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Title: A Sensitive in Vitro High-Throughput Screen To Identify Pan-filoviral Replication Inhibitors Targeting the VP35–NP Interface

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. Here, 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.

Authors:
 [1];  [2];  [3];  [4];  [3];  [4];  [5];  [2];  [2];  [3]; ORCiD logo [3]
  1. Washington Univ. School of Medicine, St. Louis, MO (United States); Microbiotix Inc., Worcester, MA (United States)
  2. Microbiotix Inc., Worcester, MA (United States)
  3. Washington Univ. School of Medicine, St. Louis, MO (United States)
  4. Univ. of Texas Medical Branch, Galveston, TX (United States)
  5. Georgia State Univ., Atlanta, GA (United States)
Publication Date:
Research Org.:
Argonne National Laboratory (ANL), Argonne, IL (United States)
Sponsoring Org.:
National Institutes of Health (NIH)
OSTI Identifier:
1351401
Grant/Contract Number:  
R01AI123926; R01 AI114654; U19AI109945; P01AI120943
Resource Type:
Journal Article: Accepted Manuscript
Journal Name:
ACS Infectious Diseases
Additional Journal Information:
Journal Volume: 3; Journal Issue: 3; Journal ID: ISSN 2373-8227
Publisher:
American Chemical Society (ACS)
Country of Publication:
United States
Language:
ENGLISH
Subject:
60 APPLIED LIFE SCIENCES; Ebola virus; nucleoprotein; VP35; fluorescence polarization assay; high-throughput screening; DNA replication; peptides and proteins; fluorescence; assays; inhibitors

Citation Formats

Liu, Gai, Nash, Peter J., Johnson, Britney, Pietzsch, Colette, Ilagan, Ma. Xenia G., Bukreyev, Alexander, Basler, Christopher F., Bowlin, Terry L., Moir, Donald T., Leung, Daisy W., and Amarasinghe, Gaya K. A Sensitive in Vitro High-Throughput Screen To Identify Pan-filoviral Replication Inhibitors Targeting the VP35–NP Interface. United States: N. p., 2017. Web. doi:10.1021/acsinfecdis.6b00209.
Liu, Gai, Nash, Peter J., Johnson, Britney, Pietzsch, Colette, Ilagan, Ma. Xenia G., Bukreyev, Alexander, Basler, Christopher F., Bowlin, Terry L., Moir, Donald T., Leung, Daisy W., & Amarasinghe, Gaya K. A Sensitive in Vitro High-Throughput Screen To Identify Pan-filoviral Replication Inhibitors Targeting the VP35–NP Interface. United States. https://doi.org/10.1021/acsinfecdis.6b00209
Liu, Gai, Nash, Peter J., Johnson, Britney, Pietzsch, Colette, Ilagan, Ma. Xenia G., Bukreyev, Alexander, Basler, Christopher F., Bowlin, Terry L., Moir, Donald T., Leung, Daisy W., and Amarasinghe, Gaya K. 2017. "A Sensitive in Vitro High-Throughput Screen To Identify Pan-filoviral Replication Inhibitors Targeting the VP35–NP Interface". United States. https://doi.org/10.1021/acsinfecdis.6b00209. https://www.osti.gov/servlets/purl/1351401.
@article{osti_1351401,
title = {A Sensitive in Vitro High-Throughput Screen To Identify Pan-filoviral Replication Inhibitors Targeting the VP35–NP Interface},
author = {Liu, Gai and Nash, Peter J. and Johnson, Britney and Pietzsch, Colette and Ilagan, Ma. Xenia G. and Bukreyev, Alexander and Basler, Christopher F. and Bowlin, Terry L. and Moir, Donald T. and Leung, Daisy W. and Amarasinghe, Gaya K.},
abstractNote = {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. Here, 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.},
doi = {10.1021/acsinfecdis.6b00209},
url = {https://www.osti.gov/biblio/1351401}, journal = {ACS Infectious Diseases},
issn = {2373-8227},
number = 3,
volume = 3,
place = {United States},
year = {Thu Feb 02 00:00:00 EST 2017},
month = {Thu Feb 02 00:00:00 EST 2017}
}

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Works referenced in this record:

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Works referencing / citing this record:

The Natural Product Eugenol Is an Inhibitor of the Ebola Virus In Vitro
journal, May 2019


Ebolaviruses: New roles for old proteins
journal, May 2018


Inhibitory Effects of Antiviral Drug Candidates on Canine Parvovirus in F81 cells
journal, August 2019