title: Inhibitors of foot and mouth disease virus targeting a novel pocket of the RNA-dependent RNA polymerase.
abstract: 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.
  
  
  RESULTS
  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 (IC(50)s 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 EC(50) = 12 µM and EC(90) = 20 µM).
  
  
  CONCLUSIONS
  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.
authors.affiliation: Christopher Bond Life Sciences Center, Department of Molecular Microbiology and Immunology, University of Missouri School of Medicine, Columbia, Missouri, United States of America.
authors.forename: Ryan C
authors.initials: RC
authors.lastname: Durk
authors.forename: Kamalendra
authors.initials: K
authors.lastname: Singh
authors.forename: Ceili A
authors.initials: CA
authors.lastname: Cornelison
authors.forename: Devendra K
authors.initials: DK
authors.lastname: Rai
authors.forename: Kayla B
authors.initials: KB
authors.lastname: Matzek
authors.forename: Maxwell D
authors.initials: MD
authors.lastname: Leslie
authors.forename: Elizabeth
authors.initials: E
authors.lastname: Schafer
authors.forename: Bruno
authors.initials: B
authors.lastname: Marchand
authors.forename: Adeyemi
authors.initials: A
authors.lastname: Adedeji
authors.forename: Eleftherios
authors.initials: E
authors.lastname: Michailidis
authors.forename: Christopher A
authors.initials: CA
authors.lastname: Dorst
authors.forename: Jennifer
authors.initials: J
authors.lastname: Moran
authors.forename: Christie
authors.initials: C
authors.lastname: Pautler
authors.forename: Luis L
authors.initials: LL
authors.lastname: Rodriguez
authors.forename: Mark A
authors.initials: MA
authors.lastname: McIntosh
authors.forename: Elizabeth
authors.initials: E
authors.lastname: Rieder
authors.forename: Stefan G
authors.initials: SG
authors.lastname: Sarafianos
mesh_terms: D000255:Adenosine Triphosphate / Q000737:chemistry; D000818:Animals; D000998:Antiviral Agents / Q000494:pharmacology*; D001665:Binding Sites; D002384:Catalysis; D002417:Cattle; D004791:Enzyme Inhibitors; D029544:Foot-and-Mouth Disease Virus / Q000378:metabolism*; D007700:Kinetics; D008024:Ligands; D008156:Luciferases / Q000378:metabolism; D008956:Models, Chemical; D016297:Mutagenesis, Site-Directed; D012324:RNA-Dependent RNA Polymerase / Q000737:chemistry / Q000378:metabolism*; D014779:Virus Replication / Q000187:drug effects
chemical_list: D000998:Antiviral Agents; D004791:Enzyme Inhibitors; D008024:Ligands; D000255:Adenosine Triphosphate; D008156:Luciferases; D012324:RNA-Dependent RNA Polymerase