Pharmaceutical composition composed of GC376 and GS-441524, and application of pharmaceutical composition in inhibiting novel coronavirus

A technology of GS-441524 and GC376, which is applied in the direction of drug combinations, medical preparations containing active ingredients, antiviral agents, etc., can solve problems such as coronaviruses that have not yet been seen, to reduce viral drug resistance, reduce drug side effects, good curative 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

[0010] The above two drugs are currently mainly used to treat feline infectious peritonitis (FIP) clinically. The applicant previously applied for a patent of "a feline coronavirus inhibitor composition composed of GC376 and GS-441524", publication number CN110215456A , the study found that the composition can inhibit the proliferation of feline coronavirus in cells and has a significant synergistic effect, but there is no relevant application of it for inhibiting human susceptible coronavirus and the COVID-19 pneumonia it causes to report

Method used

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  • Pharmaceutical composition composed of GC376 and GS-441524, and application of pharmaceutical composition in inhibiting novel coronavirus
  • Pharmaceutical composition composed of GC376 and GS-441524, and application of pharmaceutical composition in inhibiting novel coronavirus
  • Pharmaceutical composition composed of GC376 and GS-441524, and application of pharmaceutical composition in inhibiting novel coronavirus

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Experimental program
Comparison scheme
Effect test

Embodiment 1

[0022] The toxicity test of embodiment 1 pharmaceutical composition to cell

[0023] experiment method:

[0024] 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;

[0025] 2) After 16 hours, the medium in the well was discarded, washed three times with 1×PBS, dried and then added to the cell maintenance solution, and the composition was added at the same time (molar ratio 1:1), so that the final concentration of the composition in the well was 10000 μM, 1000μM, 500μM, 100μM, 50μM, 25μM, 12.5μM, 6.25μM, set cell control at the same time, place at 37°C, 5% CO 2 incubator cultivation;

[0026] 3) After 72h, use Reagents for cell viability assays.

[0027] test results:

[0028] see results figure 1 . Determination of cell viability can reflect ...

Embodiment 2

[0029] Example 2 The inhibitory effect of the pharmaceutical composition on the virus at the cellular level

[0030] experiment method:

[0031] 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;

[0032] 2) After 16 hours, discard the culture medium in the well, wash with 1×PBS three times, add 50 μL of GC376 and GS-441524 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, The final concentration of the composition was 50 μM. Set up cell control and virus control at the same time, place at 37°C, 5% CO 2 incubator cultivation;

[0033] 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.

[0034] test results:

[0035] see r...

Embodiment 3

[0036] Embodiment 3 pharmaceutical composition antiviral activity (EC 50 )test

[0037] experiment method:

[0038] 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;

[0039]2) After 16 hours, dilute the composition of GC376 and GS-441524 with the cell maintenance solution at a molar ratio of 1:1, so that the concentration of the composition is 200 μM, 100 μM, 50 μM, 25 μM, 12.5 μM, 6.25 μM, 3.125 μM;

[0040] 3) Make a mark on the cover of the 96-well plate, discard the medium in the well, wash with 1×PBS three times, dry it, add 100 μL of the diluted composition to each well in the order of marking, and then add 100 μL of 0.01 MOI virus solution to each well , so that the final concentration of the composition is 100 μM, 50 μM, 25 μM, 12.5 μM, 6.25 μM, 3.125 μM, 1.5625 μM, set th...

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PUM

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Abstract

The invention discloses a pharmaceutical composition. The pharmaceutical composition is composed of GC376 and GS-441524. In-vitro cell test results show that the composition can inhibit proliferationof novel coronavirus SARS-CoV-2 in cells, has a better drug effect than a single drug, has a significant synergistic effect, can be used as a novel coronavirus SARS-CoV-2 inhibitor, is used for treating COVID-19 pneumonia, and has the advantages of good curative effect, high safety, no toxic or side effects, capability of reducing drug resistance of viruses and toxic and side effects of drugs, etc.

Description

technical field [0001] The invention belongs to the field of pharmacy, and in particular relates to a pharmaceutical composition for inhibiting novel coronavirus SARS-CoV-2 and its application. 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...

Claims

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Application Information

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Patent Type & Authority Applications(China)
IPC IPC(8): A61K31/4015A61K31/53A61P31/14A61P11/00
CPCA61K31/4015A61K31/53A61P11/00A61P31/14A61K2300/00
Inventor 彭贵青刘紫微陈西田大勇
Owner HUAZHONG AGRI UNIV
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