Open AccessA Sensitive in Vitro High-Throughput Screen To Identify Pan-filoviral Replication Inhibitors Targeting the VP35–NP Interface
Author(s) -
Liu Gai,
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
Publication year - 2017
Publication title -
acs infectious diseases
Language(s) - English
Resource type - Journals
SCImago Journal Rank - 1.324
H-Index - 39
ISSN - 2373-8227
DOI - 10.1021/acsinfecdis.6b00209
Subject(s) - ebola virus , virology , nucleoprotein , viral replication , drug discovery , ebolavirus , biology , antiviral drug , marburg virus , computational biology , virus , bioinformatics
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.