TMC647055 is a potent and selective non-nucleoside inhibitor of the hepatitis C virus NS5B polymerase with EC50 value of 82 nM . Hepatitis C virus (HCV) NS5B polymerase is a RNA-dependant RNA-polymerase that is responsible for viral replication and plays an important role in HCV life cycle. HCV is a major cause of chronic liver disease and acute hepatitis, ultimately leading to liver failure, cirrhosis and hepatocellular carcinoma . TMC647055 is a selective and cell-permeating HCV NS5B inhibitor. In RNA polymerase primer-dependent transcription assay, TMC647055 inhibited HCV NS5B polymerase with IC50 value of 34 nM. In the Huh7-Luc cell line, TMC647055 inhibited the replication of genotype 1b replicon with EC50 values of 77 and 139 nM measured with luciferase readout and qRT-PCR readout, respectively. TMC647055 exhibited high-affinity interaction with NS5B across all genotypes (except for genotype 2b) with median KD value < 35 nM. In Huh7-Luc replicon cells, TMC647055 (750 nM) reduced 30 cell colonies formation. Also, TMC647055 (1.5 μM and 3.75 μM) reduced HCV replicon RNA .