1. A carbazole-based compound characterized in that: the structural general formula is shown as formula (I):

wherein the content of the first and second substances,

R₁selected from halogen, hydroxy or C₁-C₆An alkoxy group;

R₂and R₃Each of which is independent of the otherSelected from

(a) Hydrogen or formyl;

(b)C₅-C₈heterocyclyl or aryl optionally substituted with 0-4R₄Substitution; wherein R is₄Is selected from C₁-C₆Alkoxy radical, C₂-C₆Alkenyloxy radical, C₂-C₆Alkynyloxy or halogen;

X₁and X₂Each of which is independently selected from hydrogen or halogen;

Z₁and Z₂Each of which is independently selected from hydrogen, cyano or-SOp-R₅(ii) a Wherein p is selected from 0, 1 or 2; r₅Is selected from C₁-C₆Alkyl radical, C₂-C₆Alkenyl radical, C₂-C₆An alkynyl group;

each of n and m is independently selected from 1, 2, 3 or 4.

2. The carbazole-based compound according to claim 1, characterized in that:

R₁selected from halogen or hydroxy;

R₂and R₃Each of which is independently selected from

(a) Hydrogen;

(b) aryl optionally substituted with 0-4R₄Substitution; wherein R is₄Is selected from C₁-C₆Alkoxy or halogen;

X₁and X₂Each of which is independently selected from hydrogen or halogen;

Z₁and Z₂Each of which is independently selected from hydrogen, cyano or-SO₂-R₅(ii) a Wherein R is₅Is selected from C₁-C₆An alkyl group;

each of n and m is independently selected from 1, 2, 3 or 4.

3. The carbazole-based compound according to claim 2, characterized in that:

R₁selected from halogen or hydroxy;

R₂selected from hydrogen; r₃Selected from aryl, optionally substituted by 0-4R₄Is substituted in which R₄Is selected from C₁-C₃An alkoxy group;

X₁selected from halogen, X₂Selected from hydrogen;

Z₁is selected from-SO₂-R₅Wherein R is₅Is selected from C₁-C₃An alkyl group; z₂Selected from hydrogen;

n and m are both 1.

4. The carbazole-based compound according to claim 2, characterized in that:

R₁selected from halogen or hydroxy;

R₂selected from hydrogen; r₃Selected from aryl, optionally substituted with 0-4R₄Is substituted in which R₄Is selected from C₁-C₃An alkoxy group;

X₁selected from halogen, X₂Selected from hydrogen;

Z₁is selected from cyano; z₂Selected from hydrogen;

n and m are both 1.

5. The carbazole-based compound according to claim 2, characterized in that: the carbazole-based compound is selected from:

1- (3-bromo-6-methanesulfonyl-9H-carbazol-9-yl) -3- (3-methoxyphenylamino) propan-2-ol;

n- (3- (3-bromo-6-methanesulfonyl-9H-carbazol-9-yl) -2-fluoropropyl) -3-methoxyaniline;

1- (3-bromo-6-methanesulfonyl-9H-carbazol-9-yl) -3- (phenylamino) propan-2-ol;

n- (3- (3-bromo-6-cyano-9H-carbazol-9-yl) -2-fluoropropyl) -3-methoxyaniline;

1- (3-bromo-6-cyano-9H-carbazol-9-yl) -3- (phenylamino) propan-2-ol.

6. The pharmaceutically acceptable salt of the carbazole-based compound of claim 1.

7. The pharmaceutically acceptable salt of a carbazole-based compound according to claim 6, characterized in that: the pharmaceutically acceptable salt is an inorganic acid salt or an organic acid salt.

8. The pharmaceutically acceptable salt of a carbazole-based compound according to claim 7, characterized in that: the inorganic acid comprises hydrochloric acid, sulfuric acid, phosphoric acid, diphosphoric acid, hydrobromic acid or nitric acid; the organic acid includes acetic acid, maleic acid, fumaric acid, tartaric acid, succinic acid, lactic acid, p-toluenesulfonic acid, salicylic acid or oxalic acid.

