title: High-throughput screening of low molecular weight NS3-NS4A protease inhibitors using a fluorescence resonance energy transfer substrate.
abstract: Hepatitis C virus (HCV) NS3-NS4A protease is an attractive target for anti-HCV agents because of its important role in replication. An optimized fluorescence resonance energy transfer (FRET) substrate for NS3-NS4A protease, based on the sequence of the NS5A-5B cleavage site, was designed and synthesized. High-throughput screening of in-house compound libraries was performed using a FRET substrate FS10 (MOCAcDKIVPC-SMSYK-Dnp) and MBP-NS3-NS4A fusion protein. Several hit compounds were found, including YZ-9577 (2-oxido-1,2,5-oxadiazole-3,4-diyl) bis (phenylmethanone) with potent inhibitory activity (IC50=1.6 microM) and good selectivity against other human serine proteases.
authors.affiliation: Institute for Drug Discovery Research, Yamanouchi Pharmaceutical Co. Ltd., Ibaraki, Japan.
authors.forename: Kenji
authors.initials: K
authors.lastname: Sudo
authors.forename: Kayo
authors.initials: K
authors.lastname: Yamaji
authors.forename: Kouich
authors.initials: K
authors.lastname: Kawamura
authors.forename: Tomoko
authors.initials: T
authors.lastname: Nishijima
authors.forename: Naoko
authors.initials: N
authors.lastname: Kojima
authors.forename: Kazuhiko
authors.initials: K
authors.lastname: Aibe
authors.forename: Kunitada
authors.initials: K
authors.lastname: Shimotohno
authors.forename: Yasuaki
authors.initials: Y
authors.lastname: Shimizu
mesh_terms: D000998:Antiviral Agents / Q000138:chemical synthesis / Q000494:pharmacology*; D015195:Drug Design; D031541:Fluorescence Resonance Energy Transfer; D016174:Hepacivirus / Q000187:drug effects*; D006801:Humans; D010455:Peptides / Q000737:chemistry / Q000378:metabolism; D011480:Protease Inhibitors / Q000494:pharmacology*; D014760:Viral Fusion Proteins / Q000494:pharmacology
chemical_list: D000998:Antiviral Agents; D010455:Peptides; D011480:Protease Inhibitors; D014760:Viral Fusion Proteins