CN113440517A

CN113440517A - Application of licarin B in preparation of medicine for preventing or/and treating non-alcoholic fatty liver disease

Info

Publication number: CN113440517A
Application number: CN202110632201.3A
Authority: CN
Inventors: 王刚; 赵文宇; 王琳琳; 赵建新; 张灏; 陈卫; 顾震南
Original assignee: Jiangnan University
Current assignee: Jiangnan University
Priority date: 2021-06-07
Filing date: 2021-06-07
Publication date: 2021-09-28

Abstract

The invention discloses the application of Likaling B in the preparation of medicines for preventing or/and treating non-alcoholic fatty liver disease, and belongs to the technical field of medicine application. The invention provides a new application of Likaling B in preventing or treating non-alcoholic fatty liver. Using Likaling B by gavage can make mice lose weight, reduce blood lipids, significantly reduce liver lipids, and improve blood sugar levels. , the liver function level is significantly improved, which can have a positive effect on the prevention and/or treatment of non-alcoholic fatty liver by regulating liver lipid metabolism. Compared with chemical or biosynthetic drugs, Likaline B extracted from natural Chinese herbal medicines has lower cost and less toxic and side effects, and has an extremely wide range of applications in the preparation of liver-protecting and liver-protecting products.

Description

Application of licarin B in preparation of medicine for preventing or/and treating non-alcoholic fatty liver disease

Technical Field

The invention relates to application of licarin B in preparation of a medicine for preventing or/and treating non-alcoholic fatty liver disease, belonging to the technical field of medicine application.

Background

Non-alcoholic fatty liver disease is a disease characterized by excessive accumulation of fat in liver cells, and is closely related to severe obesity, insulin resistance and chronic low-grade inflammation. The clinicopathological syndrome of nonalcoholic fatty liver disease is characterized by diffuse hepatocyte with hepatocyte globulocystic, intrahepatic inflammatory response and progressive fibrosis. The pathological changes can be classified into simple steatosis, non-alcoholic steatohepatitis and steatocirrhosis, and fibrosis and even liver cancer can be developed. Persistent liver fatty infiltration and inflammatory responses can increase the incidence of cirrhosis and liver cancer. In addition, nonalcoholic fatty liver can lead to the development of dyslipidemia and significantly increase the incidence of cardiovascular disease.

The mechanism by which the non-alcoholic fatty liver process occurs is not clear, although it is generally believed that it consists of at least two components, the "secondary strike" theory. Liver fat accumulation caused by disorders of fat metabolism is a common mechanism among the various causes causing liver steatosis, followed by immune cell activation and inflammatory cytokine production. In recent years, the theory of "multiple hits" has been widely proposed, including genetic factors, insulin resistance, lipid metabolism disorder, hepatic steatosis, oxidative stress, inflammation, endoplasmic reticulum stress, and intestinal flora imbalance, and these important factors jointly promote the development of non-alcoholic fatty liver through different stages of the onset of non-alcoholic fatty liver. Among them, the factors of lipid metabolism disorder and hepatic cell steatosis appear in various theories.

Studies have shown a decrease in lipid metabolism in obese patients with non-alcoholic fatty liver. Sustained excess of free fatty acids is involved in the pathogenesis of steatosis and leads to non-alcoholic fatty liver-related metabolic complications, such as inflammatory reactions. Prolonged exposure of the liver to excess free fatty acids can lead to accumulation of intracellular triglycerides and impair insulin function, further affecting blood glucose levels. Excessive fatty acids promote secretion of TG and increase liver neolipogenesis, leading to hepatotoxicity and inducing liver to continuously produce inflammatory factors, affecting liver function. By inhibiting fatty acid synthesis, the proliferation and growth of liver cancer cells can be inhibited, and the survival rate is reduced. Therefore, the effective means for regulating liver lipid metabolism to reduce weight and fat, inhibiting inflammatory reaction, recovering liver function and improving non-alcoholic fatty liver is provided.

