl) oxymethyl) -3- (4-hydroxyphenyl) -4H-benzopyran and ethyl bromoacetate in DMF, adding potassium carbonate and potassium iodide, and reacting at 60 ℃ for 6H. After the reaction, the reaction mixture was cooled to room temperature, washed with ethyl acetate and saturated brine, and dried over anhydrous sodium sulfate. The solvent was spin dried and column chromatographed to give 650mg of white solid in 86.3% yield.¹H NMR(300MHz，CDCl₃)δ8.14 (d，J＝8.9Hz，1H)，7.90(s，1H)，7.49(d，J＝8.7Hz，2H)，7.45-7.37(m，2H)，7.32(dd，J＝9.3，6.6Hz，1H)，6.97 (d，J＝8.7Hz，2H)，6.84(dd，J＝8.9，2.2Hz，1H)，6.75(d，J＝2.1Hz，1H)，4.85(s，2H)，4.65(s，2H)，4.28(q，J＝ 7.1Hz，2H)，3.36(dt，J＝14.0，7.0Hz，1H)，1.45(d，J＝7.0Hz，6H)，1.31(t，J＝7.1Hz，3H).

Example 20

Preparation of 2- (4- (7- ((3- (2, 6-dichlorophenyl) -5-isopropylisoxazol-4-yl) oxymethyl) -4 oxo-4H-benzopyran-3-yl) phenoxyacetic acid (A6)

Dissolving ethyl 2- (4- (7- ((3- (2, 6-dichlorophenyl) -5-isopropyl isoxazol-4-yl) oxymethyl) -4 oxo-4H-benzopyran-3-yl) phenoxyacetate in dioxane, dropwise adding concentrated hydrochloric acid, performing reflux reaction for 10 hours, after the reaction is finished, performing spin-drying on the solution, extracting by using ethyl acetate, washing by using saturated saline, drying by using anhydrous sodium sulfate, performing spin-drying on the solvent, performing column chromatography to obtain a light yellow solid, wherein the yield is 78.2%.¹H NMR(300MHz，d₆-DMSO)δ 8.41(s，1H)，7.95(d，J＝8.9Hz，1H)，7.62(d，J＝7.2Hz，2H)，7.58-7.52(m，1H)，7.49(d，J＝8.6Hz，2H)，7.09 (s，1H)，6.96(d，J＝8.6Hz，2H)，6.84(d，J＝8.6Hz，1H)，5.01(s，2H)，4.67(s，2H)，3.51(dt，J＝14.2，7.0Hz， 2H)，1.36(d，J＝6.9Hz，6H).

Example 21

Preparation of 4- (7- ((3- (2, 6-dichlorophenyl) -5-isopropylisoxazol-4-yl) oxymethyl) -4 oxo-4H-benzopyran-3-yl) benzoic acid (A8)

The same synthesis as that of the compound (a6) was carried out to obtain a yellow solid with a yield of 90.0%.¹H NMR(300MHz，d₆-DMSO)δ12.94(s， 1H)，8.50(s，1H)，7.91(d，J＝6.8Hz，3H)，7.64(d，J＝7.4Hz，2H)，7.54(s，2H)，7.49(s，1H)，7.06(s，1H)，6.79 (d，J＝7.9Hz，1H)，4.95(s，2H)，3.44(s，1H)，1.28(d，J＝6.3Hz，6H).

Example 22

Preparation of 4- (7- ((3- (2, 6-dichlorophenyl) -5-isopropylisoxazol-4-yl) oxymethyl) -4 oxo-4H-benzopyran-3-yl) -N-methylbenzamide (A9)

4- (7- ((3- (2, 6-dichlorophenyl) -5-isopropylisoxazol-4-yl) oxymethyl) -4 oxo-4H-benzopyran-3-yl) benzoic acid, EDCI, HOBT, triethylamine were dissolved in DMF, methylamine hydrochloride was added, and the reaction was carried out at room temperature for 10 hours. After completion of the reaction, the mixture was poured into water, extracted with ethyl acetate, washed with saturated brine, and dried over anhydrous sodium sulfate. The solvent was spin dried and column chromatographed to give a pale yellow solid in 83.2% yield.¹H NMR(300 MHz，d₆-DMSO)δ8.51(s，1H)，8.48(d，J＝4.5Hz，1H)，7.94(d，J＝8.9Hz，1H)，7.85(d，J＝8.3Hz，2H)，7.63 (dd，J＝10.0，5.0Hz，3H)，7.58(s，1H)，7.54(s，1H)，7.10(d，J＝2.0Hz，1H)，6.83(dd，J＝8.9，2.1Hz，1H)，4.99 (s，2H)，3.49(dd，J＝13.9，6.9Hz，3H)，2.77(d，J＝4.3Hz，1H)，1.33(d，J＝6.9Hz，6H).

