Application of DNA topoisomerase inhibitor in preparation of medicine for preventing and/or treating novel coronavirus infection

A technology of topoisomerase and coronavirus, which is applied in antiviral agents, drug combinations, pharmaceutical formulas, etc., can solve the problem of lack of effective prevention and treatment drugs for infection, and achieve the effects of enhancing human immunity, inhibiting replication, and strengthening curative effect

Active Publication Date: 2020-12-15
SUN YAT SEN UNIV
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  • Summary
  • Abstract
  • Description
  • Claims
  • Application Information

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Problems solved by technology

[0010] The present invention aims at the problem that the current infection caused by the new coronavirus SARS-CoV-2 lacks effective prevention and treatment drugs, and provides DNA topoisomerase inhibitors su...

Method used

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  • Application of DNA topoisomerase inhibitor in preparation of medicine for preventing and/or treating novel coronavirus infection
  • Application of DNA topoisomerase inhibitor in preparation of medicine for preventing and/or treating novel coronavirus infection
  • Application of DNA topoisomerase inhibitor in preparation of medicine for preventing and/or treating novel coronavirus infection

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Embodiment 1

[0036] The SARS-CoV-2 safe replicon can simulate the RNA transcription and replication of SARS-CoV-2 in cells, and the luciferase activity in cells can directly reflect the degree of RNA transcription and replication of SARS-CoV-2, while not The infectious novel coronavirus SARS-CoV-2 was produced. Therefore, the present invention uses the new coronavirus SARS-CoV-2 safe replicon to verify the inhibitory effect of mitoxantrone and doxorubicin on the RNA synthesis of SARS-CoV-2.

[0037] Relevant experiments can be carried out in the P2 laboratory, and the specific experimental steps are as follows:

[0038] The SARS-CoV-2 safe replicon constitutes plasmid ps2V (SEQ ID NO: 1), ps2AN (SEQ ID NO: 2), ps2AC (SEQ ID NO: 3), ps2B (SEQ ID NO: 4), according to certain Ratio Use Thermo Fisher’s Lipo2000 transfection reagent and Opti-MEM to transfect into HEK293T cells (cell density is about 6.5×10 4 / cm 2 ), see Table 1 below for reagent consumption. Specifically, 100 μL Opti-MEM w...

Embodiment 2

[0046] The present invention uses the isolated wild-type novel coronavirus SARS-CoV-2 to verify the inhibitory effect of mitoxantrone and doxorubicin on virus replication, and the specific experimental methods are as follows:

[0047] HEK293T cells in good growth state were evenly spread in 48-well culture plates treated with poly-lysine (cell density was about 6.5×10 4 / cm 2 ). After 16 hours of cell growth (the cell density is about 1.6×10 5 / mL), transfected with 0.1 μg of pCMV-ACE2-FLAG plasmid expressing SARS-CoV-2 receptor ACE2 purchased from Beijing Yiqiao Shenzhou Company. 24 hours after transfection, the cells were washed with PBS, and then infected with wild-type SARS-CoV-2 (MOI=0.1, 37° C., 1 hour). Subsequently, DMEM (2% FBS) containing mitoxantrone and doxorubicin at different concentration gradients (20 μM, 5 μM, 1.25 μM, 0.3125 μM, 0.078125 μM, 0.01953125 μM) were replaced. After 24 hours of drug treatment, cellular RNA was extracted using TRIZOL. The RNA c...

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Abstract

The invention belongs to the field of antiviral drugs, and discloses an application of a DNA topoisomerase inhibitor in preparation of medicines for preventing and/or treating novel coronavirus infection, and the DNA topoisomerase inhibitor comprises mitoxantrone, doxorubicin or pharmaceutically acceptable salts of above small molecule compounds. DNA topoisomerase inhibitors such as mitoxantrone and doxorubicin can extremely strongly inhibit the synthesis of novel coronavirus virus SARS-CoV-2 replicon RNA and reduce the RNA copy amount of wild novel coronavirus virus SARS-CoV-2 in cells, so that the replication of novel coronavirus SARS-CoV-2 is inhibited, and the effect of preventing and/or treating novel coronavirus SARS-CoV-2 infection is achieved.

Description

technical field [0001] The invention relates to the field of antiviral drugs, in particular to the application of DNA topoisomerase inhibitors in the preparation of drugs for preventing and / or treating novel coronavirus infections. Background technique [0002] Since the outbreak of the novel coronavirus disease COVID-19 at the end of 2019, the disease has spread rapidly around the world, causing a huge global economic downturn and medical burden. As of August 5, 2020, according to data from Johns Hopkins University in the United States, a total of 18,543,662 confirmed cases and 700,714 deaths have been reported worldwide. [0003] At present, there is no specific drug for COVID-19, and the current treatment is mostly symptomatic, which cannot effectively reverse the progression of the disease in severe patients. Although the new coronavirus is partially similar to SARS-CoV and MERS-CoV, they are completely different. Some patients do not respond well after antiviral and an...

Claims

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Application Information

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IPC IPC(8): A61K31/136A61K31/704A61P31/14A61P37/04A61P11/00
CPCA61K31/136A61K31/704A61P31/14A61P37/04A61P11/00
Inventor 张辉罗越文周末
Owner SUN YAT SEN UNIV
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