title: A novel benzonitrile analogue inhibits rhinovirus replication.
abstract: OBJECTIVE
  To study the characteristics and the mode of action of the anti-rhinovirus compound 4-[1-hydroxy-2-(4,5-dimethoxy-2-nitrophenyl)ethyl]benzonitrile (LPCRW_0005).
  
  
  METHODS
  The antiviral activity of LPCRW_0005 was evaluated in a cytopathic effect reduction assay against a panel of human rhinovirus (HRV) strains. To unravel its precise molecular mechanism of action, a time-of-drug-addition study, resistance selection and thermostability assays were performed. The crystal structure of the HRV14/LPCRW_0005 complex was elucidated as well.
  
  
  RESULTS
  LPCRW_0005 proved to be a selective inhibitor of the replication of HRV14 (EC(50) of 2 ± 1 μM). Time-of-drug-addition studies revealed that LPCRW_0005 interferes with the earliest stages of virus replication. Phenotypic drug-resistant virus variants were obtained (≥30-fold decrease in susceptibility to the inhibitory effect of LPCRW_0005), which carried either an A150T or A150V amino acid substitution in the VP1 capsid protein. The link between the mutant genotype and drug-resistant phenotype was confirmed by reverse genetics. Cross-resistance studies and thermostability assays revealed that LPCRW_0005 has a similar mechanism of action to the capsid binder pleconaril. Elucidation of the crystal structure of the HRV14/LPCRW_0005 complex revealed the existence of multiple hydrophobic and polar interactions between the VP1 pocket and LPCRW_0005.
  
  
  CONCLUSIONS
  LPCRW_0005 is a novel inhibitor of HRV14 replication that acts as a capsid binder. The compound has a chemical structure that is markedly smaller than that of other capsid binders. Structural studies show that LPCRW_0005, in contrast to pleconaril, leaves the toe end of the pocket in VP1 empty. This suggests that extended analogues of LPCRW_0005 that fill the full cavity could be more potent inhibitors of rhinovirus replication.
authors.affiliation: Laboratory of Virology and Experimental Chemotherapy, Rega Institute for Medical Research, KU Leuven, Leuven, Belgium.
authors.forename: Céline
authors.initials: C
authors.lastname: Lacroix
authors.affiliation: Molecular Biology Institute of Barcelona (IBMB), Consejo Superior de Investigaciones Científicas, Barcelona, Spain.
authors.forename: Jordi
authors.initials: J
authors.lastname: Querol-Audí
authors.affiliation: Aix-Marseille Université, Institut de Chimie Radicalaire UMR 7273 CNRS 27, Boulevard Jean Moulin, Marseille, France.
authors.forename: Manon
authors.initials: M
authors.lastname: Roche
authors.affiliation: Laboratory of Virology and Experimental Chemotherapy, Rega Institute for Medical Research, KU Leuven, Leuven, Belgium.
authors.forename: David
authors.initials: D
authors.lastname: Franco
authors.affiliation: Laboratory of Medicinal Chemistry, Rega Institute for Medical Research, KU Leuven, Leuven, Belgium.
authors.forename: Mathy
authors.initials: M
authors.lastname: Froeyen
authors.affiliation: Molecular Biology Institute of Barcelona (IBMB), Consejo Superior de Investigaciones Científicas, Barcelona, Spain.
authors.forename: Pablo
authors.initials: P
authors.lastname: Guerra
authors.affiliation: Aix-Marseille Université, Institut de Chimie Radicalaire UMR 7273 CNRS 27, Boulevard Jean Moulin, Marseille, France.
authors.forename: Thierry
authors.initials: T
authors.lastname: Terme
authors.affiliation: Aix-Marseille Université, Institut de Chimie Radicalaire UMR 7273 CNRS 27, Boulevard Jean Moulin, Marseille, France.
authors.forename: Patrice
authors.initials: P
authors.lastname: Vanelle
authors.affiliation: Molecular Biology Institute of Barcelona (IBMB), Consejo Superior de Investigaciones Científicas, Barcelona, Spain.
authors.forename: Núria
authors.initials: N
authors.lastname: Verdaguer
authors.affiliation: Laboratory of Virology and Experimental Chemotherapy, Rega Institute for Medical Research, KU Leuven, Leuven, Belgium johan.neyts@rega.kuleuven.be.
authors.forename: Johan
authors.initials: J
authors.lastname: Neyts
authors.affiliation: Laboratory of Virology and Experimental Chemotherapy, Rega Institute for Medical Research, KU Leuven, Leuven, Belgium.
authors.forename: Pieter
authors.initials: P
authors.lastname: Leyssen
mesh_terms: D000818:Animals; D000998:Antiviral Agents / Q000737:chemistry / Q000494:pharmacology*; D001665:Binding Sites; D002460:Cell Line; D003588:Cytopathogenic Effect, Viral / Q000187:drug effects; D024882:Drug Resistance, Viral / Q000235:genetics; D005838:Genotype; D006801:Humans; D057927:Hydrophobic and Hydrophilic Interactions; D008958:Models, Molecular; D008968:Molecular Conformation; D009154:Mutation; D009570:Nitriles / Q000737:chemistry / Q000494:pharmacology*; D011485:Protein Binding; D012367:RNA, Viral; D012229:Rhinovirus / Q000187:drug effects* / Q000235:genetics; D014764:Viral Proteins / Q000737:chemistry / Q000378:metabolism; D014779:Virus Replication / Q000187:drug effects*
chemical_list: C000598011:4-(1-hydroxy-2-(4,5-dimethoxy-2-nitrophenyl)ethyl)benzonitrile; D000998:Antiviral Agents; D009570:Nitriles; D012367:RNA, Viral; D014764:Viral Proteins; C074995:viral protein 1, rhinovirus; C014356:benzonitrile
keywords: VP1; capsid binder; pleconaril; rhinovirus 14