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Application of FHPI for preparing medicine for treatingcfeline infectious peritonitis

A technology of inhibitors and active ingredients, applied in the field of preparation of medicines for treating feline infectious peritonitis, can solve problems such as no FHPI report

Active Publication Date: 2019-07-30
HUAZHONG AGRI UNIV +1
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  • Summary
  • Abstract
  • Description
  • Claims
  • Application Information

AI Technical Summary

Problems solved by technology

[0006] After reviewing relevant literature, there is no relevant report on FHPI in the treatment of feline infectious peritonitis

Method used

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  • Application of FHPI for preparing medicine for treatingcfeline infectious peritonitis
  • Application of FHPI for preparing medicine for treatingcfeline infectious peritonitis
  • Application of FHPI for preparing medicine for treatingcfeline infectious peritonitis

Examples

Experimental program
Comparison scheme
Effect test

Embodiment 1

[0017] The toxicity test of embodiment 1FHPI to cell

[0018] experiment method:

[0019] 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;

[0020] 2) After 16 hours, the culture medium in the well was discarded, washed three times with 1×PBS, and after drying, FHPI was diluted 2-fold with cell maintenance solution, so that the final concentration of FHPI was 200 μM, 100 μM, 50 μM, 25 μM, 12.5 μM, 6.25μM, 3.125μM, and set the cell control at the same time;

[0021] 3) After 72h, use CellTiter- Reagents for cell viability assays.

[0022] test results:

[0023] see results figure 1 . Determination of cell viability can reflect the toxic effect of FHPI on CRFK cells. It can be seen from the figure that FHPI has less toxic effect on CRFK cells, and can promote cell growt...

Embodiment 2

[0024] Embodiment 2FHPI is to the inhibitory effect of virus at cell level

[0025] experiment method:

[0026] 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;

[0027] 2) After 16 hours, the medium in the well was discarded, washed three times with 1×PBS, and after drying, FHPI was diluted 2-fold with cell growth medium, and 0.01 MOI of feline coronavirus (FIPV) was added to each well. Make the final concentration of FHPI 100 μM, 50 μM, 25 μM, 12.5 μM, and set cell control and virus control at the same time;

[0028] 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.

[0029] test results:

[0030] see results figure 2 . From the microscope pictures ...

Embodiment 3

[0031] Embodiment 3FHPI antiviral activity (EC 50 )test

[0032] experiment method:

[0033] 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;

[0034] 2) After 16 hours, discard the medium in the wells, wash with 1×PBS three times, and add 100 μL cell maintenance solution to the first horizontal row of each group after drying, and add 50 μL cell maintenance solution to the second, third, fourth, fifth, sixth, and seventh rows. liquid;

[0035] 3) Make a mark on the cover, add FHPI to the first horizontal row of each group according to the marking order, perform 2-fold dilution, mix gently with a row gun for ten times, suck out 50 μL and add it to the second column, and so on, and finally mix Evenly discard 50 μL, so that the final concentration of FHPI is 100 μM, 50 μM, 25 μM, 12.5 μ...

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Abstract

The invention discloses application of FHPI for preparing a medicine for treating feline infectious peritonitis. FHPI has the little toxic effect on normal feline cells, has the obvious inhibition effect on feline coronaviruses, and can obviously lower the virulence of viruses in cells; indirect immunofluorescence light and Western Blot determination results show that FHPI can inhibit proliferation of feline coronaviruses in cells, and has the dose dependence. Accordingly, the FHPI has the activity for resisting feline coronaviruses, can be used for preparing a feline coronavirus inhibitor, can be used for treating feline infectious peritonitis and has the advantages of being good in curative effect, high in safety and the like.

Description

technical field [0001] The invention belongs to the field of pharmacy, and in particular relates to a new application of FHPI in the preparation of medicaments 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] FHPI is a p38MAPK inhibitor that targets p38α / β and can activate the p38(MAPK) ...

Claims

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Application Information

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IPC IPC(8): A61K31/4439A61P31/14
CPCA61K31/4439A61P31/14
Inventor 彭贵青毛军福刘紫微
Owner HUAZHONG AGRI UNIV
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