title: Identification of novel inhibitors of HCV RNA-dependent RNA polymerase by pharmacophore-based virtual screening and in vitro evaluation.
abstract: Hepatitis C virus (HCV) is the major etiological agent of non-A, non-B hepatitis where no effective treatment is available. The HCV NS5B with RNA-dependent RNA polymerase (RdRp) activity is a key target for the treatment of HCV infection. Here we report novel NS5B polymerase inhibitors identified by virtual screening and in vitro evaluation of their inhibitory activities. On the basis of a newly identified binding pocket of NS5B, distinct from the nucleotide binding site but highly conserved among various HCV isolates, we performed virtual screening of compounds that fit this binding pocket from the available chemical database of 3.5 million compounds. The inhibitory activities of the in silico selected 119 compounds were estimated with in vitro RdRp assay. Three compounds with IC50 values of about 20 microM were identified, and their kinetic analyses suggest that these compounds are noncompetitive inhibitors with respect to the ribonucleotide substrate. Furthermore, the single-point mutations of the conserved residues in the binding pocket of NS5B resulted in the significant decrease of the RdRp activity, indicating that the binding pocket presented here might be important for the therapeutic intervention of HCV. These novel inhibitors would be useful for the development of effective anti-HCV agents.
authors.affiliation: Department of Biotechnology, College of Engineering, Yonsei University, Seoul 120-749, Republic of Korea.
authors.forename: Kisun
authors.initials: K
authors.lastname: Ryu
authors.forename: Nam Doo
authors.initials: ND
authors.lastname: Kim
authors.forename: Seong Il
authors.initials: SI
authors.lastname: Choi
authors.forename: Cheol Kyu
authors.initials: CK
authors.lastname: Han
authors.forename: Jeong Hyeok
authors.initials: JH
authors.lastname: Yoon
authors.forename: Kyoung Tai
authors.initials: KT
authors.lastname: No
authors.forename: Kyun-Hwan
authors.initials: KH
authors.lastname: Kim
authors.forename: Baik L
authors.initials: BL
authors.lastname: Seong