9. Use of the carbazole-based compound of claim 1 or a pharmaceutically acceptable salt of the carbazole-based compound of claim 6 for the preparation of a medicament for the treatment of glycolipid metabolic diseases and complications thereof.

10. Use according to claim 9, characterized in that: the glycolipid metabolic diseases and complications thereof are fatty liver, diabetes, hyperlipidemia, obesity or arteriosclerotic cardiovascular and cerebrovascular diseases.

Background

With the global epidemic of obesity and metabolic syndrome thereof, metabolic diseases such as obesity, diabetes mellitus and fatty liver disease become important diseases in European and American countries and affluent areas of China, and are important health problems. Fatty liver includes simple fatty liver and fatty hepatitis. Some simple fatty liver patients gradually progress to fatty hepatitis, and the liver produces severe inflammation and fibrosis, resulting in liver fibrosis, cirrhosis and even hepatocellular carcinoma. Treatment for fatty liver disease is primarily lifestyle intervention, including weight loss, diet improvement, and exercise. No drug is currently approved by the national food and drug administration (CFDA), the U.S. FDA (food and drug administration), or the European medicine agency EMA (European Medicines agency) for the treatment of fatty liver disease. Diabetes mellitus is the absolute or relative insufficiency of insulin secretion. Metabolic-related diseases such as fatty liver and type 2 diabetes have complex etiology, but have similar pathogenesis, and have many common points.

The patent CN108619137B of the inventor's earlier application discloses the application of carbazole compounds 3, 6-dibromo-beta-fluoro-N- (3-methoxyphenyl) -9H-carbazole-9-propylamine (P7C3-A20) in the preparation of drugs for treating metabolic diseases and complications thereof, and the animal models of hyperlipidemia, obesity and fatty liver induced by high-fat and high-sugar diet are used for proving that P7C3-A20 can obviously reduce blood fat and blood sugar, relieve obesity and fatty liver pathological changes, and the myocardial ischemia and anoxia model is used for simulating ischemic myocardial injury to prove that P7C3-A20 can obviously reduce myocardial cell injury caused by ischemia and anoxia. The patent provides a new medicine research and development direction for metabolic diseases and complications thereof. Therefore, the chemical modification of the P7C3-A20 compound is of great significance in research and development of the analogue with more remarkable effect.

Disclosure of Invention

The invention aims to overcome the defects in the prior art and provides a carbazole compound and pharmaceutically acceptable salts thereof, and application and a preparation method thereof.

In a first aspect, the present invention provides a carbazole compound, which has a structural general formula shown in formula (I):

wherein the content of the first and second substances,

R₁selected from halogen, hydroxy or C₁-C₆An alkoxy group;

R₂and R₃Each of which is independently selected from

(a) Hydrogen or formyl;

(b)C₅-C₈heterocyclyl or aryl optionally substituted with 0-4R₄Substitution; wherein R is₄Is selected from C₁-C₆Alkoxy radical, C₂-C₆Alkenyloxy radical, C₂-C₆Alkynyloxy or halogen;

X₁and X₂Each of which is independently selected from hydrogen or halogen;

Z₁and Z₂Each of which is independently selected from hydrogen, cyano or-SOp-R₅(ii) a Wherein p is selected from 0, 1 or 2; r₅Is selected from C₁-C₆Alkyl radical, C₂-C₆Alkenyl radical, C₂-C₆An alkynyl group;

each of n and m is independently selected from 1, 2, 3 or 4.