At present, non-alcoholic fatty liver disease can be prevented to some extent by limiting dietary energy intake (e.g. low sugar, low fat, etc.) or by exercising in moderate amounts, but the extent of intervention is limited. With the acceleration of modern life rhythm and the increase of social working pressure, irregular diet and sleep increase the incidence of the non-alcoholic fatty liver disease. Common non-alcoholic fatty liver treatment drugs comprise simvastatin, pentoxifylline, pioglitazone, obeticholic acid, vitamin E supplement and the like, but have the defects of high price, large side effect and the like of the drugs (for example, pioglitazone can cause the common adverse reactions of thiazolidinedione drugs, namely weight increase of high risk groups, foot edema, bone loss and congestive heart failure, and vitamin E supplement can obviously increase the risk of healthy men suffering from prostate cancer). Therefore, there is a need to develop a product extracted from natural plant compounds for treating non-alcoholic fatty liver disease to solve the problem.

Disclosure of Invention

The invention relates to the technical field of medicine application, in particular to a natural Chinese herbal medicine extract component licarin B (Licarin B), and provides a product which can reduce weight, blood fat and liver lipid accumulation, has small toxic and side effects and can be used for preventing or/and treating non-alcoholic fatty liver.

The invention provides an application of licarin B in preparing a medicament for preventing/treating non-alcoholic fatty liver, wherein the chemical structural formula of the licarin B is as follows:

in one embodiment, the prevention and/or treatment of non-alcoholic fatty liver disease is characterized by at least one of (a) to (d):

(a) reducing blood lipid of non-alcoholic fatty liver individuals;

(b) reducing lipid synthesis in the liver of a non-alcoholic fatty liver subject;

(c) regulating liver lipid metabolism and/or reducing liver lipid accumulation in a non-alcoholic fatty liver subject;

(d) improving liver function of non-alcoholic fatty liver individual.

In one embodiment, the improvement of liver function in a non-alcoholic fatty liver subject is a reduction in the abnormal elevation of alanine aminotransferase and/or aspartate aminotransferase in a non-alcoholic fatty liver subject.

In one embodiment, the subject with non-alcoholic fatty liver disease is a mammal with symptoms of non-alcoholic fatty liver disease.

In one embodiment, the mammal includes, but is not limited to, a human.

In one embodiment, the oral Dose of licarin B mice is not less than 100 mg/kg-d, as determined by the conversion factor given in the optimization of the Maximum Safe Start Dose in the Initial Clinical laboratories for Therapeutics in the oral health volumes of FDA, for adults is not less than 8.1 mg/kg-d.

In one embodiment, the medicament further comprises a pharmaceutically acceptable carrier.

In one embodiment, the carrier is one or more of a pharmaceutically acceptable filler, wetting agent, disintegrant, binder, or lubricant.

In one embodiment, the filler is one or more of microcrystalline cellulose, lactose, mannitol, starch, or dextrin; the wetting agent is one or more of ethanol or glycerol; the disintegrant is one or more of sodium carboxymethyl starch, cross-linked povidone or low-substituted hydroxypropyl cellulose; the adhesive is one or more of starch paste, syrup, maltose, refined honey or liquid glucose; the lubricant is one or more of magnesium stearate, sodium fumarate stearate, talcum powder or silicon dioxide.

In one embodiment, the medicament contains licarin B and sodium carboxymethyl cellulose.

In one embodiment, the medicament contains licarin B at a final concentration of 100mg/L and sodium carboxymethyl cellulose (CMC) at a final concentration of 5 g/L.

In one embodiment, the medicament is a formulation that is ingestible into the gastrointestinal tract, such as an oral formulation, including but not limited to a tablet, capsule, or oral liquid.

The invention also claims the use of the licarin B for the preparation of a non-medical purpose health product.

Has the advantages that:

(1) the licarin B is extracted from nutmeg, so that the weight of a mouse can be reduced, the blood fat is reduced, the liver lipid is obviously reduced, the blood sugar level is improved, and the liver function level is obviously improved.

(2) Compared with the prior art, the invention has the beneficial effects that: licarin B is widely contained in various Chinese herbal medicines such as nutmeg, antifeverile dichroa root, magnolia flower (magnolia), Chinese actinidia root, saururus chinensis and the like, is used for treating/preventing non-alcoholic fat of mice, and can obtain good effect; compared with chemical or biological synthetic medicines, the licarin B of natural Chinese herbal medicine components has lower cost and small toxic and side effects; the licarin B is used for treating the non-alcoholic fatty liver, can obviously improve the blood fat and liver function level of mice and improve the survival quality of experimental animals, so the licarin B and the liver protection product thereof have extremely wide application space.

