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New use of compound i-bet-762 for preparing medicine for preventing or treating African swine fever

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

Active Publication Date: 2021-06-08
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 use of compound i-bet-762 for preparing medicine for preventing or treating African swine fever
  • New use of compound i-bet-762 for preparing medicine for preventing or treating African swine fever
  • New use of compound i-bet-762 for preparing medicine for preventing or treating African swine fever

Examples

Experimental program
Comparison scheme
Effect test

Embodiment 1

[0038] The impact of embodiment 1 compound I-BET-762 on the transcriptional expression of African swine fever virus gene

[0039] Porcine alveolar macrophages (PAM, 2 × 10 5 / well), the experimental group treated the cells with different concentrations of I-BET-762 (0.5 μM, 1 μM, 2 μM, 5 μM, 8 μM, 10 μM) for 16 h, and the infection control group treated the cells with DMSO (1%) for 16 h; Serial 10-fold dilution of ASFV CN / SC / 2019 strain (MOI=0.1), 8 dilutions, each dilution repeated 8 wells, inoculated into PAM cells for culture, while adding pig erythrocytes; At 37°C, 5% CO 2 The conditions were cultured for 3-6 days, and the hematocyte adsorption reaction (HAD) in each cell culture well was observed every day.

[0040] The hematocyte adsorption reaction (HAD) is based on the phenomenon that porcine erythrocytes will adsorb around monocytes and macrophages infected with African swine fever virus, thereby generating hematocyte adsorption. Experimental results such as figure...

Embodiment 2

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

[0052] Porcine alveolar macrophages (PAM, 1 × 10 6 / well), the experimental group treated the cells with I-BET-762 (2μM) for 16h, and then infected the ASFV CN / SC / 2019 strain (MOI=0.1); the blank control group did not receive any treatment; the compound I-BET-762 alone The treatment group treated the cells with I-BET-762 (2μM) for 16h; the virus infection group treated the cells with DMSO (1%) for 16h, and then directly infected 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. Total RNA was extracted, and after cDNA was reverse transcribed, the expression difference of host inflammation-related factors was detected by Q-PCR method (same as above).

[0053] Wherein the TNF-α primer sequence is: upstream primer 5'...

Embodiment 3

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

[0059] By constructing a stable in vitro cell screening system, the cytotoxicity of the small molecule compound I-BET-762 was detected by the CCK-8 method. Porcine alveolar macrophages (PAM, 2 × 10 5 / well), culture overnight, add different concentrations of I-BET-762 (0.5μM, 1μM, 5μM, 10μM, 20μM, 40μM, 80μM, 160μM, 240μM) to the wells, and set blank wells (only containing medium) , the control well (containing cells and medium), after the culture plate was incubated in the incubator for 16h, 10 μL of CCK-8 solution was added to each well of the plate, and the culture plate was incubated in the incubator for 1-4h. Mix gently on a shaker before reading the plate. Then read the microplate reader to measure the absorbance at 450nm, and calculate the cell viability.

[0060] The result is as Figure 5 As shown, compound I-BET-762 has little toxicity to cells, and even when the dosage reaches 40 μM, the cell activity can sti...

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Abstract

The invention belongs to the technical field of treating African swine fever, and in particular relates to a new application of compound I-BET-762 for preventing or treating African swine fever. The present invention unexpectedly finds that compound I-BET-762 can significantly inhibit the RNA and protein expression levels of p30 and p72 in ASFV, prevent the virus from invading host cells, and can be used to inhibit the early infection of ASFV; and compound I-BET-762 can significantly increase The expression levels of TNF-α, NF-κB, IL-1β, IL-8 and other genes after ASFV infection can up-regulate the transcription levels of TNF-α, NF-κB, IL-1β, IL-8, and enhance the immune response; therefore, the compound I‑BET‑762 can be used to prevent or treat African swine fever.

Description

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

Claims

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Application Information

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Patent Type & Authority Patents(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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