As a preferred embodiment of the present invention,

R₁selected from halogen or hydroxy;

R₂and R₃Each of which is independently selected from

(a) Hydrogen;

(b) aryl optionally substituted with 0-4R₄Substitution; wherein R is₄Is selected from C₁-C₆Alkoxy or halogen;

X₁and X₂Each of which is independently selected from hydrogen or halogen;

Z₁and Z₂Each of which is independently selected from hydrogen, cyano or-SO₂-R₅(ii) a Wherein R is₅Is selected from C₁-C₆An alkyl group;

each of n and m is independently selected from 1, 2, 3 or 4.

As another preferred embodiment of the present invention,

R₁selected from halogen or hydroxy;

R₂selected from hydrogen; r₃Selected from aryl, optionally substituted with 0-4R₄Is substituted in which R₄Is selected from C₁-C₃An alkoxy group;

X₁selected from halogen, X₂Selected from hydrogen;

Z₁is selected from-SO₂-R₅Wherein R is₅Is selected from C₁-C₃An alkyl group; z₂Selected from hydrogen;

n and m are both 1.

As another preferred embodiment of the present invention,

R₁selected from halogen or hydroxy;

R₂selected from hydrogen; r₃Selected from aryl, optionally substituted with 0-4R₄Is substituted in which R₄Is selected from C₁-C₃An alkoxy group;

X₁selected from halogen, X₂Selected from hydrogen;

Z₁is selected from cyano; z₂Selected from hydrogen;

n and m are both 1.

As another preferred embodiment of the present invention, the carbazole-based compound is selected from:

1- (3-bromo-6-methanesulfonyl-9H-carbazol-9-yl) -3- (3-methoxyphenylamino) propan-2-ol;

n- (3- (3-bromo-6-methanesulfonyl-9H-carbazol-9-yl) -2-fluoropropyl) -3-methoxyaniline;

1- (3-bromo-6-methanesulfonyl-9H-carbazol-9-yl) -3- (phenylamino) propan-2-ol;

n- (3- (3-bromo-6-cyano-9H-carbazol-9-yl) -2-fluoropropyl) -3-methoxyaniline;

1- (3-bromo-6-cyano-9H-carbazol-9-yl) -3- (phenylamino) propan-2-ol.

In a second aspect, the present invention provides pharmaceutically acceptable salts of the carbazole-based compounds described above.

As a preferred embodiment of the present invention, the pharmaceutically acceptable salt is an inorganic acid salt or an organic acid salt.

More preferably, the inorganic acid comprises hydrochloric acid, sulfuric acid, phosphoric acid, diphosphoric acid, hydrobromic acid, or nitric acid; the organic acid includes acetic acid, maleic acid, fumaric acid, tartaric acid, succinic acid, lactic acid, p-toluenesulfonic acid, salicylic acid or oxalic acid.

In a third aspect, the present invention provides the use of the carbazole-based compound or the pharmaceutically acceptable salt thereof as described above in the preparation of a medicament for treating glycolipid metabolic diseases and complications thereof.

In a preferred embodiment of the present invention, the glycolipid metabolic disease and its complications are fatty liver, diabetes, hyperlipidemia, obesity or arteriosclerotic cardiovascular and cerebrovascular diseases.

In a fourth aspect, the present invention provides a preparation method of the carbazole-based compound as described above:

when the carbazole compound is a carbazole amino alcohol compound, the carbazole amino alcohol compound can be prepared through the following reaction route:

when the carbazole compound is a carbazole fluoroalkyl compound, the carbazole fluoroalkyl compound can be prepared through the following reaction route:

herein, the term "C₁-C₆Alkoxy "means a saturated carbon chain of 1 to 6 carbon atoms in length, which may be straight or branched, attached to oxygen. For example, "C₁-C₆Alkoxy "includes, but is not limited to, methoxy, ethoxy, propoxy, and butoxy.

The term "C₂-C₆Alkenyloxy "means an unsaturated carbon chain of 2 to 6 carbon atoms in length, which may be straight or branched, attached to oxygen.

The term "C₂-C₆Alkynyloxy "denotes an unsaturated carbon chain of 2 to 6 carbon atoms in length, which may be straight or branched, containing at least one carbon-carbon triple bond, linked to oxygen.

