Pharmaceutical composition and application thereof in acute liver injury protection

Abstract

The invention is applicable to the technical field of biological medicine, and provides a pharmaceutical composition and application thereof in acute liver injury protection, the pharmaceutical composition comprises the following raw materials by weight: 45-6% of ginsenoside Rh, 2-4% of ginsenoside Rk3, 20-30% of ginsenoside Rg3, 3-15% of ginsenoside Rg513, 18-10% of ginsenoside Rk8, and the balance of ginseng extract, totaling 100%. According to the pharmaceutical composition provided by the embodiment of the invention, ginsenoside Rh4, ginsenoside Rk3, ginsenoside Rg3, ginsenoside Rg5, ginsenoside Rk1 and the ginseng extract are taken as effective components, so that the pharmaceutical composition has an obvious improvement effect on the body weight and liver index of a mouse with liver injury, can inhibit the rise of ALT and AST levels in serum of the mouse and reduce the injury degree of liver tissues, and can be used for preparing a medicine for treating liver injury. A new thought is provided for preparation of medicines for preventing and treating acute liver injury.

Description

Technical Field

[0001] The present invention belongs to the field of biomedicine technology, and in particular relates to a pharmaceutical composition and its application in protecting acute liver damage. Background Art

[0002] As one of the most important solid organs in the human body, the liver participates in a variety of physiological processes, such as metabolism, immune response, hematopoiesis, and hemostasis. Normally, the liver has the ability to metabolize a variety of exogenous substances into low-toxic products and excrete them from the body. However, the liver is sensitive to toxic substances or drugs, and liver cells may suffer varying degrees of damage, especially under specific physical and chemical conditions or external environmental factors. Furthermore, the pathogenesis of liver injury is complex and diverse, often resulting from a single factor or the combined action of multiple factors. Over time and through a series of complex pathological changes, liver injury can progress to liver fibrosis, cirrhosis, liver cancer, and liver failure.

[0003] Drugs are considered a contributing factor to acute liver failure. Acetaminophen (APAP) is a commonly used anti-inflammatory, analgesic, and antipyretic drug that is safe and highly effective when used within the recommended dosage range. However, overdose can easily cause severe liver damage, even leading to acute liver failure and death. Currently, N-acetylcysteine (NAC) is the only approved treatment for APAP-induced acute liver injury. It treats liver injury by promptly replenishing glutathione (GSH) to eliminate APAP's toxic metabolite (NAPQI), inhibiting the formation of NAPQI-protein adducts and the resulting pathological reactions, such as oxidative stress. However, adverse reactions and a narrow therapeutic window limit its use. Therefore, new strategies for the prevention and treatment of drug-induced liver injury are urgently needed. Summary of the Invention

[0004] The purpose of the embodiments of the present invention is to provide a pharmaceutical composition, aiming to solve the problems raised in the above background technology.

[0005] The embodiment of the present invention is implemented as follows: a pharmaceutical composition includes the following raw materials in weight percentage: ginsenoside Rh45-6%, ginsenoside Rk32-4%, ginsenoside Rg320-30%, ginsenoside Rg513-15%, ginsenoside Rk18-10%, and the remainder of ginseng extract is supplemented to 100%.

[0006] Preferably, the raw materials are included in the following weight percentages: ginsenoside Rh 46%, ginsenoside Rk 33%, ginsenoside Rg 320%, ginsenoside Rg 514%, ginsenoside Rk 19%, and ginseng extract 48%.

[0007] Preferably, the raw materials are included in the following weight percentages: ginsenoside Rh 45%, ginsenoside Rk 33%, ginsenoside Rg 330%, ginsenoside Rg 513%, ginsenoside Rk 19%, and ginseng extract 40%.

[0008] Another object of the present invention is to provide a method for preparing a pharmaceutical composition, comprising the following steps:

[0009] Preparation of ginseng extract: Ginseng powder obtained by crushing ginseng is mixed with ethanol, the extract is heated and filtered to obtain a first filtrate and a first filter residue, the first filter residue is mixed with ethanol, the extract is heated and filtered to obtain a second filtrate and a second filter residue, the first filtrate and the second filtrate are combined, concentrated, and dried to obtain the ginseng extract;

[0010] Ginsenoside Rh4, ginsenoside Rk3, ginsenoside Rg3, ginsenoside Rg5, and ginsenoside Rk1 are mixed with ginseng extract.

