title: Novel small-molecule inhibitors of hepatitis C virus entry block viral spread and promote viral clearance in cell culture.
abstract: Combinations of direct-acting anti-virals offer the potential to improve the efficacy, tolerability and duration of the current treatment regimen for hepatitis C virus (HCV) infection. Viral entry represents a distinct therapeutic target that has been validated clinically for a number of pathogenic viruses. To discover novel inhibitors of HCV entry, we conducted a high throughput screen of a proprietary small-molecule compound library using HCV pseudoviral particle (HCVpp) technology. We independently discovered and optimized a series of 1,3,5-triazine compounds that are potent, selective and non-cytotoxic inhibitors of HCV entry. Representative compounds fully suppress both cell-free virus and cell-to-cell spread of HCV in vitro. We demonstrate, for the first time, that long term treatment of an HCV cell culture with a potent entry inhibitor promotes sustained viral clearance in vitro. We have confirmed that a single amino acid variant, V719G, in the transmembrane domain of E2 is sufficient to confer resistance to multiple compounds from the triazine series. Resistance studies were extended by evaluating both the fusogenic properties and growth kinetics of drug-induced and natural amino acid variants in the HCVpp and HCV cell culture assays. Our results indicate that amino acid variations at position 719 incur a significant fitness penalty. Introduction of I719 into a genotype 1b envelope sequence did not affect HCV entry; however, the overall level of HCV replication was reduced compared to the parental genotype 1b/2a HCV strain. Consistent with these findings, I719 represents a significant fraction of the naturally occurring genotype 1b sequences. Importantly, I719, the most relevant natural polymorphism, did not significantly alter the susceptibility of HCV to the triazine compounds. The preclinical properties of these triazine compounds support further investigation of entry inhibitors as a potential novel therapy for HCV infection.
authors.affiliation: Research and Development, Progenics Pharmaceuticals, Inc., Tarrytown, New York, United States of America. gcoburn@progenics.com
authors.forename: Glen A
authors.initials: GA
authors.lastname: Coburn
authors.forename: Danielle N
authors.initials: DN
authors.lastname: Fisch
authors.forename: Sameer M
authors.initials: SM
authors.lastname: Moorji
authors.forename: Jean-Marc
authors.initials: JM
authors.lastname: de Muys
authors.forename: Jose D
authors.initials: JD
authors.lastname: Murga
authors.forename: Dorothy
authors.initials: D
authors.lastname: Paul
authors.forename: Kathleen P
authors.initials: KP
authors.lastname: Provoncha
authors.forename: Yakov
authors.initials: Y
authors.lastname: Rotshteyn
authors.forename: Amy Q
authors.initials: AQ
authors.lastname: Han
authors.forename: Dapeng
authors.initials: D
authors.lastname: Qian
authors.forename: Paul J
authors.initials: PJ
authors.lastname: Maddon
authors.forename: William C
authors.initials: WC
authors.lastname: Olson
mesh_terms: D000998:Antiviral Agents / Q000737:chemistry* / Q000494:pharmacology* / Q000627:therapeutic use; D002460:Cell Line; D024882:Drug Resistance, Viral; D005838:Genotype; D016174:Hepacivirus / Q000187:drug effects* / Q000235:genetics; D006526:Hepatitis C / Q000188:drug therapy* / Q000235:genetics; D006801:Humans; D054852:Small Molecule Libraries / Q000737:chemistry* / Q000494:pharmacology* / Q000627:therapeutic use; D014227:Triazines / Q000737:chemistry / Q000494:pharmacology / Q000627:therapeutic use; D014759:Viral Envelope Proteins / Q000235:genetics; D053586:Virus Internalization / Q000187:drug effects
chemical_list: D000998:Antiviral Agents; D054852:Small Molecule Libraries; D014227:Triazines; D014759:Viral Envelope Proteins