Application of mebendazole in preparation of medicine for treating metabolism-related fatty liver disease

Abstract

The invention relates to the technical field of biological medicines, and particularly discloses application of mebendazole in preparation of a medicine for treating metabolism-related fatty liver diseases. Mebendazole has a remarkable blood fat reducing effect and can remarkably reduce the level of total cholesterol and / or triglyceride and / or low-density lipoprotein in serum; the weight can be reduced, the fatty liver can be improved, and wide application prospects are achieved for obese people. The medicine is simple and convenient to take and easy to absorb by oral administration. In addition, mebendazole is easy to obtain, and compared with other imported drugs, the mebendazole is low in price, high in cost performance, easy to accept by patients and convenient to transport and store.

Description

Technical Field

[0001] The present application relates to the field of biomedicine technology, and in particular to the use of mebendazole in the preparation of a drug for treating metabolism-related fatty liver disease. Background Art

[0002] Metabolic-associated fatty liver disease (MASLD), previously known as nonalcoholic fatty liver disease (NAFLD), is defined as fatty liver disease (SLD) in the presence of at least one cardiometabolic risk factor and in the absence of harmful alcohol consumption. The renaming of NAFLD to MASLD more accurately describes the characteristics of the MASLD disease, emphasizing the relationship between fatty liver and metabolic dysfunction rather than nonalcoholic factors as the etiology. MASLD has become the most common chronic liver disease, and its prevalence is likely to continue to rise. The presence of MASLD is strongly associated with type 2 diabetes, obesity, and other cardiometabolic risk factors. MASLD is associated with an increased risk of cardiovascular events, chronic kidney disease, liver and extrahepatic malignancies, and liver-related outcomes including liver failure and hepatocellular carcinoma (HCC). Therefore, the high socioeconomic burden of MASLD constitutes a global health challenge that needs to be addressed by the medical community and policy makers. Although the hallmark of MASLD, hepatic steatosis, is reversible, it can also develop into the more severe metabolic-associated steatohepatitis (MASH), which is characterized by liver necrosis, inflammation, and fibrosis, and can gradually evolve into cirrhosis and even hepatocellular carcinoma (HCC). MASLD is also becoming a major cause of liver failure.

[0003] In terms of drug treatment, it is currently divided into two categories: metabolic disorders targeting the pathogenesis of the disease and physiological manifestations targeting the occurrence of the disease. Common ones include insulin sensitizers (metformin and thiazolidinediones), lipid-lowering drugs (fibrates or statins) and drugs for liver disease (vitamin E, ursodeoxycholic acid, etc.). However, long-term use of these drugs will cause corresponding side effects to the liver. Although the US FDA has approved the drug Rezdiffra for the treatment of non-alcoholic fatty liver disease, its actual clinical application effect and potential side effects still need to be closely monitored. The current problems in drug research and development include: drug safety and tolerability issues, unknown long-term effects, limited efficacy, lack of individualized treatment and high treatment costs. Therefore, it is of great significance to research and develop safe and effective drugs for the treatment of metabolic-related fatty liver disease. Summary of the invention

[0004] In view of the above-mentioned deficiencies in the prior art, the purpose of the present application is to provide an application of mebendazole in the preparation of a drug for treating metabolic-related fatty liver disease, aiming to solve the problems of strong side effects, unknown safety and high cost of existing related drugs.

[0005] The technical solution of this application is as follows:

[0006] In a first aspect of the present application, a method for preparing a drug for treating metabolic-related fatty liver disease is provided.

[0007] Optionally, the mebendazole can reduce the level of one or more of total cholesterol, triglyceride, and low-density lipoprotein in the serum and liver of animals induced by a high-fat diet.

[0008] In a second aspect of the present application, a drug for treating metabolism-related fatty liver disease is provided, comprising mebendazole.

[0009] Optionally, the medicament comprises a pharmaceutically acceptable excipient.

[0010] Optionally, the auxiliary material includes at least one of a carrier, an osmotic pressure regulator, a pH regulator, a diluent, a disintegrant, an excipient, a solubilizer, a stabilizer, and a preservative.

