Application of sanguinarine in preparation of medicine for preventing and treating fatty liver or obesity

Abstract

The invention discloses application of sanguinarine in preparation of a medicine for preventing and treating fatty liver or obesity, and belongs to the technical field of medicines. The invention is implemented in vitro and in vivo, and aims to solve the problem of lack of low-toxicity natural small molecule drugs for effectively preventing and treating fatty liver or obesity at present. The embodiment of the invention proves that sanguinarine can obviously reduce high-fat diet induced mouse fat deposition, reduce in-vivo and in-vitro liver TG content and lipid droplet accumulation, enhance glucose tolerance of the high-fat diet induced mouse and improve insulin resistance of the high-fat diet induced mouse. As a natural antioxidant, sanguinarine has an obvious relieving effect on energy metabolism disorder and oxidative stress induced by high fat. As a low-toxicity natural small molecule, sanguinarine can improve glucose and lipid metabolism disorder induced by high fat, is low in price, and has an application prospect in preparation of medicines for preventing and treating fatty liver or obesity.

Description

Technical Field

[0001] This invention belongs to the field of pharmaceutical technology, and in particular relates to the application of sanguinarine in the preparation of drugs for the prevention and treatment of fatty liver or obesity. Background Technology

[0002] Obesity is a major health problem in our time. The fast pace of modern society and unhealthy eating habits and lifestyles lead to metabolic disorders, resulting in excessive fat accumulation and weight gain, causing overweight or obesity. Obese patients often experience visceral fat accumulation, which can lead to cardiovascular disease and metabolic disorders such as diabetes and fatty liver. Epidemiological surveys show that obesity is even closely related to the development of cancers such as thyroid cancer, kidney cancer, and liver cancer, seriously affecting human physical and mental health.

[0003] Metabolic dysfunction-associated steatotic liver disease (MASLD) is a new name for non-alcoholic fatty liver disease (NAFLD) adopted in 2020. It refers to a clinical syndrome characterized by lipid deposition in hepatocytes caused by factors other than alcohol, resulting in metabolic dysregulation due to fat accumulation in the liver. More than 5% of hepatocytes exhibiting steatosis can be defined as MASLD. Oxidative stress, mitochondrial dysfunction, insulin resistance, endoplasmic reticulum stress, and chronic inflammation play important roles in MASLD, and these factors always interact, ultimately leading to the development of fatty liver.

[0004] In recent years, the global prevalence of MASLD is estimated at 25%, and its incidence is increasing, making it the second most common liver disease after viral hepatitis. Metabolic fatty liver disease ranges from simple hepatic steatosis to metabolic dysfunction-associated steatohepatitis (MASH) and cirrhosis, and is often accompanied by obesity, diabetes, and cardiovascular disease. Severe fatty liver can even develop into liver cancer, seriously endangering human health. Although steady progress has been made in understanding the epidemiology, pathogenesis, and identifying therapeutic targets, progress in treatment has been slow, and there are currently no officially approved methods or drugs for the prevention or treatment of fatty liver or obesity. Therefore, the search for drugs that can prevent and alleviate fatty liver or obesity is urgent. In recent years, an increasing number of studies have shown that natural small molecule compounds have safe and effective alleviating effects on MASLD; therefore, the search for drugs from natural small molecules for the prevention or treatment of fatty liver or obesity holds great promise.

[0005] Sanguinarine (SAN) is a benigneous-phenanthrene-isoquinoline alkaloid extracted from plants such as *Gnaphalium affine*. The purified solid form is a reddish-yellow powder with a relative molecular mass of 332. It possesses antibacterial, anti-inflammatory, antioxidant, and performance-enhancing properties, as well as insecticidal and maggot-killing effects, immune-boosting, antitussive, and analgesic properties. *Gnaphalium affine* extract (Sangrovit), a plant-derived feed additive containing sanguinarine, affects the growth performance of weaned piglets, specifically by increasing body weight and average daily weight gain, and decreasing feed conversion ratio.

