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Application of Flammulina velutipes polysaccharide extract in the preparation of antiviral drugs and its extraction method

A technology of Flammulina velutipes polysaccharides and extracts, which can be applied in the directions of antiviral agents, pharmaceutical formulations, and medical preparations containing active ingredients, etc., can solve the problems of limited preventive effect, limited clinical application, and influenza hysteresis.

Active Publication Date: 2021-09-24
GUANGDONG INST OF MICROBIOLOGY GUANGDONG DETECTION CENT OF MICROBIOLOGY
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  • Summary
  • Abstract
  • Description
  • Claims
  • Application Information

AI Technical Summary

Problems solved by technology

[0003] Vaccination is the most effective way to prevent and control the spread of influenza, but the main antigens of influenza virus, hemagglutinin (HA) and neuraminidase (neuraminidase, NA) are prone to antigenic shift and antigenic drift, leading to human Emerging strains lack immunity, causing influenza virus outbreaks
Therefore, there is a certain lag in preventing influenza through vaccination, and the preventive effect is limited
The drugs used to treat influenza viruses are mainly amantadines (such as amantadine and rimantadine) and neuraminidase inhibitors (such as oseltamivir and zanamivir), but these drugs are due to The emergence of drug resistance and its inherent toxic side effects limit clinical application

Method used

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  • Application of Flammulina velutipes polysaccharide extract in the preparation of antiviral drugs and its extraction method
  • Application of Flammulina velutipes polysaccharide extract in the preparation of antiviral drugs and its extraction method
  • Application of Flammulina velutipes polysaccharide extract in the preparation of antiviral drugs and its extraction method

Examples

Experimental program
Comparison scheme
Effect test

Embodiment 1

[0044] The preparation of embodiment 1 Flammulina velutipes polysaccharide extract

[0045](1) Take the fruiting bodies of Flammulina velutipes, pulverize them with a pulverizer (25000r / min, 5s), use the No. 1 Pharmacopoeia inspection sieve to screen, collect the filtrate, add it to 15 times the weight of water to soak until it is completely wet, and heat and extract for 1.5h , filter, and collect the filtrate; re-add water to the filter residue 10 times, heat and extract for 1 hour, filter, collect the filtrate, combine the obtained filtrate, concentrate under reduced pressure at 50°C to about 20L, and further concentrate to about 60% of the barley degree to obtain a concentrated solution;

[0046] (2) Add dehydrated ethanol to the concentrated solution obtained in step (1) so that the volume fraction of dehydrated alcohol is 85%, let it stand overnight, take the precipitate and transfer it to a baking tray, and dry it in vacuum at 45°C to obtain the extract of Flammulina velu...

Embodiment 2

[0047] Cytotoxicity research of embodiment 2 Flammulina velutipes polysaccharide

[0048] The polysaccharide extract of Flammulina velutipes obtained in Example 1 was diluted with PBS to prepare a 3 mg / mL drug stock solution, and stored at 4°C.

[0049] MDCK cells (canine kidney cells) in good growth state were digested with EDTA-trypsin and counted at 2×10 4 / 100μL / well seeded cells in 96-well plate, 37°C, 5% CO 2 Cell culture incubator for 24 hours. When the cell confluence is nearly 90%, the cell culture medium is removed, and the cells are washed twice with 1×PBS (phosphate buffered saline). The drug mother solution was serially diluted by 2 times in DMEM containing 2% fetal bovine serum, and the concentrations of the drug dilutions were 100 μg / mL, 200 μg / mL, 400 μg / mL, 800 μg / mL, 1000 μg / mL, and 2000 μg / mL. Add the drug solution to the above-mentioned cell culture plate at 100 μL / well, add 100 μL / well of the blank control well and the normal cell control well to the ME...

Embodiment 3

[0054] Example 3 Anti-influenza Virus Research of Flammulina velutipes Polysaccharides in Vitro

[0055] MDCK cells in good growth state were divided into about 2 × 10 per well 4 Density seeded into 96-well cell culture plates at 37°C, 5% CO 2 The cells were cultured in the incubator for about 24 hours, and the degree of cell confluence was nearly 100%. The cell culture medium was removed, the cells were washed twice with 1×PBS, and different drug stock solutions were serially diluted by 2 times with DMEM containing 2% fetal bovine serum. The Flammulina velutipes polysaccharide extract obtained in Example 1 was diluted with PBS to make a medicinal solution and added to the above-mentioned cell culture plate at 50 μL / well, and 50 μL / well of virus control wells and normal cell control wells were added with 2% fetal bovine serum. DMEM basal medium, 37°C, 5% CO 2 Continue culturing in the cell culture incubator. At the same time, 25 μL of the drug solution was mixed with 25 μL...

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Abstract

The present invention relates to the medical application of polysaccharide extract, in particular, relates to the application of the polysaccharide extract of Flammulina velutipes in the preparation of anti-influenza virus medicine and its extraction method. The invention provides the application of the polysaccharide extract of Flammulina velutipes in the preparation of antiviral drugs, food or health care products. The present invention proposes for the first time that Flammulina velutipes polysaccharide extract has clear anti-influenza virus effect, and the obtained Flammulina velutipes polysaccharide extract is subjected to in vitro anti-influenza virus drug efficacy research, and the results show that Flammulina velutipes polysaccharide extract can significantly inhibit influenza virus infection. Pathological changes; the extracted Flammulina velutipes polysaccharide extract is carried out animal experiment (H1N1 virus infection mouse experiment), the result confirms that Flammulina velutipes polysaccharide extract can effectively relieve the respiratory symptoms caused by H1N1 virus infection, illustrates that Flammulina velutipes polysaccharide extract is used in the preparation of anti-influenza virus The drug holds promise.

Description

technical field [0001] The present invention relates to the medical application of polysaccharide extract, in particular, relates to the application of the polysaccharide extract of Flammulina velutipes in the preparation of anti-influenza virus medicine and its extraction method. Background technique [0002] Influenza is an acute respiratory disease caused by influenza virus (Influenza Virus), which has the characteristics of strong infectivity, rapid spread, and widespread prevalence, which seriously threatens human life and health and poses a huge economic threat. Human influenza virus is a kind of influenza virus, which is divided into three types: A (A), B (B), and C (C). Among them, type A influenza virus is the most threatening to humans because of its antigenicity that is most likely to mutate. The 2009 influenza A (H1N1) pandemic occurred worldwide, highlighting the severity of influenza A virus infection. [0003] Vaccination is the most effective way to prevent ...

Claims

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Application Information

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Patent Type & Authority Patents(China)
IPC IPC(8): A61K31/715A61P31/18A61K36/07
CPCA61K31/715A61K36/07A61P31/18
Inventor 苏冀彦吕淑媚谢意珍吴清平
Owner GUANGDONG INST OF MICROBIOLOGY GUANGDONG DETECTION CENT OF MICROBIOLOGY
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