A class of compounds IMB-C52, and preparation method and application thereof

Abstract

The application provides a kind of compound IMB-C52 and its preparation method and application, belong to the field of biotechnology.The compound IMB-C52 provided by the application can significantly improve the body weight, liver steatosis, glucose and lipid metabolism disorder and insulin resistance of mice with metabolic dysfunction associated steatosis liver disease (MASLD) induced by high-fat diet (HFD), and shows its potential in preventing or treating fatty liver disease, diabetes and obesity.The results of examples show that the compound IMB-C52 provided by the application can significantly reduce the body weight of MASLD mice induced by HFD, improve liver histopathological abnormalities (steatosis, ballooning and intralobular inflammation), glucose and lipid metabolism disorder and insulin resistance, and show its potential in preventing or treating fatty liver disease, diabetes and obesity.

Description

Technical Field

[0001] This invention relates to the field of biotechnology, and in particular to a class of compounds, IMB-C52, their preparation methods, and applications. Background Technology

[0002] Obesity, metabolic dysfunction-associated fatty liver disease (MASLD, which can progress to metabolic dysfunction-associated steatohepatitis (MASH) and liver fibrosis / cirrhosis), and diabetes, among other disorders of glucose and lipid metabolism, have become a major public health challenge posing serious threats to health. These diseases do not exist in isolation but constitute a tightly intertwined and mutually reinforcing "metabolic disorder syndrome," with highly overlapping and interactive core pathophysiological mechanisms (such as insulin resistance, chronic inflammation, oxidative stress, mitochondrial dysfunction, endoplasmic reticulum stress, adipose tissue dysfunction, and gut microbiota dysbiosis). Patients often suffer from multiple metabolic diseases simultaneously or sequentially, forming a complex "multi-disease coexistence" state, significantly increasing the risk of cardiovascular and cerebrovascular events, liver and kidney failure, and even the severity of diseases caused by sudden infectious diseases.

[0003] These diseases are complex and difficult to treat, with a significant gap in new drug development. Taking MASLD as an example, its natural course is complex and dynamically evolving, encompassing a gradual progression from metabolic dysfunction-associated fatty liver (MAFL) to MASH, liver fibrosis, cirrhosis, and even hepatocellular carcinoma. Currently, investigational drugs targeting MASLD / MASH commonly act on metabolism, fibrosis, and inflammation. Among these, research targeting metabolism is the most popular, involving targets including THR-β, FXR, GLP-1R, PPAR, and FGF21. The first new MASH drug, the THR-β agonist Resmetirom (MGL-3196), has been marketed; however, its overall efficacy and safety require further data validation. Prior to this, the PPARα / γ dual agonist Saroglitazar showed limited market response for the treatment of MAFLD and MASH, and its safety profile remains to be investigated. Therefore, exploring and developing a highly effective and safe drug for treating fatty liver diseases such as MASLD / MASH and their common comorbidities such as obesity and diabetes is a pressing technical challenge in this field. Summary of the Invention

[0004] The purpose of this invention is to provide a compound IMB-C52, its preparation method, and its applications. The compound IMB-C52 provided by this invention can significantly reduce the body weight of MASLD model mice induced by a high-fat diet, improve liver function, glucose and lipid metabolism disorders, insulin resistance indicators, and pathological abnormalities such as hepatic steatosis. It has great potential in the prevention or treatment of diseases with glucose and lipid metabolism disorders such as fatty liver disease, diabetes, and obesity. At the same time, no toxic side effects were observed after 6 weeks of gavage at a dose of 100 mg / kg / day, and acute toxicity tests also showed that it has extremely high safety.

[0005] To achieve the above-mentioned objectives, the present invention provides the following technical solution:

[0006] This invention provides a class of compounds, IMB-C52, comprising the following structures:

[0007]

[0008] Where X is , , , , and Any one of them.

[0009] This invention provides a method for preparing the compound IMB-C52 described above, comprising the following steps:

[0010] (1) Berberine hydrochloride was heated under vacuum to obtain intermediate product 1;

[0011] (2) The intermediate product 1 obtained in step (1), the protected amino acid derivative, the polar solvent, the iodide and the strong base weak acid salt are mixed and subjected to a nucleophilic substitution reaction to obtain intermediate product 2;

[0012] (3) The intermediate product 2 obtained in step (2), the first organic solvent and trifluoroacetic acid are mixed and subjected to a deBoc reaction to obtain intermediate product 3;

[0013] (4) The bicyclic alcohol, the second organic solvent, 4-nitrophenyl chloroformate and anhydrous pyridine were mixed and subjected to an active esterification reaction to obtain intermediate product 4;

[0014] (5) The intermediate product 3 obtained in step (3), the third organic solvent, the intermediate product 4 obtained in step (4) and N,N-diisopropylethylamine are mixed and subjected to a nucleophilic acyl substitution reaction to obtain compound IMB-C52;

[0015] In the compound IMB-C52, X is... .

[0016] Preferably, in step (1), the vacuum degree of vacuum heating is 20~30 mmHg, the vacuum heating temperature is 200~230℃, and the vacuum heating holding time is 30~60 min.

[0017] Preferably, in step (2), the protected amino acid derivative is chloromethyl 3-((tert-butoxycarbonyl)amino)propionate, the polar solvent is anhydrous acetonitrile, the iodide is sodium iodide, and the strong base-weak acid salt is anhydrous potassium carbonate.

[0018] Preferably, in step (2), the molar ratio of the protected amino acid derivative and intermediate product 1 is (1~1.5):1.

[0019] Preferably, in step (3), the ratio of the amount of intermediate product 2 to the volume of trifluoroacetic acid is (4~6) mmol: 4 mL.

[0020] This invention provides a method for preparing the compound IMB-C52 described above, comprising the following steps:

[0021] (1) Berberine hydrochloride was heated under vacuum to obtain intermediate product 1;

[0022] (2) The intermediate product 1 obtained in step (1), methyl bromoacetate and anhydrous dichloromethane are mixed and subjected to a nucleophilic substitution reaction to obtain intermediate product 2;

[0023] (3) The intermediate product 2 obtained in step (2), the mixed solvent and lithium hydroxide are mixed and subjected to ester hydrolysis reaction to obtain intermediate product 3;

[0024] (4) The intermediate product 3 obtained in step (3), DMF, 2-(7-azabenzotriazole)-N,N,N',N'-tetramethylurea hexafluorophosphate, triethylamine and mono-Boc protected diamine compound are mixed and subjected to amide condensation reaction to obtain intermediate product 4.

[0025] (5) The intermediate product 4 obtained in step (4), anhydrous dichloromethane and trifluoroacetic acid are mixed and subjected to deBoc reaction to obtain intermediate product 5;

[0026] (6) The bicyclic alcohol, the second organic solvent, 4-nitrophenyl chloroformate and anhydrous pyridine were mixed and subjected to an active esterification reaction to obtain intermediate product 6;

[0027] (7) The intermediate product 5 obtained in step (5), the third organic solvent, the intermediate product 6 obtained in step (6) and N,N-diisopropylethylamine are mixed and subjected to a nucleophilic acyl substitution reaction to obtain compound IMB-C52.

[0028] In the compound IMB-C52, X is... , , and Any one of them.

[0029] This invention provides a method for preparing the compound IMB-C52 described above, comprising the following steps:

[0030] (1) Berberine hydrochloride was heated under vacuum to obtain intermediate product 1;

[0031] (2) The bicyclic alcohol, the second organic solvent, 4-nitrophenyl chloroformate and anhydrous pyridine were mixed and subjected to an active esterification reaction to obtain intermediate product 2;

[0032] (3) The intermediate product 2 obtained in step (2), the amino acid derivative, DMF and N,N-diisopropylethylamine are mixed and subjected to a nucleophilic acyl substitution reaction to obtain intermediate product 3;

[0033] (4) The intermediate product 3 obtained in step (3), the intermediate product 1 obtained in step (1), anhydrous dichloromethane and dicyclohexylcarbodiimide are mixed and subjected to ester condensation reaction to obtain compound IMB-C52.

[0034] In the compound IMB-C52, X is... .

[0035] This invention provides the use of the compound IMB-C52 described in the above-described technical solution or the compound IMB-C52 prepared by the preparation method described in the above-described technical solution in the preparation of a medicament for the prevention or treatment of metabolic syndrome accompanied by glucose and lipid metabolism disorders.

[0036] Preferably, the metabolic syndrome accompanied by glucose and lipid metabolism disorders includes any one or more of fatty liver disease, diabetes, and obesity.

[0037] The compound IMB-C52 provided by this invention can significantly improve body weight, hepatic steatosis, glucose and lipid metabolism disorders, and insulin resistance in HFD-induced MASLD mice, demonstrating its potential in the prevention or treatment of fatty liver disease, diabetes, and obesity. The results of the examples show that the compound IMB-C52 provided by this invention can significantly reduce body weight, improve liver histopathological abnormalities (steatosis, ballooning degeneration, and intralobular inflammation), glucose and lipid metabolism disorders, and insulin resistance in HFD-induced MASLD mice, demonstrating its potential in the prevention or treatment of fatty liver disease, diabetes, and obesity. Attached Figure Description

[0038] Figure 1 The initial group body weight of mice after 10 weeks of induction;

[0039] Figure 2 The final body weight of mice 6 weeks after drug administration;

[0040] Figure 3 The average weekly food intake per mouse during the 6-week drug administration period;

[0041] Figure 4 The body fat percentage of mice after 6 weeks of drug administration;

[0042] Figure 5 The lean meat percentage of mice after 6 weeks of drug administration;

[0043] Figure 6 The content of white adipose tissue in the epididymis of mice after 6 weeks of drug administration;

[0044] Figure 7 This represents the fasting blood glucose level in mice.

[0045] Figure 8 This refers to the fasting serum insulin content in mice.

[0046] Figure 9 This represents the insulin resistance index in mice.

[0047] Figure 10 The alanine aminotransferase (ALT) level in mouse serum;

[0048] Figure 11 The aspartate aminotransferase (AST) content in mouse serum;

[0049] Figure 12 This refers to the triglyceride content in mouse serum;

[0050] Figure 13 This represents the total cholesterol content in mouse serum;

[0051] Figure 14 This refers to the triglyceride content in mouse liver;

[0052] Figure 15 This refers to the total cholesterol content in mouse liver;

[0053] Figure 16 The results of gross examination, H&E staining, and Oil Red staining of mouse liver tissue are shown.

[0054] Figure 17 The results of NAS scoring for mouse liver tissue pathology;

[0055] Figure 18 The body weight curve of mice in the acute toxicity test of IMB-C52;

[0056] Figure 19 Serum biochemical indicators of mice in the acute toxicity test of IMB-C52;

[0057] Figure 20 H&E staining pathological images of the heart, liver, spleen, lungs, and kidneys of mice in the IMB-C52 acute toxicity experiment. Detailed Implementation

[0058] This invention provides a class of compounds, IMB-C52, comprising the following structures:

[0059]

[0060] Where X is , , , , and Any one of them.

[0061] The compound IMB-C52 provided by this invention can significantly improve body weight, hepatic steatosis, glucose and lipid metabolism disorders, and insulin resistance in HFD-induced MASLD mice, showing its potential in the prevention or treatment of fatty liver disease, diabetes, and obesity.

