Resolution of the interaction mechanisms and characteristics of non-nucleoside inhibitors of hepatitis C virus polymerase

► The interaction details for HCV polymerase inhibitors were determined by an SPR biosensor. ► Tegobuvir did not interact with the enzyme, consistent with a requirement for activation. ► Filibuvir interacts in a single step while VX-222 inhibition involves induced fit. ► The induced fit mechanism of VX-222 gives it a longer residence time than filibuvir. ► Filibuvir and VX-222 interfere with the interaction between the polymerase and RNA.