title: Antiviral stilbene 1,2-diamines prevent initiation of hepatitis C virus RNA replication at the outset of infection.
abstract: The recent development of a cell culture model of hepatitis C virus (HCV) infection based on the JFH-1 molecular clone has enabled discovery of new antiviral agents. Using a cell-based colorimetric screening assay to interrogate a 1,200-compound chemical library for anti-HCV activity, we identified a family of 1,2-diamines derived from trans-stilbene oxide that prevent HCV infection at nontoxic, low micromolar concentrations in cell culture. Structure-activity relationship analysis of ~ 300 derivatives synthesized using click chemistry yielded compounds with greatly enhanced low nanomolar potency and a > 1,000:1 therapeutic ratio. Using surrogate models of HCV infection, we showed that the compounds selectively block the initiation of replication of incoming HCV RNA but have no impact on viral entry, primary translation, or ongoing HCV RNA replication, nor do they suppress persistent HCV infection. Selection of an escape variant revealed that NS5A is directly or indirectly targeted by this compound. In summary, we have identified a family of HCV inhibitors that target a critical step in the establishment of HCV infection in which NS5A translated de novo from an incoming genomic HCV RNA template is required to initiate the replication of this important human pathogen.
authors.affiliation: Department of Immunology and Microbial Science, The Scripps Research Institute, La Jolla, California 9203, USA. pgastaminza@cnb.csic.es
authors.forename: Pablo
authors.initials: P
authors.lastname: Gastaminza
authors.forename: Suresh M
authors.initials: SM
authors.lastname: Pitram
authors.forename: Marlene
authors.initials: M
authors.lastname: Dreux
authors.forename: Larissa B
authors.initials: LB
authors.lastname: Krasnova
authors.forename: Christina
authors.initials: C
authors.lastname: Whitten-Bauer
authors.forename: Jiajia
authors.initials: J
authors.lastname: Dong
authors.forename: Josan
authors.initials: J
authors.lastname: Chung
authors.forename: Valery V
authors.initials: VV
authors.lastname: Fokin
authors.forename: K Barry
authors.initials: KB
authors.lastname: Sharpless
authors.forename: Francis V
authors.initials: FV
authors.lastname: Chisari
mesh_terms: D000998:Antiviral Agents / Q000737:chemistry / Q000302:isolation & purification / Q000494:pharmacology* / Q000633:toxicity; D002460:Cell Line; D002470:Cell Survival / Q000187:drug effects; D003959:Diamines / Q000737:chemistry / Q000302:isolation & purification / Q000494:pharmacology* / Q000633:toxicity; D004353:Drug Evaluation, Preclinical / Q000379:methods; D024882:Drug Resistance, Viral; D016174:Hepacivirus / Q000187:drug effects*; D022781:Hepatocytes / Q000187:drug effects / Q000821:virology; D006801:Humans; D008826:Microbial Sensitivity Tests; D012367:RNA, Viral / Q000378:metabolism; D013267:Stilbenes / Q000737:chemistry / Q000302:isolation & purification / Q000494:pharmacology* / Q000633:toxicity; D013329:Structure-Activity Relationship; D017361:Viral Nonstructural Proteins / Q000235:genetics / Q000378:metabolism; D014779:Virus Replication / Q000187:drug effects*; D012324:RNA-Dependent RNA Polymerase
chemical_list: D000998:Antiviral Agents; D003959:Diamines; D012367:RNA, Viral; D013267:Stilbenes; D017361:Viral Nonstructural Proteins; C087036:NS-5 protein, hepatitis C virus; D012324:RNA-Dependent RNA Polymerase