Identification of inhibitors of yellow fever virus replication using a replicon-based high-throughput assay.
Flaviviruses cause severe disease in humans and are a public health priority worldwide. However, no effective therapies or drugs are commercially available yet. Several flavivirus replicon-based assays amenable to high-throughput screening of inhibitors have been reported recently. We developed and performed a replicon-based high-throughput assay for screening small-molecule inhibitors of yellow fever virus (YFV) replication. This assay utilized packaged pseudoinfectious particles containing a YFV replicon that expresses Renilla luciferase in a replication-dependent manner. Several small-molecule compounds with inhibitory activity at micromolar concentrations were identified in the high-throughput screen. These compounds were subsequently tested for their inhibitory activities against YFV replication and propagation in low-throughput assays. Furthermore, YFV mutants that escaped inhibition by two of the compounds were isolated, and in both cases, the mutations were mapped to the NS4B coding region, suggesting a novel inhibitory target for these compounds. This study opens up new avenues for pursuing the nonenzymatic nonstructural proteins as targets for antivirals against YFV and other flaviviruses.
Patkar
Chinmay G
CG
Markey Center for Structural Biology, Department of Biological Sciences, 915 W. State St., Purdue University, West Lafayette, IN 47907-2054, USA.
Larsen
Martha
M
Owston
Michael
M
Smith
Janet L
JL
Kuhn
Richard J
RJ
D000818:Animals; D000998:Antiviral Agents / Q000494:pharmacology*; D002460:Cell Line; D006224:Cricetinae; D005455:Fluorescent Antibody Technique; D016679:Genome, Viral / Q000235:genetics*; D012093:Replicon / Q000235:genetics*; D020133:Reverse Transcriptase Polymerase Chain Reaction; D014779:Virus Replication / Q000187:drug effects*; D015005:Yellow fever virus / Q000187:drug effects* / Q000235:genetics
D000998:Antiviral Agents