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Application of Inauhzin in preparation of drug for preventing foot-and-mouth disease virus infection

A technology of foot-and-mouth disease virus and medicine, applied in the field of application

Active Publication Date: 2019-07-19
LANZHOU INST OF VETERINARY SCI CHINESE ACAD OF AGRI SCI
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  • Summary
  • Abstract
  • Description
  • Claims
  • Application Information

AI Technical Summary

Problems solved by technology

Foot-and-mouth disease virus (FMDV), the causative agent of foot-and-mouth disease, belongs to the genus FMDV of the family Picornaviridae. There are 7 serotypes including O, A, C, Asia I and SAT1, SAT2, and SAT3. Each type also has multiple subtypes. Types and even subtypes of viruses have their own special immunity, mutation and transmission capabilities, which brings certain challenges to the prevention and control of foot-and-mouth disease

Method used

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  • Application of Inauhzin in preparation of drug for preventing foot-and-mouth disease virus infection
  • Application of Inauhzin in preparation of drug for preventing foot-and-mouth disease virus infection
  • Application of Inauhzin in preparation of drug for preventing foot-and-mouth disease virus infection

Examples

Experimental program
Comparison scheme
Effect test

Embodiment 1

[0027] Toxicity test of Inauhzin on IBRS-2 cells:

[0028] The cytotoxicity of Inauhzin (purchased from MCE Company) to IBRS-2 cells was determined by MTS method (MTS detection kit was purchased from Abcam Company). After spreading the IBRS-2 cells in a 96-well plate to a full monolayer, the supernatant culture medium of the cells was discarded, washed 3 times with fresh DMEM (purchased from Gibco), and finally added with FBS containing 2% fetal bovine serum (purchased from Gibco). 100 μL of Inauhzin diluted in DMEM culture fluid from Gibco Company) was used as a negative control well with the corresponding concentration of the Inauhzin preparation solution DMSO, and as a cell control well without any treatment. Place at 37°C for continuous culture for 72 hours, discard the upper cell culture medium, wash with fresh DMEM three times, then add 100 μL of fresh DMEM, and add 20 μL of MTS solution to each well. After incubating at 37°C for 4 h, measure the absorbance value at 490...

Embodiment 2

[0031] Evaluation of the anti-foot-and-mouth disease virus activity of Inauhzin on IBRS-2 cells

[0032] Spread the well-grown IBRS-2 cells on DMEM complete medium containing 10% fetal bovine serum FBS to a 96-well plate. After the IBRS-2 cells grow to a monolayer, discard the culture medium on the upper layer of the cells and wash with fresh DMEM. 3 times, inoculated with 100TCID50O / MY98 / BY / 2010 (saved and provided by the National Foot-and-Mouth Disease Reference Laboratory). After 1 hour, remove the virus solution, wash with fresh DMEM for 3 times, add 100 μL of Inauhzin diluted with DMEM culture solution containing 2% FBS, and use the corresponding DMSO concentration of the prepared solution of Inauhzin as the virus control well. Viruses were used as cell control wells. Place at 37°C for 48 hours, discard the upper cell culture medium, wash with fresh DMEM three times, then add 100 μL of fresh DMEM, and add 20 μL of MTS solution to each well. After incubating at 37°C for ...

Embodiment 3

[0041] Indirect Immunofluorescence Detection of FMDV Protein Expression in Infected Cell Group

[0042] Set the density to 3×10 5 Spread IBRS-2 cells per well on a 12-well plate. After the IBRS-2 cells grow to a monolayer, discard the culture medium on the upper layer of the cells, wash 3 times with fresh DMEM, and inoculate 100TCID 50 O / MY98 / BY / 2010. After 1 hour, remove the virus solution, wash 3 times with fresh DMEM, add 100 μL of Inauhzin diluted with DMEM culture solution containing 2% FBS, and use the corresponding DMSO concentration of the prepared solution of Inauhzin as the virus The control wells were cultured at 37°C for 12 hours. Discard the upper cell culture medium, wash with PBS twice, fix the cells with 4% paraformaldehyde for 15 min, discard the paraformaldehyde, add methanol for 5 min, rinse with PBS for 3 times, each time for 5 min, add blocking solution (10% FBS, 0.3% Triton X-100, 89.7% PBS) to block for 10min, then add the primary antibody diluted wit...

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Abstract

The invention provides an application of Inauhzin in preparation of an inhibitor for foot-and-mouth disease virus, which relates to the technical field of veterinary drugs. The Inauhzin of the presentinvention has low toxicity, has inhibitory effects on foot-and-mouth disease virus type A and foot-and-mouth disease virus type O, and can significantly delay the lethal effect of foot-and-mouth disease virus on suckling rats.

Description

technical field [0001] The invention belongs to the technical field of veterinary medicines, and in particular relates to the application of Inauhzin in the preparation of medicines for preventing foot-and-mouth disease virus infection. Background technique [0002] Foot-and-mouth disease is an acute, febrile and highly contagious cloven-hoofed infectious disease caused by foot-and-mouth disease virus, which seriously endangers the sustainable and healthy development of local animal husbandry and the foreign trade of related products. The World Organization for Animal Health lists it as a legally notifiable animal disease, and my country lists it as a first-class animal disease. Foot-and-mouth disease virus (FMDV), the causative agent of foot-and-mouth disease, belongs to the genus FMDV of the family Picornaviridae. It has 7 serotypes including O, A, C, Asia I and SAT1, SAT2, and SAT3. Each type also has multiple subtypes. Types and even subtypes of viruses have their own s...

Claims

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Application Information

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Patent Type & Authority Applications(China)
IPC IPC(8): A61K31/5415A61P31/14
CPCA61K31/5415A61P31/14
Inventor 李世芳常惠芸龚美娇邵军军常艳燕张永光
Owner LANZHOU INST OF VETERINARY SCI CHINESE ACAD OF AGRI SCI
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