title: Identification of hepatitis C virus inhibitors targeting different aspects of infection using a cell-based assay.
abstract: With 2 to 3% of the worldwide population chronically infected, hepatitis C virus (HCV) infection continues to be a major health care burden. Unfortunately, current interferon-based treatment options are not effective in all patients and are associated with significant side effects. Consequently, there is an ongoing need to identify and develop new anti-HCV therapies. Toward this goal, we previously developed a cell-based HCV infection assay for antiviral compound screening based on a low-multiplicity-of-infection approach that uniquely allows for the identification of antiviral compounds that target cell culture-derived HCV (HCVcc) at any step of the viral infection cycle. Using this assay, here we report the screening of the NCI Diversity Set II library, containing 1,974 synthesized chemical compounds, and the identification of compounds with specific anti-HCV activity. In combination with toxicity counterscreening, we identified 30 hits from the compound library, 13 of which showed reproducible and dose-dependent inhibition of HCV with mean therapeutic indices (50% cytotoxic concentration [CC(50)]/50% effective concentration [EC(50)]) of greater than 6. Using HCV pseudotype and replicon systems of multiple HCV genotypes, as well as infectious HCVcc-based assembly and secretion analysis, we determined that different compounds within this group of candidate inhibitors target different steps of viral infection. The compounds identified not only will serve as biological probes to study and further dissect the biology of viral infection but also should facilitate the development of new anti-HCV therapeutic treatments.
authors.affiliation: Department of Medicine, The University of Illinois, Chicago, IL, USA.
authors.forename: Xuemei
authors.initials: X
authors.lastname: Yu
authors.forename: Bruno
authors.initials: B
authors.lastname: Sainz
authors.forename: Pavel A
authors.initials: PA
authors.lastname: Petukhov
authors.forename: Susan L
authors.initials: SL
authors.lastname: Uprichard
mesh_terms: D000998:Antiviral Agents / Q000494:pharmacology*; D002460:Cell Line; D002470:Cell Survival / Q000187:drug effects; D016000:Cluster Analysis; D004305:Dose-Response Relationship, Drug; D031541:Fluorescence Resonance Energy Transfer; D016174:Hepacivirus / Q000187:drug effects*; D006526:Hepatitis C / Q000188:drug therapy* / Q000821:virology; D057166:High-Throughput Screening Assays; D006801:Humans; D007202:Indicators and Reagents; D012367:RNA, Viral / Q000235:genetics / Q000302:isolation & purification; D060888:Real-Time Polymerase Chain Reaction; D012190:Retroviridae / Q000235:genetics; D012192:Retroviridae Infections / Q000235:genetics / Q000821:virology; D054852:Small Molecule Libraries; D013329:Structure-Activity Relationship
chemical_list: D000998:Antiviral Agents; D007202:Indicators and Reagents; D012367:RNA, Viral; D054852:Small Molecule Libraries