Inhibitors of Foot and Mouth Disease Virus Targeting a Novel Pocket of the RNA-Dependent RNA Polymerase

• Published in: PloS one Vol. 5; no. 12; p. e15049
• Main Authors: Durk, Ryan C, Singh, Kamalendra, Cornelison, Ceili A, Rai, Devendra K, Matzek, Kayla B, Leslie, Maxwell D, Schafer, Elizabeth, Marchand, Bruno, Adedeji, Adeyemi, Michailidis, Eleftherios, Dorst, Christopher A, Moran, Jennifer, Pautler, Christie, Rodriguez, Luis L, McIntosh, Mark A, Rieder, Elizabeth
• Format: Journal Article
• Language: English
• Published: United States Public Library of Science 21.12.2010; Public Library of Science (PLoS)
• Subjects: Adenosine Triphosphate - chemistry; Agriculture; Analysis; Animals; Antiviral agents; Antiviral Agents - pharmacology; Attorneys; Binding; Binding Sites; Biology; Catalysis; Cattle; DNA-directed RNA polymerase; Docking; Drug development; enzyme activity; enzyme inhibition; Enzyme Inhibitors; Foot & mouth disease; Foot and mouth disease; Foot-and-mouth disease virus; Foot-and-Mouth Disease Virus - metabolism; Health aspects; Immune response; Immune system; Inhibitors; Kinetics; Law schools; Ligands; Livestock; Livestock production; Luciferase; Luciferases - metabolism; Medical screening; Models, Chemical; Molecular modelling; Mutagenesis, Site-Directed; Outbreaks; Prevention; Residues; Ribonucleic acid; RNA; RNA polymerase; RNA-Dependent RNA Polymerase - chemistry; RNA-Dependent RNA Polymerase - metabolism; RNA-directed RNA polymerase; screening; Site-directed mutagenesis; Substrates; Vaccination; Virus replication; Virus Replication - drug effects; Viruses; New York; Plum Island; United States > US; Missouri
• ISSN: 1932-6203; 1932-6203
• DOI: 10.1371/journal.pone.0015049

Background: Foot-and-Mouth Disease Virus (FMDV) is a picornavirus that infects cloven-hoofed animals and leads to severe losses in livestock production. In the case of an FMD outbreak, emergency vaccination requires at least 7 days to trigger an effective immune response. There are currently no approved inhibitors for the treatment or prevention of FMDV infections. Methodology/Principal Findings: Using a luciferase-based assay we screened a library of compounds and identified seven novel inhibitors of 3Dpol, the RNA-dependent RNA polymerase of FMDV. The compounds inhibited specifically 3Dpol (IC50s from 2-17 µM) and not other viral or bacterial polymerases. Enzyme kinetic studies on the inhibition mechanism by compounds 5D9 and 7F8 showed that they are non-competitive inhibitors with respect to NTP and nucleic acid substrates. Molecular modeling and docking studies into the 3Dpol structure revealed an inhibitor binding pocket proximal to, but distinct from the 3Dpol catalytic site. Residues surrounding this pocket are conserved among all 60 FMDV subtypes. Site directed mutagenesis of two residues located at either side of the pocket caused distinct resistance to the compounds, demonstrating that they indeed bind at this site. Several compounds inhibited viral replication with 5D9 suppressing virus production in FMDV-infected cells with EC50 = 12 µM and EC90 = 20 µM). Significance: We identified several non-competitive inhibitors of FMDV 3Dpol that target a novel binding pocket, which can be used for future structure-based drug design studies. Such studies can lead to the discovery of even more potent antivirals that could provide alternative or supplementary options to contain future outbreaks of FMD.