title: Natural prevalence of NS5A polymorphisms in subjects infected with hepatitis C virus genotype 3 and their effects on the antiviral activity of NS5A inhibitors.
abstract: BACKGROUND
  Hepatitis C virus (HCV) NS5A replication complex inhibitors (RCIs) have been shown to exhibit picomolar antiviral activity against genotype 1 (GT1) in vitro. This has translated into rapid and robust declines in HCV RNA in GT1 patients. Less is known about the susceptibility of other genotypes such as GT3 to inhibition by NS5A RCIs.
  
  
  OBJECTIVE
  To detect and phenotype naturally occurring HCVGT3 NS5A polymorphisms against two NS5A RCIs (daclatasvir [DCV] and GS-5885) currently in clinical development.
  
  
  METHODS
  The NS5A region from 96 HCV GT3 treatment-naive patients spanning North America, Europe and Australia was determined.
  
  
  RESULTS
  Phylogenetic analysis revealed a broad distribution with no significant geographic clustering. GT1 DCV resistance-associated variants (RAVs) were observed in GT3 subjects; variants (and their frequencies) included 28M/V (1%), 30A/K/S/T/V (10%), 31L/M (1%), E92A (1%) and Y93H (8.3%). A consensus sequence was used to generate a JFH1/3a-NS5A hybrid replicon and employed to assess susceptibility to NS5A RCIs. Against JFH1/3a-NS5A, DCV was more potent (EC(50) = 0.52 nM) than GS-5885 (EC(50) = 141 nM). DCV sensitivity was increased against JFH1/3a-NS5A-M28V (EC50 = 0.006 nM), A30V (EC(50) = 0.012 nM), and E92A (EC(50) = 0.004 nM) while the NS5A-A30K and -Y93H variants exhibited reduced sensitivity to DCV (EC50 values of 23 nM and 1120 nM, respectively) and to GS-5885 (EC50 values of 1770 nM and 4300 nM, respectively).
  
  
  CONCLUSIONS
  Substitutions conferring resistance to NS5A RCIs pre-existed in treatment-naive patients infected with HCV GT3. The effectiveness of these NS5A RCIs to exert efficacy in the clinic may depend on which inhibitor is used in combination with other antivirals.
authors: Bristol-Myers Squibb, 5 Research Parkway, Wallingford, CT 06492, USA. dennis.hernandez@bms.com
authors: Dennis
authors: D
authors: Hernandez
authors: Nannan
authors: N
authors: Zhou
authors: Joseph
authors: J
authors: Ueland
authors: Aaron
authors: A
authors: Monikowski
authors: Fiona
authors: F
authors: McPhee
mesh_terms: D000595:Amino Acid Sequence; D000998:Antiviral Agents / Q000494:pharmacology*; D002219:Carbamates; D024882:Drug Resistance, Viral; D016174:Hepacivirus / Q000145:classification / Q000187:drug effects* / Q000235:genetics*; D019698:Hepatitis C, Chronic / Q000453:epidemiology / Q000821:virology*; D006801:Humans; D007093:Imidazoles / Q000494:pharmacology; D008826:Microbial Sensitivity Tests; D008969:Molecular Sequence Data; D010802:Phylogeny; D011759:Pyrrolidines; D014633:Valine / Q000031:analogs & derivatives; D017361:Viral Nonstructural Proteins / Q000037:antagonists & inhibitors* / Q000235:genetics*; D012324:RNA-Dependent RNA Polymerase
chemical_list: D000998:Antiviral Agents; D002219:Carbamates; D007093:Imidazoles; D011759:Pyrrolidines; D014633:Valine; D017361:Viral Nonstructural Proteins; C087036:NS-5 protein, hepatitis C virus; D012324:RNA-Dependent RNA Polymerase; C549273:daclatasvir