Application of macrolides in anti-filovirus infection
A technology of macrolides and filoviruses, which is applied in the field of medicine and can solve the problems that the antiviral effect has not been reported.
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Embodiment 1
[0166] Example 1. Principle of Screening Model
[0167] Filovirus entry into host cells is the first step in virus infection, and inhibiting virus entry can effectively block virus infection. The glycoprotein (Glycoprotein, GP) on the surface of the filamentous virus envelope is a key protein in the process of filamentous entry.
[0168] We synthesized the envelope GP gene of Zaire type Ebola virus (EBOV-Zaire GP, Gene AccessionNo.L11365), Sudan type Ebola virus envelope GP gene (EBOV-Sudan GP, Gene AccessionNo.FJ968794.1 ) and Marburg virus envelope GP gene (Marburg GP, Gene Accession No.NC_001608.3). Expression of GP protein plasmid and pNL4-3-Luc-R by co-transfection - E. - , the EBOV recombinant virus EBV-GP / HIV [Manicassamy B, Wang J, Jiang H, RongL.Comprehensive analysis of ebola virus GP1 in viral entry.J Virol.2005; 79: 4793-805.] and Marburg recombinant virus MGP / HIV [Manicassamy B, Wang J, et al. Characterization of Marburg virus glycoprotein in viral entry. Vi...
Embodiment 2
[0169] Embodiment 2. Experimental method
[0170] In the present invention, EBOV-Zaire (Gene Accession No.L11365), EBOV-Sudan (Gene Accession No.FJ968794.1) and Marburg (Gene Accession No.NC_001608.3) are applied to the anti-filovirus of macrolides Infection for pharmacological activity evaluation:
[0171] Preparation of recombinant virus [Manicassamy B, Wang J, Jiang H, Rong L.Comprehensive analysis of ebola virus GP1 in viral entry.J Virol.2005; 79:4793-805.]: Co-transfect 2μg pcDNA3.1 / EBOV-Zaire GP Plasmid (or 2 μg pcDNA3.1 / EBOV-Sudan GP plasmid or 2 μg pcDNA3.1 / Marburg GP plasmid) and 2 μg pNL4-3-Luc-R - E. - The plasmid was transferred to 293T cells, and the supernatant was collected 48 h after transfection, and the supernatant was filtered through a 0.45 μm filter membrane, and the supernatant contained EBOV-Zaire GP / HIV virus particles (or EBOV-Sudan GP / HIV virus particles or Marburg GP / HIV virus particles), the recombinant virus can be used for infection. Prepare...
Embodiment 3
[0180] Example 3. Cytotoxicity test
[0181] The cytotoxicity of all involved drugs to A549 and 293T cells was determined by MTS method, and the results showed that azithromycin, telithromycin, clarithromycin, carbomycin A, josamycin, beilimycin, midecamycin, mica Oleandomycin, troleandomycin, spiramycin, troleandomycin, tulamycin, fluerythromycin, tilmicosin, rotamycin, erythromycin, amphotericin B, tagram Limus, pimecrolimus, and sirolimus had no cytotoxicity at a final concentration of 10 μM.
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