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Edwards MR, Pietzsch C, Vausselin T, et al. High-Throughput Minigenome System for Identifying Small-Molecule Inhibitors of Ebola Virus Replication. ACS Infect Dis. 2015;1(8):380-7doi: 10.1021/acsinfecdis.5b00053.
Edwards, M. R., Pietzsch, C., Vausselin, T., Shaw, M. L., Bukreyev, A., & Basler, C. F. (2015). High-Throughput Minigenome System for Identifying Small-Molecule Inhibitors of Ebola Virus Replication. ACS infectious diseases, 1(8), 380-7. https://doi.org/10.1021/acsinfecdis.5b00053
Edwards, Megan R, et al. "High-Throughput Minigenome System for Identifying Small-Molecule Inhibitors of Ebola Virus Replication." ACS infectious diseases vol. 1,8 (2015): 380-7. doi: https://doi.org/10.1021/acsinfecdis.5b00053
Edwards MR, Pietzsch C, Vausselin T, Shaw ML, Bukreyev A, Basler CF. High-Throughput Minigenome System for Identifying Small-Molecule Inhibitors of Ebola Virus Replication. ACS Infect Dis. 2015 Aug 14;1(8):380-7. doi: 10.1021/acsinfecdis.5b00053. Epub 2015 Jun 24. PMID: 26284260; PMCID: PMC4537067.
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ACS Infect Dis. 2015 Aug 14;1(8):380-7. doi: 10.1021/acsinfecdis.5b00053. Epub 2015 Jun 24.

High-Throughput Minigenome System for Identifying Small-Molecule Inhibitors of Ebola Virus Replication.

ACS infectious diseases

Megan R Edwards, Colette Pietzsch, Thibaut Vausselin, Megan L Shaw, Alexander Bukreyev, Christopher F Basler

Affiliations

  1. Department of Microbiology, Icahn School of Medicine at Mount Sinai , New York, New York 10029, United States.
  2. Department of Pathology, Department of Microbiology and Immunology, and Galveston National Laboratory, University of Texas Medical Branch at Galveston , Galveston, Texas 77555, United States.

PMID: 26284260 PMCID: PMC4537067 DOI: 10.1021/acsinfecdis.5b00053

Abstract

Ebola virus (EBOV), a member of the family Filoviridae, is a nonsegmented negative-sense RNA virus that causes severe, often lethal, disease in humans. EBOV RNA synthesis is carried out by a complex that includes several viral proteins. The function of this machinery is essential for viral gene expression and viral replication and is therefore a potential target for antivirals. We developed and optimized a high-throughput screening (HTS) assay based on an EBOV minigenome assay, which assesses the function of the polymerase complex. The assay is robust in 384-well format and displays a large signal to background ratio and high Z-factor values. We performed a pilot screen of 2080 bioactive compounds, identifying 31 hits (1.5% of the library) with >70% inhibition of EBOV minigenome activity. We further identified eight compounds with 50% inhibitory concentrations below their 50% cytotoxic concentrations, five of which had selectivity index (SI) values >10, suggesting specificity against the EBOV polymerase complex. These included an inhibitor of inosine monophosphate dehydrogenase, a target known to modulate the EBOV replication complex. They also included novel classes of inhibitors, including inhibitors of protein synthesis and hypoxia inducible factor-1. Five compounds were tested for their ability to inhibit replication of a recombinant EBOV that expresses GFP (EBOV-GFP), and four inhibited EBOV-GFP growth at sub-cytotoxic concentrations. These data demonstrate the utility of the HTS minigenome assay for drug discovery and suggest potential directions for antifiloviral drug development.

Keywords: Ebola virus; RNA polymerase; antiviral; filovirus; high-throughput screen; translation

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