Genotype-dependent sensitivity of hepatitis C virus to inhibitors of the p7 ion channel.
The hepatitis C virus (HCV) p7 protein plays a critical role during particle formation in cell culture and is required for virus replication in chimpanzees. The discovery that it displayed cation channel activity in vitro led to its classification within the "viroporin" family of virus-coded ion channel proteins, which includes the influenza A virus (IAV) M2 protein. Like M2, p7 was proposed as a potential target for much needed new HCV therapies, and this was supported by our finding that the M2 inhibitor, amantadine, blocked its activity in vitro. Since then, further compounds have been shown to inhibit p7 function but the relationship between inhibitory effects in vitro and efficacy against infectious virus is controversial. Here, we have sought to validate multiple p7 inhibitor compounds using a parallel approach combining the HCV infectious culture system and a rapid throughput in vitro assay for p7 function. We identify a genotype-dependent and subtype-dependent sensitivity of HCV to p7 inhibitors, in which results in cell culture largely mirror the sensitivity of recombinant protein in vitro; thus building separate sensitivity profiles for different p7 sequences. Inhibition of virus entry also occurred, suggesting that p7 may be a virion component. Second site effects on both cellular and viral processes were identified for several compounds in addition to their efficacy against p7 in vitro. Nevertheless, for some compounds antiviral effects were specific to a block of ion channel function.


CONCLUSIONS
These data validate p7 inhibitors as prototype therapies for chronic HCV disease. (HEPATOLOGY 2008;48:1779-1790.).
Griffin
Stephen
S
Institute of Molecular and Cellular Biology, Faculty of Biological Sciences and Astbury Centre for Structural Molecular Biology, University of Leeds, Leeds, UK.
Stgelais
Corine
C
Owsianka
Ania M
AM
Patel
Arvind H
AH
Rowlands
David
D
Harris
Mark
M
D017485:1-Deoxynojirimycin / Q000031:analogs & derivatives / Q000494:pharmacology / Q000627:therapeutic use; D000547:Amantadine / Q000494:pharmacology / Q000627:therapeutic use; D000595:Amino Acid Sequence; D000998:Antiviral Agents / Q000494:pharmacology* / Q000627:therapeutic use; D006528:Carcinoma, Hepatocellular / Q000473:pathology / Q000821:virology; D045744:Cell Line, Tumor; D002470:Cell Survival / Q000187:drug effects; D004305:Dose-Response Relationship, Drug; D005838:Genotype; D016174:Hepacivirus / Q000187:drug effects* / Q000235:genetics*; D006526:Hepatitis C / Q000188:drug therapy; D006801:Humans; D008113:Liver Neoplasms / Q000473:pathology / Q000821:virology; D008969:Molecular Sequence Data; D012299:Rimantadine / Q000494:pharmacology / Q000627:therapeutic use; D014764:Viral Proteins / Q000032:analysis / Q000037:antagonists & inhibitors* / Q000235:genetics*; D014779:Virus Replication / Q000187:drug effects; D000086082:Viroporin Proteins
D017485:1-Deoxynojirimycin; D000547:Amantadine; D000998:Antiviral Agents; D012299:Rimantadine; D014764:Viral Proteins; C112150:N-nonyl-1-deoxynojirimycin; C088369:p7 protein, Hepatitis C virus; D000086082:Viroporin Proteins