Application of ginsenoside Rg3 in preparing medicine for treating or preventing enterovirus infection

Abstract

The present invention discloses the use of ginsenoside Rg3 in the preparation of a drug for treating or preventing enteroviral infection, relating to the field of biomedicine. The ginsenoside Rg3 monomer in the present invention can enrich intestinal symbiotic bacteria. The short-chain fatty acids metabolites of the symbiotic bacteria enriched in this monomer, particularly acetic acid and propionic acid, have a strong antagonistic effect on enteroviral infection, improving the pathological damage caused by enteroviral infection, and is suitable for clinical promotion.

Description

Technical Field

[0001] The present invention relates to the field of biomedicine, and in particular to the use of ginsenoside Rg3 in preparing medicines for treating or preventing enterovirus infections. Background Art

[0002] Pathogenic enteroviruses, characterized by fecal-oral transmission and intestinal replication, cause widespread morbidity and mortality worldwide. Faced with the public health pressures and economic losses caused by infections, and in the absence of effective vaccines, effective preventive and therapeutic approaches are urgently needed to control enterovirus infections.

[0003] Up to 10 14 Intestinal microbes, including commensal bacteria, play a key role in maintaining host homeostasis. Studies have shown that intestinal bacteria can regulate host immune cells through their metabolites, maintaining a basal immune activation state to counteract host damage and death caused by infection. This universal antiviral response, dependent on commensal bacteria, has significant translational value in clinical prevention, control, and treatment.

[0004] Enteroviruses infect the gastrointestinal tract through the fecal-oral route, directly interacting with the microbiome. This interaction with the microbiome has multiple impacts on enteroviral infection. Therefore, identifying bacteria and their metabolites with clear anti-enteroviral activity could effectively and conveniently modulate the patient's intestinal flora, thereby achieving the goal of clinical prevention and control of enteroviral infection.

[0005] Ginsenosides are the main active ingredients of the traditional Chinese herbal medicine ginseng and are widely used worldwide as natural tonics. For example, ginsenoside Rk1 can alleviate intestinal inflammation in mice caused by antibiotic treatment by enriching intestinal probiotics. Patent application number CN114642676A discloses the use of ginsenoside Rb1 or a composition containing ginsenoside Rb1 in the preparation of a formulation for improving a subject's intestinal flora structure, improving a subject's intestinal barrier function, and / or lowering a subject's serum endotoxin levels, or in the preparation of a drug for treating or preventing abnormal glucose and lipid metabolism, gastrointestinal diseases, mental and nervous system diseases, and immune system diseases. Patent application number CN115990173A also discloses that ginsenoside Rh1 can be used to prepare a drug for the immunotherapy of colon cancer and a drug for improving and regulating the intestinal flora of colon cancer patients.

[0006] Therefore, the discovery of new ginsenoside monomers that reshape the intestinal flora and antagonize enteroviral infection through the intestinal flora and its metabolites can provide new ideas and theoretical basis for the clinical treatment and prevention of enteroviral infection. Summary of the Invention

[0007] The purpose of the present invention is to provide the use of ginsenoside Rg3 in the preparation of a drug for treating or preventing enterovirus infection. The ginsenoside monomer Rg3 can enrich intestinal symbiotic bacteria. The short-chain fatty acids, especially acetic acid and propionic acid, the metabolites of the symbiotic bacteria enriched by the monomer can activate the host's innate immune macrophage heterologous interferon response and antagonize enterovirus infection.

[0008] To achieve the above object, the present invention adopts the following technical solutions:

[0009] The present invention provides the use of ginsenoside Rg3 in preparing a drug for treating or preventing enterovirus infection. Ginsenoside Rg3 is a ginsenoside monomer 20(S)-Ginsenoside Rg3.

[0010] The present invention also provides the use of ginsenoside Rg3 in the preparation of a drug for regulating intestinal flora.

[0011] The present invention enriches bacteria of the genus Blautia (Blautia spp.), which are symbiotic bacteria producing short-chain fatty acids, by treating mice with the ginsenoside monomer 20(S)-Ginsenoside Rg3.

[0012] The ginsenoside monomer Rg3 is treated in mice at a dosage of 50 mg / kg of mouse body weight, and is continuously administered orally for 3 days, which can effectively antagonize enterovirus infection.

