Background: Virus replication requires complex interactions between viral and cellular factors. In this study we investigated viral PPIs for Zika virus, West Nile virus (WNV), respiratory syncytial virus (RSV) and human immunodeficiency virus type 1 (HIV-1) with the goal of identifying small molecules that interfer with their association. Compounds with confirmed ability to specifically inhibit their targets will be tested in virus replication assays.
Method: NanoBIT or NanoBRET assays were employed to measure interaction for Zika NS2B and NS3pro, West Nile NS2B and NS3pro, RSV N and P, and HIV-1 RT and eEF1A. The assays were used to screen a library of small compounds. A systematic screening approach interrogated 40,000 compounds in the library. Cell viability was monitored during these live cell assays.
Results: We identified compounds that specifically inhibit the RT:eEF1A PPI in a dose dependent manner were indentified. Experiments in progress will test their effects on virus replication. Recent results have identified lead compounds that inhibit Zika and RSV PPIs, which are undergoing further evaluation. Interestingly our results show that compounds targeting Zika NS2B-NS3pro PPI have no effect on WNV NS2B-NS3pro. Recent progress will be presented.
Conclusion: NanoBIT and NanoBRET can be used to identify specific PPI inhibitors but whether these lead compounds can be developed into antiviral agent remains to be determined.