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New application of compound I-BET-762 in preventing or treating African swine fever

A technology of I-BET-762 and African swine fever, which is applied in the field of compound I-BET-762 for the prevention or treatment of African swine fever, which can solve problems such as economic loss and inability to meet large-scale pig breeding, and achieve enhanced immune response Effect

Active Publication Date: 2020-09-22
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

Since there are still no effective vaccines and specific therapeutic drugs, once the African swine fever epidemic occurs, it can only be controlled by means of culling, but this method not only leads to economic losses, but also cannot meet the needs of large-scale pig raising in my country

Method used

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  • New application of compound I-BET-762 in preventing or treating African swine fever
  • New application of compound I-BET-762 in preventing or treating African swine fever
  • New application of compound I-BET-762 in preventing or treating African swine fever

Examples

Experimental program
Comparison scheme
Effect test

Embodiment 1

[0038] Example 1 Effect of compound I-BET-762 on transcriptional expression of African swine fever virus genes

[0039] Culture porcine alveolar macrophages (PAM, 2×10) with RPMI 1640+10% FBS medium in a 96-well plate 5 / Well), the experimental group was treated with different concentrations of I-BET-762 (0.5μM, 1μM, 2μM, 5μM, 8μM, 10μM) for 16h, the infection control group was treated with DMSO (1%) after 16h; with PBS Continuously dilute the ASFV CN / SC / 2019 strain (MOI=0.1) by 10 times, make 8 dilutions, repeat 8 wells for each dilution, inoculate PAM cells for culture, and add porcine red cells at the same time; set the cell plate At 37℃, 5% CO 2 Conditioned culture for 3-6 days, observe the red blood cell adsorption reaction (HAD) in each cell culture well every day.

[0040] Red blood cell adsorption reaction (HAD) is based on the phenomenon that red blood cells of pig origin are adsorbed around mononuclear macrophages infected with African swine fever virus, resulting in red ...

Embodiment 2

[0051] Example 2 Effect of compound I-BET-762 on the expression level of host inflammation-related factors

[0052] Culture porcine alveolar macrophages (PAM, 1×10) with RPMI 1640+10% FBS medium in a 48-well plate 6 / Well), the experimental group was treated with I-BET-762 (2μM) for 16 hours and then infected with the ASFV CN / SC / 2019 strain (MOI = 0.1); the blank control group did not have any treatment; compound I-BET-762 alone The treatment group was treated with I-BET-762 (2μM) for 16h; the virus-infected group was treated with DMSO (1%) for 16h, and then directly infected with the ASFV CN / SC / 2019 strain (MOI=0.1); After the treated cells were cultured for 48 hours, the cell culture was collected, the cells were washed once with PBS, centrifuged, and the supernatant was discarded. After extracting total RNA and reverse transcribing cDNA, Q-PCR method (same as above) was used to detect the expression differences of host inflammation-related factors.

[0053] The TNF-α primer seq...

Embodiment 3

[0058] Example 3 Cytotoxicity of Compound I-BET-762

[0059] By constructing a stable in vitro cell screening system, CCK-8 method was used to detect the cytotoxicity of the small molecule compound I-BET-762. Culture porcine alveolar macrophages (PAM, 2×10) with RPMI 1640+10% FBS medium in a 96-well plate 5 / Well), overnight culture, add different concentrations of I-BET-762 (0.5μM, 1μM, 5μM, 10μM, 20μM, 40μM, 80μM, 160μM, 240μM) to the wells, and set a blank well (contains medium only) , Control wells (containing cells and culture medium), after incubating the culture plate in the incubator for 16 hours, add 10 μL of CCK-8 solution to each well of the plate, and incubate the culture plate in the incubator for 1-4 hours. Before reading the plate, you can gently mix on a shaker. Then read the microplate reader to measure the absorbance at 450nm to calculate the cell viability.

[0060] The result is Figure 5 It is shown that the compound I-BET-762 is less toxic to cells, even wh...

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Abstract

The invention belongs to the technical field of African swine fever treatment, and particularly relates to a new application of a compound I-BET-762 in preventing or treating African swine fever. Theinvention accidentally discovers that the compound I-BET-762 can obviously inhibit the RNA and protein expression level of p30 and p72 in ASFV, prevent viruses from invading host cells, and can be used for inhibiting early infection of ASFV; and the compound I-BET-762 can obviously increase the expression level of TNF-alpha, NF-kappa B, IL-1 beta, IL-8 and other genes after ASFV infection, up-regulate the transcription level of TNF-alpha, NF-kappa B, IL-1 beta and IL-8, and enhance the immune response. Therefore, the compound I-BET-762 can be used for preventing or treating African swine fever.

Description

Technical field [0001] The invention belongs to the technical field of African swine fever treatment, and specifically relates to a new use of compound I-BET-762 for preventing or treating African swine fever. Background technique [0002] African Swine Fever (ASF) is an acute and violent infectious disease caused by African swine fever virus (ASFV) infection, characterized by fever and hemorrhage from the whole body of the pig. The fatality rate for domestic pigs is as high as 100%. . The disease first broke out in Kenya in 1921 and then became widespread in domestic pigs and wild boars throughout Africa. It was introduced to Europe in the 1950s and it took 40 years to eliminate the disease throughout Europe. However, the disease was once again introduced into Georgia from East Africa in 2007, and then spread widely in Eastern Europe, and in 2017 was introduced to Irkutsk, the Russian Far East. In early August 2019, researcher Hu Rongliang took the lead in reporting the first...

Claims

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Application Information

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Patent Type & Authority Applications(China)
IPC IPC(8): A61K31/5517A61P31/14A61P29/00
CPCA61K31/5517A61P29/00A61P31/14
Inventor 牛庆丽杨吉飞赵亚茹刘志杰关贵全罗建勋殷宏
Owner LANZHOU INST OF VETERINARY SCI CHINESE ACAD OF AGRI SCI
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