TW201422230A - Pharmaceutical composition for preventing and treating liver fibrosis - Google Patents

Abstract

The present invention provides a pharmaceutical composition for preventing and treating liver fibrosis or nonalcoholic fatty liver disease, comprising 50 to 90% by weight of the Cordyceps sinensis mycelium powder, and 10 to 50% by weight of condensed astragalus powder.

Description

Medicine composition for preventing and treating liver fibrosis

The present invention relates to a pharmaceutical composition for preventing and treating liver fibrosis or nonalcoholic fatty liver, comprising 50 to 90% by weight of Cordyceps sinensis mycelium and 10 to 50% by weight of jaundice.

Chronic liver damage caused by alcohol, viruses or chemicals can activate liver stellate cells and secrete a large amount of extracellular matrix such as collagen, and excessive deposition of these extracellular matrices can cause liver fibrosis. According to the above-mentioned pathogenic mechanism, the development of drugs for treating liver fibrosis mainly aims at inhibiting the synthesis of extracellular matrix or promoting the degradation of extracellular matrix, but most of the substances found so far are biologically toxic or may have strong side effects. No one has been found to be effective in animal models.

Nonalcoholic fatty liver disease (NAFLD) is a liver fat content of more than 5% of liver weight in patients who do not consume excessive alcohol. These patients are often accompanied by diseases such as obesity, diabetes, and hyperlipidemia. Liver fibrosis and cirrhosis. At present, the treatment of nonalcoholic fatty liver is controlled by weight loss, blood sugar and blood lipids, but no drug has been proven to be effective in treating nonalcoholic fatty liver disease and further preventing its progression to liver fibrosis and cirrhosis.

It is an object of the present invention to provide a pharmaceutical composition for preventing and treating liver fibrosis or nonalcoholic fatty liver using Chinese herbal medicine. The pharmaceutical composition of the present invention mainly comprises 50 to 90% by weight of Cordyceps sinensis mycelium and 10 to 50% by weight of scutellaria.

Cordyceps sinensis is a traditional Chinese herbal medicine with antibacterial, anti-inflammatory, antipyretic, sedative, vasodilating, anti-asthmatic, anti-arrhythmia, metabolism, anti-aging, anti-tumor and immune activity. Because the wild Cordyceps sinensis is sparsely produced and expensive, the Cordyceps sinensis used in the present invention is a mycelium obtained by separating and purifying Cordyceps sinensis, which comprises the following species: Hirsutella sinensis , Paecilomyces sinensis Chrysosporium sinensis , Sporothrix insectorum , Stachybotrys sp., Tolypocladium sinensis , Paecilomyces hepiali , and bat moth Spores ( Hirsutella hepiali ) and the like.

According to a preferred embodiment of the present invention, the Cordyceps sinensis mycelium system is isolated, cultured and fermented by Paecilomyces hepiali Chen et Dai mycelia, and the obtained mycelium is frozen or dried. It is made into a powder as one of the main components in the pharmaceutical composition of the present invention.

The medicinal part of Astragalus membranaceus is its root and has the function of enhancing cellular immunity. The present invention is concentrated by using dried roots selected from the group consisting of Astragalus membranaceus (Fisch.) Bge. Var. mongholicus (Bge.) Hsiao or Astragalus membranaceus (Fisch.) Bge. Astragalus powder is the other major component of the pharmaceutical composition of the present invention, and any other type of xanthine is also within the scope of the present invention.

The pharmaceutical composition of the present invention may further comprise a concentrated powder of Zizyphi Sativae , and further comprising a pharmaceutically acceptable excipient. The excipient can be selected from a combination of one or more of calcium phosphate, zinc gluconate, magnesium stearate, cerium oxide, and starch.

Another object of the present invention is to provide a pharmaceutical composition for preventing or treating liver fibrosis or nonalcoholic fatty liver medicine or health food, and the medicament or health food can be an oral preparation such as a capsule, a lozenge or a powder. form.

Hereinafter, embodiments thereof will be described in detail by a preferred embodiment of the present invention, and their excellent effects for preventing and treating liver fibrosis or nonalcoholic fatty liver.

BRIEF DESCRIPTION OF THE DRAWINGS Fig. 1 is a flow chart for preparing a filamentous powder of Paecilomyces pallidum of the present invention.

Figure 2 shows the contents of (A) triglyceride, (B) cholesterol, (C) AST and (D) ALT measured under four treatments in the TAA-induced liver fibrosis test. The data in the table are mean ± SEM (n=12), indicating different letters indicates significant difference (p<0.05).

Figure 3 is a graph showing changes in body weight of mice measured in the control group and two doses (557 and 2786 mg/kg) in a high-oil feed-induced fatty liver test.

Figure 4 shows the (A) AST and (B) ALT levels measured under four treatments in a high oil feed induced fatty liver test.

Figure 5 shows the insulin content measured under four treatments in a high oil feed induced fatty liver test.