[0062] This invention also provides a method for preparing the compound IMB-C52 described in the above technical solution, comprising the following steps:

[0063] (1) Berberine hydrochloride was heated under vacuum to obtain intermediate product 1;

[0064] (2) The intermediate product 1 obtained in step (1), the protected amino acid derivative, the polar solvent, the iodide and the strong base weak acid salt are mixed and subjected to a nucleophilic substitution reaction to obtain intermediate product 2;

[0065] (3) The intermediate product 2 obtained in step (2), the first organic solvent and trifluoroacetic acid are mixed and subjected to a deBoc reaction to obtain intermediate product 3;

[0066] (4) The bicyclic alcohol, the second organic solvent, 4-nitrophenyl chloroformate and anhydrous pyridine were mixed and subjected to an active esterification reaction to obtain intermediate product 4;

[0067] (5) The intermediate product 3 obtained in step (3), the third organic solvent, the intermediate product 4 obtained in step (4) and N,N-diisopropylethylamine are mixed and subjected to a nucleophilic acyl substitution reaction to obtain compound IMB-C52;

[0068] In the compound IMB-C52, X is... .

[0069] Unless otherwise specified, all raw materials used in this invention are commercially available products well known to those skilled in the art.

[0070] In this invention, the terms "intermediate product 1", "intermediate product 2", "intermediate product 3", "intermediate product 4", "first organic solvent", "second organic solvent" and "third organic solvent" are not specifically required, but are merely used to distinguish between intermediate products and organic solvents.

[0071] This invention involves heating berberine hydrochloride to obtain intermediate product 1. By using berberine hydrochloride as a raw material, this invention can combine its efficacy against fatty liver disease, diabetes, and obesity, thereby improving the therapeutic effect of the compound. Simultaneously, berberine hydrochloride exhibits excellent safety, thus reducing the compound's side effects on organisms.

[0072] In this invention, the vacuum degree of the vacuum heating is preferably 20-30 mmHg; the vacuum heating temperature is preferably 200-230℃; and the vacuum heating holding time is preferably 30-60 min. As one embodiment of this invention, the vacuum degree of the vacuum heating can be 21 mmHg, 22 mmHg, 23 mmHg, 24 mmHg, 25 mmHg, 26 mmHg, 27 mmHg, 28 mmHg, or 29 mmHg; the vacuum heating temperature can be 205℃, 210℃, 215℃, 220℃, or 225℃; and the vacuum heating holding time can be 35 min, 40 min, 45 min, 50 min, or 55 min. This invention, by vacuum heating berberine hydrochloride, causes the loss of water of crystallization and some HCl, resulting in the loosening of the isoquinoline ring system and the breakage of the tetrahydroisoquinoline skeleton, thereby obtaining intermediate product 1.

[0073] After vacuum heating, the product is preferably purified. In this invention, purification is preferably performed using a Flash column; the mobile phase used for Flash column purification is preferably a mixture of dichloromethane and methanol; the volume ratio of dichloromethane to methanol in the mixture is preferably (80-90):(10-20), more preferably 85:15; the flow rate of the mobile phase is preferably 80-150 mL / min, more preferably 90-140 mL / min, and even more preferably 100-120 mL / min. This invention removes impurities generated during vacuum heating through purification, thereby obtaining a high-purity intermediate product 1.

[0074] After obtaining intermediate product 1, the present invention mixes intermediate product 1, protected amino acid derivative, polar solvent, iodide and strong base weak acid salt and carries out nucleophilic substitution reaction to obtain intermediate product 2.

[0075] In this invention, the protected amino acid derivative is preferably chloromethyl 3-((tert-butoxycarbonyl)amino)propionate; the polar solvent is preferably anhydrous acetonitrile; the iodide is preferably sodium iodide; the strong base-weak acid salt is preferably anhydrous potassium carbonate; the molar ratio of the protected amino acid derivative to the volume of the polar solvent is preferably (7~8) mmol:(30~50) mL, more preferably (7.5~7.8) mmol:(35~45) mL, and even more preferably 7.763 mmol:40 mL; the protected amino acid derivative and the iodide... The preferred molar ratio is 1:(0.8~1.2), more preferably 1:(0.9~1.1), and even more preferably 1:1; the preferred molar ratio of the protected amino acid derivative to intermediate 1 is (1~1.5):1, more preferably (1.2~1.3):1, and even more preferably 1.25:1; the preferred molar ratio of the protected amino acid derivative to the strong base-weak acid salt is (7~8):(10~15), more preferably (7.5~7.8):(12~13), and even more preferably 7.763:12.418. This invention improves reaction efficiency and avoids waste of raw materials by controlling the amount of each component.

[0076] In this invention, the preferred method for mixing the intermediate product 1, the protected amino acid derivative, the polar solvent, the iodide, and the strong base-weak acid salt is as follows: first, the protected amino acid derivative and the polar solvent are mixed; then, the iodide is added and heated and stirred; next, the intermediate product 1 is added; and finally, the strong base-weak acid salt is added. In this invention, the heating and stirring temperature is preferably 35-50°C, more preferably 40-45°C; and the heating and stirring time is preferably 1-3 hours, more preferably 2 hours. This invention does not impose a specific limitation on the heating and stirring rate; based on the technical knowledge of those skilled in the art, it is sufficient to avoid solution splashing and ensure uniform mixing of all components. By employing the above method, this invention ensures uniform mixing of all components.

[0077] In this invention, the temperature of the nucleophilic substitution reaction is preferably 35-50°C, more preferably 40-45°C; the holding time of the nucleophilic substitution reaction is preferably 6-10 hours, more preferably 8 hours; the nucleophilic substitution reaction is preferably carried out under stirring conditions; and the nucleophilic substitution reaction is preferably monitored by TLC. This invention does not have a specific limitation on the stirring rate, as long as it avoids solution splashing. This invention uses TLC monitoring to monitor whether the reaction is complete.

[0078] After the nucleophilic substitution reaction is completed, the product of the nucleophilic substitution reaction is preferably extracted, washed, dried, concentrated and purified in sequence to obtain intermediate product 2.

[0079] In this invention, the extractant used for extraction is preferably ethyl acetate; the number of extractions is preferably 2 to 5 times, more preferably 3 to 4 times; and the extracted organic phases are preferably combined. This invention extracts intermediate product 2 through extraction, and by multiple extractions, the extraction efficiency and yield of intermediate product 2 can be improved.

[0080] In this invention, the detergent used for washing is preferably saturated saline solution; the number of washing cycles is preferably 2 to 5 times, more preferably 3 to 4 times. This invention removes residual impurities and further improves purity through washing.

[0081] In this invention, the preferred method of drying and concentration is using anhydrous sodium sulfate. The specific amount of anhydrous sodium sulfate used is not particularly limited in this invention and can be determined based on the technical knowledge of those skilled in the art.

[0082] In this invention, the purification is preferably performed using a Flash column; the mobile phase used for Flash column purification is preferably a mixture of dichloromethane and methanol; the volume ratio of dichloromethane to methanol in the mixture is preferably 99:1; the flow rate of the mobile phase is preferably 20-40 mL / min, more preferably 30 mL / min. Through purification, this invention can further remove impurities to obtain a high-purity intermediate product 2.

[0083] After obtaining intermediate product 2, the present invention mixes intermediate product 2, the first organic solvent and trifluoroacetic acid and carries out a deBoc reaction to obtain intermediate product 3.

[0084] In this invention, the first organic solvent is preferably anhydrous dichloromethane; the ratio of the amount of the intermediate product 2 to the volume of the first organic solvent is preferably (4~6) mmol:20 mL, more preferably (4.5~5) mmol:20 mL, and even more preferably 4.718 mmol:20 mL; the ratio of the amount of the intermediate product 2 to the volume of trifluoroacetic acid is preferably (4~6) mmol:4 mL, more preferably (4.5~5) mmol:4 mL, and even more preferably 4.718 mmol:4 mL.

[0085] In this invention, the preferred method for mixing the intermediate product 2, the first organic solvent, and trifluoroacetic acid is as follows: the intermediate product 2 is first dissolved in the first organic solvent, and then the mixture is cooled to 0°C before adding trifluoroacetic acid dropwise. This invention does not impose any particular limitation on the dropping rate of the trifluoroacetic acid; it can be determined based on the technical knowledge of those skilled in the art. By employing the above-described mixing method, this invention avoids an excessively vigorous reaction.

[0086] In this invention, the Boc removal reaction is preferably carried out under ice-water bath conditions; the Boc removal reaction time is preferably 2-6 hours, more preferably 3-5 hours, and even more preferably 4 hours; the Boc removal reaction is preferably monitored by TLC. This invention ensures complete reaction by controlling the temperature and time of the Boc removal reaction.

[0087] After the Boc removal reaction is completed, the product of the Boc removal reaction is preferably concentrated under reduced pressure. The specific operation of this reduced pressure concentration is not particularly limited and can be determined based on the technical knowledge of those skilled in the art.

[0088] In this invention, a bicyclic alcohol, a second organic solvent, 4-nitrophenyl chloroformate and anhydrous pyridine are mixed and subjected to an active esterification reaction to obtain intermediate product 4.

[0089] In this invention, the second organic solvent is preferably anhydrous tetrahydrofuran or anhydrous dichloromethane; the molar ratio of the bicyclic alcohol to the volume of the second organic solvent is preferably (4.5~6) mmol:20 mL, more preferably (5~5.5) mmol:20 mL, and even more preferably 5.123 mmol:20 mL; the molar ratio of the bicyclic alcohol to 4-nitrophenyl chloroformate is preferably 1:(1.2~1.5), more preferably 1:(1.25~1.4), and even more preferably 1:1.3; the molar ratio of the bicyclic alcohol to anhydrous pyridine is preferably 1:(2.2~2.8), more preferably 1:(2.4~2.6), and even more preferably 1:2.5. By using an anhydrous organic solvent, this invention avoids the failure of 4-nitrophenyl chloroformate during the reaction process.

[0090] In this invention, the preferred method for mixing the bicyclic alcohol, the second organic solvent, 4-nitrophenyl chloroformate, and anhydrous pyridine is as follows: first, the bicyclic alcohol is dissolved in the second organic solvent, then 4-nitrophenyl chloroformate is added, and finally anhydrous pyridine is added. In this invention, the mixing is preferably carried out under stirring conditions. This invention does not impose a particular limitation on the stirring rate; based on the technical knowledge of those skilled in the art, it is sufficient to ensure that the components are mixed uniformly and that the solution does not splash. By employing the above-described mixing method, this invention ensures complete mixing of all components.

[0091] In this invention, the temperature of the active esterification reaction is preferably room temperature; the time of the active esterification reaction is preferably 6-10 hours, more preferably 7-9 hours, and even more preferably 8 hours; the active esterification reaction is preferably carried out under stirring conditions; and the active esterification reaction is preferably monitored by TLC. This invention does not impose any special limitation on the stirring rate; it only needs to be determined based on the technical knowledge of those skilled in the art to avoid solution splashing. By controlling the parameters of the active esterification reaction, this invention can ensure complete reaction.

[0092] After the active esterification reaction is completed, the product of the active esterification reaction is preferably extracted, washed, dried, concentrated and purified in sequence.

[0093] In this invention, the extractant used for extraction is preferably ethyl acetate; the number of extractions is preferably 2 to 5 times, more preferably 3 to 4 times; and the extracted organic phases are preferably combined. This invention extracts intermediate product 2 through extraction, and by multiple extractions, the extraction efficiency and yield of intermediate product 4 can be improved.

[0094] In this invention, the detergent used for washing is preferably saturated saline solution; the number of washing cycles is preferably 2 to 5 times, more preferably 3 to 4 times. This invention removes residual impurities and further improves purity through washing.