[0006] To date, there are no reports in the existing technology regarding the preparation of sanguinarine for the treatment of fatty liver or obesity. Summary of the Invention

[0007] The purpose of this invention is to provide an application of sanguinarine in the preparation of drugs for the prevention and treatment of fatty liver or obesity. This invention aims to address the current lack of effective, low-toxicity, natural small-molecule drugs for the prevention and treatment of fatty liver or obesity. Sanguinarine, as a natural antioxidant, has a significant alleviating effect on lipid accumulation induced by high cholesterol, and thus shows promise in the preparation of drugs for the prevention and treatment of fatty liver or obesity.

[0008] The objective of this invention is achieved through the following technical solution:

[0009] The embodiments of this invention demonstrate both in vitro and in vivo experiments:

[0010] Sanguisorbine has no significant effect on hepatocyte activity within 100 nM and has no obvious toxic side effects on hepatocytes, showing promise for drug application.

[0011] In in vitro experiments, sanguinarine reduced triglyceride levels in HepG2 human liver cancer cells at 100 nM; and promoted the antioxidant activity of GSH and SOD in HepG2 human liver cancer cells.

[0012] In in vivo experiments, sanguinarine can alleviate obesity induced by a high-fat diet and reduce triglyceride accumulation in the liver of mice fed a high-fat diet. Sanguinarine improves glucose tolerance and insulin sensitivity in mice fed a high-fat diet. In conclusion, sanguinarine can improve glucose and lipid metabolism disorders and alleviate the occurrence of fatty liver.

[0013] The results of in vitro and in vivo experiments in the embodiments of the present invention show that sanguinarine is used in the preparation of drugs for the prevention and treatment of fatty liver or obesity.

[0014] The aforementioned prevention and control includes one or more of prevention, mitigation, or treatment. More preferably, the prevention and control includes prevention, mitigation, or treatment of obesity and fatty liver caused by a high-fat diet.

[0015] Preferably, the obesity is caused by a high-fat diet.

[0016] Preferably, the fatty liver is metabolic-associated fatty liver disease. More preferably, the fatty liver is metabolic-associated fatty liver disease caused by a high-fat diet.

[0017] The preparation of the drug for preventing and treating fatty liver or obesity results in an oral formulation.

[0018] The oral preparation is one of the following: capsule preparation, soft capsule preparation, oral liquid preparation, drop pill preparation, or tablet preparation.

[0019] The oral formulation further contains pharmaceutically acceptable excipients and / or carriers. The excipients and / or carriers are one or more selected from saline, sterile water, Ringer's solution, buffered saline, glucose solution, maltodextrin solution, glycerol, ethanol, lactose, glucose, sucrose, sorbitol, mannitol, starch, gum arabic, calcium phosphate, alginate, gelatin, calcium silicate, microcrystalline cellulose, polyvinylpyrrolidone, cellulose, syrup, methylcellulose, methylparaben, propylparaben, talc, magnesium stearate, and mineral oil.

[0020] In a preferred embodiment, the appropriate dosage of sanguinarine is 0.05-0.1 mg / kg; the preferred dosage in this invention is 0.1 mg / kg.

[0021] Compared with the prior art, the present invention has the following advantages:

[0022] In this embodiment of the invention, it was confirmed that, in in vitro experiments, sanguinarine reduced triglyceride levels in human liver cancer cells HepG2 at 100 nM; and could also promote the antioxidant levels of GSH and SOD.

[0023] In in vivo experiments, sanguinarine can alleviate obesity induced by a high-fat diet and reduce the accumulation of triglycerides in the liver of mice fed a high-fat diet. Sanguinarine improves glucose tolerance and insulin sensitivity in mice fed a high-fat diet. In conclusion, sanguinarine can delay the onset of obesity, improve glucose and lipid metabolism disorders, and alleviate the occurrence of fatty liver.

[0024] This confirms that sanguinarine can be used in the preparation of drugs for the prevention and treatment of fatty liver or obesity; and sanguinarine does not produce side effects such as changes in cell activity. Therefore, sanguinarine provides a low-toxicity natural small molecule drug for the effective prevention or treatment of fatty liver or obesity. Furthermore, as a low-toxicity natural small molecule compound, sanguinarine is inexpensive and has broad application prospects. Attached Figure Description

[0025] Figure 1 shows the effect of different concentrations of SAN on the activity of HepG2 human liver cancer cells.

[0026] Figure 2 shows the detection results of TG in HepG2 human liver cancer cells after treatment with 100 nM SAN.