[0011] Optionally, the dosage form of the drug includes at least one of tablets, capsules, solutions, granules, pills, powders, pills, suspensions, powders, injections, sustained-release agents, controlled-release agents, and targeted preparations.

[0012] Optionally, the drug can reduce the levels of total cholesterol, and / or triglycerides, and / or low-density lipoprotein in the serum and liver of animals induced by a high-fat diet.

[0013] Beneficial effects of this application:

[0014] This application has opened up a new role for the broad-spectrum anthelmintic mebendazole, which can be used as a safe and effective drug for the treatment of metabolic-related fatty liver disease. Mebendazole has a significant lipid-lowering effect, and can significantly reduce the levels of total cholesterol, and / or triglycerides, and / or low-density lipoprotein in serum and liver; it can reduce body weight, improve fatty liver, and has broad application prospects for obese people. Through mouse experiments, it was found that mebendazole has a large tolerance dose and no obvious toxic and side effects. The medication is simple and convenient, and it is easily absorbed orally. In addition, mebendazole is easy to obtain, and compared with other imported drugs, it is cheap, cost-effective, and easy for patients to accept; it is easy to transport and store, sealed, and placed in a cool and dry place away from light. BRIEF DESCRIPTION OF THE DRAWINGS

[0015] In order to more clearly illustrate the technical solutions in the embodiments of the present application, the drawings required for describing the embodiments are briefly introduced below.

[0016] Figure 1 This is an analysis diagram of the effect of feeding mice with a high-fat diet with mebendazole on the liver provided in the examples of the present application;

[0017] Figure 2 Analysis of the effects of mebendazole on the body weight, liver weight and liver weight to body weight ratio of mice fed a high-fat diet provided in the examples of this application:

[0018] A is body weight analysis; B is liver weight analysis; C is liver weight / body weight ratio analysis;

[0019] Figure 3 Schematic diagram of Oil Red O staining of liver of C57BL / 6 mice fed with a high-fat diet using mebendazole provided in the examples of the present application:

[0020] A is a schematic diagram of Oil Red O staining; B is a quantitative analysis of the Oil Red O staining area;

[0021] Figure 4 The effect of mebendazole on serum TC, TG and LDL-C levels in C57BL / 6 mice fed a high-fat diet provided in the examples of the present application is analyzed as follows:

[0022] A is the total cholesterol level; B is the triglyceride level; C is the low-density lipoprotein level;

[0023] Figure 5 The analysis diagram of the effect of mebendazole on the TC and TG levels in the liver tissue of C57BL / 6 mice fed with high fat diet provided in the examples of this application:

[0024] A is the total cholesterol level; B is the triglyceride level;

[0025] Figure 6 Schematic diagram of H&E staining of the liver of mice fed a mebendazole high-fat diet provided in the examples of the present application. DETAILED DESCRIPTION

[0026] In order to make the purpose, technical solutions and advantages of the present application clearer, the technical solutions in the embodiments of the present application will be clearly and completely described below in combination with the drawings and embodiments of the present application. Obviously, the described embodiments are only part of the embodiments of the present application, not all of the embodiments. Based on the embodiments of the present application, all other embodiments obtained by ordinary technicians in this field without making creative work belong to the protection scope of the present application. In the absence of conflict, the following embodiments and features in the embodiments can be combined with each other.

[0027] It should be noted that if there are descriptions involving "first", "second", etc. in the implementation of this application, the descriptions of "first", "second", etc. are only used for descriptive purposes and cannot be understood as indicating or suggesting their relative importance and implicitly indicating the number of technical features indicated. Therefore, the features defined as "first" and "second" may explicitly or implicitly include at least one of the features. In addition, the technical solutions between the various embodiments can be combined with each other, but they must be based on the fact that ordinary technicians in the field can implement them. When the combination of technical solutions is contradictory or cannot be implemented, it should be deemed that such combination of technical solutions does not exist and is not within the scope of protection required by this application.