Example 23

Preparation of 4- (7- ((3- (2, 6-dichlorophenyl) -5-isopropylisoxazol-4-yl) oxymethyl) -4 oxo-4H-benzopyran-3-yl) -N-ethylbenzamide (A10)

The same synthesis method as that of the compound (a9) was employed, and a pale yellow solid was obtained with a yield of 83%.¹H NMR(300MHz，CDCl₃)δ8.13(d，J ＝8.9Hz，1H)，7.98(s，1H)，7.81(d，J＝8.0Hz，2H)，7.58(d，J＝7.9Hz，2H)，7.41(d，J＝7.1Hz，2H)，7.34(d，J ＝6.7Hz，1H)，6.86(d，J＝8.6Hz，1H)，6.77(s，1H)，6.59(s，1H)，4.86(s，2H)，3.54-3.42(t，2H)，3.37(dd，J＝ 14.0，7.0Hz，1H)，1.45(d，J＝6.9Hz，6H)，1.27-1.20(q，3H).

Example 24

Preparation of 4- (7- ((3- (2, 6-dichlorophenyl) -5-isopropylisoxazol-4-yl) oxymethyl) -4 oxo-4H-benzopyran-3-yl) -N-cyclopropylbenzamide (A11)

The same synthesis method as that of the compound (A9) was employed, and a pale yellow solid was obtained with a yield of 80.4%.¹H NMR(400MHz，CDCl₃)δ8.13(d， J＝8.9Hz，1H)，7.97(s，1H)，7.78(d，J＝8.2Hz，2H)，7.59(d，J＝8.2Hz，2H)，7.44-7.37(m，2H)，7.33(dd，J＝ 9.0，7.0Hz，1H)，6.86(dd，J＝8.9，2.3Hz，1H)，6.76(d，J＝2.2Hz，1H)，6.51(s，1H)，4.85(s，2H)，3.36(dt，J＝ 14.0，7.0Hz，1H)，2.90(dt，J＝10.3，3.4Hz，1H)，1.45(d，J＝7.0Hz，6H)，0.86(d，J＝6.3Hz，2H)，0.63(d，J＝ 2.0Hz，2H).

Example 25

Preparation of 4- (7- ((3- (2, 6-dichlorophenyl) -5-isopropylisoxazol-4-yl) oxymethyl) -4 oxo-4H-benzopyran-3-yl) -N, N-dimethylbenzamide (A12)

By reacting with compounds(A9) The same synthesis procedure gave a pale yellow solid with a yield of 85.3%.¹H NMR(400MHz，CDCl₃)δ8.06(d，J＝8.9 Hz，1H)，7.87(s，1H)，7.52(d，J＝8.3Hz，2H)，7.40(d，J＝8.3Hz，2H)，7.35-7.29(m，2H)，7.25(dd，J＝9.1，6.9 Hz，1H)，6.77(dd，J＝8.9，2.4Hz，1H)，6.68(d，J＝2.3Hz，1H)，4.77(s，2H)，3.28(dt，J＝14.0，7.0Hz，1H)，2.99 (d，J＝39.7Hz，6H)，1.37(d，J＝7.0Hz，6H).

Claims

1. A compound of the following general formula (I) or a pharmaceutically acceptable salt thereof:

in,

R ₁ and R ₂ are independently hydrogen, halogen, C _1-3 alkyl, C _1-3 alkoxy, halogenated C _1-3 alkyl, and halogenated C _1-3 alkoxy;

X is O, S, NH;

R ₃ is hydrogen, halogen, C _1-6 alkyl, C _3-6 cycloalkyl, C _1-6 alkoxy, halogenated C _1-6 alkyl, halogenated C _1-6 alkoxy;

R ₄ is hydrogen, halogen, hydroxyl, nitro, cyano, methoxy, tetrazolyl, COOR ₅ , CONR ₆ R ₇ , OCH ₂ COOR ₈ substituted at any position on the benzene ring;

R ₅ , R ₆ , R ₇ and R ₈ are each independently hydrogen, C _1-6 alkyl, and C _3-6 cycloalkyl.

2. The compound according to any one of claims 1 or a pharmaceutically acceptable salt thereof, wherein R ₁ and R ₂ are independently hydrogen, chlorine, bromine, fluorine, methoxy, monofluoromethoxy, di Fluoromethoxy, trifluoromethoxy;

X is O;

R ₃ is isopropyl, cyclopropyl;

R ₄ is hydrogen, halogen, hydroxyl, nitro, cyano, methoxy, tetrazolyl, COOR ₅ , CONR ₆ R ₇ , OCH ₂ COOR ₈ substituted at the meta or para position of the benzene ring;

R ₅ , R ₆ , R ₇ , and R ₈ are each independently hydrogen, methyl, ethyl, n-propyl, isopropyl, and cyclopropyl.

3. A pharmaceutical composition comprising a compound according to any one of claims 1-2, or a pharmaceutically acceptable salt thereof, and a pharmaceutically acceptable carrier.

4. The compound according to any one of claims 1-2 or a pharmaceutically acceptable salt thereof, for use in the preparation of a medicament for the prevention or treatment of hyperlipidemia, type II diabetes, atherosclerosis, nonalcoholic steatohepatitis the use of.