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Feline infectious peritonitis virus inhibitor composition composed of GC376 and GS-441524

A technology of GS-441524 and feline coronavirus, which is applied in the field of preparation of feline infectious peritonitis drugs and feline coronavirus inhibitor compositions, and can solve problems such as no reports of feline infectious peritonitis

Inactive Publication Date: 2019-09-10
HUAZHONG AGRI UNIV
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  • Summary
  • Abstract
  • Description
  • Claims
  • Application Information

AI Technical Summary

Problems solved by technology

[0008] After reviewing the relevant literature, there is no report on the design of the combination of GC376 and GS-441524 in the treatment of feline infectious peritonitis

Method used

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  • Feline infectious peritonitis virus inhibitor composition composed of GC376 and GS-441524
  • Feline infectious peritonitis virus inhibitor composition composed of GC376 and GS-441524
  • Feline infectious peritonitis virus inhibitor composition composed of GC376 and GS-441524

Examples

Experimental program
Comparison scheme
Effect test

Embodiment 1G

[0020] The toxicity test of embodiment 1GC376, GS-441524 to cell

[0021] experiment method:

[0022] 1) Take well-growing CRFK cells (cat kidney cells) for digestion and passage, and use cell growth medium to adjust the cell density to 1×10 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;

[0023] 2) After 16 hours, the medium in the well was discarded, washed three times with 1×PBS, and after drying, the compound was diluted 2-fold with the cell maintenance solution, so that GC376 was 1000 μM, 500 μM, 100 μM, 50 μM, 25 μM, 12.5 μM, 6.25 μM, the final concentration of GS-441524 is 10000 μM, 1000 μM, 500 μM, 100 μM, 50 μM, 25 μM, 12.5 μM, and set the cell control at the same time;

[0024] 3) After 72h, use Reagents for cell viability assays.

[0025] test results:

[0026] see results figure 1 . Determination of cell viability can reflect the toxic effect of GC376 and GS-441524 on CRFK cells. It can be s...

Embodiment 2

[0027] Example 2 The inhibitory effect of GC376 and GS-441524 on the virus at the cellular level

[0028] experiment method:

[0029] 1) Take CRFK cells in good growth state for digestion and passage, and adjust the cell density to 1×10 with cell maintenance solution 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;

[0030] 2) After 16 hours, discard the culture medium in the well, wash it three times with 1×PBS, dry it and dilute GC376 and GS-441524 by 2 times with the cell growth medium, and add 0.01MOI feline coronavirus (FIPV) to each well . Make the final concentration of GC376 and GS-441524 100 μM, 50 μM, 25 μM, 12.5 μM, and set up cell control and virus control at the same time;

[0031] 3) When the virus control lesion is 70%, observe the changes in the cell state after the compound and the virus interact with a microscope and take pictures for preservation.

[0032] test results:

[0033] see resul...

Embodiment 3

[0034] Embodiment 3GC376, GS-441524 antiviral activity (EC 50 )test

[0035] experiment method:

[0036] 1) Take CRFK 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;

[0037] 2) After 16 hours, discard the culture medium in the well and wash with 1×PBS three times;

[0038] 3) Make a mark on the lid, add GC376 and GS-441524 to the first horizontal row of each group in the order of marking, and perform 2-fold dilution so that the final concentration of GC376 is 50 μM, 25 μM, 12.5 μM, 6.25 μM, 3.125 μM , 1.5625 μM, the final concentration of GS-441524 was 100 μM, 50 μM, 25 μM, 12.5 μM, 6.25 μM, 3.125 μM, 1.5625 μM. At the same time, 0.01 MOI virus solution was added to each well. Set cell control and virus control;

[0039] 4) After 72h, use Reagents for cell viability assays.

[0040] test r...

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Abstract

The invention discloses a feline infectious peritonitis virus inhibitor composition. The feline infectious peritonitis virus inhibitor composition is composed of GC376 and GS-441524. As is shown by results of in-vitro cell experiments, through combined drug application of the GC376 and GS-441524, the proliferation of feline infectious peritonitis viruses in cells can be inhibited, and the effect of combined drug application is obviously better than that of separate drug application; meanwhile, a great inhibition effect can be achieved with a low dose, the synergistic interaction effect is obvious, and the composition can be used as a feline infectious peritonitis virus inhibitor, is used for treating feline infectious peritonitis, and has the advantages that the curative effect is good, the safety is high, the drug resistance of the viruses is lowered, and the toxic and side effects of the drug are lowered.

Description

technical field [0001] The invention belongs to the field of veterinary medicine, and in particular relates to a feline coronavirus inhibitor composition and its application in preparing a medicine for treating feline infectious peritonitis. Background technique [0002] Feline infectious peritonitis (FIP) is a chronic, progressive, and fatal disease caused by a pathogenic coronavirus. It is characterized by peritonitis and a large amount of pleural effusion. The fatality rate of sick cats is high. There is no healing effect, it can only delay its life. [0003] At present, the treatment of feline infectious peritonitis mainly uses antiviral drugs, immunomodulators and immunosuppressants for symptomatic treatment, but none of these drugs can reduce the high mortality rate of FIP, and can only temporarily delay the disease process. Therefore it is urgent to find a new drug for the treatment of feline infectious peritonitis. [0004] GC376 is a broad-spectrum 3C-like proteas...

Claims

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Application Information

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