Figure 6 is a graph showing the wet weight of the liver measured under four treatments in a high oil feed induced fatty liver test.

(Preparation of the pharmaceutical composition of the present invention)

The Cordyceps sinensis mycelium system used in a preferred embodiment of the present invention is a Paecilomyces pallidum mycelium powder. The preparation of the Paecilomyces pallidum mycelium powder is carried out according to the flow chart of Fig. 1. The strain of Paecilomyces palustris is cultured and fermented, and the obtained mycelium is frozen or dried, ground and sieved. Into a powder. In addition, Astragalus is a commercially available concentrated extract of Astragalus membranaceus which is made from dried roots of Astragalus membranaceus or Astragalus membranaceus. Further, the pharmaceutical composition of the present invention further comprises jujube concentrated powder and a pharmaceutically acceptable excipient which may be selected from the group consisting of calcium phosphate, zinc gluconate, magnesium stearate, cerium oxide and A combination of one or more of the starches. The above composition is preferably in the form of a capsule containing the ingredients and contents as follows:

The human dose (70 kg) of the composition of the present invention is 3120 mg/day; the oral dose of the rat is calculated according to the ratio of the equivalent surface dose of the experimental animal to the human body, and the ratio of the surface area of the human body of 200 g to 70 kg is 0.018. The dose of 200 grams of rats was 56.16 mg / day. The efficacy of the composition of the present invention for preventing and treating liver fibrosis or nonalcoholic fatty liver will be examined below.

(protection of liver fibrosis induced by thioacetamide (TAA))

Five-week-old male Wistar rats were divided into four groups and treated as follows:

TAA is administered with 100 mg/kg three times a week to induce liver fibrosis symptoms in rats; and the composition of the present invention is continuously administered with deionized water, one-time or five-fold doses of each group per Saturday. Eight weeks.

Eight weeks later, blood was collected to measure the biochemical index and blood lipids related to liver injury in the serum of each group, and the lipid content, cytokines, collagen, antioxidant enzyme activities and peroxidative metabolites of liver tissues were determined. The results are described as follows:

1. The effect of blood biochemical value and liver lipid content

As shown in Fig. 2A and Fig. 2B, TAA treatment resulted in a decrease in blood lipids (triglyceride (TG) and cholesterol (TC)), and TG and TC were significantly higher in individual serum supplemented with the composition of the present invention. Individuals treated with TAA, TC can even return to the level of the control group; and liver lipid analysis results show that supplementation of the composition of the invention can effectively reduce the accumulation of liver fat (as shown in Table 1).

2. The effect of liver antioxidant capacity

The antioxidant components and enzyme activities of each group were measured, and the results are shown in Table 2.

The results show that supplementing the composition of the present invention significantly reduces the lipid peroxidation products malondialdehyde (MDA), increases the total antioxidant capacity (TEAC), and increases the activity of antioxidant enzymes SOD, CAT, GPx.

3. The impact of liver damage

Alanine transaminase (ALT) and aspartate transaminase (AST) can be used as indicators of liver damage, and the results show that supplementing the composition of the present invention can effectively reduce ALT and AST in blood (such as 2C and 2D), and the low dose composition exhibits the ability to reduce ALT and AST.

Tumor necrosis factor-α (TNFα) and interleukin-1β (IL1-β) are cytokines secreted by damaged liver cells, which can aggravate cell damage and cause a strong inflammatory response. The test results are shown in Table 3. It is shown that supplementing the composition of the present invention can effectively reduce the content of TNFα and IL1-β, indicating that the degree of liver damage is slowed down. In addition, analysis of collagen in the liver and supplementation of the composition of the present invention can also significantly reduce the amount of collagen in the liver (Table 3).

From the above-mentioned pattern of rat liver fibrosis induced by TAA, it was proved that the pharmaceutical composition of the present invention can improve lipid metabolism in the liver, reduce fat accumulation, improve antioxidant capacity, and alleviate liver damage caused by TAA, and use low dose. (normal test group) can achieve the desired results.

(Initiation of chemical liver damage with carbon tetrachloride)

Spraque-Dawley rats were divided into five groups and treated as follows:

Oral administration for six weeks, feeding 1 ml/kg of 40% carbon tetrachloride dissolved in olive oil three times a week; and daily feeding of each group of deionized water, one or ten times the dose of the invention Composition and silymarin.

1. Blood biochemical value and analysis

Six weeks later, blood tests were performed to examine the various liver functions in Table 4. The results show that administration of the composition of the present invention lowers AST, ALT and bilirubin and increases serum albumin content.

2. Antioxidant enzymes and protein concentration

Detection of antioxidant molecules glutamine-based sulfur (GSH), glutamine-based thioperoxidase (GSH-Px), glutamine-based sulfur reductase (GSH-Rd), superoxide dismutase (SOD) and Activity of catalase (Catalase). The results are shown in Table 5.