[0095] In this invention, the preferred method of drying and concentration is using anhydrous sodium sulfate. The specific amount of anhydrous sodium sulfate used is not particularly limited in this invention and can be determined based on the technical knowledge of those skilled in the art.

[0096] In this invention, the purification is preferably performed using a Flash column; the mobile phase used for Flash column purification is preferably a mixture of petroleum ether and ethyl acetate; the volume ratio of petroleum ether to ethyl acetate in the mixture is preferably 40:60; the flow rate of the mobile phase is preferably 20-40 mL / min, more preferably 30 mL / min. Through purification, this invention can further remove impurities to obtain a high-purity intermediate product 4.

[0097] After obtaining intermediates 3 and 4, the present invention mixes intermediate 3, a third organic solvent, intermediate 4 and N,N-diisopropylethylamine and performs a nucleophilic acyl substitution reaction to obtain compound IMB-C52.

[0098] In this invention, the third organic solvent is preferably anhydrous N,N-dimethylformamide; the molar ratio of the intermediate product 3 to the volume of the third organic solvent is preferably (4~6) mmol:20 mL, more preferably (4.2~5) mmol:20 mL, and even more preferably 4.628 mmol:20 mL; the molar ratio of the intermediate product 3 to the intermediate product 4 is preferably 1:(0.8~1.2), more preferably 1:(0.9~1.1), and even more preferably 1:1; the molar ratio of the intermediate product 3 to N,N-diisopropylethylamine is preferably 1:(2~4), more preferably 1:(2.5~3.5), and even more preferably 1:3.

[0099] In this invention, the preferred method for mixing intermediate product 3, the third organic solvent, intermediate product 4, and N,N-diisopropylethylamine is as follows: first, intermediate product 3 is dissolved in the third organic solvent, then intermediate product 4 is added, and finally N,N-diisopropylethylamine is added. In this invention, the mixing is preferably carried out under stirring conditions. This invention does not impose a particular limitation on the stirring rate; it can be determined based on the technical knowledge of those skilled in the art, as long as it avoids solution splashing. By employing the above-described mixing method, this invention enables a more uniform mixing of the components.

[0100] In this invention, the temperature of the nucleophilic acyl substitution reaction is preferably room temperature; the holding time of the nucleophilic acyl substitution reaction is preferably 6-10 h, more preferably 7-9 h, and even more preferably 8 h; the nucleophilic acyl substitution reaction is preferably carried out under stirring conditions; and the nucleophilic acyl substitution reaction is preferably monitored by TLC. This invention does not have a specific limitation on the stirring rate, as long as it avoids solution splashing. By controlling the conditions of the nucleophilic acyl substitution reaction, this invention can ensure the completeness of the reaction; and by using TLC monitoring, it can determine whether the nucleophilic acyl substitution reaction has ended.

[0101] After the nucleophilic acyl substitution reaction is completed, the product of the nucleophilic acyl substitution reaction is preferably extracted, washed, dried, concentrated and purified in sequence.

[0102] In this invention, the extractant used for extraction is preferably ethyl acetate; the number of extractions is preferably 2 to 5 times, more preferably 3 to 4 times; and the extracted organic phases are preferably combined. This invention extracts intermediate product 2 through extraction, and multiple extractions can improve extraction efficiency and increase the yield of the compound.

[0103] In this invention, the detergent used for washing is preferably saturated saline solution; the number of washing cycles is preferably 2 to 5 times, more preferably 3 to 4 times. This invention removes residual impurities and further improves purity through washing.

[0104] In this invention, the preferred method of drying and concentration is using anhydrous sodium sulfate. The specific amount of anhydrous sodium sulfate used is not particularly limited in this invention and can be determined based on the technical knowledge of those skilled in the art.

[0105] In this invention, the purification is preferably performed using a Flash column; the mobile phase used for Flash column purification is preferably a mixture of dichloromethane and methanol; the volume ratio of dichloromethane to methanol in the mixture is preferably 98:2; the flow rate of the mobile phase is preferably 20-40 mL / min, more preferably 30 mL / min. This invention, through purification, can further remove impurities and obtain a high-purity compound.

[0106] This invention also provides a method for preparing the compound IMB-C52 described in the above technical solution, comprising the following steps:

[0107] (1) Berberine hydrochloride was heated under vacuum to obtain intermediate product 1;

[0108] (2) The intermediate product 1 obtained in step (1), methyl bromoacetate and anhydrous dichloromethane are mixed and subjected to a nucleophilic substitution reaction to obtain intermediate product 2;

[0109] (3) The intermediate product 2 obtained in step (2), the mixed solvent and lithium hydroxide are mixed and subjected to ester hydrolysis reaction to obtain intermediate product 3;

[0110] (4) The intermediate product 3 obtained in step (3), DMF, 2-(7-azabenzotriazole)-N,N,N',N'-tetramethylurea hexafluorophosphate, triethylamine and mono-Boc protected diamine compound are mixed and subjected to amide condensation reaction to obtain intermediate product 4.

[0111] (5) The intermediate product 4 obtained in step (4), anhydrous dichloromethane and trifluoroacetic acid are mixed and subjected to deBoc reaction to obtain intermediate product 5;

[0112] (6) The bicyclic alcohol, the second organic solvent, 4-nitrophenyl chloroformate and anhydrous pyridine were mixed and subjected to an active esterification reaction to obtain intermediate product 6;

[0113] (7) The intermediate product 5 obtained in step (5), the third organic solvent, the intermediate product 6 obtained in step (6) and N,N-diisopropylethylamine are mixed and subjected to a nucleophilic acyl substitution reaction to obtain compound IMB-C52.

[0114] In the compound IMB-C52, X is... , , and Any one of them.

[0115] In this invention, berberine hydrochloride is heated under vacuum to obtain intermediate product 1.

[0116] In this invention, the parameters of the vacuum heating are preferably the same as those of the aforementioned vacuum heating, and will not be repeated here.

[0117] After vacuum heating, the product of vacuum heating is preferably purified. In this invention, the specific purification operation is preferably the same as the purification operation and function described above after vacuum heating, and will not be repeated here.

[0118] After obtaining intermediate product 1, the present invention mixes intermediate product 1, methyl bromoacetate and anhydrous dichloromethane and carries out a nucleophilic substitution reaction to obtain intermediate product 2.

[0119] In this invention, the molar ratio of intermediate product 1 to methyl bromoacetate is preferably 1:(2~6), more preferably 1:(3~5), and even more preferably 1:4; the molar ratio of intermediate product 1 to the volume ratio of anhydrous dichloromethane is preferably (5.5~7) mmol:(30~100) mL, more preferably (6~6.5) mmol:(40~60) mL, and even more preferably 6.209 mmol:50 mL.

[0120] In this invention, the intermediate product 1, methyl bromoacetate, and anhydrous dichloromethane are preferably mixed by first dissolving the intermediate product 1 in anhydrous dichloromethane, and then adding the methyl bromoacetate; the mixing is preferably carried out under stirring conditions. This invention does not impose any particular limitations on the stirring rate and time for the mixing, as long as it ensures uniform mixing.

[0121] In this invention, the parameters of the nucleophilic substitution reaction are preferably the same as those of the aforementioned nucleophilic substitution reaction, and will not be repeated here.

[0122] After the nucleophilic substitution reaction is completed, the product of the nucleophilic substitution reaction is preferably subjected to filtration, heating and reflux stirring, and filtration sequentially to obtain intermediate product 2. The specific filtration operation is not particularly limited in this invention, as long as solid-liquid separation can be achieved. In this invention, the solution used for heating and reflux stirring is preferably a 95% ethanol solution. The amount of ethanol solution used is not particularly limited in this invention and can be determined based on the technical knowledge of those skilled in the art. The above operations can remove impurities.

[0123] After obtaining intermediate product 2, the present invention mixes intermediate product 2, mixed solvent and lithium hydroxide and performs ester hydrolysis reaction to obtain intermediate product 3.

[0124] In this invention, the mixed solvent is preferably a mixture of methanol and water; the volume ratio of methanol to water is preferably (2~5):1, more preferably (3~4):1; the molar ratio of intermediate product 2 to the volume ratio of the mixed solvent is preferably (5~6.5) mmol:80 mL, more preferably (5.5~6) mmol:80 mL, and even more preferably 5.835 mmol:80 mL; the molar ratio of intermediate product 2 to lithium hydroxide is preferably (5~6.5):11.67, more preferably (5.5~6):11.67, and even more preferably 5.835:11.67.

[0125] In this invention, the temperature of the ester hydrolysis reaction is preferably room temperature; the time of the ester hydrolysis reaction is preferably 5-8 hours, more preferably 6-7 hours; and the ester hydrolysis reaction is preferably monitored by TLC. This invention uses TLC monitoring to monitor whether the reaction is complete.

[0126] After the ester hydrolysis reaction is completed, the products of the ester hydrolysis reaction are preferably subjected to pH adjustment and filtration sequentially to obtain intermediate product 3. In this invention, the pH adjustment is preferably performed by adding hydrochloric acid; the concentration of the hydrochloric acid is preferably 6N; and the pH value after pH adjustment is preferably 3. This invention does not have specific limitations on the specific operation of the filtration, as long as solid-liquid separation can be achieved.

[0127] After obtaining intermediate product 3, the present invention mixes intermediate product 3, anhydrous N,N-dimethylformamide (DMF), 2-(7-azabenzotriazole)-N,N,N',N'-tetramethylurea hexafluorophosphate (HATU), triethylamine and a mono-Boc protected diamine compound and performs an amide condensation reaction to obtain intermediate product 4.

[0128] In this invention, the mono-Boc protected diamine compound is preferably N-tert-butoxycarbonyl-1,2-ethylenediamine, N-tert-butoxycarbonyl-1,3-propanediamine, or 1-tert-butoxycarbonyl-4-aminomethylpiperidine. In this invention, the molar ratio of intermediate product 3 to DMF is preferably (5~6) mmol:30 mL, more preferably (5.4~5.7) mmol:30 mL, and even more preferably 5.524 mmol:30 mL; the molar ratio of intermediate product 3 to HATU is preferably (0.5~1.5):2, more preferably (0.8~1.2):2, and even more preferably 1:2; the molar ratio of intermediate product 3 to triethylamine is preferably (0.5~2):5, more preferably (1~1.5):5, and even more preferably 1:5; the molar ratio of intermediate product 3 to mono-Boc protected diamine compound is preferably (5~6):7.181, more preferably (5.3~5.8):7.181, and even more preferably 5.524:7.181.

[0129] In this invention, the preferred method for mixing intermediate 3, DMF, HATU, triethylamine, and the mono-Boc protected diamine compound is as follows: first, intermediate 3 is dissolved in DMF, then the mono-Boc protected diamine compound is added, followed by HATU, and finally triethylamine is added. This method ensures uniform mixing.

[0130] In this invention, the temperature of the amide condensation reaction is preferably room temperature; the reaction time is preferably 6-10 hours, more preferably 8 hours; and the amide condensation reaction is preferably monitored by TLC. This invention controls the reaction parameters, which can promote reaction efficiency; and TLC monitoring can be used to monitor whether the reaction is complete.

[0131] After the amide condensation reaction is completed, the product of the amide condensation reaction is preferably subjected to vacuum distillation, solid precipitation, and filtration in sequence. The specific operations of vacuum distillation and filtration are not particularly limited in this invention and can be determined based on the technical knowledge of those skilled in the art. The preferred method for solid precipitation is the addition of anhydrous methanol followed by standing. The amount of anhydrous methanol and the standing time are not particularly limited in this invention and can be determined based on the technical knowledge of those skilled in the art.

