Identification of bisindolylmaleimides and indolocarbazoles as inhibitors of HCV replication by tube-capture-RT-PCR

Yuko Murakami, Kohji Noguchi, Satoshi Yamagoe, Tetsuro Suzuki, Takaji Wakita, Hidesuke Fukazawa

Research output: Contribution to journalArticlepeer-review

17 Citations (Scopus)

Abstract

We devised a screening method for hepatitis C virus (HCV) inhibitors by exploiting the JFH1 viral culture system. The viral RNA released in the medium was adsorbed onto PCR plates, and real-time RT-PCR was performed by directly adding the one-step RT-PCR reaction mixture to the wells. The "tube-capture-RT-PCR" method obviates the need for labor-intensive RNA isolation and should allow high-throughput screening of HCV inhibitors. To substantiate the validity of the assay for drug screening, a pilot screen of an inhibitor library composed of 95 compounds was performed. In addition to the known inhibitors of HCV replication included in the library, the assay identified the PKC inhibitor bisindolylmaleimide I (BIM I) as an HCV replication inhibitor. BIM I was also effective in reducing the viral protein level in genotype 1b and 2a subgenomic replicon cells, indicating inhibition of HCV replication. Further assays revealed that a broad range of bisindolylmaleimides and indolocarbazoles inhibit HCV, but no correlation was found between the PKC inhibition pattern and anti-HCV activity. These series of compounds represent new classes of inhibitors that may warrant further development.

Original languageEnglish
Pages (from-to)112-117
Number of pages6
JournalAntiviral Research
Volume83
Issue number2
DOIs
Publication statusPublished - 2009 Aug 1

Keywords

  • Bisindolylmaleimide
  • HCV
  • High-throughput
  • Tube-capture-RT-PCR

ASJC Scopus subject areas

  • Pharmacology
  • Virology

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