[0027] Figure 3 shows the Oil Red O staining results of HepG2 human liver cancer cells after SAN treatment.

[0028] Figure 4 shows the detection results of mtDNA levels in HepG2 human liver cancer cells after SAN treatment.

[0029] Figure 5 shows the GSH detection results of HepG2 human liver cancer cells after SAN treatment.

[0030] Figure 6 shows the results of weight and body fat percentage measurements of mice after high-fat diet and SAN treatment.

[0031] Figure 7 shows the serum TG, AST, and ALT levels in mice after high-fat diet and SAN treatment.

[0032] Figure 8 shows the results of glucose tolerance (GTT) tests and the area under the glucose tolerance curve in mice after high-fat diet and SAN treatment.

[0033] Figure 9 shows the results of the insulin sensitivity (ITT) test and the area under the insulin tolerance curve in mice after high-fat diet and SAN treatment. Detailed Implementation

[0034] The technical solutions of the embodiments of the present invention will be clearly and completely described below with reference to the accompanying drawings. Obviously, the described embodiments are only a part of the embodiments of the present invention, and not all of them. Based on the embodiments of the present invention, all other embodiments obtained by those skilled in the art without creative effort are within the scope of protection of the present invention.

[0035] Currently, there is a lack of low-toxicity natural small molecule drugs that can effectively alleviate hepatic steatosis. To address these technical problems, this invention proposes the application of sanguinarine in the preparation of drugs for the prevention and treatment of fatty liver or obesity.

[0036] The objective of this invention is achieved through the following technical solution:

[0037] This invention demonstrates the following steps: (1) establishing a mouse model of obesity induced by a high-fat diet and a model of metabolic-related fatty liver disease; (2) intervening in the induced models by adding sanguisorbin to the high-fat diet; (3) tracking various physiological indicators of mice and conducting research on lipid metabolism in mice and their fat accumulation; (4) establishing an in vitro model of lipid accumulation in HepG2 cells induced by oleic acid / palmitic acid; (5) intervening in the induced cell models by adding sanguisorbin to the culture medium; and (6) measuring various indicators of the cells after intervention. The experiments verify the application of sanguisorbin in the preparation of drugs for the prevention and treatment of obesity and fatty liver.

[0038] Experiments have verified that sanguinarine can improve weight gain and reduce fat accumulation caused by a high-fat diet, as well as reduce significant characteristics of obesity induced by a high-fat diet, and reduce the accumulation of triglycerides in the liver, as well as significant characteristics of metabolic-related fatty liver disease. This demonstrates its significant therapeutic effect in preventing and treating obesity induced by a high-fat diet and metabolic-related fatty liver disease. It also verifies that sanguinarine can improve oxidative stress and energy metabolism disorders induced by a high-fat diet and reduce lipid deposition in vitro; thus supporting the application of sanguinarine in the preparation of drugs for the prevention and treatment of obesity and fatty liver.

[0039] The aforementioned prevention and control includes one or more of prevention, mitigation, or treatment. More preferably, the prevention and control includes prevention, mitigation, or treatment of obesity and fatty liver caused by a high-fat diet.

[0040] The experimental results show that:

[0041] 1. Sanguisorbine has no significant effect on the activity of HepG2 cells.

[0042] To investigate the effect of sanguinarine on cell viability, HepG2 cells were treated with different concentrations (20 nM, 40 nM, 60 nM, 80 nM and 100 nM). It was found that sanguinarine had no toxic effect on HepG2 cells at concentrations up to 100 nM (the results are shown in Figure 1).

[0043] 2. Sanguisorbin alleviates lipid metabolism disorders in HepG2 cells.

[0044] Cell treatment with 100 nM sanguinarine revealed its effect on lipid metabolism both in vivo and in vitro (Figure 2). Oil Red O staining also showed that sanguinarine can reduce tissue fat accumulation (Results shown in Figure 3). These results indicate that sanguinarine can alleviate lipid dysregulation in HepG2 cells.

[0045] 3. Effects of sanguinarine on energy metabolism in HepG2 cells

[0046] Mitochondria are responsible for lipid metabolism and are therefore widely considered important participants in the pathogenesis and progression of fatty degeneration. Mitochondrial dysfunction is associated with energy metabolism disorders. Sanguisorbine can significantly increase the mtDNA content in HepG2 cells (Figure 4), indicating that sanguisorbine can regulate intracellular energy metabolism.