The results showed that GSH and GSH-Px were increased to be close to the normal control group after administration of the composition of the present invention, and Catalase and SOD were even higher than the positive control group administered with silymarin, and the protein concentration was also the same.

3. Pathological section

Liver damage caused by carbon tetrachloride was found to be rough on the surface of the liver by pathological sections, but the rat liver tissue to which the composition of the present invention was administered showed a slight phenomenon of fibrosis and a decreased rate of hepatic lobular deformation.

The distribution of collagen in liver tissue was observed by Sirius red tissue staining. The negative control group treated with carbon tetrachloride showed an increase in collagen area in liver tissue sections and severe liver fibrosis in hepatic lobules. The results of the quantitative analysis are shown in Table 6. It is shown that both doses of the composition of the present invention reduce collagen in the liver and have the effect of protecting the liver.

From the above model of inducing liver injury in rats by carbon tetrachloride, it was proved that the pharmaceutical composition of the present invention can reduce the damage of liver carbon index of rat carbon tetrachloride, increase the activity of antioxidant enzymes and protein concentration, in pathological sections and collagen. The protein content also indicates that the phenomenon of liver fibrosis can be reduced.

(Evaluation of the role of high oil feed in inducing fatty liver)

6-week-old C57BL/6 mice were fed with high-oil feed (D12492, Research Diets, USA) containing more than 60% of calories as oil for 18 weeks to induce obesity in mice; and normal feed containing 10% calories as oil (D12450B, Research Diets, USA) C57BL/6 mice of the same age were housed as a control group.

Each group was orally administered with sterile water (10 ml/kg) or the composition of the present invention (557 mg/kg or 2786 mg/kg) every 10 weeks for ten consecutive weeks. Body weight was measured weekly, and after ten weeks, blood was collected to measure serum AST, ALT, and insulin, and the liver was weighed.

The body weight of the mice in the blank group fed the normal diet did not change significantly throughout the experiment; the mice in the control group fed the high oil diet were statistically significantly higher than the blank group. This hair The composition of 557 mg/kg of the composition was not significantly different from the control group; the weight of the mice orally administered at the high dose of 2786 mg/kg decreased from the 22nd day; the 36th to 57th days were clearly observed. The weight loss of the mice (Fig. 3).

After the oral administration of the high dose of 2786 mg / kg for eight weeks, the composition of the present invention reduced the serum AST value (73 ± 6.6 U / L) by about 58%, especially the significant decrease compared with the control group (174 ± 46.7 U / L). Serum ALT values (9 ± 3.7 U / L) were about 84% (control group was 56 ± 13.2 U / L, p < 0.05) (Figure 4A and Figure 4B), with statistical differences.

The insulin (9.69±2.04 ng/ml) fed to the high-oil feed mice was 2.7 times higher than the normal feed (3.55±0.65 ng/ml), and the composition of the present invention was 557 mg/kg or 2786 mg/kg. Oral doses were increased by 24% and 36% insulin (12.00 ± 1.83 ng / dl and 13.17 ± 2.31 ng / dl), respectively, compared to the control group (9.69 ± 2.04 ng / dl) (Figure 5). According to the literature, the secretion of insulin contributes to the protective function of liver function.

The liver wet weight of mice fed high oil diet was higher than that of the blank group. After 10 weeks of oral administration of the composition of the present invention at a dose of 557 mg/kg, the liver wet weight of the mice was statistically indistinguishable from that of the control group. However, oral administration of the composition of the present invention at a dose of 2786 mg/kg for 8 weeks reduced the liver wet weight of the mice by about 29% (1.16 ± 0.1 g vs. 1.64 ± 0.1 g of the control group) compared with the control group (Fig. 6).

The present invention has been described above with reference to preferred embodiments and drawings, which are not intended to limit the invention. Various modifications, omissions and changes to the details of the specific embodiments described above are the scope of the invention.

Claims (6)

A pharmaceutical composition for preventing and treating liver fibrosis comprising 50 to 90% by weight of Cordyceps sinensis mycelium and 10 to 50% by weight of jaundice. The pharmaceutical composition of claim 1, wherein the Cordyceps sinensis mycelium system is Paecilomyces hepiali Chen et Dai mycelia. The pharmaceutical composition of claim 1, wherein the Cordyceps sinensis mycelium system is in the form of a dry powder. The pharmaceutical composition of claim 1, wherein the scutellaria is selected from the group consisting of Astragalus membranaceus (Fisch.) Bge. Var. mongholicus (Bge.) Hsiao or Astragalus membranaceus ( Astragalus membranaceus ). Concentrated sassafras powder made from dried roots of Fisch.) Bge.). For example, the pharmaceutical composition of claim 1 is used for the manufacture of a medicament or a health food. The pharmaceutical composition of claim 5, wherein the pharmaceutical or health food is an oral preparation of a capsule, a lozenge or a powder.