title: Potent antiviral agents fail to elicit genetically-stable resistance mutations in either enterovirus 71 or Coxsackievirus A16.
abstract: Enterovirus 71 (EV71) and Coxsackievirus A16 (CVA16) are the two major causative agents of hand, foot and mouth disease (HFMD), for which there are currently no licenced treatments. Here, the acquisition of resistance towards two novel capsid-binding compounds, NLD and ALD, was studied and compared to the analogous compound GPP3. During serial passage, EV71 rapidly became resistant to each compound and mutations at residues I113 and V123 in VP1 were identified. A mutation at residue 113 was also identified in CVA16 after passage with GPP3. The mutations were associated with reduced thermostability and were rapidly lost in the absence of inhibitors. In silico modelling suggested that the mutations prevented the compounds from binding the VP1 pocket in the capsid. Although both viruses developed resistance to these potent pocket-binding compounds, the acquired mutations were associated with large fitness costs and reverted to WT phenotype and sequence rapidly in the absence of inhibitors. The most effective inhibitor, NLD, had a very large selectivity index, showing interesting pharmacological properties as a novel anti-EV71 agent.
authors.affiliation: School of Molecular and Cellular Biology, University of Leeds, Leeds, UK.
authors.forename: James T
authors.initials: JT
authors.lastname: Kelly
authors.affiliation: Division of Structural Biology, University of Oxford, Oxford, UK.
authors.forename: Luigi
authors.initials: L
authors.lastname: De Colibus
authors.affiliation: School of Molecular and Cellular Biology, University of Leeds, Leeds, UK.
authors.forename: Lauren
authors.initials: L
authors.lastname: Elliott
authors.affiliation: Division of Structural Biology, University of Oxford, Oxford, UK.
authors.forename: Elizabeth E
authors.initials: EE
authors.lastname: Fry
authors.affiliation: Division of Structural Biology, University of Oxford, Oxford, UK; Diamond Light Source, Didcot, UK.
authors.forename: David I
authors.initials: DI
authors.lastname: Stuart
authors.affiliation: School of Molecular and Cellular Biology, University of Leeds, Leeds, UK. Electronic address: d.j.rowlands@leeds.ac.uk.
authors.forename: David J
authors.initials: DJ
authors.lastname: Rowlands
authors.affiliation: School of Molecular and Cellular Biology, University of Leeds, Leeds, UK. Electronic address: n.j.stonehouse@leeds.ac.uk.
authors.forename: Nicola J
authors.initials: NJ
authors.lastname: Stonehouse
mesh_terms: D000818:Animals; D000998:Antiviral Agents / Q000494:pharmacology*; D002213:Capsid / Q000187:drug effects; D036022:Capsid Proteins / Q000737:chemistry / Q000235:genetics; D002460:Cell Line; D018360:Crystallography, X-Ray; D024882:Drug Resistance, Viral; D004770:Enterovirus / Q000187:drug effects* / Q000235:genetics; D029821:Enterovirus A, Human / Q000187:drug effects* / Q000235:genetics; D006232:Hand, Foot and Mouth Disease / Q000188:drug therapy / Q000821:virology*; D006801:Humans; D008958:Models, Molecular; D009154:Mutation / Q000187:drug effects*; D011487:Protein Conformation; D014709:Vero Cells
chemical_list: D000998:Antiviral Agents; D036022:Capsid Proteins
keywords: Capsid binding; Drug resistance; Fitness; Hand foot and mouth disease; VP1 pocket