[0047] 4. Effects of sanguinarine on oxidative stress in HepG2 cells

[0048] Oxidative stress is a key factor in the formation of fatty liver. After treatment of HepG2 cells with sanguinarine, GSH increased significantly and MDA decreased (Figure 5). These results indicate that sanguinarine can alleviate the occurrence of oxidative stress induced by high lipids.

[0049] 5. Sanguisorbin alleviates obesity induced by a high-fat diet.

[0050] Twenty-seven six-week-old male C57BL / 6J mice were randomly divided into three groups of nine each: a standard diet group (CK), a high-fat diet group (HFD), and a high-fat diet group treated with 0.1 mg / kg sanguinarine (HFD+SAN). Treatment lasted for 10 weeks. The study found that, unlike the rapidly gaining HFD group, SAN significantly slowed the increase in body weight and fat percentage induced by the high-fat diet (results shown in Figure 6). Serum TG, AST, and ALT levels in mice after high-fat diet and SAN treatment were as follows: Figure 7 As shown above, sanguinarine can reduce fat deposition in mice induced by a high-fat diet.

[0051] 6. Sanguisorbic acid can alleviate glucose metabolism disorders induced by a high-fat diet.

[0052] To further evaluate glucose and insulin levels in mice under different treatment conditions, GTT and ITT tests were performed at weeks 9 and 10 of treatment. The GTT results are shown in Figure 8. Mice in the HFD group showed decreased glucose tolerance after glucose injection. Compared with the HFD group, HFD+SAN significantly reduced the rise in blood glucose after glucose injection and promoted the recovery of blood glucose to normal levels. The ITT results are shown in Figure 9. After insulin injection, the blood glucose level in the HFD group decreased slowly, indicating insulin resistance. The blood glucose decrease trend in the HFD+SAN group was significantly lower than that in the HFD group, indicating that adding SAN to the diet can improve insulin resistance induced by HFD. The experimental results suggest that sanguinarine can alleviate glucose metabolism disorders caused by high-fat diets.

[0053] In summary, the experimental results are as follows:

[0054] Sanguisorbine has no significant effect on hepatocyte activity within 100 nM and has no obvious toxic side effects on hepatocytes, showing promise for drug application.

[0055] In in vitro experiments, sanguinarine can reduce TG levels and lipid droplet accumulation in human hepatocellular carcinoma cells HepG2; sanguinarine can increase mtDNA in human hepatocellular carcinoma cells HepG2 to promote hepatocyte energy metabolism; and it can also promote the increase of GSH and decrease of MDA in human hepatocellular carcinoma cells HepG2 to alleviate oxidative stress.

[0056] In in vivo experiments, MRI scans of mice revealed that sanguinarine significantly slowed the increase in body weight and fat percentage induced by a high-fat diet and increased lean meat percentage. Sanguinarine also reduced the accumulation of triglycerides in the mouse liver, indicating that it can improve fat deposition induced by a high-fat diet. Measurements of glucose and insulin levels in mice under different treatment conditions showed that sanguinarine enhanced glucose tolerance and improved insulin resistance induced by a high-fat diet, indicating that it can alleviate glucose metabolism disorders caused by a high-fat diet. These results demonstrate that sanguinarine can improve glucose and lipid metabolism disorders induced by a high-fat diet.

[0057] Based on the results of in vitro and in vivo experiments in the embodiments of the present invention, it can be seen that sanguinarine is used in the preparation of drugs for the prevention and treatment of fatty liver or obesity.

[0058] The aforementioned prevention and control includes one or more of prevention, mitigation, or treatment. More preferably, the prevention and control includes prevention, mitigation, or treatment of obesity and fatty liver caused by a high-fat diet.

[0059] The obesity mentioned refers to obesity caused by a high-fat diet.

[0060] The fatty liver described is metabolic-associated fatty liver disease. More preferably, the fatty liver is metabolic-associated fatty liver disease caused by a high-fat diet. The drug prepared for the prevention and treatment of fatty liver or obesity is an oral formulation.