[0132] After obtaining intermediate product 4, the present invention mixes intermediate product 4, anhydrous dichloromethane and trifluoroacetic acid and carries out a deBoc reaction to obtain intermediate product 5.

[0133] In this invention, the preferred ratio of the amount of intermediate product 4 to the volume of anhydrous dichloromethane is (3.5~5) mmol:20 mL, more preferably (4~4.5) mmol:20 mL, and even more preferably 4.159 mmol:20 mL; the preferred ratio of the amount of intermediate product 4 to the volume of trifluoroacetic acid is (3.5~5) mmol:4 mL, more preferably (4~4.5) mmol:4 mL, and even more preferably 4.159 mmol:4 mL.

[0134] In this invention, the preferred method for mixing intermediate product 4, anhydrous dichloromethane, and trifluoroacetic acid is to first dissolve intermediate product 4 in anhydrous dichloromethane, then cool to 0°C and add trifluoroacetic acid dropwise. This invention does not impose any special limitations on the specific operations of cooling and dropwise addition; these can be determined based on the technical knowledge of those skilled in the art.

[0135] In this invention, the Boc removal reaction is preferably carried out under ice-water bath conditions; the Boc removal reaction time is preferably 2-6 hours, more preferably 3-5 hours, and even more preferably 4 hours; the Boc removal reaction is preferably monitored by TLC. This invention ensures complete reaction by controlling the temperature and time of the Boc removal reaction.

[0136] After the Boc removal reaction is completed, the product of the Boc removal reaction is preferably concentrated under reduced pressure. The present invention does not have specific limitations on the specific operation of the reduced pressure concentration; any reduced pressure concentration method well known to those skilled in the art can be used.

[0137] In this invention, a bicyclic alcohol, a second organic solvent, 4-nitrophenyl chloroformate and anhydrous pyridine are mixed and subjected to an active esterification reaction to obtain intermediate product 6.

[0138] In this invention, the specific types, amounts, and mixing methods of the bicyclic alcohol, the second organic solvent, 4-nitrophenyl chloroformate, and anhydrous pyridine are the same as described above, and will not be repeated here. In this invention, the parameters of the active esterification reaction and the post-treatment process are the same as described above, and will not be repeated here.

[0139] After obtaining intermediates 5 and 6, the present invention mixes intermediate 5, a third organic solvent, intermediate 6 and N,N-diisopropylethylamine and performs a nucleophilic acyl substitution reaction to obtain compound IMB-C52.

[0140] In this invention, the third organic solvent is preferably anhydrous N,N-dimethylformamide; the molar ratio of the intermediate product 5 to the volume of the third organic solvent is preferably (3.5~4.8) mmol:20 mL, more preferably (4~4.5) mmol:20 mL, and even more preferably 4.263 mmol:20 mL; the molar ratio of the intermediate product 5 to the intermediate product 6 is preferably 1:(0.5~1.5), more preferably 1:(0.8~1.2), and even more preferably 1:1; the molar ratio of the intermediate product 5 to N,N-diisopropylethylamine is preferably 1:(2.5~4), more preferably 1:(3~3.5).

[0141] In this invention, the temperature of the nucleophilic acyl substitution reaction is preferably room temperature; the holding time of the nucleophilic acyl substitution reaction is preferably 6-10 hours, more preferably 7-9 hours, and even more preferably 8 hours; the nucleophilic acyl substitution reaction is preferably carried out under stirring conditions; and the nucleophilic acyl substitution reaction is preferably monitored by TLC. This invention does not impose any special limitation on the stirring rate; it can be determined based on the technical knowledge of those skilled in the art, as long as it avoids solution splashing. By controlling the conditions of the nucleophilic acyl substitution reaction, this invention can ensure the completeness of the reaction; and by using TLC monitoring, it can determine whether the nucleophilic acyl substitution reaction has ended.

[0142] After the nucleophilic acyl substitution reaction is completed, the product of the nucleophilic acyl substitution reaction is preferably extracted, washed, dried, concentrated and purified in sequence.

[0143] In this invention, the extractant used for extraction is preferably ethyl acetate; the number of extractions is preferably 2 to 5 times, more preferably 3 to 4 times; and the extracted organic phases are preferably combined. This invention extracts intermediate product 2 through extraction, and multiple extractions can improve extraction efficiency and increase the yield of the compound.

[0144] In this invention, the detergent used for washing is preferably saturated saline solution; the number of washing cycles is preferably 2 to 5 times, more preferably 3 to 4 times. This invention removes residual impurities and further improves purity through washing.

[0145] In this invention, the preferred method of drying and concentration is using anhydrous sodium sulfate. The specific amount of anhydrous sodium sulfate used is not particularly limited in this invention and can be determined based on the technical knowledge of those skilled in the art.

[0146] In this invention, the purification is preferably performed using a Flash column; the mobile phase used for Flash column purification is preferably a mixture of dichloromethane and methanol; the volume ratio of dichloromethane to methanol in the mixture is preferably 98:2; the flow rate of the mobile phase is preferably 20-40 mL / min, more preferably 30 mL / min. This invention, through purification, can further remove impurities and obtain a high-purity compound.

[0147] This invention also provides a method for preparing the compound IMB-C52 described in the above technical solution, comprising the following steps:

[0148] (1) Berberine hydrochloride was heated under vacuum to obtain intermediate product 1;

[0149] (2) The bicyclic alcohol, the second organic solvent, 4-nitrophenyl chloroformate and anhydrous pyridine were mixed and subjected to an active esterification reaction to obtain intermediate product 2;

[0150] (3) The intermediate product 2 obtained in step (2), the amino acid derivative, DMF and N,N-diisopropylethylamine are mixed and subjected to a nucleophilic acyl substitution reaction to obtain intermediate product 3;

[0151] (4) The intermediate product 3 obtained in step (3), the intermediate product 1 obtained in step (1), anhydrous dichloromethane and dicyclohexylcarbodiimide are mixed and subjected to ester condensation reaction to obtain compound IMB-C52.

[0152] In the compound IMB-C52, X is... .

[0153] In this invention, berberine hydrochloride is heated under vacuum to obtain intermediate product 1.

[0154] In this invention, the parameters of the vacuum heating are preferably the same as those of the aforementioned vacuum heating, and will not be repeated here.

[0155] After vacuum heating, the product of vacuum heating is preferably purified. In this invention, the specific purification operation is preferably the same as the purification operation and function described above after vacuum heating, and will not be repeated here.

[0156] In this invention, a bicyclic alcohol, a second organic solvent, 4-nitrophenyl chloroformate and anhydrous pyridine are mixed and subjected to an active esterification reaction to obtain intermediate product 2.

[0157] In this invention, the specific types, amounts, and mixing methods of the bicyclic alcohol, the second organic solvent, 4-nitrophenyl chloroformate, and anhydrous pyridine are the same as described above, and will not be repeated here. In this invention, the parameters of the active esterification reaction and the post-treatment process are the same as described above, and will not be repeated here.

[0158] After obtaining intermediate product 2, the present invention mixes intermediate product 2, amino acid derivative, anhydrous N,N-dimethylformamide (DMF) and N,N-diisopropylethylamine and performs a nucleophilic acyl substitution reaction to obtain intermediate product 3.

[0159] In this invention, the amino acid derivative is preferably 4-aminobutyric acid; the molar ratio of the intermediate product 2 to the amino acid derivative is preferably 1:(1~1.3), more preferably 1:1.1; the molar ratio of the intermediate product 2 to the volume of DMF is preferably (30~40) mmol:80 mL, more preferably (35~38) mmol:80 mL, and even more preferably 36.750 mmol:80 mL; the molar ratio of the intermediate product 2 to N,N-diisopropylethylamine is preferably 1:(2~4), more preferably 1:(2.5~3.5), and even more preferably 1:3.

[0160] In this invention, the preferred method for mixing intermediate product 2, the amino acid derivative, anhydrous N,N-dimethylformamide (DMF), and N,N-diisopropylethylamine is as follows: first, intermediate product 2 is dissolved in DMF, then the amino acid derivative is added, and finally, N,N-diisopropylethylamine is added; the mixing is preferably carried out under stirring conditions. This invention does not impose any special limitations on the specific mixing operation, as long as it ensures uniform mixing.

[0161] In this invention, the temperature of the nucleophilic acyl substitution reaction is preferably room temperature; the holding time of the nucleophilic acyl substitution reaction is preferably 6-10 hours, more preferably 7-9 hours, and even more preferably 8 hours; the nucleophilic acyl substitution reaction is preferably carried out under stirring conditions; and the nucleophilic acyl substitution reaction is preferably monitored by TLC. This invention does not impose any special limitation on the stirring rate; it can be determined based on the technical knowledge of those skilled in the art, as long as it avoids solution splashing. By controlling the conditions of the nucleophilic acyl substitution reaction, this invention can ensure the completeness of the reaction; and by using TLC monitoring, it can determine whether the nucleophilic acyl substitution reaction has ended.

[0162] After the nucleophilic acyl substitution reaction is completed, the product of the nucleophilic acyl substitution reaction is preferably extracted, washed, dried, concentrated and purified in sequence.

[0163] In this invention, the extractant used for extraction is preferably ethyl acetate; the number of extractions is preferably 2 to 5 times, more preferably 3 to 4 times; and the extracted organic phases are preferably combined. This invention extracts intermediate product 2 through extraction, and multiple extractions can improve extraction efficiency and increase the yield of the compound.

[0164] In this invention, the detergent used for washing is preferably saturated saline solution; the number of washing cycles is preferably 2 to 5 times, more preferably 3 to 4 times. This invention removes residual impurities and further improves purity through washing.

[0165] In this invention, the preferred method of drying and concentration is using anhydrous sodium sulfate. The specific amount of anhydrous sodium sulfate used is not particularly limited in this invention and can be determined based on the technical knowledge of those skilled in the art.

[0166] In this invention, the purification is preferably performed using a Flash column; the mobile phase used for Flash column purification is preferably a mixture of petroleum ether and ethyl acetate; the volume ratio of petroleum ether to ethyl acetate in the mixture is preferably 40:60; the flow rate of the mobile phase is preferably 20-40 mL / min, more preferably 30 mL / min. This invention, through purification, can further remove impurities and obtain a high-purity compound.

[0167] After obtaining intermediate product 1 and intermediate product 3, the present invention mixes intermediate product 3, intermediate product 1, anhydrous dichloromethane and dicyclohexylcarbodiimide and performs an ester condensation reaction to obtain compound IMB-C52.

[0168] In this invention, the molar ratio of intermediate product 3 to intermediate product 1 is preferably (2~3):1, more preferably 2.5:1; the molar ratio of intermediate product 3 to the volume of anhydrous dichloromethane is preferably (25~35) mmol:100mL, more preferably (28~30) mmol:100mL, and even more preferably 29.396 mmol:100mL; the molar ratio of intermediate product 3 to dicyclohexylcarbodiimide is preferably 1:(1~2), more preferably 1:1.5.

[0169] In this invention, the preferred method for mixing intermediate product 3, intermediate product 1, anhydrous dichloromethane, and dicyclohexylcarbodiimide is as follows: first, intermediate product 3 is dissolved in anhydrous dichloromethane, then the temperature is lowered to 0°C, followed by the addition of dicyclohexylcarbodiimide and stirring for 0.5-1.5 hours, and finally intermediate product 1 is added. In this invention, the mixing is preferably carried out under stirring conditions. This invention does not impose any particular limitations on the stirring rate and time, as long as the mixture is homogeneous.