[0011] Preferably, the ginseng is one or more of white ginseng, fresh ginseng and red ginseng.

[0012] Preferably, the mass volume ratio of the ginseng powder to ethanol is 1:10-20.

[0013] Preferably, the mass volume ratio of the filter residue 1 to ethanol is 1:10-20.

[0014] Another object of an embodiment of the present invention is to provide a pharmaceutical composition for use in preparing drugs for preventing and treating acute liver injury.

[0015] A pharmaceutical composition provided in an embodiment of the present invention contains ginsenoside Rh4, ginsenoside Rk3, ginsenoside Rg3, ginsenoside Rg5, ginsenoside Rk1 and ginseng extract as active ingredients, which can significantly improve the body weight and liver index of mice with liver damage, inhibit the increase of ALT and AST levels in mouse serum and reduce the degree of liver tissue damage. At the same time, as a traditional Chinese medicine, the composition has extremely strong stability, no toxic side effects, and is suitable for people at all stages, providing new ideas for the preparation of drugs for the prevention and treatment of acute liver injury. BRIEF DESCRIPTION OF THE DRAWINGS

[0016] Figure 1 This is a graph showing the ALT and AST levels in mice provided in Example 5 of the present invention;

[0017] Figure 2 This is a diagram of the mouse tissue pathology results provided in Example 6 of the present invention. DETAILED DESCRIPTION

[0018] In order to make the purpose, technical solutions and advantages of the present invention more clearly understood, the present invention will be further described in detail below with reference to the accompanying drawings and embodiments. It should be understood that the specific embodiments described herein are only used to explain the present invention and are not intended to limit the present invention.

[0019] The sources of ginsenosides Rh4, Rk3, Rg3, Rg5 and Rk1 can be naturally extracted, and can be extracted from wild ginseng, garden ginseng, American ginseng, etc., and the extraction method preferably includes one or more of organic solvent extraction, water extraction, ultrasonic impregnation, extraction, distillation, osmosis, and macroporous adsorption resin separation methods. In addition, these saponins can also be obtained by converting other diol or triol ginsenosides, for example, Rk1 and Rg5 can be converted from Rb1, and Rh4 and Rk3 can be converted from Rg1. The conversion method is preferably a bioconversion method. They can also be purchased directly.

[0020] Ginsenoside Rh4, ginsenoside Rk3, ginsenoside Rg3, ginsenoside Rg5, and ginsenoside Rk1 used in the examples of the present invention were all purchased from Shanghai Yuanye Biotechnology Co., Ltd., and their purity met the pharmaceutical standards.

[0021] The method for extracting the ginseng extract used in the embodiment of the present invention comprises the following steps:

[0022] Accurately weigh a certain amount of ginseng powder, put it into an extraction container, add 75% ethanol according to a certain material-liquid ratio (volume ratio of ginseng powder to ethanol), the common material-liquid ratio is 1:10 to 1:20, and more preferably 1:15, heat at 75°C for 3 hours (water bath heating method), filter the extract, and store the filtrate in a 4°C refrigerator. Add new ethanol to the remaining residue for a second extraction, repeat twice, and store the filtrate in a 4°C refrigerator; finally, combine the filtrates obtained from the two times, concentrate the filtered extract at normal pressure, remove ethanol, obtain a concentrated solution, and further dry the concentrated solution to obtain a ginseng extract; the ginseng extract includes ginsenoside components (Rg1, Re, Rf, Rb1, Rc and Rd), ginseng polysaccharide components, ginseng polypeptide components, ginseng polyphenol components and ginseng flavonoid components.

[0023] The specific implementation of the present invention is described in detail below with reference to specific embodiments.