The term "heterocyclyl" denotes a 4, 5 or 6 membered fully or partially saturated monocyclic ring containing 1, 2 or 3 heteroatoms selected from nitrogen, oxygen or sulfur, connected via a ring carbon atom or a ring nitrogen atom. 4. Examples of 5 or 6 membered heterocyclic rings include azetidine, tetrahydrofuran, tetrahydropyran, pyrroline, pyrrolidine, thiazolidine, morpholine, piperidine, piperazine, dihydropyridine, dihydropyrimidine and azepane.

The term "-SOp-R₅", when p is selected as 0, it represents alkylmercapto; when p is selected to be 1, it represents R₅A substituted sulfoxide; when p is selected to be 2, R is represented₅A substituted sulfonyl group.

The compound and the pharmaceutically acceptable salt thereof can be prepared into granules, powder, capsules, tablets or oral liquid according to the conventional pharmaceutical preparation method. Can be applied to individuals needing treatment by oral administration, intraperitoneal injection, subcutaneous injection, intravenous injection, intramuscular injection, mucosal administration and other ways according to dosage forms. The dosage of the invention is generally 1-1000 mg/kg body weight/day, and can be changed according to the age, illness condition and the like of an individual.

The invention has the advantages that:

the inventor carries out chemical transformation on P7C3-A20 compounds to synthesize a large class of chloropropyl carbazole compounds with a brand-new structure; subsequently, whether the new compound has potential protective effect is tested by using a fatty liver cell model (fatty toxicity and inflammation induction of palmitic acid on liver cells) and a type 2 diabetes cell model (insulin resistance is induced on liver cells by using high-concentration sucrose, and the insulin resistance of peripheral metabolic organs, which is the core pathogenic link of type 2 diabetes is simulated). The results show that the compound has obvious treatment effect on glycolipid metabolic diseases, has prominent effect and has clinical application potential.

Drawings

FIG. 1: compound I₁Potential therapeutic effect on fatty liver on cell model.^##P<0.01PA vs blank<0.01vs PA alone. N-8. PA, palmitic acid.

FIG. 2: compound I₂Potential therapeutic effect on fatty liver on cell model.^##P<0.01PA vs blank P<0.01vs PA alone. N-8. PA, palmitic acid.

FIG. 3: compound I₃Potential therapeutic effect on fatty liver on cell model.^##P<0.01PA vs blank P<0.01vs PA alone. N-8. PA, palmitic acid.

FIG. 4: compound I₄Potential therapeutic effect on fatty liver on cell model.^##P<0.01PA vs blank P<0.01vs PA alone. N-8. PA, palmitic acid.

FIG. 5: oil red O stained picture.

FIG. 6: oil red O staining statistical histogram.

FIG. 7: i is₁-I₄Potential therapeutic effect of compounds on type 2 diabetes on cell models.

FIG. 8:compound I₁And the effect of a20 on the triglyceride content of liver tissue in an animal model of fatty liver.

Detailed Description

The technical solutions of the present invention will be described clearly and completely with reference to the following detailed description, but those skilled in the art will understand that the following described examples are some, not all, of the examples of the present invention, and are only used for illustrating the present invention, and should not be construed as limiting the scope of the present invention. All other embodiments, which can be derived by a person skilled in the art from the embodiments given herein without making any creative effort, shall fall within the protection scope of the present invention. The examples, in which specific conditions are not specified, were conducted under conventional conditions or conditions recommended by the manufacturer. The reagents or instruments used are not indicated by the manufacturer, and are all conventional products available commercially.

Examples 1-5 are synthetic methods for the preparation of novel compounds. Examples 6-10 are tests for the protective effect (activity) of the novel compounds.

