Applications of azithromycin and telithromycin in anti-Ebola virus infection
A technology of Ebola virus and azithromycin, applied in the field of medicine, can solve the problems of no reports of antiviral activity of azithromycin and telithromycin, no reports of anti-Ebola virus effects, etc.
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Embodiment 1
[0042] Example 1. Principle of Screening Model
[0043] The entry of Ebola virus into the host cell is the first step of virus infection, and inhibiting the entry of the virus can effectively block the virus infection. The glycoprotein (Glycoprotein, GP) on the surface of EBV envelope is the key protein in the process of EBV entry.
[0044] We synthesized the envelope GP gene of Zaire type Ebola virus (EBV-ZaireGP, GeneAccessionNo.L11365). By co-transfection of EBV-GP plasmid and pNL4-3-Luc-R - E. - , EBV recombinant virus EBV-GP / HIV can be obtained with EBV-GP as the outer shell wrapping the HIV core [16] . The virus particle has the following characteristics: 1) the selectivity of the virus to the host cell depends on the characteristics of EBVGP; 2) due to the deletion of env, nef and vpr genes on the HIV vector, the virus can only enter the host cell once and cannot replicate, So the virus is safe; 3) the HIV vector has a luciferase reporter gene, so the infected cell...
Embodiment 2
[0045] Embodiment 2. Experimental method
[0046] In the present invention, EBV-Zaire (GeneAccessionNo.L11365) is used to evaluate the pharmacological activity of azithromycin and telithromycin against Ebola virus infection:
[0047] Recombinant virus preparation [16] : Co-transfect 2 μg pcDNA3.1 / EBV-GP plasmid and 2 μg pNL4-3-Luc-R-E-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 EBV - GP / HIV virions, the recombinant virus being used for infection. Prepare VSV-G / HIV recombinant virus in the same way.
[0048] Infect [16] : One day before infection, press 6×10 per well 4 A549 cells were seeded on a 24-well plate at a density of 1 cell. The positive control compound or the compound to be screened was dissolved in DMSO, and added to the cell culture medium 15 minutes before infection, and the DMSO solvent was used as a blank control. Add appropriate dilutio...
Embodiment 3
[0051] Example 3. Cytotoxicity test
[0052] The cytotoxicity of all involved drugs to A549 and 293ET cells was measured by MTS method, and the results showed that neither azithromycin nor telithromycin had cytotoxicity at a final concentration of 10 μM.
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