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Application of GC376 in preparation of novel coronavirus SARS-CoV-2 inhibitor

A technology of 1. GC376 and coronavirus, which is applied in antiviral agents, respiratory diseases, medical preparations containing active ingredients, etc., can solve the problems of no vaccine and the new coronavirus is in the initial stage of exploration, and achieve less toxic and side effects , high safety effect

Inactive Publication Date: 2020-05-12
HUAZHONG AGRI UNIV +1
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  • Summary
  • Abstract
  • Description
  • Claims
  • Application Information

AI Technical Summary

Problems solved by technology

Since the outbreak of the epidemic, research on the infectious disease has mainly focused on epidemiological investigations, sequence comparison and analysis of clinically isolated strains, virus detection methods, etc. Research on the invasion, replication, and immune regulation mechanisms of new coronaviruses Research on antiviral drugs for SARS-CoV-2 is of great importance as it is still in the early stages of exploration and no vaccine can provide sufficient protection

Method used

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  • Application of GC376 in preparation of novel coronavirus SARS-CoV-2 inhibitor
  • Application of GC376 in preparation of novel coronavirus SARS-CoV-2 inhibitor
  • Application of GC376 in preparation of novel coronavirus SARS-CoV-2 inhibitor

Examples

Experimental program
Comparison scheme
Effect test

Embodiment 1

[0016] Example 1 Toxicity test of GC376 to cells

[0017] experiment method:

[0018] 1) Take Vero-E6 cells (African green monkey kidney cells) in good growth state for digestion and passage, and adjust the cell density to 1×10 with cell growth medium. 5 / mL to inoculate a 96-well plate, 100 μL / well, placed at 37°C, 5% CO 2 Cultivate in an incubator for 16 hours;

[0019] 2) After 16 hours, discard the culture medium in the well, wash three times with 1×PBS, dry it, add cell maintenance solution, and add GC376 at the same time, so that the final concentration of GC376 in the well is 10000 μM, 1000 μM, 500 μM, 100 μM, 50 μM, 25 μM, 12.5 μM, 6.25μM, set cell control, place at 37°C, 5% CO 2 incubator cultivation;

[0020] 3) After 72h, use Reagents for cell viability assays.

[0021] test results:

[0022] see results figure 1 . Determination of cell viability can reflect the toxic effect of GC376 on Vero-E6 cells. As can be seen from the figure, GC376 has less toxic ef...

Embodiment 2

[0023] Example 2 The inhibitory effect of GC376 on the virus at the cellular level

[0024] experiment method:

[0025] 1) Take Vero-E6 cells in good growth state for digestion and passage, and adjust the cell density to 1×10 5 / mL inoculated in a 96-well plate, 100 μL / well, placed at 37°C, 5% CO 2 Cultivate in an incubator for 16 hours;

[0026] 2) After 16 hours, discard the culture medium in the well, wash with 1×PBS three times, add 100 μL of GC376 with a concentration of 100 μM to each well after drying, and add 100 μL of 0.01 MOI of SARS-CoV-2 virus to each well, so that the final concentration of GC376 is 50μM, set cell control and virus control at the same time, place at 37°C, 5% CO 2 incubator cultivation;

[0027] 3) After 48 hours of inoculation, observe the changes in the cell state after the compound and the virus interact with a microscope and take pictures for preservation.

[0028] test results:

[0029] see results figure 2 . From the microscope pictu...

Embodiment 3

[0030] Embodiment 3 GC376 antiviral activity (EC 50 )test

[0031] experiment method:

[0032] 1) Take Vero-E6 cells in good growth state for digestion and passage, and adjust the cell density to 1×10 with cell growth medium. 5 / mL inoculated in a 96-well plate, 100 μL / well, placed at 37°C, 5% CO 2 Cultivate in an incubator for 16 hours;

[0033] 2) After 16 hours, dilute GC376 with cell maintenance solution so that the concentrations are 200 μM, 100 μM, 50 μM, 25 μM, 12.5 μM, 6.25 μM, 3.125 μM;

[0034] 3) Mark the lid of the 96-well plate, discard the culture medium in the well, wash with 1×PBS three times, and dry it. Add 100 μL of diluted GC376 to each well in the order of marking, and add 100 μL of 0.01 MOI virus solution to each well. Make the final concentration of GC376 100 μM, 50 μM, 25 μM, 12.5 μM, 6.25 μM, 3.125 μM, 1.5625 μM, set the cell control group and the virus control group, and place them at 37°C, 5% CO 2 incubator cultivation;

[0035] 4) After 72h, u...

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Abstract

The invention discloses an application of GC376 in preparation of a novel coronavirus SARS-CoV-2 inhibitor. In-vitro cell test results prove that GC376 can notably inhibit infection of SARS-CoV-2 on normal cells, has an inhibition function on SARS-CoV-2 in terms of cell level, can notably reduce toxicity of the virus in cells and inhibit proliferation of the virus in the cells and is dose-dependent. Therefore, GC376 can serve as the novel coronavirus SARS-CoV-2 inhibitor and has the potential of treating COVID-19 caused by the virus or other diseases caused after animals are infected.

Description

technical field [0001] The invention belongs to the field of pharmacy, and in particular relates to the new application of GC376 in preparing novel coronavirus SARS-CoV-2 inhibitors. Background technique [0002] Corona Virus Disease 2019 (COVID-19), referred to as "New Coronary Pneumonia", is a disease caused by the new coronavirus SARS-CoV-2, characterized by fever, dry cough, fatigue and a certain lethality. Contact infectious diseases. [0003] Since the outbreak of COVID-19 in December 2019, the number of confirmed cases of infection has surpassed that of SARS, and the number of deaths has continued to increase, causing serious social harm and economic losses. public health event". Since the outbreak of the epidemic, research on the infectious disease has mainly focused on epidemiological investigations, sequence comparison and analysis of clinically isolated strains, virus detection methods, etc. Research on the invasion, replication, and immune regulation mechanisms...

Claims

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Application Information

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Patent Type & Authority Applications(China)
IPC IPC(8): A61K31/4015A61P31/14A61P11/00
CPCA61K31/4015A61P11/00A61P31/14
Inventor 彭贵青刘紫微陈西田大勇
Owner HUAZHONG AGRI UNIV
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