Some of the reagents and the like used in the embodiments of the present invention may be as follows:

example 1: 1- (3-bromo-6-methanesulfonyl-9H-carbazol-9-yl) -3- (3-methoxyphenylamino) propan-2-ol (Compound I)₁) Preparation of

Compounds I of the present example₁The preparation method comprises the following steps:

a. dissolving 6g of 3, 6-dibromocarbazole in 60mL of N, N-dimethylformamide, adding 1.2g of potassium hydroxide powder, stirring at room temperature for 1h, dropwise adding 6.2g of bromoepoxypropane, and reacting at room temperature for 3 h. The reaction was diluted with 100mL of water, filtered, the solid collected and dried to give Compound 1 as a white solid (6g, 85.7%).

b. Compound 1(6g), m-anisidine (2.2g) and bismuth chloride (2.6g) were added to cyclohexane (120mL) and reacted at 90 ℃ for 2 days. The reaction mixture was cooled to room temperature, ethyl acetate (200mL) and water (200mL) were added, the mixture was allowed to stand, the organic phase was separated, washed once with saturated brine, dried over anhydrous sodium sulfate, filtered, concentrated, and subjected to column chromatography to give compound 2(1.9g, 23.7%) as a white solid.

c. Compound 2(500mg), sodium methanesulfinate (151mg), cuprous iodide (37mg) and L-proline (23mg) were added to dimethylsulfoxide (10mL), and reacted at 150 ℃ for 4h under nitrogen. Cooling the reaction solution to room temperature, adding water (40mL), extracting with ethyl acetate (50mL × 2), combining organic phases, washing with saturated brine, drying over anhydrous sodium sulfate, filtering, concentrating, and separating by column chromatography to obtain compound I₁(210mg，46％)。

Conventionally separating and purifying the compound obtained in each step, and characterizing the pure product, wherein the structural characterization data are respectively as follows:

compound 1:¹H NMR(CDCl₃)δ:8.16(d,J＝1.8Hz,2H),7.59(dd,J＝8.7,1.9Hz,2H),7.35-7.37(m，2H),4.67(dd,J＝15.9,2.7Hz,1H),4.31(dd,J＝15.9,5.1Hz,1H),3.32-3.36(m,1H),2.82-2.85(m,1H),2.51-2.53(m,1H)；ESI-MS:m/z C₁₅H₁₁Br₂NO[M+H]⁺calculated values: 381.9; measured value: 381.9;

compound 2:¹H NMR(CDCl₃)δ:8.17(d,J＝1.8Hz,2H),7.58(dd,J＝8.7,1.9Hz,2H),7.36-7.38(m,2H),7.11(t,J＝8.1Hz,1H),6.36(dd,J＝8.1,2.1Hz,1H),6.26(dd,J＝8.0,1.9Hz,1H),6.17(t,J＝2.2Hz,1H),4.38-4.45(m,3H),4.02(m,1H),3.76(s,3H),3.34-3.38(m,1H),3.20-3.24(m,1H),2.18(d,J＝3.3Hz,1H)；ESI-MS:m/z C₂₂H₂₀Br₂N₂O₂[M+H]⁺calculated values: 504.9, respectively; found 504.9;

chemical combinationSubstance I₁：¹H NMR(CDCl₃)δ:8.66(d,J＝1.6Hz,1H),8.28(d,J＝1.8Hz,1H),8.00(dd,J＝8.7,1.8Hz,1H),7.61-7.66(m,2H),7.45(d,J＝8.7Hz,1H),7.12(t,J＝8.1Hz,1H),6.36(dd,J＝8.2,2.1Hz,1H),6.27(dd,J＝8.0,1.9Hz,1H),6.18-6.19(m,1H),4.41-4.56(m,3H),4.05-4.07(m,1H),3.77(s,3H),3.37-3.41(m,1H),3.21-3.26(m,1H),3.14(s,3H),2.35(d,J＝3.3Hz,1H)；ESI-MS:m/z C₂₃H₂₃BrN₂O₄S[M+H]⁺Calculated values: 505.1/503.1; found 505.1/503.1.

