Application of cepharanthine in preparation of anti-influenza virus drugs

An anti-influenza virus, fenugreek technology, applied in antiviral agents, pharmaceutical formulations, plant raw materials, etc., can solve the problem that the application of fenugreek in anti-influenza virus has not been reported, and achieve broad-spectrum antiviral activity, good Application prospects, the effect of suppressing copying

Active Publication Date: 2022-04-08
SOUTHERN MEDICAL UNIVERSITY
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  • Summary
  • Abstract
  • Description
  • Claims
  • Application Information

AI Technical Summary

Problems solved by technology

The above research results all suggest that the clinical application value of stepherin is very great, but the application of stepherin in anti-influenza virus has not been reported yet.

Method used

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  • Application of cepharanthine in preparation of anti-influenza virus drugs
  • Application of cepharanthine in preparation of anti-influenza virus drugs
  • Application of cepharanthine in preparation of anti-influenza virus drugs

Examples

Experimental program
Comparison scheme
Effect test

Embodiment 1

[0045] Example 1: Detection of Stephalin on Cytotoxicity

[0046] The cytotoxicity of cephaglidin is detected by MTT method, so as to determine the concentration of the drug. The specific method is as follows:

[0047] MDCK or A549 cells by 1×10 4 / well seeded in 96-well plate at 37°C, 5% CO 2 Cultured in a constant temperature cell culture incubator until monolayer, stepwise dilute stepherin with serum-free DMEM or 1640 (DMEM medium for MDCK cells, 1640 medium for A549 cells) and add it to a 96-well plate, 200 μL per well , continue to cultivate for 48h. Discard the culture supernatant, add 100 μL of 1640 or DMEM medium containing 0.5 mg / mL MTT to each well, and incubate at 37 °C for 4 h. DMSO was used as the control group. A multifunctional microplate reader (Genios Pro, Tecan, US) was used to detect the absorbance at 570nm, and the cell survival rate was used as an indicator of the toxicity of stephalin to MDCK or A549 cells.

[0048] Cell viability (%)=E / N×100

[004...

Embodiment 2

[0052] Example 2: Anti-Influenza A Virus Activity Detection of Stephalin in Vitro

[0053] In the in vitro antiviral experiment of the present invention, various subtypes of influenza A viruses are involved, including H1N1 and H3N2, and the specific methods are as follows:

[0054] MDCK cells by 2 x 10 4 / well seeded in 96-well plate at 37°C, 5% CO 2 cultured to a monolayer in a constant temperature cell incubator. Use 100TCID 50 H3N2, PR8 or WSN-infected cells, 100 μL per well, incubated at 37°C for 1 h, discarded the virus solution, and added stepped stepherin diluted in DMEM (containing 1 μg / mL TPCK), 200 μL per well, continued to culture for 48 h, combined with MTT method and plaque assay to detect the antiviral activity of stepherin. Calculate the half effective concentration IC by observing the cytopathic phenomenon (CPE) caused by the virus and detecting the survival rate of the cells 50 .

[0055] Virus inhibition rate (%)=1-(E / N) / (P-N)×100

[0056] E is the abs...

Embodiment 3

[0061] Example 3: Inhibitory Experiment of Stephalin on the Replication of Influenza A Virus

[0062] In order to evaluate the inhibitory effect of stepherin on the replication of influenza virus, the present invention uses methods such as q-PCR and Western blotting to detect the influence of stepherin on virus replication from the expression levels of genes and proteins respectively. The specific methods are as follows:

[0063] q-PCR and Western blot: A549 cells by 4×10 5 / well seeded in a 6-well plate at 37 °C, 5% CO 2 cultured to a monolayer in a constant temperature cell incubator. Infect the cells with 100TCID50 of WSN, 1mL per well, incubate at 37°C for 1h, discard the virus solution, add Stephalogenin gradiently diluted in DMEM (containing 1μg / mL TPCK), 1mL per well, continue to culture for 24h, then use RNA extraction reagent Kit (provided by Fuji Bio) to fully lyse cells and extract total RNA, samples for q-PCR; or use 75 μL RIPA lysate containing protease and phos...

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Abstract

The invention discloses an application of cepharanthine in preparation of anti-influenza virus drugs in the technical field of medicines, and particularly relates to influenza A viruses, such as influenza viruses H1N1, H3N2, H5N1, H7N9, H9N2, H10N8 and the like. The invention aims to provide an application of a leukocyte hyperplasia drug cepharanthine in preparation of an anti-influenza virus infection drug, and discovers that cepharanthine has anti-influenza virus activity and can specifically destroy the activity of an influenza virus ribonucleoprotein complex vRNP so as to influence replication of influenza viruses for the first time. As a novel anti-influenza virus drug and a lead compound, cepharanthine has the characteristics of low toxicity, high activity, low price, easiness in obtaining and clear mechanism, can be used for developing novel anti-influenza virus drugs under existing application conditions, and has a relatively good application prospect.

Description

technical field [0001] The invention belongs to the technical field of medicine, and in particular relates to the application of paternin in the preparation of anti-influenza virus drugs. Background technique [0002] Influenza is one of the major infectious diseases that threaten human health. It has always attracted the attention of people in the field of disease prevention and control. However, due to the characteristics of easy change and drift of influenza virus antigens, the situation of influenza prevention and control is not optimistic. Among them, influenza A virus Because of its strong infectivity, rapid mutation, high morbidity and mortality, it has become one of the viruses that seriously threaten human health and social stability. Vaccination is currently the main means of preventing influenza virus infection, but the update speed of the vaccine lags behind the mutation of influenza virus, which makes conventional seasonal vaccines unable to effectively prevent ...

Claims

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Application Information

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Patent Type & Authority Applications(China)
IPC IPC(8): A61K31/4741A61K36/59A61P31/16
Inventor 杨洁桑会婷陈柳蓉李银燕
Owner SOUTHERN MEDICAL UNIVERSITY
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