1. 1,5-Benzodiazepines, a Novel Class of Hepatitis C Virus Polymerase Nonnucleoside Inhibitors
- Author
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Kenneth Simmen, Pierre Raboisson, Frederic Delouvroy, Maxwell D. Cummings, Wendy Mostmans, Tania Ivens, Pascale Dehertogh, Liesbet van der Helm, Geneviève Vandercruyssen, Origène Nyanguile, Walter Van den Broeck, Carlo Boutton, Jean Francois Bonfanti, Frederik Pauwels, Ludo Maria Marcel Quirynen, and Analytical Chemistry and Pharmaceutical Technology
- Subjects
Models, Molecular ,Plus ribavirin ,replication ,Genotype ,viruses ,Hepatitis C virus ,Molecular Sequence Data ,RNA-dependent RNA polymerase ,Hepacivirus ,Viral Nonstructural Proteins ,Biology ,Crystallography, X-Ray ,Virus Replication ,medicine.disease_cause ,Antiviral Agents ,Virus ,Benzodiazepines ,Structure-Activity Relationship ,chemistry.chemical_compound ,Cell Line, Tumor ,RNA polymerase ,medicine ,Humans ,Viral-RNA ,Pharmacology (medical) ,Enzyme Inhibitors ,NS5B ,Polymerase ,Pharmacology ,Binding Sites ,IN-VITRO ,RNA-Dependent RNA Polymerase ,Virology ,Potent inhibitors ,Infectious Diseases ,NS5B polymerase ,Protease inhibitor ,chemistry ,Viral replication ,Dependent-RNA-polymerase ,Mutation ,biology.protein ,De-novo initiation ,DNA Polymerase Inhibitor ,Mutations conferring resistance - Abstract
The exogenous control of hepatitis C virus (HCV) replication can be mediated through the inhibition of the RNA-dependent RNA polymerase (RdRp) activity of NS5B. Small-molecule inhibitors of NS5B include nucleoside and nonnucleoside analogs. Here, we report the discovery of a novel class of HCV polymerase nonnucleoside inhibitors, 1,5-benzodiazepines (1,5-BZDs), identified by high-throughput screening of a library of small molecules. A fluorescence-quenching assay and X-ray crystallography revealed that 1,5-BZD 4a bound stereospecifically to NS5B next to the catalytic site. When introduced into replicons, mutations known to confer resistance against chemotypes that bind at this site were detrimental to inhibition by 1,5-BZD 7a. Using a panel of enzyme isolates that covered genotypes 1 to 6, we showed that compound 4a inhibited genotype 1 only. In mechanistic studies, 4a was found to inhibit the RdRp activity of NS5B noncompetitively with GTP and to inhibit the formation of the first phosphodiester bond during the polymerization cycle. The specificity for the HCV target was evaluated by profiling the 1,5-BZDs against other viral and human polymerases, as well as BZD receptors.
- Published
- 2008
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