[0028] The present application provides an application of mebendazole in the preparation of a drug for treating metabolic-related fatty liver disease. Mebendazole, chemical name 5-benzoylbenzimidazole carbamate methyl ester, molecular formula: 16 H 13 N3O3, molecular weight is 295.293. Its chemical structure is as follows:

[0029]

[0030] Metabolic-associated fatty liver disease (MASLD) is defined as excess triglyceride storage in the liver in the presence of at least one cardiometabolic risk factor. MASLD includes isolated fatty liver (metabolic dysfunction-associated fatty liver, MASL), metabolic-associated steatohepatitis (MASH), fibrosis, and cirrhosis.

[0031] In some embodiments, mebendazole can reduce the levels of total cholesterol, and / or triglycerides, and / or low-density lipoprotein in the serum and liver of animals induced by a high-fat diet.

[0032] The present application also provides a drug for treating metabolic-related fatty liver disease, which can reduce the levels of total cholesterol, and / or triglycerides, and / or low-density lipoprotein in the serum and liver of animals induced by a high-fat diet, and includes mebendazole.

[0033] In some embodiments, the drug for treating metabolic-related fatty liver disease includes ingredients with synergistic compatibility. Synergistic compatibility means that when two or more ingredients are used in combination, the effect produced by their cooperation is greater than the sum of the effects of each ingredient when used alone. Synergistic effects can be reflected in many aspects, such as enhancing efficacy, reducing toxicity, improving stability or optimizing product performance.

[0034] In some embodiments, the drug for treating metabolic-related fatty liver disease includes pharmaceutically acceptable excipients. Pharmaceutically acceptable excipients refer to all ingredients added in addition to the main drug (active ingredient that exerts a therapeutic effect) during the production process of the drug preparation. These ingredients themselves generally do not have major therapeutic activity, but they play a vital role in the preparation, stability, suitability of administration routes, patient acceptance, etc. of the drug preparation.

[0035] In some embodiments, the pharmaceutically acceptable excipients include at least one of a carrier, an osmotic pressure regulator, a pH regulator, a diluent, a disintegrant, an excipient, a solubilizer, a stabilizer, and a preservative.

[0036] In some embodiments, the dosage form of the drug for treating metabolism-related fatty liver disease includes at least one of tablets, capsules, solutions, granules, pills, powders, pills, suspensions, powders, injections, sustained-release agents, controlled-release agents, and targeted preparations.

[0037] The following is further described by specific examples. The experimental methods in the examples are conventional methods unless otherwise specified. If no specific conditions are specified in the examples, they are carried out according to conventional conditions or conditions recommended by the manufacturer. If the manufacturer of the reagents or instruments is not specified, they are all conventional products that can be obtained commercially.

[0038] Example 1

[0039] 1. Experimental Materials

[0040] Experimental animals: 60 animals of uniform weight.

[0041] Test substance: Broad-spectrum anthelmintic mebendazole.

[0042] High-fat feed: Add 20.0% sucrose, 15% lard, 1.2% cholesterol, 0.2% sodium cholate, appropriate amount of casein, calcium hydrogen phosphate, stone powder, etc. to the maintenance feed. In addition to crude fat, the moisture, crude protein, crude fat, crude fiber, crude ash, calcium and phosphorus of the model feed must meet the national standards for maintenance feed. The feed is clean grade, vacuum packed and stored at 4℃.

[0043] 2. Experimental Principle

[0044] Feeding animals with a high-fat diet containing cholesterol, sucrose, lard, and sodium cholate can form an animal model of lipid metabolism disorder. Then, giving the animals drugs can detect the effects of the test substances on high fat and determine the effects of the test substances on the animals' lipid accumulation, fatty degeneration, and inflammatory response.

[0045] Determination of MASLD model: After the modeling period, the serum triglycerides, total cholesterol or low-density lipoprotein cholesterol in the high-fat diet model group increased compared with the blank control group, and the difference was significant, indicating that the model was established.