[0170] In this invention, the temperature of the ester condensation reaction is preferably room temperature; the time of the ester condensation reaction is preferably 6-10 hours, more preferably 8 hours. By controlling the temperature and time of the ester condensation reaction, this invention ensures complete reaction.

[0171] After the ester condensation reaction is completed, the present invention preferably concentrates and purifies the product of the ester condensation reaction in sequence.

[0172] The present invention does not impose any special limitation on the concentration method, which can be determined based on the technical common sense of those skilled in the art.

[0173] In this invention, the purification is preferably performed using a Flash column; the mobile phase used for Flash column purification is preferably a mixture of dichloromethane and methanol; the volume ratio of dichloromethane to methanol in the mixture is preferably 98:2; the flow rate of the mobile phase is preferably 20-40 mL / min, more preferably 30 mL / min. This invention, through purification, can further remove impurities and obtain a high-purity compound.

[0174] This invention involves vacuum heating berberine hydrochloride to obtain a berberine metabolite (intermediate product 1, denoted as M1). This berberine metabolite is then linked to the hepatoprotective and anti-inflammatory drug bicyclol (Bic) via different linkers to synthesize the desired compound. Berberine has a well-defined efficacy in treating metabolic diseases caused by abnormal glucose and lipid metabolism, including fatty liver disease, diabetes, and obesity. Its main metabolite, M1, plays a crucial role in its pharmacological effects against metabolic diseases. Bicyclol is a class I hepatoprotective and anti-inflammatory drug, clinically used to reduce elevated transaminase levels from various causes, and also has some effect in improving fatty liver disease. Therefore, this invention combines the beneficial effects of M1 and bicyclol to improve metabolic syndrome, and overcomes the problem of rapid in vivo metabolism of both M1 and bicyclol by linking them into a single compound via linkers, thus allowing the compound to exert a better effect.

[0175] This invention provides the use of the compound IMB-C52 described in the above-described technical solution or the compound IMB-C52 prepared by the preparation method described in the above-described technical solution in the preparation of a medicament for the prevention or treatment of metabolic syndrome accompanied by glucose and lipid metabolism disorders.

[0176] In this invention, the active ingredient of the drug preferably includes compound IMB-C52. This invention does not impose any specific limitations on the specific composition of the drug; it can be determined based on the technical knowledge of those skilled in the art. The drug provided by this invention can be used to prevent or treat metabolic syndrome accompanied by disorders of glucose and lipid metabolism.

[0177] In this invention, the disease accompanied by metabolic syndrome with glucose and lipid metabolism disorders preferably includes any one or more of fatty liver disease, diabetes, and obesity.

[0178] In this invention, the compound IMB-C52 can reduce body weight and liver fat accumulation, reduce transaminase levels, and improve liver tissue pathological abnormalities (fatty degeneration, ballooning degeneration, and inflammatory response), thereby achieving the purpose of preventing and / or treating fatty liver disease.

[0179] In this invention, the compound IMB-C52 can reduce fasting blood glucose and / or alleviate insulin resistance, thereby achieving the purpose of preventing and / or treating diabetes.

[0180] In this invention, the compound IMB-C52 can reduce weight and body fat percentage, thereby achieving the purpose of preventing and / or treating obesity.

[0181] The technical solutions of this invention will be clearly and completely described below with reference to the embodiments thereof. Obviously, the described embodiments are only a part of the embodiments of this invention, and not all of them. All other embodiments obtained by those skilled in the art based on the embodiments of this invention without creative effort are within the scope of protection of this invention.

[0182] Example 1

[0183] Compound IMB-C52 has the following structure:

[0184]

[0185] The preparation method of the compound IMB-C52 includes the following steps:

[0186] (1) 20g of berberine hydrochloride was heated under vacuum to produce a dark purple solid, which was then purified by Flash column to obtain intermediate product 1 (intermediate product 1 is berberine, yield is 11.30g, yield is 58%, dark purple solid); the vacuum degree of the vacuum heating was 20mmHg, the temperature was 230℃, and the holding time was 40min; the mobile phase of the Flash column purification was a mixture of dichloromethane and methanol, and the volume ratio of dichloromethane to methanol in the mixture was 85:15; the flow rate of the mobile phase was 110mL / min;

[0187] (2) Under stirring conditions, chloromethyl 3-((tert-butyloxycarbonyl)amino)propionate (7.763 mmol) and anhydrous acetonitrile (40 mL) were first mixed, then sodium iodide (7.763 mmol) was added and the mixture was heated and stirred at 40 °C for 2 h. Next, intermediate product 1 (2.0 g, 6.209 mmol) obtained in step (1) was added, and finally anhydrous potassium carbonate (12.418 mmol) was added. The nucleophilic substitution reaction was carried out at 40 °C for 8 h. After the reaction was completed by TLC monitoring, extraction was performed sequentially (ethyl acetate extraction). The organic phases extracted three times were combined, washed (three times with saturated brine), dried and concentrated (dried and concentrated using anhydrous sodium sulfate), and purified. The purification was carried out using a Flash column with a mobile phase of a mixture of dichloromethane and methanol at a volume ratio of 99:1 and a flow rate of 30 mL / min, yielding intermediate 2 (intermediate 2 is berberine-9-yloxymethyl 3-((tert-butyloxycarbonyl)amino)propionate, 2.47 g, yield 76%, yellow solid).

[0188] (3) First, the intermediate product 2 (2.47 g, 4.718 mmol) obtained in step (2) was dissolved in anhydrous dichloromethane (20 mL), and then the temperature was lowered to 0 °C and trifluoroacetic acid (4 mL) was added dropwise. The de-Boc reaction was carried out for 4 h under ice-water bath (0 °C). After the de-Boc reaction was completed by TLC monitoring, the product was concentrated under reduced pressure to obtain intermediate product 3 (intermediate product 3 is berberine-9-yloxymethyl 3-aminopropionate, 1.96 g, yield 98%, yellow solid).

[0189] (4) Under stirring conditions, bicyclol (5.123 mmol) was first dissolved in anhydrous tetrahydrofuran (20 mL), then 4-nitrophenyl chloroformate (6.660 mmol) was added, and finally anhydrous pyridine (12.808 mmol) was added. The active esterification reaction was carried out at room temperature for 8 h. After the active esterification reaction was monitored by TLC, the following steps were performed: extraction (3 times with ethyl acetate and the extracted organic phases were combined), washing (3 times with saturated brine), drying and concentration (drying and concentration with anhydrous sodium sulfate), and purification by Flash column. The mobile phase for purification by Flash column was a mixture of petroleum ether and ethyl acetate with a volume ratio of 40:60 and a flow rate of 30 mL / min. Intermediate product 4 (intermediate product 4 is bicyclol-4-nitrophenyl carbonate, 2.42 g, yield 85%, white solid) was obtained.

[0190] (5) Under stirring conditions, the intermediate product 3 (1.96 g, 4.628 mmol) obtained in step (3) was first dissolved in anhydrous N,N-dimethylformamide (20 mL), then the intermediate product 4 (2.40 g, 4.628 mmol) obtained in step (4) was added, and finally N,N-diisopropylethylamine (13.886 mmol) was added. The nucleophilic acyl substitution reaction was carried out at room temperature for 8 h. After the nucleophilic acyl substitution reaction was completed by TLC monitoring, the following steps were performed in sequence: extraction (3 times with ethyl acetate and the extracted organic phases were combined), washing (3 times with saturated brine), drying and concentration (drying and concentration using anhydrous sodium sulfate) and purification. The purification was carried out by Flash column purification. The mobile phase for Flash column purification was a mixture of dichloromethane and methanol with a volume ratio of 98:2 and a flow rate of 30 mL / min. Compound IMB-C52 (1.68 g, yield 43%, yellow) was obtained.

[0191] The NMR data for compound IMB-C52 provided in Example 1 are as follows:

[0192] 1H NMR (500 MHz, DMSO) δ 9.76 (s, 1H), 8.98 (s, 1H), 8.16 (dd, J =71.4, 9.2 Hz, 2H), 7.80 (s, 1H), 7.24 (s, 1H), 7.08 (d, J = 37.3 Hz, 2H), 6.69 (s, 1H), 6.18 (s, 2H), 6.06 (s, 1H), 5.99 (s, 1H), 5.96 – 5.71 (m, 5H), 4.91 (s, 2H), 4.65 (q, J = 12.6 Hz, 2H), 4.06 (s, 3H), 3.84 (d, J = 24.5 Hz, 6H), 3.57 (s, 3H), 3.15 (q, J = 6.4 Hz, 4H), 2.81 (d, J = 80.2 Hz, 1H).

[0193] 13 C NMR (126 MHz, DMSO) δ 170.67, 165.43, 159.64, 155.75, 150.52, 149.93, 147.71, 147.50, 146.39, 144.84, 142.29, 142.16, 139.83, 139.43,137.85, 133.77, 132.97, 130.72, 129.30, 126.31, 125.01, 123.90, 121.71,120.36, 110.74, 109.65, 109.41, 108.39, 108.03, 105.44, 102.37, 102.15,101.33, 88.71, 63.21, 57.18, 56.40, 56.32, 55.32, 51.86, 35.97, 33.88, 26.26. HR-MS (ESI): calcd for C 43 H 39 N2O 16 + [M+H] + 839.2300, found 839.2599.

[0194] Example 2

[0195] The compound IMB-C52 has the following structural formula:

[0196] ;

[0197] The preparation method of the compound IMB-C52 is as follows:

[0198] (1) Same as step (1) in Example 1, to obtain intermediate product 1;

[0199] (2) Under stirring conditions, the intermediate product 1 (2.0 g, 6.209 mmol) obtained in step (1) was dissolved in 50 mL of anhydrous dichloromethane, and then methyl bromoacetate (24.836 mmol) was added. The reaction was heated to 40 °C for 8 h of nucleophilic substitution. The reaction was monitored by TLC until it was complete. The reaction system was filtered to obtain a brownish-yellow solid. The brownish-yellow solid was transferred to 70 mL of 95% ethanol solution and heated under reflux for 4 h. The reaction system was filtered to obtain a yellow solid intermediate product 2 (2.30 g, yield 94%).

[0200] (3) Dissolve the intermediate product 2 (2.3g, 5.835mmol) obtained in step (2) in 60 mL of methanol and 20 mL of water, add 11.670mmol of lithium hydroxide, stir the reaction at room temperature for 6h, monitor the reaction by TLC until it is complete, add 6N hydrochloric acid dropwise to the reaction system until the pH value of the solution is 3, and filter to obtain yellow solid intermediate product 3 (2.12g, yield 94%).

[0201] (4) The intermediate product 3 (2.1 g, 5.524 mmol) obtained in step (3) was dissolved in 30 mL of anhydrous N,N-dimethylformamide, 7.181 mmol of N-tert-butoxycarbonyl-1,2-ethylenediamine was added, followed by 11.048 mmol of 2-(7-azobenzotriazole)-N,N,N',N'-tetramethylurea hexafluorophosphate (HATU), and finally 27.620 mmol of triethylamine was added. The mixture was stirred at room temperature for 8 h to carry out the condensation reaction. The reaction was monitored by TLC to complete. The solvent N,N-dimethylformamide was removed by vacuum distillation. 20 mL of anhydrous methanol was added to the system and allowed to stand for 30 min. A solid precipitated and was filtered to obtain a yellow solid intermediate product 4 (2.31 g, yield 80%).

