Application of epigallocatechin gallate (EGCG) in preparing drug for resisting respiratory syncytial virus

A technology of epigallocatechin and gallate, applied in the field of application of epigallocatechin gallate in the preparation of anti-respiratory syncytial virus, can solve toxic and side effects, no safe and effective preventive vaccine, curative effect It is not ideal enough to achieve the effect of reducing the pathological damage of the lungs

Inactive Publication Date: 2015-11-11
WUHAN UNIV
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  • Summary
  • Abstract
  • Description
  • Claims
  • Application Information

AI Technical Summary

Problems solved by technology

At present, ribavirin and interferon are used clinically to treat respiratory syncytial virus infection, which can relieve symptoms, shorten the course of disease and detoxification time, but both have unsatisfactory curative effect and obvious side effects, and there is still no safe and effective prevention vaccine

Method used

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  • Application of epigallocatechin gallate (EGCG) in preparing drug for resisting respiratory syncytial virus
  • Application of epigallocatechin gallate (EGCG) in preparing drug for resisting respiratory syncytial virus
  • Application of epigallocatechin gallate (EGCG) in preparing drug for resisting respiratory syncytial virus

Examples

Experimental program
Comparison scheme
Effect test

Embodiment 1

[0027] Embodiment 1MTT method detects the cytotoxic effect of EGCG

[0028] HEp-2 cells cultured in DMEM (containing 10% fetal bovine serum, V / V) were digested with trypsin and diluted to 1-3×10 5 cells / mL, 100 μL per well was inoculated in a 96-well plate. After the cells grow into a single layer, add 100 μL of DMEM (containing 2% fetal bovine serum, V / V) to dilute to different concentrations (2 μg / mL, 4 μg / mL, 8 μg / mL, 16 μg / mL, 32 μg / mL, 64 μg / mL mL and 128 μg / mL) of EGCG drug (drug group), cultivated for 48h. Cells without EGCG treatment (adding 100 μL of DMEM containing 2% fetal calf serum) were normal control group. The absorbance values ​​of the normal control group and the drug group were detected by the MTT method, and the cell survival rate was calculated (HEp-2 cell survival rate=average absorbance value of the drug group / average absorbance value of the normal control group×100%), thereby evaluating the cytotoxic effect of EGCG.

[0029] The results showed that t...

Embodiment 2E

[0033] The blocking effect of embodiment 2EGCG on virus invasion into cells

[0034] Normal control group: Add EGCG-free DMEM (containing 2% fetal calf serum, V / V) to a 96-well plate that has grown to 80% monolayer HEp-2 cells, 100 μL per well, incubate at 37°C for 2 hours, discard Wash the supernatant 3 times with PBS, then add 20 μL / well of virus-free DMEM (containing 2% fetal bovine serum, V / V), incubate at 35°C for 1 hour, discard the supernatant, wash 3 times with PBS, and then add cell maintenance solution (DMEM containing 2% fetal bovine serum, V / V) 100 μL / well, incubate at 35°C, observe CPE, until ≥75% of the cells in the virus control wells have CPE (+++~++++) and are normal When the cells in the control group were normal, the virus inhibition rate was detected by MTT method.

[0035] Virus control group: Add EGCG-free DMEM (containing 2% fetal calf serum, V / V) to a 96-well plate that has grown to 80% monolayer HEp-2 cells, 100 μL per well, incubate at 37°C for 2 hou...

Embodiment 3E

[0042] Embodiment 3EGCG directly inactivates the virus

[0043] Normal control group: Mix 100 μL of DMEM (containing 2% fetal bovine serum, V / V) without EGCG and 20 μL of DMEM (containing 2% fetal bovine serum, V / V), act at 35°C for 1 hour, and then mix the The solution was added to a 96-well plate filled with 80% monolayer HEp-2 cells, placed in an incubator at 35°C for 1 h, discarded the supernatant, washed 3 times with PBS, and finally added DMEM (containing 2% fetal bovine serum, V / V) 100 μL / well, cultivate at 35°C, observe CPE, and when ≥75% of the cells in the virus control wells have CPE (+++~++++) and the cells in the normal control group are normal, use the MTT method to detect Virus suppression rate.

[0044] Virus control group: 100 μL of DMEM (containing 2% fetal calf serum, V / V) without EGCG and 20 μL of 100 TCID 50 (10 -2.6 / mL) of RSV virus solution was mixed and reacted at 35°C for 1h, and then the mixture was added to a 96-well plate that had grown to 80% m...

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Abstract

The invention discloses an application of epigallocatechin gallate (EGCG) in preparing a drug for resisting a respiratory syncytial virus (RSV), and belongs to the field of antiviral drugs. According to the application, it is discovered that the EGCG can inhibit RSV infection both in vitro and in vivo and is free of toxic and side effects; in vitro, the EGCG can prevent cells from being infected by the RSV and inhibit biosynthesis of the RSV in the cells; in vivo, the EGCG can alleviate lung tissue pathological lesion caused by RSV infection and lower the virus titer of the RSV in the lung tissue. The discovery proves that the EGCG can be used for preparing the drug for resisting the RSV and also can be used for preparing a drug for treating and / or preventing pulmonary diseases caused by the RSV.

Description

technical field [0001] The invention belongs to the field of antiviral drugs, and in particular relates to the application of epigallocatechin gallate (EGCG) in preparing anti-respiratory syncytial virus (RSV). Background technique [0002] Respiratory syncytial virus (RSV) belongs to the Pneumovirus genus of the Paramyxoviridae family. It is an enveloped single-stranded negative-sense RNA virus with one serotype, which can be divided into two subtypes, A and B. Respiratory syncytial virus seriously endangers the health of infants and young children. It is a common respiratory infectious pathogen in hospitalized children. About 40% to 50% of infantile bronchiolitis and 25% of infantile pneumonia hospitalized each year are caused by RSV infection. . Respiratory syncytial virus infection for the first time is mostly infants younger than 1 year old, mainly invading the bronchioles, bronchi and alveoli, leading to respiratory syncytial virus pneumonia. The main clinical manifes...

Claims

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Application Information

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Patent Type & Authority Applications(China)
IPC IPC(8): A61K31/353A61P31/14A61P11/00
Inventor 侯炜王晓昆罗凡曲川张文斐
Owner WUHAN UNIV
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