[0046] 3. Experimental methods

[0047] 3.1 Animal grouping

[0048] First random grouping: After the animals are received, they are fed adaptively for 5-7 days. During the acclimation period, the appearance and general condition of the animals are observed. Only animals that pass the inspection can enter this experiment. After the adaptation period, the animals are weighed and randomly divided into a blank control group and a high-fat model group.

[0049] Second random grouping: After the animal hyperlipidemia model was established, the animal blood lipids were tested, and the hyperlipidemia model group was randomly divided into the positive drug group and the other groups, namely, the hyperlipidemia model group (HFD), the low-dose group of the test substance (HFD+MBZ-L), the high-dose group of the test substance (HFD+MBZ-H), and the positive control drug group (atorvastatin calcium group, HFD+ATV).

[0050] 3.2 MASLD model establishment period

[0051] As shown in Table 1, in each experimental group, the blank control group was fed with maintenance feed, and the remaining groups were fed with high-fat feed, and the levels of triglyceride (TC), total cholesterol (TG) and low-density lipoprotein (LDL-C) in the animal serum were measured.

[0052] The high-fat model group was given a high-fat diet, and the animals in the blank control group and high-fat model group were fasted and blood was collected from the canthus. After blood collection, serum was separated and serum TC, TG and LDL-C levels were measured. The high-fat model group was randomly divided into groups according to TC, TG and LDL-C levels, and TC, TG and LDL-C were compared with the blank control group.

[0053] Table 1 Drug administration and diet of animals in each group

[0054]

[0055] 3.3 Dosing Period

[0056] The animals in the successfully grouped test substance (high and low dose) groups and the positive control drug group (atorvastatin calcium group) were given drugs by oral gavage every day, and the blank control group and the high-fat model group were given drugs of corresponding doses. The dosage of each group is shown in Table 1.

[0057] The feeding conditions remained unchanged. After 8 weeks of administration, blood was collected from the canthus of the eyes to determine the levels of TC, TG and LDL-C in the animal serum and the levels of TC and TG in the liver tissue homogenate. The effects of the test substances on TC, TG, LDL-C in the animal serum and TC and TG in the liver were observed. The lipid droplet size and inflammatory infiltration of the liver tissue were observed by Oil Red O and H&E staining.

[0058] 3.4 Observation period

[0059] General vital signs were observed during the experiment.

[0060] 3.5 Main testing indicators

[0061] (1) Serum TC, TG and LDL-C levels.

[0062] (2) Oil red O staining and H&E staining.

[0063] (3) After sampling, the levels of TC and TG in liver tissue homogenate were measured.

[0064] 4. Experimental data and results

[0065] 4.1 Data processing

[0066] Variance analysis was used, but the variance homogeneity test must be performed first according to the procedure of variance analysis. If the variance is homogeneous, the F value is calculated. If the F value is <0.05, the conclusion is that there is no significant difference in the means of each group; F value ≥ 0.05, P ≤ 0.05, and the statistics are performed using the pairwise comparison method of the means between multiple experimental groups and a control group; for non-normal data or data with uneven variance, appropriate variable transformation is performed, and after meeting the requirements of normality or homogeneity of variance, the transformed data is used for statistics; if the purpose of normality or homogeneity of variance is still not achieved after variable transformation, the rank sum test is used for statistics instead.

[0067] 4.2 Determination of Animal Experimental Results

[0068] Results of lipid-lowering function: Compared with the blank control group, the high-fat model control group showed significant increases in serum triglycerides, total cholesterol and low-density lipoprotein, and the model was established.

[0069] (1) Compared with the model control group, the serum total cholesterol, triglyceride, alanine aminotransferase, aspartate aminotransferase and low-density lipoprotein in each dose group were significantly reduced, which can be judged as positive for the animal experiment results of the test sample's lipid-lowering function.

[0070] (2) Compared with the model control group, the cholesterol and triglyceride levels in the liver tissue homogenate of each dose group were significantly lower, and the cholesterol and triglyceride levels in each dose group were not significantly higher than those in the model control group. It can be determined that the test sample has positive results in the cholesterol-lowering and triglyceride-lowering animal experiment.