[0202] (5) The intermediate product 4 (2.31 g, 4.159 mmol) obtained in step (4) was dissolved in 20 mL of anhydrous dichloromethane, cooled to 0 °C, and 4 mL of trifluoroacetic acid was added dropwise. The Boc removal reaction was carried out in an ice-water bath (0 °C) for 4 h. The reaction was monitored by TLC until it was completed. The reaction system was concentrated under reduced pressure to obtain yellow solid intermediate product 5 (1.80 g, yield 97%).

[0203] (6) Same as step (4) in Example 1, to obtain intermediate product 6 (corresponding to intermediate product 4 in Example 1).

[0204] (7) The intermediate product 5 (1.8 g, 4.263 mmol) obtained in step (5) was dissolved in 20 mL of anhydrous N,N-dimethylformamide, and the intermediate product 6 (2.4 g, 4.263 mmol) obtained in step (6) was added. Finally, 12.789 mmol of N,N-diisopropylethylamine was added. The mixture was stirred at room temperature for 8 h. The reaction was monitored by TLC until it was complete. The reaction system was extracted three times with ethyl acetate. The organic phases were combined and washed three times with saturated brine. The organic phases were dried over anhydrous sodium sulfate and concentrated. The mixture was purified by Flash column chromatography. The mobile phase was a mixture of dichloromethane and methanol with a volume ratio of 98:2 and a flow rate of 30 mL / min. The yellow compound IMB-C52 (1.52 g, yield 42%) was obtained.

[0205] The NMR data for compound IMB-C52 provided in Example 2 are as follows:

[0206] 1 H NMR (600 MHz, DMSO-) d 6) δ 9.96 (s, 1H), 8.93 (s, 1H), 8.24 (t, J =5.8 Hz, 1H), 8.20 (d, J = 9.2 Hz, 1H), 8.01 (d, J = 9.1 Hz, 1H), 7.80 (s, 1H), 7.23 (s, 1H), 7.13 (t, J = 5.9 Hz, 1H), 7.08 (s, 1H), 6.71 (s, 1H), 6.18 (s, 2H), 6.07 (d, J = 1.1 Hz, 1H), 5.99 (s, 1H), 5.90 (d, J = 1.0 Hz, 1H), 5.87 (d, J = 0.9 Hz, 1H), 4.89 (t, J = 6.4 Hz, 2H), 4.75 (s, 2H), 4.71–4.63 (m, 2H), 4.04 (s, 3H), 3.88 (s, 3H), 3.83 (s, 3H), 3.56 (s, 3H), 3.18 (dt, J = 16.6, 6.5 Hz, 4H), 3.07 (q, J = 6.4 Hz, 2H).

[0207] 13 C NMR (151 MHz, DMSO- d6) δ 167.83, 165.40, 156.01, 149.87, 149.82,147.69, 147.47, 146.36, 145.71, 142.26, 142.13, 141.85, 137.79, 137.52,133.73, 132.89, 130.59, 129.31, 126.64, 123.89, 123.73, 121.17, 120.37,120.09, 110.74, 109.60, 109.38, 108.39, 108.06, 105.40, 102.30, 102.09(2),101.26, 71.61, 63.17, 57.08, 56.38, 56.32, 55.38, 51.78, 38.44, 26.33. HR-MS (ESI): calcd for C 43 H 40 N3O 15 + [M+H] + 838.2459, found 838.2745.

[0208] Example 3

[0209] The compound IMB-C52 has the following structural formula:

[0210] ;

[0211] In the preparation method: N-tert-butoxycarbonyl-1,2-ethylenediamine in step (4) of Example 2 is replaced with N-tert-butoxycarbonyl-1,3-propanediamine, and other parameters are the same as in Example 2.

[0212] The NMR data for compound IMB-C52 provided in Example 3 are as follows:

[0213] 1 H NMR (500 MHz, DMSO-) d6) δ 9.97 (s, 1H), 8.93 (s, 1H), 8.20 (d, J = 9.3 Hz, 2H), 8.00 (d, J = 9.1 Hz, 1H), 7.80 (s, 1H), 7.25 (s, 1H), 7.09 (s, 1H), 7.05 (t, J = 5.8 Hz, 1H), 6.71 (s, 1H), 6.18 (s, 2H), 6.08 (s, 1H), 6.00 (s, 1H), 5.91 (s, 1H), 5.87 (s, 1H), 4.89 (s, 2H), 4.77 (s, 2H), 4.66 (q, J = 12.6 Hz, 2H), 4.04 (s, 3H), 3.88 (s, 3H), 3.83 (s, 3H), 3.57 (s, 3H), 3.15 (d, J = 6.6 Hz, 4H), 2.96 (d, J = 6.9 Hz, 2H), 1.58–1.51 (m, 2H).

[0214] 13 C NMR (101 MHz, DMSO- d 6) δ 167.65, 165.44, 155.90, 149.89, 149.86, 147.72, 147.50, 146.39, 145.76, 142.29, 142.17, 141.92, 137.82, 137.54, 133.74, 132.91, 130.65, 129.46, 126.64, 123.94, 123.76, 121.31, 120.43, 120.15, 110.74, 109.68, 109.43, 108.44, 108.02, 105.45, 102.35, 102.12, 101.30, 71.70, 63.10, 57.12, 56.40, 56.32, 55.43, 51.​​​​​​​​​​​The compound IMB-C52 has the following structural formula:

[0217] ;

[0218] In the preparation method: N-tert-butoxycarbonyl-1,2-ethylenediamine in step (4) of Example 2 is replaced with 1-tert-butoxycarbonyl-4-aminomethylpiperidine, and other parameters are the same as in Example 2.

[0219] The NMR data for compound IMB-C52 provided in Example 4 are as follows:

[0220] 1 H NMR (500 MHz, DMSO-) d 6) δ 9.98 (s, 1H), 8.93 (s, 1H), 8.24–8.19 (m, 2H), 8.00 (d, J = 9.1 Hz, 1H), 7.80 (s, 1H), 7.28 (s, 1H), 7.10 (s, 1H), 6.73 (s, 1H), 6.18 (s, 2H), 6.07 (s, 1H), 6.03 (s, 1H), 5.92 (s, 1H), 5.89 (s, 1H), 4.92 (d, J = 6.4 Hz, 2H), 4.80 (s, 2H), 4.73 (d, J = 12.3 Hz, 1H), 4.04 (s, 4H), 3.87 (d, J = 17.5 Hz, 8H), 3.57 (s, 3H), 3.21 (d, J = 6.4 Hz, 2H), 3.02 (s, 2H), 1.56 (s, 3H), 1.21–0.75 (m, 2H).

[0221] 13 C NMR (101 MHz, DMSO-) d6) δ 167.78, 165.46, 154.01, 149.90, 149.82,147.73, 147.19, 146.59, 145.86, 142.17, 142.11, 142.02, 137.93, 137.53,133.95, 132.89, 130.66, 129.10, 126.61, 125.41, 123.75, 123.70, 121.36,120.43, 120.13, 110.86, 110.17, 109.81, 108.77, 108.46, 105.44, 102.34,102.12, 101.35, 71.68, 64.76, 59.31, 57.11, 56.45, 56.33, 55.45, 51.80,43.63, 43.03,35.52, 29.21, 26.38. HR-MS (ESI): calcd for C 47 H 46 N3O 15 + [M+H] + 892.2923, found 892.3227.

[0222] Example 5

[0223] The compound IMB-C52 has the following structural formula:

[0224] ;

[0225] In the preparation method: N-tert-butoxycarbonyl-1,2-ethylenediamine in step (4) of Example 2 is replaced with 1,1-dimethyl ethyl 13-amino-5,8,11-trioxa-2-azatridecanoic acid, and other parameters are the same as in Example 2.

[0226] The NMR data for compound IMB-C52 provided in Example 5 are as follows:

[0227] 1 H NMR (500 MHz, DMSO- d6) δ 9.98 (s, 1H), 8.93 (s, 1H), 8.23 ​​(d, J =6.1 Hz, 1H), 8.20 (d, J = 9.1 Hz, 1H), 8.00 (d, J = 9.1 Hz, 1H), 7.80 (s, 1H), 7.26 (s, 1H), 6.72 (s, 1H), 6.18 (s, 2H), 6.09–5.88 (m, 3H), 4.91 (d, J= 6.6 Hz, 2H), 4.80 (s, 2H), 4.72 –4.62 (m, 1H), 4.09–3.77 (m, 10H), 3.58 (s, 3H), 3.54–3.43 (m, 12H), 3.39––3.28 (m, 7H), 3.06–2.71 (m, 2H).

[0228] 13 C NMR (101 MHz, DMSO-) d 6) δ 167.81, 165.46, 155.86, 149.88(2), 147.73(2), 146.38, 145.83, 142.29, 142.17, 141.97, 137.84, 137.53, 133.73, 132.91,130.67, 129.43, 126.61, 123.93, 123.76, 121.32, 120.43, 120.13, 110.74,109.67, 108.46, 108.03, 105.44, 102.38, 102.12, 101.30, 71.70, 69.72, 69.68,69.54, 69.51, 69.07, 68.91, 63.15, 57.12, 56.41, 56.32, 55.44, 51.85, 49.09,40.20, 38.36, 26.38. HR-MS (ESI): calcd for C 49 H 52 N3O 18 + [M+H] + 970.3319, found970.3316.

[0229] Example 6

[0230] The compound IMB-C52 has the following structural formula:

[0231] ;

[0232] The preparation method of the compound IMB-C52 is as follows:

[0233] (1) Same as step (1) in Example 1, to obtain intermediate product 1;

[0234] (2) Same as step (4) in Example 1, to obtain intermediate product 2 (corresponding to intermediate product 4 in Example 1).

[0235] (3) Dissolve intermediate product 2 (20.4 g, 36.727 mmol) obtained in step (4) of Example 1 in 80 mL of anhydrous N,N-dimethylformamide (the molar ratio of intermediate product 2 to anhydrous N,N-dimethylformamide is 36.750 mmol: 80 mL), add 40.425 mmol of 4-aminobutyric acid (the molar ratio of intermediate product 2 to 4-aminobutyric acid is 1:1.1), and finally add 110.250 mmol. N,N-Diisopropylethylamine (the molar ratio of intermediate 2 to N,N-diisopropylethylamine was 1:3) was stirred at room temperature for 8 h. The reaction was monitored by TLC until completion. The reaction system was extracted three times with ethyl acetate, and the organic phases were combined. The organic phases were washed three times with saturated brine, dried over anhydrous sodium sulfate, and concentrated. The solution was purified by Flash column chromatography using a mixture of petroleum ether and ethyl acetate in a volume ratio of 40:60 at a flow rate of 30 mL / min to obtain a light yellow oily intermediate 3 (15.27 g, yield 80%).

[0236] (4) Dissolve the intermediate product 3 (15.27 g, 29.396 mmol) obtained in step (3) in 100 mL of anhydrous dichloromethane (the molar ratio of intermediate product 3 to anhydrous dichloromethane is 29.396 mmol: 100 mL), cool to 0 °C and add 44.094 mmol of dicyclohexylcarbodiimide (the molar ratio of intermediate product 3 to dicyclohexylcarbodiimide is 1:1.5), stir at 0 °C for 1 h and add 11.758 mmol of intermediate product 1 obtained in step (1) (the molar ratio of intermediate product 3 to intermediate product 1 is 2.5:1), stir at room temperature for 8 h and filter to collect the filtrate. Concentrate the filtrate and purify it by Flash column. The mobile phase is a mixture of dichloromethane and methanol with a volume ratio of 98:2 and a flow rate of 30 mL / min to obtain a pale yellow solid compound IMB-C52 (1.34 g, yield 5%).