[0024] Example 1: A pharmaceutical composition, the preparation method of which comprises the following steps:

[0025] (1) Preparation of ginseng extract: Weigh 20 g of ginseng powder, mix the crushed ginseng with 300 ml of ethanol (75%), heat at 75°C for 3 h, and filter the extract to obtain filtrate 1 and filter residue 1. Add 150 ml of ethanol to the filter residue 1 for a second extraction, heat at 75°C for 3 h, and filter the extract to obtain filtrate 2 and filter residue 2. Combine the two filtrates, concentrate the filtered extract at normal pressure to remove ethanol, and obtain a concentrated solution. The concentrated solution is further dried to obtain ginseng extract.

[0026] (2) Weigh the ingredients according to the weight percentage of ginsenoside Rh 46%, ginsenoside Rk 33%, ginsenoside Rg 320%, ginsenoside Rg 514%, ginsenoside Rk 19%, and ginseng extract 48%, and mix them.

[0027] Example 2: A pharmaceutical composition, the preparation method of which comprises the following steps:

[0028] (1) Preparation of ginseng extract: Weigh 20 g of ginseng powder, mix the crushed ginseng with 300 ml of ethanol (75%), heat at 75°C for 3 h, and filter the extract to obtain filtrate 1 and filter residue 1. Add 150 ml of ethanol to the filter residue 1 for a second extraction, heat at 75°C for 3 h, and filter the extract to obtain filtrate 2 and filter residue 2. Combine the two filtrates, concentrate the filtered extract at normal pressure to remove ethanol, and obtain a concentrated solution. The concentrated solution is further dried to obtain ginseng extract.

[0029] (2) Weigh the ingredients according to the weight percentage of ginsenoside Rh 45%, ginsenoside Rk 33%, ginsenoside Rg 330%, ginsenoside Rg 513%, ginsenoside Rk 19%, and ginseng extract 40%, and mix them.

[0030] Example 3, Biosafety Analysis of Pharmaceutical Compositions:

[0031] Mice (BALB / C mice, purchased from Liaoning Changsheng Biotechnology Co., Ltd.) were randomly divided into a negative control group (male), a pharmaceutical composition group A (male), a pharmaceutical composition group B (male), a negative control group (female), a pharmaceutical composition group A (female), and a pharmaceutical composition group B (female) according to sex. They were fasted for 16 hours but not water. Starting at 8:00 a.m., pharmaceutical composition A (Example 1) and pharmaceutical composition B (Example 2) were gavaged into the mice that had fasted for 16 hours, and the male and female mice in the negative control group were gavaged with an equal volume of purified water. Immediately after exposure, the mice were observed and their various poisoning symptoms were recorded; starting at 16:00 p.m., the mice were gavaged once more. After exposure, the various poisoning symptoms of the mice were continuously observed and recorded. The observation was continued for 14 days. The results of the acute toxicity test are shown in Table 1:

[0032] Table 1

[0033]

[0034] As can be seen from Table 1, when the maximum dosage concentration and maximum dosage volume were administered by gavage twice within 24 hours, that is, at the maximum dosage of 24.0 g / kg, there were no obvious abnormalities in the body shape, limb activity, respiration, coat color, and feces and bowel movements of the mice compared with the control group; and the drug composition had no obvious effect on the body weight of both male and female mice.

[0035] Example 4: Effects of the pharmaceutical composition on body weight and liver index in mice with acute liver injury:

[0036] Mice were randomly divided into a PBS control group, an APAP model group, a pharmaceutical composition A group, and a pharmaceutical composition B group. Pharmaceutical composition A (Example 1) and pharmaceutical composition B (Example 2) were administered to the mice by gavage for 7 days. After the end of gavage on the 7th day, the mice in the PBS control group were administered an equal volume of PBS by gavage, and the mice in the APAP model group and the pharmaceutical composition A and pharmaceutical composition B groups were administered an equal concentration of APAP solution by intraperitoneal injection. The mice were sacrificed at 24 hours to collect whole blood and obtain liver tissue for future use. In order to study the effects of the pharmaceutical compositions on the body weight and liver index of mice with acute liver injury, the body weight of the mice was measured at the beginning and end of the experiment, and the weight of the mouse liver was weighed to calculate the liver index. The results are shown in Table 2:

[0037] Table 2

[0038]

[0039] It can be seen that the body weight of mice in the model group was significantly lower than that in the PBS control group, while the body weight of mice in the two drug composition treatment groups was significantly higher than that in the APAP model group. The liver index of mice in the model group was significantly higher than that in the PBS control group, while the liver index of mice in the two drug composition treatment groups was significantly lower than that in the APAP model group.