[0071] (3) When comparing each dose group with the model control group, the difference in the Oil Red O staining area of ​​the liver in any dose group was significant. Meanwhile, the Oil Red O staining area of ​​the liver in each dose group was not significantly higher than that in the model control group. This indicates that the animal experiment results of the test sample in lowering triglyceride were positive.

[0072] 4.3 Experimental Results

[0073] The data are expressed as mean ± standard deviation, ND: normal diet group, n = 12; HFD: high-fat model group, n = 12; MBZ (25, 50, 100 mg / kg): low (L) and high (H) dose groups of the test substance, n = 12; ATV: atorvastatin calcium (20 mg / kg); Oil red O staining and H&E staining, n = 4.

[0074] like Figure 1 As shown in the figure, after sampling, the liver of the model group had severe granularity, while the liver of the mebendazole group had significantly reduced granularity. Figure 2 As shown in A, B, and C, compared with the control group, the body weight, liver weight, and liver weight to body weight ratio of the model group increased significantly, while the body weight, liver weight, and liver weight to body weight ratio of the mebendazole group decreased significantly. Figure 3 As shown in A and B, low and high doses of mebendazole and atorvastatin calcium can reduce the Oil Red O staining area in mice induced by a high-fat diet. Figure 4 As shown in A, B, and C, after 8 weeks of administration, the serum total cholesterol, triglyceride, and low-density lipoprotein levels of mice induced by a high-fat diet were significantly increased compared with those of the blank control group, indicating that the model was successfully established. Low and high doses of mebendazole and atorvastatin calcium can significantly reduce serum TC, TG, and LDL-C in mice induced by a high-fat diet. Figure 5 As shown in Figures A and B, low and high doses of mebendazole can reduce TC and TG in liver tissue homogenates of mice induced by a high-fat diet. Figure 6 As shown, compared with the control group, the area of ​​fat vacuoles in liver tissue of the model group was significantly increased, while the area of ​​fat vacuoles in the liver of the mebendazole-treated group was significantly reduced.

[0075] In summary, through the lipid-lowering pharmacodynamic evaluation of mebendazole, it was found that mebendazole has the potential to be used to prepare drugs for the treatment of metabolic-related fatty liver disease, and atorvastatin calcium was used as the main control drug for the lipid-lowering experiment to observe the pharmacological effects of mebendazole on high-fat diet mice, proving that mebendazole can reduce the body weight, liver weight and body weight to liver weight ratio of high-fat diet mice, and can also reduce the levels of total cholesterol, triglycerides and low-density lipoprotein in the serum and liver of high-fat diet mice. This application has opened up a new role for the broad-spectrum anthelmintic mebendazole, which can be used as a safe and effective drug for the treatment of metabolic-related fatty liver disease.

[0076] It should be understood that the application of the present application is not limited to the above examples. For ordinary technicians in this field, improvements or changes can be made based on the above description. All these improvements and changes should fall within the scope of protection of the claims attached to this application.

Claims

1. The application of mebendazole in the preparation of drugs for treating metabolic-related fatty liver disease.

2. The use according to claim 1, characterized in that: The mebendazole can reduce the level of one or more of total cholesterol, triglyceride and low-density lipoprotein in the serum and liver of animals induced by a high-fat diet.

3. A drug for treating metabolic-related fatty liver disease, characterized in that: Includes mebendazole.

4. The drug according to claim 3, characterized in that The drug includes pharmaceutically acceptable excipients.

5. The drug according to claim 4, characterized in that The auxiliary materials include at least one of a carrier, an osmotic pressure regulator, a pH regulator, a diluent, a disintegrant, an excipient, a solubilizer, a stabilizer, and a preservative.

6. The drug according to claim 4, characterized in that The dosage form of the drug includes at least one of tablets, capsules, solutions, granules, pills, powders, pills, suspensions, powders, injections, sustained-release agents, controlled-release agents, and targeted preparations.

7. The drug according to claim 4, characterized in that The drug can reduce the level of one or more of total cholesterol, triglyceride and low-density lipoprotein in the serum and liver of animals induced by a high-fat diet.

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