[0013] Furthermore, the short-chain fatty acid-producing symbiotic bacteria are Blautia coccoides and Blautia Obeum.

[0014] Furthermore, the major metabolites of Blautia coccoides and Blautia Obeum were acetic acid and propionic acid;

[0015] Preferably, treating mice with 200 mM / ml acetic acid and propionic acid in drinking water for 2 weeks has the same function of antagonizing enterovirus infection as ginsenoside monomer Rg3.

[0016] Preferably, the enterovirus is encephalomyocarditis virus (EMCV).

[0017] The medicine comprises ginsenoside Rg3 and pharmaceutically acceptable excipients.

[0018] The dosage form of the drug is granules, pills, tablets, capsules, oral liquids, injections or infusions. When used, the dosage of the drug is 50 mg / kg.

[0019] The present invention also provides the use of acetic acid and propionic acid, metabolites of intestinal symbiotic bacteria enriched in the ginsenoside monomer Rg3, in the preparation of a drug for treating or preventing enterovirus infection.

[0020] Compared with the prior art, the present invention has the following beneficial technical effects:

[0021] The ginsenoside monomer Rg3 in the present invention can enrich intestinal symbiotic bacteria. The short-chain fatty acids, especially acetic acid and propionic acid, the metabolites of the symbiotic bacteria enriched by the monomer, have a good effect of antagonizing enteroviral infection, improving the pathological damage caused by enteroviral infection, and are suitable for clinical promotion. BRIEF DESCRIPTION OF THE DRAWINGS

[0022] Figure 1 Survival curves of mice in the ginsenoside monomer Rg3-treated and control groups (A) and viral genome copy numbers in the brain and spleen (B);

[0023] Figure 2 Figure 3. Intestinal bacterial abundance (A) and short-chain fatty acid levels (B) in mice treated with ginsenoside monomer Rg3 and control groups.

[0024] Figure 3 Survival curves of mice in the acetic acid / propionic acid treatment group and the control group (A) and the viral genome copy numbers in the brain and spleen (B). DETAILED DESCRIPTION

[0025] The present invention will be further described below with reference to specific examples, but the present invention is not limited thereto.

[0026] Example 1

[0027] Experimental study on the antagonism of enterovirus infection using ginsenoside monomer Rg3.

[0028] Control group setting:

[0029] PBS was used to gavage 6-8 week old C57 / B6J mice at a dose of 100 μL / mouse for 3 consecutive days, once a day, followed by intraperitoneal injection of 200 TCID 50 Encephalomyocarditis virus, infecting mice.

[0030] Treatment group settings:

[0031] 6-8 week old C57 / B6J mice were treated with ginsenoside monomer Rg3 at a dose of 50 mg / kg mouse body weight, 100 μL / mouse, once a day for 3 consecutive days, and then the mice were infected with encephalomyocarditis virus by intraperitoneal injection;

[0032] Main indicators: Mouse survival curve (Proportion of survival): Record the number of mice surviving within 2 weeks after infection with encephalomyocarditis virus.

[0033] Secondary indicator: Viral genome copy number in mice: Three days after mice were infected with encephalomyocarditis virus, the viral genome copy number in the brain and spleen was detected by qRT-PCR.

[0034] Statistical analysis methods:

[0035] Statistical analyses were performed using Prism GraphPad software v.8.0. Error bars represent the standard error (SD) in all figures, and p-values were determined by two-tailed Student's t-test. Survival curves were analyzed using the log-rank test. A p-value < 0.05 was considered statistically significant. *p < 0.05, **p < 0.01, ***p < 0.001, ****p < 0.0001.

[0036] result:

[0037] (1) Survival curve of mice: Compared with the control group, the survival rate of mice infected with encephalomyocarditis virus increased by more than 40% ( Figure 1 A).

[0038] (2) Viral genome copy number in mice: Compared with the control group, the viral genome copy number in the brain and spleen of the treated group was significantly reduced 3 days after infection with encephalomyocarditis virus ( Figure 1 B).

[0039] In summary, ginsenoside monomer Rg3 can effectively antagonize enteroviral encephalomyocarditis virus infection in mice.

[0040] Example 2

[0041] Experimental study on enrichment of metabolites produced by symbiotic bacteria using ginsenoside monomer Rg3.

