Study on the Antagonism of Reboxetine Mesylate Against Senecavirus in vitro
[Objective]The aim of this study was to screen compounds with antagonistic Senecavirus A(SVA)activity from an FDA-approved drug library containing 127 compounds and studying their pathways of action.[Method]An in vitro active compounds screening platform for SVA was established using luciferase recombinant Senecavirus(rSVA-NLuc)combined with Flue high-throughput screening technology.The compounds were screened from the FDA-approved drug library for their luciferase inhibitory activity at a concentration of 10 μmol/L.The inhibitory activity was further verified by Real-time quantitative RT-PCR,and the maximum non-toxic concentration was determined by a cytotoxicity assay which was measured by the released lactate dehydrogenase(LDH)from cell.According to the four main processes of the virus infection cycle,such as adsorption,endocytosis,replication,assembly and release,different cell treatment methods and Real-time quantitative RT-PCR,indirect immunofluorescence assay(IFA),Western blotting and viral titer assay(TCID50)were used to study the antagonistic mechanism of the screeded compound.[Result]Eight candidate anti-SVA active molecules were screened from the FDA drug library containing 127 molecules,and one safe and effective compound,reboxetine mesylate,was identified by Real-time quantitative RT-PCR and cytotoxicity assay.Within 36 hours of virus infection in cells,reboxetine mesylate significantly reduced the expression of SVA VP3 protein and the SVA titer(P＜0.01).The treatment of golden hamster kidney cells(BSR-T7/5)with reboxetine mesylate could reduce the adsorption and entry of SVA.Real-time quantitative RT-PCR also showed that reboxetine mesylate could inhibit the assembly stage of SVA,but it had no effect on the replication and release stage of SVA.[Conclusion]This experiment screened a compound with low cytotoxicity and excellent SVA antagonistic effect from the FDA-approved drug library:Riboxetine mesylate.This compound fighted SVA infection by inhibiting the adsorption,entry,and assembly stages of SVA.This study provided important references for the further development of anti-SVA drugs.
Senecavirus A(SVA)FDA-approved drug libraryhigh-throughput screening
国家科技期刊平台
NSTL
万方数据
维普
