A Sensitive in Vitro High-Throughput Screen to Identify Pan-filoviral Replication Inhibitors Targeting the VP35-NP Interface

Gai Liu, Peter J. Nash, Britney Johnson, Colette Pietzsch, Ma Xenia G. Ilagan, Alexander Bukreyev, Christopher F. Basler, Terry L. Bowlin, Donald T. Moir, Daisy W. Leung, Gaya K. Amarasinghe

    Research output: Contribution to journalArticle

    10 Scopus citations

    Abstract

    The 2014 Ebola outbreak in West Africa, the largest outbreak on record, highlighted the need for novel approaches to therapeutics targeting Ebola virus (EBOV). Within the EBOV replication complex, the interaction between polymerase cofactor, viral protein 35 (VP35), and nucleoprotein (NP) is critical for viral RNA synthesis. We recently identified a peptide at the N-terminus of VP35 (termed NPBP) that is sufficient for interaction with NP and suppresses EBOV replication, suggesting that the NPBP binding pocket can serve as a potential drug target. Here we describe the development and validation of a sensitive high-throughput screen (HTS) using a fluorescence polarization assay. Initial hits from this HTS include the FDA-approved compound tolcapone, whose potency against EBOV infection was validated in a nonfluorescent secondary assay. High conservation of the NP-VP35 interface among filoviruses suggests that this assay has the capacity to identify pan-filoviral inhibitors for development as antivirals.

    Original languageEnglish (US)
    Pages (from-to)190-198
    Number of pages9
    JournalACS Infectious Diseases
    Volume3
    Issue number3
    DOIs
    StatePublished - Mar 10 2017

    Keywords

    • Ebola virus
    • VP35
    • fluorescence polarization assay
    • high-throughput screening
    • nucleoprotein

    ASJC Scopus subject areas

    • Infectious Diseases

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  • Cite this

    Liu, G., Nash, P. J., Johnson, B., Pietzsch, C., Ilagan, M. X. G., Bukreyev, A., Basler, C. F., Bowlin, T. L., Moir, D. T., Leung, D. W., & Amarasinghe, G. K. (2017). A Sensitive in Vitro High-Throughput Screen to Identify Pan-filoviral Replication Inhibitors Targeting the VP35-NP Interface. ACS Infectious Diseases, 3(3), 190-198. https://doi.org/10.1021/acsinfecdis.6b00209