Example 2: n- (3- (3-bromo-6-methanesulfonyl-9H-carbazol-9-yl) -2-fluoropropyl) -3-methoxyaniline (Compound I)₂) Preparation of

Compounds I of the present example₂The preparation method comprises the following steps:

d. P7C3-A20(200mg), sodium methanesulfinate (60mg), cuprous iodide (14.8mg) and L-proline (8.9mg) were added to dimethyl sulfoxide (4mL) and reacted at 150 ℃ for 5h under nitrogen. Cooling the reaction solution to room temperature, adding water (20mL) for dilution, extracting with ethyl acetate (20mL multiplied by 2), combining organic phases, washing with saturated saline solution, drying with anhydrous sodium sulfate, filtering, concentrating, and separating by column chromatography to obtain the compound I₂(85mg，43.3％)。

Compound I₂：¹H NMR(CDCl₃)δ:8.63(d,J＝1.6Hz,1H),8.25(d,J＝1.7Hz,1H),7.99(dd,J＝8.7,1.7Hz,1H),7.62(dd,J＝8.7,1.8Hz,1H),7.53(d,J＝8.7Hz,1H),7.36(d,J＝8.7Hz,1H),7.11(t,J＝8.1Hz,1H),6.36(dd,J＝8.2,2.0Hz,1H),6.25(dd,J＝8.0,1.8Hz,1H),6.15-6.17(m,1H),5.04-5.18(m,1H),4.61-4.69(m,2H),3.91-4.03(m,1H),3.76(s,3H),3.28-3.53(m,2H),3.12(s,3H)；ESI-MS:m/z C₂₃H₂₂FBrN₂O₃S[M+H]⁺Calculated values: 507.1/505.1; measured value: 507.1/505.1.

Example 3: n- (3- (3-bromo-6-cyano-9H-carbazol-9-yl) -2-fluoropropan-23-methoxy-phenylamine (Compound I)₃) Preparation of

Compounds I of the present example₃The preparation method comprises the following steps:

e. P7C3-A20(200mg) and cuprous cyanide (35mg) were added to N-methylpyrrolidone (4mL) and reacted at 150 ℃ for 6h under nitrogen. Cooling the reaction solution to room temperature, adding water (10mL) for dilution, extracting with ethyl acetate (10mL multiplied by 2), combining organic phases, washing with saturated saline solution, drying with anhydrous sodium sulfate, filtering, concentrating, and separating by column chromatography to obtain the compound I₃(14mg，7.8％)。

Compound I₃：¹H NMR(CDCl₃)δ:8.35(d,J＝1.0Hz,1H),8.22(d,J＝1.8Hz,1H),7.71(dd,J＝8.6,1.5Hz,1H),7.62(dd,J＝8.7,1.9Hz,1H),7.48(d,J＝8.6Hz,1H),7.35(d,J＝8.7Hz,1H),7.12(t,J＝8.1Hz,1H),6.36(dd,J＝8.1,2.1Hz,1H),6.24(dd,J＝8.1,1.9Hz,1H),6.16(m,1H),5.05-5.19(m,1H),4.60-4.68(m,2H),3.96(t,J＝6.4Hz,1H),3.76(s,3H),3.30-3.53(m,2H)；ESI-MS:m/z C₂₃H₁₉FBrN₃O[M+H]⁺Calculated values: 452.1/454.1; found 452.1/454.1.