[0040] Example 5: Effect of the pharmaceutical composition on ALT and AST levels in mice with acute liver injury:

[0041] The levels of liver injury markers ALT and AST in the serum of the four groups of mice were detected using a kit, and the results were as follows: Figure 1 As shown, it can be seen that the ALT and AST levels in the serum of the model group were significantly higher than those in the control group, while the ALT and AST levels in the serum of the mice in the two drug composition treatment groups were significantly lower than those in the model group.

[0042] Example 6: Effect of the pharmaceutical composition on histopathology of mice with acute liver injury:

[0043] The liver sections of the four groups of mice were stained with hematoxylin-eosin to assess the extent of liver damage. Figure 2As shown, it can be seen that 24 hours after the injection of APAP solution, severe necrosis had occurred in the central area of the lobule around the central vein of the liver of the mice in the APAP model group; and the necrosis in the central area of the liver lobule of the mice in the two drug combination treatment groups was also significantly reduced compared with the model group mice.

[0044] In summary, the pharmaceutical compositions provided in the embodiments of the present invention have good biosafety and can inhibit the weight loss and increase of liver index in mice; at the same time, in terms of protecting liver damage, the pharmaceutical compositions can significantly reduce the levels of liver injury biomarkers ALT and AST and the degree of liver tissue damage, which is statistically significant.

[0045] 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 in the scope of protection of the present invention.

Claims

1. A pharmaceutical composition, characterized in that The invention comprises the following raw materials in percentage by weight: ginsenoside Rh45-6%, ginsenoside Rk32-4%, ginsenoside Rg320-30%, ginsenoside Rg513-15%, ginsenoside Rk18-10%, and the remainder of ginseng extract is supplemented to 100%.

2. The pharmaceutical composition according to claim 1, characterized in that The invention comprises the following raw materials in percentage by weight: ginsenoside Rh 46%, ginsenoside Rk 33%, ginsenoside Rg 320%, ginsenoside Rg 514%, ginsenoside Rk 19%, and ginseng extract 48%.

3. The pharmaceutical composition according to claim 1, characterized in that The invention comprises the following raw materials in percentage by weight: ginsenoside Rh 45%, ginsenoside Rk 33%, ginsenoside Rg 330%, ginsenoside Rg 513%, ginsenoside Rk 19%, and ginseng extract 40%.

4. A method for preparing the pharmaceutical composition according to any one of claims 1 to 3, characterized in that: The following steps are involved: Preparation of ginseng extract: Ginseng powder obtained by crushing ginseng is mixed with ethanol, the extract is heated and filtered to obtain a first filtrate and a first filter residue, the first filter residue is mixed with ethanol, the extract is heated and filtered to obtain a second filtrate and a second filter residue, the first filtrate and the second filtrate are combined, concentrated, and dried to obtain the ginseng extract; Ginsenoside Rh4, ginsenoside Rk3, ginsenoside Rg3, ginsenoside Rg5, and ginsenoside Rk1 are mixed with ginseng extract.

5. The method for preparing the pharmaceutical composition according to claim 4, wherein The ginseng is one or more of white ginseng, fresh ginseng and red ginseng.

6. The method for preparing the pharmaceutical composition according to claim 4, wherein The mass volume ratio of the ginseng powder to ethanol is 1:10-20.

7. The method for preparing the pharmaceutical composition according to claim 4, wherein The mass volume ratio of the filter residue 1 to ethanol is 1:10-20.

8. Use of the pharmaceutical composition according to any one of claims 1 to 3 in the preparation of drugs for preventing and treating acute liver injury.

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