[0042] Control group settings:

[0043] 6-8 week old C57 / B6J mice were gavaged with PBS at a dose of 100 μL / mouse for 3 consecutive days, once a day. Feces were then collected for GC-MS / MS analysis.

[0044] Treatment group settings:

[0045] 6-8 week old C57 / B6J mice were treated with ginsenoside Rg3 at a dose of 50 mg / kg, 100 μL / mouse, once a day for 3 consecutive days. The feces of the mice were then collected for GC-MS / MS analysis.

[0046] Main outcome measure: Abundance of intestinal flora in mouse feces.

[0047] Secondary indicator: Short-chain fatty acid levels in mouse feces.

[0048] result:

[0049] The composition of intestinal flora changed in the ginsenoside monomer Rg3-treated group compared with the control group, and Blautia spp. were significantly enriched in the treated group ( Figure 2 A), acetate, propionate and butyrate levels were significantly increased ( Figure 2 B).

[0050] In summary, treatment with ginsenoside monomer Rg3 can enrich the short-chain fatty acid-producing symbiotic bacteria Blautia spp. and increase the levels of acetic acid and propionic acid.

[0051] Example 3

[0052] Experimental study on the antagonism of enterovirus infection by symbiotic bacterial metabolites acetic acid and propionic acid enriched in ginsenoside monomer Rg3.

[0053] Control group settings:

[0054] 6-8 week old C57 / B6J mice were given sterile water or sterile water with 200 mM / mL butyric acid for 2 weeks, after which they were intraperitoneally injected with 200 TCID 50 Encephalomyocarditis virus, infecting mice.

[0055] Treatment group settings:

[0056] 6-8 week old C57 / B6J mice were given sterile water containing 200 mM / mL acetic acid or propionic acid for 2 consecutive weeks, after which they were intraperitoneally injected with 200 TCID 50 Encephalomyocarditis virus, infecting mice.

[0057] Main indicators: Mouse survival curve (Proportion of survival): Record the number of mice surviving within 2 weeks after infection with encephalomyocarditis virus.

[0058] Secondary indicator: Viral genome copy number in mice: Three days after mice were infected with encephalomyocarditis virus, the viral genome copy number in the brain and spleen was detected by qRT-PCR.

[0059] Statistical analysis methods:

[0060] Statistical analyses were performed using Prism GraphPad software v.8.0. Error bars represent the standard error (SD) in all figures, and p-values were determined by two-tailed Student's t-test. Survival curves were analyzed using the log-rank test. A p-value < 0.05 was considered statistically significant. *p < 0.05, **p < 0.01, ***p < 0.001, ****p < 0.0001.

[0061] result:

[0062] (1) Survival curve of mice: Compared with the control group, the survival rate of mice infected with encephalomyocarditis virus increased by more than 30% ( Figure 3 A).

[0063] (2) Viral genome copy number in mice: Compared with the control group, the viral genome copy number in the brain and spleen of the treated group was significantly reduced 3 days after infection with encephalomyocarditis virus ( Figure 3 B).

[0064] In summary, ginsenoside monomer Rg3 can enrich intestinal commensal bacteria, and its metabolites, acetic acid and propionic acid, can effectively antagonize enteroviral encephalomyocarditis virus infection in mice. It deserves extensive application and further research in the clinical prevention and treatment of enterovirus.

Claims

1. Use of ginsenoside Rg3 in the preparation of a medicament for treating or preventing enterovirus infection, wherein the enterovirus is encephalomyocarditis virus.

2. The use according to claim 1, characterized in that The ginsenoside Rg3 can enrich the short-chain fatty acid-producing symbiotic bacteria Blautia genus ( Blautia spp.) bacteria.

3. The use according to claim 2, characterized in that The short-chain fatty acid-producing symbiotic bacteria is Blautiae ( Blautia coccoides ) and Blautia ovata ( Blautia Obeum ).

4. The use according to claim 1, characterized in that The medicine comprises ginsenoside Rg3 and pharmaceutically acceptable excipients.

5. The use according to claim 4, characterized in that The dosage form of the medicine is granules, pills, tablets, capsules, oral liquid or injection.

6. The use according to claim 1, characterized in that When used, the dosage of the drug is 50 mg / kg.

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