[0061] The oral preparation is one of the following: capsule preparation, soft capsule preparation, oral liquid preparation, drop pill preparation, or tablet preparation.

[0062] The oral formulation further contains pharmaceutically acceptable excipients and / or carriers. The excipients and / or carriers are one or more selected from saline, sterile water, Ringer's solution, buffered saline, glucose solution, maltodextrin solution, glycerol, ethanol, lactose, glucose, sucrose, sorbitol, mannitol, starch, gum arabic, calcium phosphate, alginate, gelatin, calcium silicate, microcrystalline cellulose, polyvinylpyrrolidone, cellulose, syrup, methylcellulose, methylparaben, propylparaben, talc, magnesium stearate, and mineral oil.

[0063] In a preferred embodiment, the dosage of sanguinarine is preferably 0.1 mg / kg.

[0064] Compared with the prior art, the present invention has the following advantages:

[0065] In this embodiment of the invention, it was confirmed that, in in vitro experiments, sanguinarine reduced triglyceride levels in human liver cancer cells HepG2 at 100 nM; sanguinarine can promote the antioxidant effects of GSH and SOD.

[0066] In in vivo experiments, sanguinarine can alleviate obesity induced by a high-fat diet and reduce triglyceride accumulation in the liver of mice fed a high-fat diet. Sanguinarine also improved glucose tolerance and insulin sensitivity in mice induced by a high-fat diet. In conclusion, sanguinarine can improve glucose and lipid metabolism disorders and alleviate the occurrence of fatty liver, and can be used in the preparation of drugs for the prevention and treatment of fatty liver or obesity.

[0067] The above description is only a preferred embodiment of the present invention and is not intended to limit the present invention. Any modifications, equivalent substitutions, and improvements made within the spirit and principles of the present invention should be included within the protection scope of the present invention.

Claims

1. Sanguisorbin is used in the preparation of drugs for the prevention and treatment of fatty liver or obesity.

2. The use of sanguinarine according to claim 1 in the preparation of drugs for the prevention and treatment of fatty liver or obesity, characterized in that: The aforementioned prevention and treatment include one or more of prevention, relief, or treatment.

3. The use of sanguinarine according to claim 1 in the preparation of drugs for the prevention and treatment of fatty liver or obesity, characterized in that: The obesity mentioned refers to obesity caused by a high-fat diet.

4. The use of sanguinarine according to claim 1 in the preparation of drugs for the prevention and treatment of fatty liver or obesity, characterized in that: The fatty liver mentioned is metabolic-related fatty liver disease.

5. The use of sanguinarine according to claim 4 in the preparation of drugs for the prevention and treatment of fatty liver or obesity, characterized in that: The fatty liver mentioned refers to metabolic-related fatty liver disease caused by a high-fat diet.

6. The use of sanguinarine according to claim 1 in the preparation of drugs for the prevention and treatment of fatty liver or obesity, characterized in that: The preparation of the drug for preventing and treating fatty liver or obesity results in an oral formulation.

7. The use of sanguinarine according to claim 6 in the preparation of drugs for the prevention and treatment of fatty liver or obesity, characterized in that: The oral preparation is one of the following: capsule preparation, soft capsule preparation, oral liquid preparation, drop pill preparation, or tablet preparation.

8. The use of sanguinarine according to claim 6 in the preparation of drugs for the prevention and treatment of fatty liver or obesity, characterized in that: The oral formulation also contains pharmaceutically acceptable excipients and / or carriers.

9. The use of sanguinarine according to claim 8 in the preparation of drugs for the prevention and treatment of fatty liver or obesity, characterized in that: The excipients and / or carriers are one or more of the following: saline, sterile water, Ringer's solution, buffered saline, glucose solution, maltodextrin solution, glycerol, ethanol, lactose, glucose, sucrose, sorbitol, mannitol, starch, gum arabic, calcium phosphate, alginate, gelatin, calcium silicate, microcrystalline cellulose, polyvinylpyrrolidone, cellulose, syrup, methylcellulose, methylparaben, propylparaben, talc, magnesium stearate, and mineral oil.

10. The use of sanguinarine according to claim 1 in the preparation of drugs for the prevention and treatment of fatty liver or obesity, characterized in that: The dosage of sanguinarine is 0.05-0.1 mg / kg.

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