Drawings

FIG. 1 is a liver and epididymal adipose pathological tissue section (HE staining).

FIG. 2 shows the serum TC content of each group of mice.

FIG. 3 shows the serum TG levels of mice in each group.

FIG. 4 shows the serum LDL-c content of each group of mice.

FIG. 5 shows the aspartate Aminotransferase (AST) levels in the sera of the groups of mice.

FIG. 6 is serum alanine Aminotransferase (ALT) from various groups of mice.

Detailed Description

The drug licarin B is purchased from Beijing Waokkai Biotech limited, has a molecular weight of 324.370 and a purity of greater than or equal to 98% by HPLC, and is extracted from dried nutlet of Myristica fragrans of Myristicaceae. According to Chinese herbal medicine data ETCM (http:// www.tcmip.cn), herbal group identification (http:// herb. ac. cn) and literature research, licarin B widely exists in a plurality of Chinese herbal medicines such as nutmeg, dichroa febrifuga, magnolia flower (magnolia), Chinese actinidia root, saururus chinensis and the like, and has great medicinal potential.

The mouse strain C57BL/6J referred to in the examples was purchased from Experimental animals technology, Inc. of Wei Tongli, Beijing;

the 60% high fat diet referred to in the examples was purchased from south-lead Techlophene (cat # TP 23300).

Example 1: ricarin B safety evaluation

With reference to Kuk Yun Shin et al (published in Isolation of pathological drug inhibitors from the seeds of Myristica fragrans) by gavage administration, the mice were observed to have no toxicity symptoms, no decrease in exercise or weight, no dyspnea or abdominal irritation, etc. in the 72h observation period, indicating that no significant adverse reaction was observed when the mice were gavage administered at a dose of 200 mg/kg.

Example 2: effect of licarin B on lipid accumulation in liver and adipose tissue of non-alcoholic fatty liver mice

Taking 5-week-old female C57BL/6 mice, feeding the mice at the room temperature of 20-26 ℃, with the humidity of 40-70% and the day and night alternation of 12h/12h, adaptively feeding the mice for 1 week under the conditions of free food and drinking water, dividing the mice into 3 groups according to the average weight, feeding 8 mice in each group, feeding one group with common low-fat feed, feeding the other two groups with 60% high-fat feed, and determining the model establishment after feeding for 10-12 weeks. The 3 groups were set as a blank control group (CON), a model group (HF), and a Licarin B group (Licarin B), respectively. And diluting the Licarin B with 5g/L sodium carboxymethyl cellulose solution to the concentration of 100mg/kg to obtain a Licarin B-sodium carboxymethyl cellulose solution, and preparing the gavage B group. The model group and the blank control group were perfused with only 5g/L sodium carboxymethylcellulose solution.

The experimental time was 5 weeks: the CON group mice were gavaged with 5g/L sodium carboxymethylcellulose solution at 200. mu.L per day for observation of changes in body weight and food intake, the HF group mice were gavaged with 5g/L sodium carboxymethylcellulose solution at 200. mu.L per day for observation of changes in body weight and food intake, and the Licarin B group mice were gavaged with Licarin B-sodium carboxymethylcellulose solution at 200. mu.L per day for observation of changes in body weight and food intake. Before the experiment is finished, the serum is taken to measure biochemical indexes by adopting a spectrophotometry method, and after the mouse is sacrificed, the liver, epididymal fat, fat and other organ tissues are dissected and taken to be pathological sections.

As can be seen from FIG. 1, by means of liver and epididymis adipose pathological tissue section (HE staining), compared with CON group (left), the liver of the mice in HF group (middle) has steatosis, that is, vacuoles (lipid droplets) with different sizes appear, the boundary is clear, the cell nucleus is squeezed to one side, the volume of the liver cell is increased, the swelling is caused, the cell nucleus is obvious, and the local inflammatory reaction appears. In the mouse liver tissue vacuole (lipid droplet) area in the Licarin B group (right) is reduced, the inflammatory reaction is reduced, and the liver lipid accumulation is obviously relieved. The adipocytes in the HF group (middle) were larger, swollen, and inflammatory reaction occurred to some extent between cells, with clearly visible boundaries, compared to the CON group (left). Mice in the Licarin B group (right) have smaller adipocyte area, reduced inflammatory reaction and obvious alleviation of fat accumulation. It can be seen that licarin B is effective in reducing lipid accumulation in mouse liver and adipose tissue.

