Selection and characterization of hepatitis C virus replicons dually resistant to the polymerase and protease inhibitors HCV-796 and boceprevir (SCH 503034)

Antimicrob Agents Chemother. 2009 Feb;53(2):401-11. doi: 10.1128/AAC.01081-08. Epub 2008 Oct 20.

Abstract

HCV-796 is a nonnucleoside inhibitor of the hepatitis C virus (HCV) nonstructural protein 5B (NS5B) polymerase, and boceprevir is an inhibitor of the NS3 serine protease. The emergence of replicon variants resistant to the combination of HCV-796 and boceprevir was evaluated. Combining the inhibitors greatly reduced the frequency with which resistant colonies arose; however, some resistant replicon cells could be isolated by the use of low inhibitor concentrations. These replicons were approximately 1,000-fold less susceptible to HCV-796 and 9-fold less susceptible to boceprevir. They also exhibited resistance to anthranilate nonnucleoside inhibitors of NS5B but were fully sensitive to inhibitors of different mechanisms: a pyranoindole, Hsp90 inhibitors, an NS5B nucleoside inhibitor, and pegylated interferon (Peg-IFN). The replicon was cleared from the combination-resistant cells by extended treatment with Peg-IFN. Mutations known to confer resistance to HCV-796 (NS5B C316Y) and boceprevir (NS3 V170A) were present in the combination-resistant replicons. These changes could be selected together and coexist in the same genome. The replicon bearing both changes exhibited reduced sensitivity to inhibition by HCV-796 and boceprevir but had a reduced replicative capacity.

MeSH terms

  • Antiviral Agents / pharmacology*
  • Benzofurans / pharmacology*
  • Cell Line
  • Cloning, Molecular
  • Drug Resistance, Viral / genetics*
  • Electroporation
  • Genetic Variation
  • Hepacivirus / drug effects*
  • Hepacivirus / genetics*
  • Humans
  • Interferons / pharmacology
  • Mutagenesis / drug effects
  • Mutation / genetics
  • Nucleic Acid Synthesis Inhibitors*
  • Proline / analogs & derivatives*
  • Proline / pharmacology
  • Protease Inhibitors / pharmacology*
  • RNA, Viral / biosynthesis
  • RNA, Viral / genetics
  • Replicon / genetics*
  • Reverse Transcriptase Polymerase Chain Reaction
  • Sulfonamides / pharmacology*

Substances

  • Antiviral Agents
  • Benzofurans
  • HCV 796
  • Nucleic Acid Synthesis Inhibitors
  • Protease Inhibitors
  • RNA, Viral
  • Sulfonamides
  • N-(3-amino-1-(cyclobutylmethyl)-2,3-dioxopropyl)-3-(2-((((1,1-dimethylethyl)amino)carbonyl)amino)-3,3-dimethyl-1-oxobutyl)-6,6-dimethyl-3-azabicyclo(3.1.0)hexan-2-carboxamide
  • Interferons
  • Proline