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New application of OGG1 small-molecule inhibitor in treating African swine fever

A small molecule inhibitor, African swine fever technology, applied in the field of African swine fever, OGG1 small molecule inhibitors, can solve the problems of vaccines and antiviral drugs without African swine virus, affecting pork supply and export trade, etc., reaching Effects of inhibiting early infection and inhibiting proliferation

Active Publication Date: 2021-07-16
LANZHOU INST OF VETERINARY SCI CHINESE ACAD OF AGRI SCI
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  • Summary
  • Abstract
  • Description
  • Claims
  • Application Information

AI Technical Summary

Problems solved by technology

Its severity has had a huge impact on the world economy and trade and the domestic pig breeding industry. After the African swine fever epidemic first appeared in my country in 2018, it quickly spread to more than 30 provinces in China, seriously affecting my country's pork supply and export trade. Effective vaccines and antiviral drugs against African swine virus, so the development of vaccines and antiviral drugs is a key issue that needs to be solved urgently in our country

Method used

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  • New application of OGG1 small-molecule inhibitor in treating African swine fever
  • New application of OGG1 small-molecule inhibitor in treating African swine fever
  • New application of OGG1 small-molecule inhibitor in treating African swine fever

Examples

Experimental program
Comparison scheme
Effect test

Embodiment 1

[0051] Example 1 Verification of siRNA interference effect of OGG1

[0052] 1. Protein level detection process

[0053] (1) PAM cells with 2×10 6 Spread in 6-well plates and culture for 24 hours;

[0054] (2) Collect the cells, discard the supernatant by centrifugation, and wash once with PBS;

[0055] (3) Resuspend the interference group cells in 2ml of transfection reagent containing siRNA; the specific components are as follows:

[0056] Double-antibody-free 1640 medium, 1863 μL; Buffer, 120 μL; Reagent, 12 μL; siRNA, 5 μL; siRNA final concentration 50 nM; control group cells were resuspended in 2 mL double-antibody-free medium containing siNC, then inoculated into a 6-well plate, and interfered for 24 hours ;

[0057] (4) Collect the cells, discard the supernatant by centrifugation, add RIPA and lyse on ice for 30 min;

[0058] (5) Centrifuge the lysate at 4°C, 12000r for 15min, and collect the supernatant as the whole protein;

[0059] (6) To measure the protein con...

Embodiment 2

[0075] Example 2 Interfering with OGG1 inhibits ASFV proliferation

[0076] 1. Protein level detection process

[0077] The specific steps refer to the "1. Protein level detection process" in the above Example 1, wherein the step (8) antibody incubation is as follows: take out the nitrocellulose membrane and place it in TBST containing 5% skimmed milk powder for blocking for 1 hour, discard the blocking solution Then wash 3 times with TBST, 5 minutes each time, add p30 and p72 antibodies after membrane shearing, and incubate overnight at 4°C. The next day, wash 3 times with TBST, 5 minutes each time, incubate the secondary antibody of the corresponding species in the dark for 45 minutes, wash 3 times with TBST, 5 minutes each time, add the chromogenic solution in the dark, and use the system to scan the membrane. Band grayscale analysis.

[0078] 2. mRNA level detection process:

[0079] The specific steps refer to the "2.mRNA level detection process" in the above-mentioned...

Embodiment 3

[0083] Inhibition of adhesion to ASFV virus after embodiment 3 interferes with OGG1

[0084] 1. mRNA level detection process:

[0085] The specific steps refer to the "2.mRNA level detection process" in the above-mentioned embodiment 2, wherein, in the step (3) described in 2 in the embodiment 2, ASFV (MO=0.5) is added to the cells of the interference group and the control group Infection was performed at 4°C for 1 h.

[0086] 2. Results

[0087] After ASFV infected cells that interfered with the expression of OGG1, the results of ASFV virus adhesion were as follows: Image 6 As shown, the results show that RNA interference OGG1 can significantly inhibit the adhesion of ASFV virus.

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Abstract

The invention belongs to the technical field of African swine fever treatment, and particularly relates to a new application of an OGG1 small-molecule inhibitor for treating African swine fever. Firstly, accidental discoveries show that the mRNA level and protein level of ASFV structural proteins p30 and p72 can be remarkably reduced and the replication and adhesion of ASFV can be inhibited by using RNA to interfere the expression of a host OGG1; secondly, discoveries show that the OGG1 small-molecule inhibitor can remarkably inhibit expression of the structural protein p30 related to ASFV early-stage expression, and early-stage infection of ASFV is inhibited; and meanwhile, the OGG1 small-molecule inhibitor can remarkably inhibit expression of the structural protein p72 related to ASFV advanced expression, can remarkably inhibit ASFV proliferation and can be used for preventing or treating African swine fever.

Description

technical field [0001] The invention belongs to the technical field of treating African swine fever, and in particular relates to a new application of an OGG1 small molecule inhibitor for treating African swine fever. Background technique [0002] African swine fever (African swine fever, ASF) is a disease caused by highly pathogenic African swine fever virus (African swine fever virus, ASFV) after infection of pigs, which spreads quickly and has a high fatality rate. Disorders, visceral bleeding, and depression are the main features. Its severity has had a huge impact on the world economy and trade and the domestic pig breeding industry. After the African swine fever epidemic first appeared in my country in 2018, it quickly spread to more than 30 provinces in China, seriously affecting my country's pork supply and export trade. Effective vaccines and antiviral drugs against African swine virus, so the research and development of vaccines and antiviral drugs is a key issue t...

Claims

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Application Information

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Patent Type & Authority Applications(China)
IPC IPC(8): A61K31/454A61K31/47A61K31/381A61K31/7105A61P31/20
CPCA61K31/454A61K31/47A61K31/381A61K31/7105A61P31/20Y02A50/30
Inventor 牛庆丽刘志杰杨吉飞樊洁吕欣倩关贵全殷宏
Owner LANZHOU INST OF VETERINARY SCI CHINESE ACAD OF AGRI SCI
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