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Ebola virus small molecular inhibitor

A virus inhibitor, Ebola virus technology, applied in antiviral agents, medical preparations containing active ingredients, organic active ingredients, etc., can solve problems such as limiting the research progress of antiviral drugs

Inactive Publication Date: 2018-11-23
INST PASTEUR OF SHANGHAI CHINESE ACADEMY OF SCI
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  • Summary
  • Abstract
  • Description
  • Claims
  • Application Information

AI Technical Summary

Problems solved by technology

Because the operation of using live Ebola virus needs to be carried out in the BSL-4 laboratory, the research progress of antiviral drugs is limited

Method used

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Examples

Experimental program
Comparison scheme
Effect test

Embodiment 1

[0086] Example 1 Optimization of screening conditions

[0087] The screening process was performed in 96-well white plates. In the BSRT7 / 5 cells cultured for 16-24 hours, after washing twice with Opti-MEM, transfer 50 μl of Ebola minigenome, NP, VP35, VP30, L 5 kinds of plasmid mixture, and transfect for 2 hours. By comparison, 20,000 BSR T7 / 5 cells per well in a 96-well plate were transformed into minigenome (25ng), NP (25ng), VP35 (25ng), VP30 (15ng), L (150ng) with the best effect Well, the maximum signal-to-background ratio (S / B) and better Z-factor (Z-factor) will be obtained. Select the above conditions to screen the natural product compound library. In order to ensure that the drug does not affect the transfection, 5 μl of the drug was added 2 hours after the transfection, and the concentration of all the drugs was 5 mg / ml. The experiment without L plasmid was used as the control group, and 5 μl of 5% DMSO was added. Finally, 45 μl of medium was added to each well s...

Embodiment 2

[0088] Example 2 9-cis retinoic acid inhibits Ebola virus replication

[0089] By screening the natural product compound library, 69 drugs with an inhibition rate of more than 50% were obtained in the first screening. By further testing the toxicity of these drugs, 5 drugs with toxicity less than 10% and inhibition rate greater than 80% were screened out, and the detection rate was 0.99%. For further verification, the plasmid RL-SV40 transfected with Renilla luciferase was used for dual fluorescence detection in 24-well plates. The verification results found that 9-cis retinoic acid inhibited the replication of Ebola virus. After removing the effect of cytotoxicity, the inhibition rate reached 94% when the concentration was 25uM, and the EC50 was 8.18uM. See the results figure 1 .

Embodiment 3

[0090] Example 3 9-cis retinoic acid inhibits the replication of respiratory syncytial virus

[0091] The present inventor detected this drug with double fluorescence in the respiratory syncytial virus (RSV) minigenome detection system, and found that 9-cis retinoic acid has an inhibitory effect on the replication of respiratory syncytial virus (RSV), with an EC50 of 24.34uM. Then we did a virus titer reduction experiment in live RSV virus, which confirmed that 9-cis retinoic acid has an inhibitory effect on RSV. see results figure 2 and image 3 .

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Abstract

The invention provides an Ebola virus small molecular inhibitor. Specifically, the invention provides a small molecular inhibitor of Ebola fake virus. The inhibitor is selected from 9-cis-retinoic acid, 13-cis-retinoic acid, all-trans retinoic acid, or a combination thereof. Experimental results show that the inhibitor can effectively inhibit Ebola virus from replicating.

Description

technical field [0001] The invention belongs to the field of biotechnology, in particular, the invention relates to small molecule inhibitors of Ebola virus. Background technique [0002] Ebola virus is a virulent virus that causes Ebola hemorrhagic fever in humans and primates. It is highly contagious and can cause fatal Ebola virus disease (EVD), with symptoms including nausea, vomiting, diarrhea, skin color changes, body aches, internal bleeding, external bleeding, and fever. Ebola broke out in West Africa in 2014, with high morbidity and high fatality rate. As of March 30, 2016, 28,646 people became ill and 11,323 died (http: / / apps.who.int / ebola / ebola-situation-reports). [0003] At present, there are no specific therapeutic drugs or vaccines approved in the world. Now the main research direction is to develop anti-viral small molecules, antisense technology and monoclonal antibody cocktail therapy (such as ZMapp). Broad-spectrum virus inhibitors show obvious defects...

Claims

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Application Information

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IPC IPC(8): A61K31/203A61P31/14
CPCA61K31/203
Inventor 王莉莉李桂明邹罡
Owner INST PASTEUR OF SHANGHAI CHINESE ACADEMY OF SCI
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