Binding-site identification and genotypic profiling of hepatitis C virus polymerase inhibitors.
The search for hepatitis C virus polymerase inhibitors has resulted in the identification of several nonnucleoside binding pockets. The shape and nature of these binding sites differ across and even within diverse hepatitis C virus genotypes. These differences confront antiviral drug discovery with the challenge of finding compounds that are capable of inhibition in variable binding pockets. To address this, we have established a hepatitis C virus mutant and genotypic recombinant polymerase panel as a means of guiding medicinal chemistry through the elucidation of the site of action of novel inhibitors and profiling against genotypes. Using a genotype 1b backbone, we demonstrate that the recombinant P495L, M423T, M414T, and S282T mutant enzymes can be used to identify the binding site of an acyl pyrrolidine analog. We assess the inhibitory activity of this analog and other nonnucleoside inhibitors with our panel of enzyme isolates generated from clinical sera representing genotypes 1a, 1b, 2a, 2b, 3a, 4a, 5a, and 6a.
Pauwels
Frederik
F
HCV Research, Tibotec BVBA, Generaal de Wittelaan L11B 3, 2800 Mechelen, Belgium.
Mostmans
Wendy
W
Quirynen
Ludo M M
LM
van der Helm
Liesbet
L
Boutton
Carlo W
CW
Rueff
Anne-Stéphanie
AS
Cleiren
Erna
E
Raboisson
Pierre
P
Surleraux
Dominique
D
Nyanguile
Origène
O
Simmen
Kenneth A
KA
D019943:Amino Acid Substitution; D001483:Base Sequence; D001665:Binding Sites / Q000235:genetics; D004791:Enzyme Inhibitors / Q000737:chemistry*; D005838:Genotype; D016174:Hepacivirus / Q000201:enzymology* / Q000235:genetics; D006801:Humans; D008969:Molecular Sequence Data; D020125:Mutation, Missense; D017434:Protein Structure, Tertiary; D011759:Pyrrolidines / Q000737:chemistry*; D012324:RNA-Dependent RNA Polymerase / Q000737:chemistry* / Q000235:genetics
D004791:Enzyme Inhibitors; D011759:Pyrrolidines; D012324:RNA-Dependent RNA Polymerase; C032519:pyrrolidine