Example 4: 1- (3-bromo-6-methanesulfonyl-9H-carbazol-9-yl) -3- (phenylamino) propan-2-ol (Compound I)₄) Preparation of

Compounds I of the present example₄The preparation method comprises the following steps:

f. P7C3(150mg), sodium methanesulfinate (48.4mg), cuprous iodide (12mg) and L-proline (7.2mg) were added to dimethyl sulfoxide (3mL) and reacted at 150 ℃ for 4h under nitrogen. The reaction was cooled to room temperature, diluted with water (10mL), extracted with ethyl acetate (10 mL. times.2), and the organics combinedWashing with saturated saline, drying with anhydrous sodium sulfate, filtering, concentrating, and separating by column chromatography to obtain compound I₄(25mg，16.7％)。

Compound I₄：¹H NMR(MeOD)δ:8.68(d,J＝1.5Hz,1H),8.34(d,J＝1.6Hz,1H),7.94(dd,J＝8.7,1.8Hz,1H),7.71(d,J＝8.8Hz,1H),7.50-7.57(m,2H),7.09-7.13(m,2H),6.63-6.66(m,3H),4.56-4.60(m,1H),4.41-4.46(m,1H),4.24-4.30(m,1H),3.27-3.28(m,1H),3.22-3.27(m,1H),3.16(s,3H).；ESI-MS:m/z C₂₂H₂₁BrN₂O₃S[M+H]⁺Calculated values: 475.1/473.1; found 475.1/473.1.

Example 5: 1- (3-bromo-6-cyano-9H-carbazol-9-yl) -3- (phenylamino) propan-2-ol (Compound I)₅) Preparation of

Compounds I of the present example₅The preparation method comprises the following steps:

g. P7C3(150mg) and cuprous cyanide (28.3mg) were added to N-methylpyrrolidone (3mL) and reacted at 150 ℃ for 5h under nitrogen. Cooling the reaction solution to room temperature, adding water (10mL) for dilution, extracting with ethyl acetate (10mL multiplied by 3), combining organic phases, washing with saturated saline solution, drying with anhydrous sodium sulfate, filtering, concentrating, and separating by column chromatography to obtain the compound I₅(25mg，18.9％)。

Compound I₅：¹H NMR(MeOD)δ:8.50(s,1H),8.33(d,J＝1.6Hz,1H),7.67(m,2H),7.49-7.57(m,2H),7.12(t,J＝7.9Hz,2H),6.64-6.67(m,3H),4.55-4.61(m,1H),4.39-4.45(m,1H),4.24-4.29(m,1H),3.27-3.28(m,1H),3.18-3.23(m,1H)；ESI-MS:m/z C₂₂H₁₈BrN₃O[M+H]⁺Calculated values: 420.1/422.1; found 420.1/422.1.

Example 6: i is₁-I₄Potential therapeutic effect of compound on fatty liver on cell model

We used palm on cultured HepG2 hepatocytesAcid (PA) induces liver cells to establish a cell fatty liver model. PA and I were added at a concentration of 0.3mM₁-I₄The compound (concentration 0.1, 1, 10 mu mol/L) is used for treating cells for 48h, and the degree of cell injury is judged by detecting the level of Malondialdehyde (MDA) which is a lipid oxidative stress marker. In addition, the degree of accumulation of lipid in cells was also judged by using oil red O staining as follows: adding the concentrations PA (0.3mM) and I₁-I₄After 48h treatment of the cells with compounds (total of 5, concentration 1. mu. mol/L), the more red areas stained with oil Red O, indicating more lipid deposition.

Cellular injury index 1: lipid oxidative stress marker-Malondialdehyde (MDA)

(1) Compound I₁The therapeutic effect of (2) is shown in figure 1, 0.1, 1, 10. mu.M of Compound I₁Has obvious inhibition effect on lipid oxidative stress induced by PA on liver cells.

(2) Compound I₂The therapeutic effect of (2) is shown in figure 2, 10. mu.M of Compound I₃Has obvious inhibition effect on lipid oxidative stress induced by PA on liver cells, and has no statistical significance at two lower concentrations.

(3) Compound I₃The therapeutic effect of (2) is shown in FIG. 3, 10. mu.M Compound I₃Has obvious inhibition effect on lipid oxidative stress induced by PA on liver cells, has certain effect at two lower concentrations, but has no statistical significance.

