title: In vitro resistance profile of the hepatitis C virus NS3/4A protease inhibitor TMC435.
abstract: TMC435 is a small-molecule inhibitor of the NS3/4A serine protease of hepatitis C virus (HCV) currently in phase 2 development. The in vitro resistance profile of TMC435 was characterized by selection experiments with HCV genotype 1 replicon cells and the genotype 2a JFH-1 system. In 80% (86/109) of the sequences from genotype 1 replicon cells analyzed, a mutation at NS3 residue D168 was observed, with changes to V or A being the most frequent. Mutations at NS3 positions 43, 80, 155, and 156, alone or in combination, were also identified. A transient replicon assay confirmed the relevance of these positions for TMC435 inhibitory activity. The change in the 50% effective concentrations (EC(50)s) observed for replicons with mutations at position 168 ranged from <10-fold for those with the D168G or D168N mutation to approximately 2,000-fold for those with the D168V or D168I mutation, compared to the EC(50) for the wild type. Of the positions identified, mutations at residue Q80 had the least impact on the activity of TMC435 (<10-fold change in EC(50)s), while greater effects were observed for some replicons with mutations at positions 43, 155, and 156. TMC435 remained active against replicons with the specific mutations observed after in vitro or in vivo exposure to telaprevir or boceprevir, including most replicons with changes at positions 36, 54, and 170 (<3-fold change in EC(50)s). Replicons carrying mutations affecting the activity of TMC435 remained fully susceptible to alpha interferon and NS5A and NS5B inhibitors. Finally, combinations of TMC435 with alpha interferon and NS5B polymerase inhibitors prevented the formation of drug-resistant replicon colonies.
authors.affiliation: Tibotec BVBA, Mechelen, Belgium. olenz@its.jnj.com
authors.forename: Oliver
authors.initials: O
authors.lastname: Lenz
authors.forename: Thierry
authors.initials: T
authors.lastname: Verbinnen
authors.forename: Tse-I
authors.initials: TI
authors.lastname: Lin
authors.forename: Leen
authors.initials: L
authors.lastname: Vijgen
authors.forename: Maxwell D
authors.initials: MD
authors.lastname: Cummings
authors.forename: Jimmy
authors.initials: J
authors.lastname: Lindberg
authors.forename: Jan Martin
authors.initials: JM
authors.lastname: Berke
authors.forename: Pascale
authors.initials: P
authors.lastname: Dehertogh
authors.forename: Els
authors.initials: E
authors.lastname: Fransen
authors.forename: Annick
authors.initials: A
authors.lastname: Scholliers
authors.forename: Katrien
authors.initials: K
authors.lastname: Vermeiren
authors.forename: Tania
authors.initials: T
authors.lastname: Ivens
authors.forename: Pierre
authors.initials: P
authors.lastname: Raboisson
authors.forename: Michael
authors.initials: M
authors.lastname: Edlund
authors.forename: Susan
authors.initials: S
authors.lastname: Storm
authors.forename: Lotta
authors.initials: L
authors.lastname: Vrang
authors.forename: Herman
authors.initials: H
authors.lastname: de Kock
authors.forename: Gregory C
authors.initials: GC
authors.lastname: Fanning
authors.forename: Kenneth A
authors.initials: KA
authors.lastname: Simmen
mesh_terms: D000998:Antiviral Agents / Q000494:pharmacology; D002460:Cell Line; D024882:Drug Resistance, Viral / Q000187:drug effects / Q000235:genetics; D004357:Drug Synergism; D005838:Genotype; D016174:Hepacivirus / Q000187:drug effects* / Q000201:enzymology / Q000235:genetics; D006526:Hepatitis C / Q000188:drug therapy* / Q000821:virology; D006575:Heterocyclic Compounds, 3-Ring / Q000494:pharmacology*; D006801:Humans; D066298:In Vitro Techniques; D016898:Interferon-alpha / Q000494:pharmacology; D016296:Mutagenesis; D011480:Protease Inhibitors / Q000494:pharmacology*; D000069616:Simeprevir; D013449:Sulfonamides / Q000494:pharmacology*; D017361:Viral Nonstructural Proteins / Q000037:antagonists & inhibitors* / Q000235:genetics; D014779:Virus Replication / Q000187:drug effects; D012697:Serine Endopeptidases; D043583:Nucleoside-Triphosphatase; D053487:DEAD-box RNA Helicases; D000085242:Viral Proteases
chemical_list: D000998:Antiviral Agents; D006575:Heterocyclic Compounds, 3-Ring; D016898:Interferon-alpha; D011480:Protease Inhibitors; D000069616:Simeprevir; D013449:Sulfonamides; D017361:Viral Nonstructural Proteins; C084422:NS3 protein, hepatitis C virus; D012697:Serine Endopeptidases; D043583:Nucleoside-Triphosphatase; D053487:DEAD-box RNA Helicases; D000085242:Viral Proteases