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Application of a small molecular compound in the preparation of anti-Zika virus drugs

A small molecular compound, Zika virus technology, applied in the field of animal virology, can solve the problems of drugs that have not entered clinical trials and limited compounds, and achieve good development value, good specificity, and clear targets

Active Publication Date: 2021-09-21
HUAZHONG AGRI UNIV
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  • Summary
  • Abstract
  • Description
  • Claims
  • Application Information

AI Technical Summary

Problems solved by technology

[0004] At present, research on antiviral drugs for Zika virus has been carried out extensively. Although many compounds with inhibitory activity against the virus have been discovered, compounds with good activity and clear targets are very limited, and no drugs have entered clinical trials.

Method used

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  • Application of a small molecular compound in the preparation of anti-Zika virus drugs
  • Application of a small molecular compound in the preparation of anti-Zika virus drugs
  • Application of a small molecular compound in the preparation of anti-Zika virus drugs

Examples

Experimental program
Comparison scheme
Effect test

Embodiment 1

[0035] Embodiment 1: Determination of the inhibitory effect of the compound of formula 1 on Zika virus NS3 protease

[0036] Add the compound of formula 1 into the established screening system (see Table 1), and dilute it into five gradients according to 50 μM-3.2 μM. Three repetitions were set up for each experiment. The mixture of buffer and NS3 protease mixed in advance was added to the black 384-well plate, and then the compound of formula 1 was added and diluted to 50 μM, 25 μM, 12.5 μM, 6.3 μM, Five concentration gradients of 3.2 μM were added to the fluorescent substrate (GenScript Biotechnology Co., Ltd.) after acting for 30 minutes at room temperature, shaken for 1 minute, and immediately placed in a microplate reader to detect the fluorescence value. Fluorescence intensity at zero time (F0), and then put the 384-well plate in a 37°C incubator, react for 60 minutes, and detect the fluorescence intensity (F1) of the system in a microplate reader again. In the test, co...

Embodiment 2

[0041] Example 2: The plaque reduction effect of the compound of formula 1 on the proliferation of Zika virus

[0042] (1) Take Vero cells in good growth state (preserved in this laboratory, common cells) for subculture, count the cells, and adjust the cell suspension density to 5x10 with cell growth medium 5 individual / mL. Inoculate 12-well cell culture plate, 1ml / well. Only the cell growth solution was added to the surrounding wells. 37°C 5% CO 2 Incubator for 12h. After 12 hours, the old cell growth solution was discarded and replaced with cell maintenance solution.

[0043] (2) Add the compound to 10 -4 In the diluted Zika virus liquid, the final concentrations were 60, 50, 40, 30, 20, and 10 μmol / L respectively. Discard the cell maintenance liquid in the 12-well plate, and add diluted compounds with different concentrations in turn, 37°C, 5% CO 2 The incubator continued to cultivate for 48h.

[0044] (3) After 48 hours, the 12-well plate was frozen at -80°C for la...

Embodiment 3

[0059] Example 3: Inhibitory effect of the compound of formula 1 on protein expression of Zika virus in cells

[0060] (1) Take Vero cells in a good growth state for subculture, count the cells, and adjust the cell suspension density to 5x10 using cell growth medium. 5 individual / mL.

[0061] (2) Inoculate 24-well cell culture plate, 500 μL / well. 37°C 5% CO 2 Incubator for 12h.

[0062] (3) After 12 hours, discard the old cell growth solution and replace with the cell maintenance solution.

[0063] (4) Add compound to 10 -4 The final concentrations in the diluted Zika virus liquid were 5 μmol / L and 20 μmol / L, respectively.

[0064] (5) Discard the cell maintenance solution in the 24-well plate, add 300 μL of the mixture of the compound of formula 1 and the Zika virus solution at different concentrations in turn, and set the cell and Zika virus control group at the same time, 37 ° C 5% CO 2 Cultivate in the incubator for 24h.

[0065] (6) After 24 hours, discard the supe...

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Abstract

The invention belongs to the technical field of animal virus prevention and control, and in particular relates to the application of a small molecular compound in the preparation of anti-Zika virus drugs. The invention includes the screening of an anti-Zika virus drug target, the drug target is a small molecule compound, and the small molecule compound can be used in preparing anti-Zika virus drugs. The application thereof includes the application in the preparation of anti-Zika virus protease inhibitors. The small molecular compound provided by the present invention can inhibit the activity of NS3 protease encoded by Zika virus, and has a good inhibitory effect on the proliferation of the virus, and the compound has a certain protection rate against Zika virus-challenged mice, which can significantly improve The pathological symptoms of the challenged mouse tissue can be applied to the development of animal anti-Zika virus drugs.

Description

technical field [0001] The invention belongs to the technical field of animal virology, and in particular relates to the screening of an anti-Zika virus drug target, the drug target is a small molecular compound, and the small molecular compound can be used in the preparation of an anti-Zika virus drug . The application thereof includes the application in the preparation of anti-Zika virus protease inhibitors. Background technique [0002] Zika virus is a member of the flavivirus genus of the family Flaviviridae. The virus first broke out in Uganda, Africa in 1952, and only sporadically occurred in the world in 2007. However, the Zika virus first broke out in Micronesia in 2007, and then broke out in French Polynesia in 2013-2014. In 2015, the Zika virus broke out in Brazil, causing more than 30,000 people to be infected. The virus spreads rapidly and has spread to more than 80 countries and regions in the past 1-2 years, causing a huge panic to global public health. [0...

Claims

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Application Information

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Patent Type & Authority Patents(China)
IPC IPC(8): A61K31/122A61P31/14
CPCA61K31/122A61P31/14Y02A50/30
Inventor 宋云峰苗娟邹佳辉曹胜波
Owner HUAZHONG AGRI UNIV
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