[0237] The NMR data for compound IMB-C52 in Example 6 are as follows:

[0238] 1H NMR (600 MHz, DMSO) δ 9.91 (s, 1H), 9.03 (s, 1H), 8.32–8.18 (m, 2H), 7.81 (s, 1H), 7.31–7.02 (m, 4H), 6.75 (s, 1H), 6.20–5.86 (m, 9H), 4.90 (t, J = 6.3 Hz, 2H), 4.03 (s, 3H), 3.92–3.82 (m, 10H), 3.59 (d, J = 3.4 Hz, 5H).

[0239] 13 C NMR (151 MHz, DMSO) δ 170.40, 165.46, 156.12, 150.38, 150.01,147.73, 147.53, 146.42, 142.33, 142.19, 138.12, 137.84, 133.77, 133.48,132.92, 130.80, 126.72, 125.91, 123.99, 121.16, 120.60, 120.33, 110.77,109.70, 109.44, 108.41, 108.05, 105.53, 102.38, 102.14, 101.30, 63.19, 57.20,56.38, 55.29, 51.85, 51.82, 51.22, 48.58, 30.86, 30.48, 26.17, 24.77. HR-MS (ESI): calcd for C 43 H 39 N2O 15 + [M+H] + 823.2350, found 823.2656.

[0240] Application Example 1

[0241] Taking the compound IMB-C52 provided in Example 1 as an example, male C57BL / 6J mice (20-22g) aged 6-8 weeks were purchased from Beijing Yaokang Biotechnology Co., Ltd. The mice were housed in an SPF-grade animal facility under specialized care. After one week of acclimatization, the formal experiment began. The results are expressed as mean ± standard error (Mean ± SEM) or representative plots. GraphPad Prism 9.0 software was used for one-way ANOVA and multiple comparisons to determine differences. # P<0.05, ##P<0.01, HFD vs. LFD group; *P<0.05, **P<0.01 vs. HFD model group; a, P<0.05 vs. Group M1; b, P<0.05 vs. Group Bic; c, P<0.05 vs. M1+Bic group.

[0242] After a week of acclimatization, mice were fed either a low-fat diet (LFD, 10kcal% Fat, D12450J, Research Diets, USA) or a high-fat diet (HFD, 60kcal% Fat, D12492, Research Diets, USA) for 10 weeks. Then, the mice were divided into groups of 6 mice each, for a total of 8 groups. The initial group weights were as follows: Figure 1 As shown. By Figure 1 It can be seen that the mice in the LFD group fed a low-fat diet weighed approximately 30g, while the mice fed a high-fat diet weighed approximately 40g, indicating that HFD feeding for 10 weeks successfully induced obesity in mice. After grouping, the mice in the 8 groups were treated as follows:

[0243] LFD group: mice were fed a low-fat diet and were gavaged with control solvent daily for a total of 6 weeks.

[0244] HFD group: mice were fed a high-fat diet and were also given control solvent by gavage daily for a total of 6 weeks;

[0245] M1 group: each mouse was fed a high-fat diet and was also given 45.2 mg / kg of M1 by gavage daily (the dosage was calculated based on the molecular weight ratio of M1 and Bic in IMB-C52) for a total of 6 weeks.

[0246] Bic group: Each mouse was fed a high-fat diet and was also given 54.8 mg / kg of bicyclol by gavage daily (the dosage was calculated based on the molecular weight ratio of M1 and Bic in IMB-C52) for a total of 6 weeks.

[0247] M1+Bic group: each mouse was fed a high-fat diet and was also given 45.2 mg / kg of M1 and 54.8 mg / kg of bicyclol by gavage daily for a total of 6 weeks;

[0248] IMB-C52 group: each mouse was fed a high-fat diet and administered 100 mg / kg of IMB-C52 by gavage daily for a total of 6 weeks.

[0249] MGL-3196 group: Each mouse was fed a high-fat diet and simultaneously administered 3 mg / kg of the positive control drug THR-β agonist MGL-3196 by gavage daily for a total of 6 weeks.

[0250] Saroglitazar group: Each mouse was fed a high-fat diet and administered 3 mg / kg of the positive control drug PPARα / γ dual agonist Saroglitazar by gavage daily for a total of 6 weeks.

[0251] The solvent for preparing the above drugs was 5% DMSO + 30% PEG300 + 5% Tween-80 + 60% sterile water by volume. These components were added sequentially and mixed thoroughly before adding the next component. The LFD and HFD groups were administered the above solvents by gavage, while the other groups were administered the corresponding concentration of the drug solution by gavage. All groups were ultimately administered a dose of 5 μL / g body weight by gavage.

[0252] During the administration period, mouse body weight and food intake were monitored weekly, and the results were as follows: Figure 2 and Figure 3 As shown. Figure 2 The final body weight of mice 6 weeks after administration ( Figure 2 The 16th week in the middle refers to the sum of 10 weeks of feeding and 6 weeks of drug administration; therefore, 16 weeks is the date of the 6th week of drug administration. Figure 3 This represents the average weekly food intake per mouse during the 6-week drug administration period. Figure 2 and Figure 3It can be seen that the body weight of mice in the LFD group was basically the same as that of the initial group; the average body weight of mice in the HFD group increased from 40g to about 47g, a significant increase, indicating that the control solvent did not have the effect of reducing mouse body weight; the food intake of mice in the M1 group decreased slightly, and the average body weight increased from 40g to about 43g, indicating that compared with the HFD group, M1 inhibited the increase in mouse body weight to some extent by reducing the amount of food intake; the average body weight of mice in the Bic group increased from 40g to about 47g, and the food intake and body weight were basically the same as those in the HFD group, indicating that bicyclol did not have the effect of inhibiting the increase in mouse body weight; the average body weight of mice in the M1+Bic group increased from 40g to about 45g, while the food intake was similar to that of mice given M1 by gavage, indicating that compared with the HFD group, the combined gavage of M1 and bicyclol had only a slight effect. The IMB-C52 group showed a weak inhibition of weight gain in mice. The average weight of mice in the IMB-C52 group decreased from 40g to approximately 38g, indicating a significant decrease in body weight and a substantial reduction in food intake. This suggests that the IMB-C52 provided by this invention significantly reduces food intake and body weight in mice. The MGL-3196 group showed an increase in food intake, with the average body weight rising from 40g to approximately 45g, indicating that compared to the HFD group, the positive control drug THR-β agonist MGL-3196 only weakly inhibited weight gain in mice. The average body weight of mice in the Saroglitazar group decreased from 40g to approximately 34g, and food intake also decreased significantly, indicating that the PPARα / γ dual agonist Saroglitazar can significantly reduce food intake and body weight in mice. The above analysis suggests that the compounds IMB-C52 and Saroglitazar provided by this invention may inhibit food intake behavior in mice by affecting appetite or energy metabolism.

[0253] The day before the end of the experiment, the body fat and lean meat content of the mice were measured using a nuclear magnetic resonance imaging (NMR) analyzer, and the body fat percentage and lean meat percentage were calculated. The epididymal white adipose tissue was collected and weighed the following day. The results are as follows: Figures 4-6 As shown. Figure 4 The body fat percentage of mice after 6 weeks of drug administration; Figure 5 The lean meat percentage of mice after 6 weeks of drug administration; Figure 6 This represents the content of white adipose tissue in the epididymis of mice 6 weeks after drug administration. Figures 4-6It can be seen that the mice fed a low-fat diet in the LFD group had the lowest body fat percentage, the highest lean meat percentage, and the lowest epididymal white adipose tissue content. The mice in the HFD group, M1 group, Bic group, M1+Bic group, and MGL-3196 group had higher body fat percentage and epididymal white adipose tissue content, and lower lean meat percentage. The IMB-C52 group had significantly lower body fat percentage and epididymal white adipose tissue content, and higher lean meat percentage, suggesting that IMB-C52 effectively reduces weight without reducing muscle mass. Its effect is better than that of single drug, combination drug, and positive control drug MGL-3196, and is generally similar to that of Saroglitazar.

[0254] Twelve hours after the last administration, mice were starved for 12 hours. Blood samples were collected, and fasting blood glucose was measured using a glucometer. Serum was separated, and fasting insulin (80-INSMSU-E01, ALPCO) was measured using a kit. The insulin resistance index (HOMA2-IR) was calculated using the open-source software HOMA2Calculator (https: / / www.dtu.ox.ac.uk / homacalculator / ). The results are as follows: Figures 7-9 As shown. Figure 7 This represents the fasting blood glucose level in mice. Figure 8 This refers to the fasting serum insulin content in mice. Figure 9 This represents the insulin resistance index in mice. (From...) Figures 7-9 It can be seen that, compared with the LFD group, the fasting blood glucose level, serum insulin level, and insulin resistance index of mice in the HFD group were significantly increased. The fasting blood glucose level of mice in the Bic group and MGL-3196 group was only slightly decreased compared with the HFD group, while the serum insulin level and insulin resistance index decreased significantly. In contrast, the fasting blood glucose level, serum insulin level, and insulin resistance index of mice in the M1 group, M1+Bic group, IMB-C52 group, and Saroglitazar group were significantly decreased compared with the HFD group, with the IMB-C52 group and Saroglitazar group showing the largest decreases, approaching the levels of the LFD group. In summary, regarding glucose metabolism, compared with the HFD group, the IMB-C52 group significantly reduced non-fasting blood glucose in mice, and also significantly reduced serum insulin levels and the insulin resistance index HOMA2-IR, with effects comparable to single-drug, combination, and two positive control drugs.

[0255] Regarding blood biochemical indicators, blood samples were left to stand at room temperature for 3 hours, then centrifuged at 2500g for 10 minutes to separate serum. A biochemical analyzer was used to detect liver function and blood lipid levels in mice. The serum alanine aminotransferase (ALT), serum aspartate aminotransferase (AST), and blood lipid indicators such as serum triglycerides (TG) and total cholesterol (CHO) were obtained as follows: Figures 10-13 As shown. Figure 10The alanine aminotransferase (ALT) level in mouse serum; Figure 11 The aspartate aminotransferase (AST) content in mouse serum; Figure 12 This refers to the triglyceride content in mouse serum; Figure 13 This represents the total cholesterol content in mouse serum. Figures 10-11 It can be seen that the serum levels of liver damage markers ALT and AST were elevated in the HFD group mice, while the IMB-C52 group combined the advantages of the hepatoprotective drug bicyclol (Bic) in lowering ALT and compound M1 in lowering AST, significantly reducing ALT and AST levels compared to the HFD group; the two positive control drugs were less effective in alleviating these two liver function indicators, while the IMB-C52 group showed better transaminase-lowering effects than the two positive control drugs. Figures 12-13 It can be seen that IMB-C52 can significantly reduce blood lipid indicators TG and CHO, with better effects than single drugs and combination drugs, and comparable to positive control drugs MGL-3196 or Saroglitazar. These results indicate that the compound IMB-C52 provided by this invention can significantly alleviate liver damage and hyperlipidemia in MASLD mice, combining the advantages of single drugs, with a better transaminase-lowering effect than positive control drugs, and a blood lipid-lowering effect comparable to positive control drugs.

