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Application of R848 in preparation of medicine for inhibiting novel coronavirus SARS-CoV-2

A technology of 5.R848, sars-cov-2, which is applied in the application field of preparing medicines for inhibiting novel coronavirus SARS-CoV-2, and can solve problems such as undiscovered anti-SARS-CoV-2 virus.

Inactive Publication Date: 2020-12-29
HUAZHONG AGRI UNIV
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  • Summary
  • Abstract
  • Description
  • Claims
  • Application Information

AI Technical Summary

Problems solved by technology

But at present, no relevant reports about R848 anti-SARS-CoV-2 virus have been found

Method used

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  • Application of R848 in preparation of medicine for inhibiting novel coronavirus SARS-CoV-2
  • Application of R848 in preparation of medicine for inhibiting novel coronavirus SARS-CoV-2
  • Application of R848 in preparation of medicine for inhibiting novel coronavirus SARS-CoV-2

Examples

Experimental program
Comparison scheme
Effect test

Embodiment 1

[0055] Toxicity test of R848 to cells

[0056] In the present invention, Caco-2 cells are used to inoculate a 96-well plate, and when the cell density reaches 80%, the cells are incubated with R848 (final concentration: 0 μM, 5 μM, 10 μM, 20 μM, 40 μM, 80 μM) for 24 and 48 hours, respectively. Cell viability was measured using TransDetect cell counting box, and the absorbance at OD450nm was measured according to the instructions of the kit. The specific implementation process is as follows:

[0057] 1. Cell Culture

[0058] The frozen and revived Caco-2 cells were subcultured twice, and expanded with DMEM medium containing 10% fetal bovine serum and double antibodies (penicillin 100 U / ml, streptomycin 100 ug / ml).

[0059] 2. Toxicity test of R848 to cells

[0060] Well-grown Caco-2 cells were taken for digestion and passage, and the cell density was adjusted to 2×10 with cell growth medium (DMEM medium + 10% fetal bovine serum + double antibody). 6 / ml, to inoculate a 96-w...

Embodiment 2

[0065] Inhibitory effect of R848 on novel coronavirus at the cellular level

[0066] Antiviral Infection Effect of R848 Cell Level

[0067] The effect of R848 on the proliferation of the SARS-COV-2 strain (wuhan-hu-1, NC_045512.2) at the cellular level was detected by plaque forming unit, and the specific steps were as follows:

[0068] 1. Proliferation of SARS-COV-2 strain (wuhan-hu-1, NC_045512.2) on cells under the action of R848

[0069] 1) The Caco-2 cells in good growth state were digested and passaged, and the cell density was adjusted to 1×10 with cell culture medium. 5 / ml, 1ml per well was inoculated in a 12-well plate to grow to a monolayer.

[0070] 2) Adding different amounts of R848, the final concentrations of R848 were 0 μM, 0.2 μM, 2 μM, and 20 μM, respectively.

[0071] 3) After being treated with different concentrations of R848 for 24 hours, the cells were infected with the new coronavirus, the virus (MOI was 0.1) was incubated for 1 hour, the virus solu...

Embodiment 3

[0086] Evaluation of the antiviral effect of R848 in a mouse lethal infection model

[0087] Acquisition of mouse-adapted SARS-CoV-2 strain and construction of mouse model:

[0088] Acquisition of mouse-adapted SARS-CoV-2 strains:

[0089] The SARS-CoV-2 strain (wuhan-hu-1, NC_045512.2) isolated from human samples was inoculated intranasally into 10-month-old female BALB / c mice, the infection dose was 50ul, and the virus content was 2.3× 10 5 PFU. Three days after infection, the mice were dissected to collect lung tissue, homogenized, centrifuged at 12,000 rpm for 5 minutes, and the supernatant was collected as the first generation of mouse adaptive passage of the new coronavirus. The first-generation lung tissue homogenate was inoculated into 4-6 week-old mice, and the lungs were harvested and homogenized 3 days later to obtain the second-generation virus liquid. In this way, the 4-6 week old mice were continuously passaged to obtain the 11th generation virus liquid. Dur...

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Abstract

The invention belongs to the field of biological agents, and discloses application of R848 in preparation of a medicine for inhibiting novel coronavirus SARS-CoV-2, an applicant finds that R848 has asmall toxic effect on Caco-2 cells through an in vitro cell test and a mouse test, infection of the Caco2 cells by the SARS-CoV-2 can be remarkably inhibited at the in vitro cell level, and proliferation of the Caco-2 cells is inhibited. The characteristics of safety and high efficiency are realized; meanwhile, by constructing a mouse SARS-CoV-2 infection model, it is found that R848 can significantly reduce the weight loss degree of the mouse, reduce the virus content in the lung tissue of the mouse and improve the survival rate of the mouse, so that R848 has very large clinical treatment potential.

Description

technical field [0001] The invention belongs to the field of biological preparations, and in particular relates to the application of R848 in the preparation of drugs for inhibiting novel coronavirus SARS-CoV-2. Background technique [0002] The novel coronavirus pneumonia (coronavirus disease 19, COVID-19) caused by severe acute respiratory syndrome coronavirus 2 (severe acute respiratory syndrome coronavirus 2, SARS-CoV-2) appeared at the end of 2019 and presented a global outbreak, seriously endangering human beings Health and public health security have brought huge burdens to the world society and economy, and have aroused great concern worldwide. As of the end of August 2020, there were more than 24.5 million confirmed cases of new coronary pneumonia worldwide, and more than 830,000 deaths. Many domestic and foreign scholars speculate that the new coronavirus may be similar to influenza virus and coexist with humans for a long time. Moreover, existing research report...

Claims

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Application Information

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Patent Type & Authority Applications(China)
IPC IPC(8): A61K31/437A61P31/14
CPCA61K31/437A61P31/14
Inventor 金梅林黄坤张宇飞杨丽邹忠回显锋赵亚孙小美张强陈西
Owner HUAZHONG AGRI UNIV
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