(4) Compound I₄The therapeutic effect of (2) is shown in FIG. 4, 10. mu.M Compound I₄Has obvious inhibition effect on lipid oxidative stress induced by PA on liver cells, has certain effect at two lower concentrations, but has no statistical significance.

In combination: compound I_1-4All have effects of compound I₁The effect is best.

Cellular injury index 2: oil red O staining (lipid deposition amount)

Representative pictures of oil red O staining are shown in fig. 5, and statistical results are shown in fig. 6. The PA-treated group showed a lot of red color compared to the control group, indicating that the color appeared after PA-treatmentIntracellular lipid deposition is observed. And PA treatment group + Compound I₁Treatment group, PA treatment group + Compound I₃Treatment group, PA treatment group + Compound I₄The red areas of the treated groups were all significantly reduced, indicating Compound I₁、I₃、I₄Has obvious inhibition effect on lipid deposition in cells caused by PA on liver cells. Compound I₂Has certain therapeutic effect at the concentration of 1 mu M, but is less than I₁、I₃、I₄Is strong.

Example 7I₁-I₄Potential therapeutic effect of compound on type 2 diabetes on cell model

High concentration glucose induced insulin resistance model on hepatocytes: glucose (product of Shanghai Biotechnology company) and dexamethasone (product of Sigma company in America, the product number D4902) are dissolved in a HepG2 liver cell line culture medium, filtration and sterilization are carried out, so that the final concentration of the glucose in the culture medium reaches 60mM, the final concentration of the dexamethasone reaches 1 mu M, and the cultured cells count for 48 hours, thus successfully preparing the type 2 diabetes model for simulating the insulin resistance phenomenon of peripheral metabolic tissues.

Subsequently, we used compound I on cells₁、I₂、I₃、I₄Or a solvent control (0.1% final concentration of dimethylsulfoxide, DMSO) for 8 hours, then transferred to a phenol red-free, serum-free medium of normal glucose concentration (25mM), stimulated with insulin for 4 hours, then the medium was decanted, and the glucose level of the medium was measured using a glucose assay kit from Nanjing manufacturer's company (Cat. F006-1-1) to determine the amount of consumption, and insulin sensitivity was indirectly determined, thereby preliminarily evaluating Compound I₁、I₂、I₃、I₄Potential therapeutic effect on type 2 diabetes.

The results are shown in FIG. 7. The decrease in glucose concentration in the culture medium of the insulin resistance model was significantly lower compared to the control cells, indicating that the cells were still unable to efficiently take up glucose under insulin stimulation. And the compound I₁、I₂、I₃、I₄For this kind ofThe insulin resistance has obvious inhibition effect. This suggests that Compound I₁、I₂、I₃、I₄Has certain antagonistic effect on type 2 diabetes.

Example 8: i is₁Potential therapeutic effect of compound on fatty liver in animal model and comparison with compound A20

We induced an animal fatty liver model on C57 normal mice on a 60% fat-powered, high-fat diet for 4 months, and then tested compound I₁Compound a20 in a whole animal disease model. The specific administration information is as follows: the dose is 20mg/kg, and the administration is performed by gavage once in 1 day for 2 weeks.

The results are shown in FIG. 8. Compound I in comparison with the control group₁The triglyceride content of the liver tissues of the compound A20 administration group is obviously reduced, which indicates that the 2 compounds can effectively reverse liver cell lipid deposition caused by high fat diet. Furthermore, compounds I₁Is stronger than compound a 20.

Compound I₁Compound a20, showed no significant effect on the reduction of cholesterol levels in liver tissue, and may be associated with a shorter administration time.

The above description is only a preferred embodiment of the present invention, and it should be noted that, for those skilled in the art, several modifications and additions can be made without departing from the method of the present invention, and these modifications and additions should also be regarded as the protection scope of the present invention.