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3C-like protease inhibitor of porcine epidemic diarrhea virus, and screening method and application of 3C-like protease inhibitor

A technology of protease inhibitors and porcine epidemic diarrhea, which is applied in the field of anti-virus, can solve the problems of no therapeutic drugs and inability to protect piglets, etc.

Pending Publication Date: 2021-11-09
WUHAN POLYTECHNIC UNIVERSITY
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  • Summary
  • Abstract
  • Description
  • Claims
  • Application Information

AI Technical Summary

Problems solved by technology

Vaccines are an effective means of preventing the disease, but vaccines do not protect infected piglets and there is currently no treatment for the disease

Method used

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  • 3C-like protease inhibitor of porcine epidemic diarrhea virus, and screening method and application of 3C-like protease inhibitor
  • 3C-like protease inhibitor of porcine epidemic diarrhea virus, and screening method and application of 3C-like protease inhibitor
  • 3C-like protease inhibitor of porcine epidemic diarrhea virus, and screening method and application of 3C-like protease inhibitor

Examples

Experimental program
Comparison scheme
Effect test

Embodiment 1

[0077] Example 1 Screening of PEDV 3C-like protease inhibitors

[0078] 1. Virtual Screening

[0079] (1) Preparation of receptor molecules: Autodock 1.5.6 software was used to dehydrate and hydrotreat the crystal structure of 3C-like protease (www.rcsb.org, protein number 6U7K) and store the molecules as PDBQT files. Use DiscoverStudio 3.5 to open the receptor molecule, select the amino acid of the active site of the protein, and determine the molecular docking area, including the coordinates of the central site and the size of the docking area. Preparation of ligand molecules: Download the molecular structures (about 80,000 molecules) in the natural molecular library on ZINC15, and use Raccon software to convert all molecules into PDBQT files respectively. Then prepare the bat file according to the above, use Autodock Vina to run the bat file for molecular docking, extract the results after screening, sort all the molecular docking results, and select the top 10,000 molecul...

Embodiment 2

[0112] Example 2 Validation of the anti-PEDV effect of PEDV 3C-like protease inhibitors

[0113] Test method: use PEDV YN144 strain to infect Vero cells (MOI=0.001), and add different concentrations of chebulinic acid (chebulinic acid) at the same time. Samples were collected 24 hours after infection, and the effects of chebulinic acid (chebulinic acid, purchased from Chengdu Purifa Technology Co., Ltd., purity>95%) on the replication of PEDV were evaluated using TCID50 assay, western blot and indirect immunofluorescence test. The results are as follows: Figures 8 to 10 shown.

[0114] Depend on Figures 7 to 9 It can be seen that Chebulinic acid can effectively inhibit the infection of PEDV in vitro.

[0115] In summary, in the screening method for PEDV 3C-like protease inhibitors provided by the present invention, through virtual screening, PEDV 3C-like protease inhibitors can be efficiently screened out from a large number of small molecules; through the provided FRET sy...

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Abstract

The invention discloses a 3C-like protease inhibitor of a porcine epidemic diarrhea virus, and a screening method and application of the 3C-like protease inhibitor. The 3C-like protease inhibitor comprises at least one of chebulinic acid, an isomeride of the chebulinic acid, quercetin-3-O-p-coumaroyl rhamnose glucoside, troxerutin, citrate B, apigenin-7-O-(2G-rhamnose) gentian glucoside, brutieridin, Heraclenol 3'-O-[beta]-D-apiofuranosyl-(1->6)-[beta]-D-glucopyranoside, Procyanidin B23, 3'-di-O-gallate, citrate A and benzoic acid benzyl alcohol glucoside. Since the 3C-like protease plays an important role in transcription and replication of the porcine epidemic diarrhea virus and an anti-host-cell defense mechanism process, the 3C-like protease inhibitor provided by the invention can prevent the porcine epidemic diarrhea virus from infecting host cells by inhibition of the activity of the porcine PEDV3C-like protease, and an inhibition effect is relatively good. Furthermore, the 3C-like protease inhibitor provided by the invention can be used for preparing a medicine for resisting the porcine epidemic diarrhea virus, and a new application way is developed for the above molecule.

Description

technical field [0001] The invention relates to the field of antiviral technology, in particular to a 3C-like protease inhibitor of porcine epidemic diarrhea virus, a screening method thereof, and an application thereof. Background technique [0002] Porcine epidemic diarrhea (PED) is a highly contagious porcine intestinal infectious disease caused by porcine epidemic diarrhea virus (PEDV), characterized by vomiting, watery diarrhea, and dehydration in pigs. main features. PED affects pigs of all ages, especially for suckling piglets within 7 days of age, with a mortality rate as high as 100%. It is an important factor affecting the survival rate of piglets and pig production. Vaccines are an effective means of preventing the disease, but vaccines do not protect infected piglets, and there are currently no therapeutic drugs against the disease. Contents of the invention [0003] The main purpose of the present invention is to propose a 3C-like protease inhibitor of porci...

Claims

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Application Information

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Patent Type & Authority Applications(China)
IPC IPC(8): C12Q1/37C12N15/85C12N15/62A61K45/00A61K31/7048A61P31/14G16B50/00
CPCC12Q1/37C12N15/85C12N9/506A61K45/00A61K31/7048A61P31/14G16B50/00C12N2800/107C07K2319/61
Inventor 李中华侯永清吴涛张倩易丹赵迪王蕾
Owner WUHAN POLYTECHNIC UNIVERSITY
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