Example 3: effect of licarin B on blood lipid levels in the liver of non-alcoholic fatty liver mice

As shown in FIGS. 2-4, it can be seen from the results of biochemical indicator analysis of mouse serum that the levels of LDL-c, TC and TG in the serum of HF group were all higher than those in the control group at week 16 of feeding. Licarin B effectively reduces the content levels of Total Cholesterol (TC) and Triglyceride (TG) in mouse serum by 32.14 percent and 30.86 percent respectively; meanwhile, the licarin B effectively reduces the level of low-density lipoprotein (LDL-c) by 49.08 percent compared with the HF group, and the effect is obvious. Licarin B can effectively regulate the blood lipid level of the non-alcoholic fatty liver disease mouse.

Example 4: effect of licarin B on liver function level of non-alcoholic fatty liver disease mice

The animal model was constructed as in example 2, and serum was taken before the end of the experiment and spectrophotometrically measured for biochemical markers, and the results shown in fig. 5-6 show that alanine Aminotransferase (ALT) and aspartate Aminotransferase (AST) in the mouse liver function markers were elevated in the HF group to different extents, indicating that a certain degree of impaired liver function and chronic inflammatory response occurred, and that the Licarin B group was effective in restoring liver function compared to the HF group, and ALT in the Licarin B group was reduced from 198.33 ± 33.25U/L to 75.35 ± 24.17U/L and AST was reduced from 130.99 ± 18.12U/L to 99.99 ± 14.47U/L.

Example 5: licarin B for preparing medicine

Licarin B is compatible with a pharmaceutically acceptable carrier or combined with a plurality of solid or liquid pharmaceutical excipients and/or auxiliary agents to prepare a proper administration form or dosage form which can be used for human.

One or more of the pharmaceutically acceptable filler, wetting agent, disintegrant, binder or lubricant.

The filler is one or more of microcrystalline cellulose, lactose, mannitol, starch or dextrin; the wetting agent is one or more of ethanol or glycerol; the disintegrant is one or more of sodium carboxymethyl starch, cross-linked povidone or low-substituted hydroxypropyl cellulose; the adhesive is one or more of starch paste, syrup, maltose, refined honey or liquid glucose; the lubricant is one or more of magnesium stearate, sodium fumarate stearate, talcum powder or silicon dioxide.

Optionally, the medicament contains licarin B at a final concentration of 100mg/L and sodium carboxymethyl cellulose (CMC) at a final concentration of 5 g/L.

Although the present invention has been described with reference to the preferred embodiments, it should be understood that various changes and modifications can be made therein by those skilled in the art without departing from the spirit and scope of the invention as defined in the appended claims.

Claims

1. Application of licarin B in preparing a medicament for preventing/treating non-alcoholic fatty liver disease.

2. The use according to claim 1, wherein the prevention and/or treatment of non-alcoholic fatty liver disease is at least one of (a) to (d):

(a) reducing blood lipid of non-alcoholic fatty liver individuals;

(d) improving liver function of non-alcoholic fatty liver individual.

3. The use according to claim 2, wherein said improvement in liver function in a non-alcoholic fatty liver subject is a reduction in the abnormal elevation of alanine aminotransferase and/or aspartate aminotransferase in a non-alcoholic fatty liver subject.

4. The use according to any one of claims 1 to 3, wherein the medicament further comprises a pharmaceutically acceptable carrier.

5. The use according to claim 4, wherein the carrier is one or more of a pharmaceutically acceptable filler, wetting agent, disintegrant, binder or lubricant.

6. The use according to any one of claims 1 to 5, wherein the medicament comprises licarin B and sodium carboxymethyl cellulose.

7. The use according to claim 6, wherein the medicament comprises licarin B at a final concentration of 100mg/L and sodium carboxymethyl cellulose at a final concentration of 5 g/L.

8. Use according to any one of claims 4 to 7, wherein the medicament is a gastrointestinal ingestible formulation.

9. The use of claim 8, wherein the pharmaceutical dosage form comprises but is not limited to a tablet, a capsule, or an oral liquid.

10. The use of licarin B in the preparation of a non-therapeutic health product.