[0256] Regarding liver biochemical indicators, according to the kit instructions, protein quantification was performed on homogenized mouse liver tissue. The kit was then used to detect the levels of TG and CHO in the mouse liver tissue using a microplate reader. The results are as follows: Figures 14-15 As shown. Figure 14 This refers to the triglyceride content in mouse liver; Figure 15 This represents the total cholesterol content in mouse liver. (From...) Figure 14 It can be seen that the liver TG content of mice in the HFD group was significantly increased, and this index was reduced by single drug and combination therapy. IMB-C52 showed better results, comparable to the positive control drug Saroglitazar, and better than MGL-3196. Figure 15 It can be seen that IMB-C52 is more effective than single drugs in reducing liver CHO content, and comparable to the two positive control drugs. These results further demonstrate that the compound IMB-C52 provided by this invention can significantly alleviate intrahepatic lipid accumulation in MASLD mice.

[0257] Mouse liver tissue was collected and fixed with 4% paraformaldehyde. Hematoxylin-eosin (H&E) and Oil Red staining were used to detect pathological changes and for photographic analysis. The severity of hepatic steatosis, ballooning degeneration, and inflammation was assessed using the NAFLD activity score (NAS) according to the NASH Clinical Research Network (NASH-CRN) criteria. The remaining liver tissue was flash-frozen in liquid nitrogen and stored at -80°C. Subsequent assay kits were used to detect the levels of TG (A110-1-1, Nanjing Jiancheng) and CHO (A111-1-1, Nanjing Jiancheng) in the liver. The results are as follows: Figures 16-17As shown. Figure 16 The results of gross examination, H&E staining, and Oil Red staining of mouse liver tissue are shown. Figure 17 The results show the NAS score for mouse liver tissue pathology. Figures 16-17 As can be seen, compared with the LFD group, the livers of mice in the HFD group were significantly enlarged and yellowish, exhibiting typical characteristics of hepatic steatosis; while the liver morphology and color of mice in the IMB-C52 treatment group were significantly improved. It is worth noting that although the liver color in the positive control group (Saroglitazar) almost returned to normal, all mice showed significant liver tissue enlargement, suggesting potential hepatotoxicity. Simultaneously, the liver tissue of mice in the HFD group showed obvious steatosis, hepatocyte ballooning degeneration, and intralobular inflammation, with numerous red-stained lipid droplets appearing in Oil Red staining. IMB-C52 significantly improved these pathological symptoms, with effects slightly better than MGL-3196 and comparable to Saroglitazar, and superior to single-drug and combination therapies.

[0258] In summary, the compound IMB-C52 provided by this invention significantly reduced body weight and improved pathological abnormalities such as glucose and lipid metabolism disorders, insulin resistance, and hepatic steatosis in the classic MASLD mouse model, with overall efficacy superior to single-agent and combination therapies. More importantly, based on comprehensive evaluation of body weight, liver function, blood lipids, blood glucose, insulin resistance, and liver pathological indicators, the overall efficacy of IMB-C52 is not inferior to the two clinically used drugs, Saroglitazar and MGL-3196, especially in lowering transaminase levels, which far exceeds the two positive control drugs, and in reducing body weight, which far surpasses MGL-3196. Therefore, IMB-C52 shows excellent potential for the prevention or treatment of fatty liver disease, diabetes, obesity, and other diseases accompanied by metabolic syndrome.

[0259] Application Example 2

[0260] Acute toxicity test of IMB-C52 in mice

[0261] SPF-grade Kunming mice (18-20g) were purchased from Spiford (Beijing) Biotechnology Co., Ltd. After purchase, the mice were housed in an SPF-grade animal facility under specialized care. The mice were allowed to acclimatize for one day before the formal experiments began. Results are expressed as mean ± standard error (Mean ± SEM) or representative plots. Differences were determined using GraphPad Prism 9.0 software via one-way ANOVA or t-tests. *P < 0.05, **P < 0.01 vs. Control group.

[0262] Mice were randomly divided into groups of six (half male and half female) based on body weight. After fasting for 6 hours, each group received a single intragastric (ig) administration of the drug, with a dosage of 0.2 mL per mouse. Mice were observed and their weight recorded before administration and on days 1, 3, 5, and 7 post-administration. After 7 days, mice were weighed, and blood samples were collected for biochemical analysis to determine alanine aminotransferase (ALT), aspartate aminotransferase (AST), alkaline phosphatase (ALP), urea, creatinine (Cre), and lactate dehydrogenase (LDH). Heart, liver, spleen, lung, and kidney samples were collected for Hematologic and Escherichia coli (H&E) staining to evaluate potential toxicity. Experimental groupings and treatments are as follows:

[0263] Control (ig) group: mice were given a single oral gavage of the control solvent;

[0264] C52-4 (500 mg / kg, ig): Mice were given a single oral gavage of 500 mg / kg of IMB-C52;

[0265] C52-4 (1000 mg / kg, ig) group: Mice were given a single oral gavage of 1000 mg / kg of IMB-C52;

[0266] Control (ip) group: Mice were injected intraperitoneally with the control solvent once;

[0267] C52-4 (100 mg / kg, ip): Mice were given a single intraperitoneal injection of 100 mg / kg of IMB-C52.

[0268] The solvent for preparing the above drugs was 10% DMSO + 30% PEG300 + 5% Tween-80 + 55% sterile water by volume. The same solvent was used for the control group via gavage and intraperitoneal injection.

[0269] Results of acute toxicity experiments in mice are as follows Figures 18-20 As shown. Figure 18 The body weight curve of mice in the acute toxicity test of IMB-C52; Figure 19 Serum biochemical indicators of mice in the acute toxicity test of IMB-C52; Figure 20 These are H&E staining pathological images of the heart, liver, spleen, lungs, and kidneys of mice in an IMB-C52 acute toxicity experiment. (Source: [Original Source Name]) Figures 18-20 It can be seen that, compared with the control group, neither gavage nor intraperitoneal injection affected the body weight of the mice. Figure 18 ) and various biochemical indicators ( Figure 19 Similarly, pathological results showed that IMB-C52 did not have a significant impact on the pathology of the heart, liver, spleen, lungs, and kidneys. Figure 20 The fact that in Example 1, mice were administered 100 mg / kg of IMB-C52 by gavage daily for 6 consecutive weeks without any abnormalities in survival or behavior demonstrates that the compound IMB-C52 provided by this invention has excellent safety.

[0270] In summary, this invention demonstrates that compound IMB-C52 can significantly reduce body weight in HFD-induced MASLD mice, improve liver histopathological abnormalities (steatodegeneration, ballooning degeneration, and intralobular inflammation), glucose and lipid metabolism disorders, and insulin resistance, showing its potential in the prevention or treatment of fatty liver disease, diabetes, and obesity.

[0271] The above description is only a preferred embodiment of the present invention. It should be noted that for those skilled in the art, several improvements and modifications can be made without departing from the principle of the present invention, and these improvements and modifications should also be considered within the scope of protection of the present invention.

Claims

1. A class of compounds, IMB-C52, characterized in that, It has the following structure: Where X is , , , , and Any one of them.

2. The method for preparing the compound IMB-C52 according to claim 1, characterized in that, Includes the following steps: (1) Berberine hydrochloride was heated under vacuum to obtain intermediate product 1; (2) The intermediate product 1 obtained in step (1), the protected amino acid derivative, the polar solvent, the iodide and the strong base weak acid salt are mixed and subjected to a nucleophilic substitution reaction to obtain intermediate product 2; (3) The intermediate product 2 obtained in step (2), the first organic solvent and trifluoroacetic acid are mixed and subjected to a deBoc reaction to obtain intermediate product 3; (4) The bicyclic alcohol, the second organic solvent, 4-nitrophenyl chloroformate and anhydrous pyridine were mixed and subjected to an active esterification reaction to obtain intermediate product 4; (5) The intermediate product 3 obtained in step (3), the third organic solvent, the intermediate product 4 obtained in step (4) and N,N-diisopropylethylamine are mixed and subjected to a nucleophilic acyl substitution reaction to obtain compound IMB-C52; In the compound IMB-C52, X is... ; In step (2), the protected amino acid derivative is chloromethyl 3-((tert-butoxycarbonyl)amino)propionate, the polar solvent is anhydrous acetonitrile, the iodide is sodium iodide, and the strong base-weak acid salt is anhydrous potassium carbonate.

3. The preparation method according to claim 2, characterized in that, In step (1), the vacuum degree of vacuum heating is 20~30 mmHg, the temperature of vacuum heating is 200~230℃, and the holding time of vacuum heating is 30~60 min.

4. The preparation method according to claim 2, characterized in that, In step (2), the molar ratio of the protected amino acid derivative and intermediate product 1 is (1~1.5):

1.

5. The preparation method according to claim 2, characterized in that, In step (3), the ratio of the amount of intermediate product 2 to the volume of trifluoroacetic acid is (4~6) mmol: 4 mL.

6. The method for preparing the compound IMB-C52 according to claim 1, characterized in that, Includes the following steps: (1) Berberine hydrochloride was heated under vacuum to obtain intermediate product 1; (2) The intermediate product 1 obtained in step (1), methyl bromoacetate and anhydrous dichloromethane are mixed and subjected to a nucleophilic substitution reaction to obtain intermediate product 2; (3) The intermediate product 2 obtained in step (2), the mixed solvent and lithium hydroxide are mixed and subjected to ester hydrolysis reaction to obtain intermediate product 3; (4) The intermediate product 3 obtained in step (3), DMF, 2-(7-azabenzotriazole)-N,N,N',N'-tetramethylurea hexafluorophosphate, triethylamine and mono-Boc protected diamine compound are mixed and subjected to amide condensation reaction to obtain intermediate product 4. (5) The intermediate product 4 obtained in step (4), anhydrous dichloromethane and trifluoroacetic acid are mixed and subjected to deBoc reaction to obtain intermediate product 5; (6) The bicyclic alcohol, the second organic solvent, 4-nitrophenyl chloroformate and anhydrous pyridine were mixed and subjected to an active esterification reaction to obtain intermediate product 6; (7) The intermediate product 5 obtained in step (5), the third organic solvent, the intermediate product 6 obtained in step (6) and N,N-diisopropylethylamine are mixed and subjected to a nucleophilic acyl substitution reaction to obtain compound IMB-C52. In the compound IMB-C52, X is... , , and Any one of them.

7. The method for preparing the compound IMB-C52 according to claim 1, characterized in that, Includes the following steps: (1) Berberine hydrochloride was heated under vacuum to obtain intermediate product 1; (2) The bicyclic alcohol, the second organic solvent, 4-nitrophenyl chloroformate and anhydrous pyridine were mixed and subjected to an active esterification reaction to obtain intermediate product 2; (3) The intermediate product 2 obtained in step (2), the amino acid derivative, DMF and N,N-diisopropylethylamine are mixed and subjected to a nucleophilic acyl substitution reaction to obtain intermediate product 3; (4) The intermediate product 3 obtained in step (3), the intermediate product 1 obtained in step (1), anhydrous dichloromethane and dicyclohexylcarbodiimide are mixed and subjected to ester condensation reaction to obtain compound IMB-C52. In the compound IMB-C52, X is... .

8. The use of the compound IMB-C52 of claim 1 or the compound IMB-C52 prepared by any one of claims 2 to 7 in the preparation of a medicament for the prevention or treatment of metabolic syndrome with disorders of glucose and lipid metabolism.

9. The application according to claim 8, characterized in that, The metabolic syndrome accompanied by glucose and lipid metabolism disorders includes any one or more of fatty liver disease, diabetes, and obesity.

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