Application of azithromycin to anti-coronavirus infection
A coronavirus and azithromycin technology, applied in the field of medicine, can solve problems such as no reports on the anti-coronavirus activity of azithromycin
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Embodiment 1
[0039] Example 1. Principle of Screening Model
[0040] The entry of coronavirus into the host cell is the first step in virus infection, and inhibiting the entry of the virus can effectively block the virus infection. The spike protein (Spike, S) on the surface of the coronavirus envelope is a key protein in the entry process of the coronavirus.
[0041] We applied the SARS-CoV envelope S gene (SARS-CoV S, Gene Bank: AY278741.1) and the MERS-CoV envelope S gene (MERS-CoV S, Gene Bank: JX869059.2) respectively. A plasmid expressing the S protein and the HIV core plasmid (pNL4-3-Luc-R) were co-transfected - E. - ), the SARS-CoV recombinant virus SARS-S / HIV [Ying Guo, Jennifer Tisoncik, Susanna McReynolds, Michael Farzan, Bellur S. Prabhabar, Thomas Gallagher, Lijun Rong and Michael Caffrey. New Region of SARS-CoV S Protein Critical for Viral Entry.Journal of Molecular Biology.2009,394:600-605.] and MERS-CoV recombinant virus MERS-S / HIV[Grehan K,Ferrara F,Temperton N.An optim...
Embodiment 2
[0042] Embodiment 2. Anti-SARS-CoV infection pharmacodynamics experimental method
[0043] The SARS coronavirus Urbani (Gene Bank: AY278741.1) strain is used in the present invention. Recombinant virus preparation [Ying Guo, Jennifer Tisoncik, Susanna McReynolds, Michael Farzan, Bellur S. Prabhabar, Thomas Gallagher, Lijun Rong and Michael Caffrey. Identification of a New Region of SARS-CoV S Protein Critical for Viral Entry. Journal of Molecular Biology.2009 ,394:600-605.]: Co-transfection of pcDNA3.1 / SARS-S plasmid and pNL4-3-Luc-R - E. - Put the plasmid into 293T cells, collect the supernatant 48 hours after transfection, and filter the supernatant through a 0.45 μm filter membrane. The supernatant contains SARS-S / HIV virus particles, and the recombinant virus can be used for infection. Prepare VSV-G / HIV recombinant virus in the same way.
[0044]Infection [Ying Guo, Jennifer Tisoncik, Susanna McReynolds, Michael Farzan, Bellur S. Prabhabar, Thomas Gallagher, Lijun Rong ...
Embodiment 3
[0047] Embodiment 3. Anti-MERS-CoV infection pharmacodynamics experimental method
[0048] Human betacoronavirus 2c EMC / 2012 (Gene Bank: JX869059.2) strain was used in the present invention. Recombinant virus preparation [Grehan K, Ferrara F, Temperton N.An optimized method for the production of MERS-CoV spike expressing viral pseudotypes.MethodsX.2015,2:379-384.]: Co-transfection of pCMV3 / MERS-S plasmid and pNL4- 3-Luc-R - E. - Put the plasmid into 293T cells, collect the supernatant 48 hours after transfection, and filter the supernatant through a 0.45 μm filter membrane. The supernatant contains MERS-S / HIV virus particles, and the recombinant virus can be used for infection.
[0049] Infection: The plasmid expressing human CD26 (Gene Bank: NM_001935.3) was transferred to 293T cells by the modified calcium phosphate method. The day before infection, press 6×10 per well 4 293T-CD26 cells were seeded on a 24-well plate at a density of 1 cell. The positive control compound...
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