Mechanism of Inhibition for BMS-791325, a Novel Non-nucleoside Inhibitor of Hepatitis C Virus NS5B Polymerase

Rigat K, et al., 289(48):33456-68, J Biol Chem, 2014

By using several biochemical and biophysical techniques, the authors were able to dissect the mechanism of action of BMS-791325 (a non-nucleoside inhibitor) on HCV (hepatitis C virus) NS5B, an RNA-dependent RNA polymerase. Particularly, this study revealed a non-competitive time-dependent inhibition mechanism for BMS-791325. In addition, it uncovered all parameters pertaining to inhibitor binding kinetics. The BLI kinetic measurements obtained from an Octet RED system eliminated the discrepancies shown by the kinetic parameters generated from the activity assay and stopped flow studies. Biotinylated NS5B enzymes were immobilized onto SSA biosensors. Although complementary, the stopped-flow and BLI techniques uncovered different steps of the mechanism giving rise to a full characterization of the interaction between BMS-791325 and NS5B polymerase.

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