Identification of West Nile virus RNA-dependent RNA polymerase non-nucleoside inhibitors by real-time high throughput fluorescence screening.

Autor: García-Zarandieta M; Facultad de Farmacia, Universidad San Pablo-CEU, CEU Universities, Urbanización Montepríncipe, 28660, Boadilla del Monte, Spain., Quesada E; Instituto de Química Médica (IQM, CSIC), 28006, Madrid, Spain., Martínez-Jiménez MI; Centro de Biología Molecular 'Severo Ochoa' (CSIC-UAM), Cantoblanco, E-28049, Madrid, Spain., Newnes CV; Facultad de Farmacia, Universidad San Pablo-CEU, CEU Universities, Urbanización Montepríncipe, 28660, Boadilla del Monte, Spain., Fernández-Cabello V; Instituto de Química Médica (IQM, CSIC), 28006, Madrid, Spain., Sáez-Álvarez Y; Facultad de Farmacia, Universidad San Pablo-CEU, CEU Universities, Urbanización Montepríncipe, 28660, Boadilla del Monte, Spain., Blázquez AB; Departamento de Biotecnología, Instituto Nacional de Investigación y Tecnología Agraria y Alimentaria, (INIA, CSIC), 289040, Madrid, Spain., Escribano-Romero E; Departamento de Biotecnología, Instituto Nacional de Investigación y Tecnología Agraria y Alimentaria, (INIA, CSIC), 289040, Madrid, Spain., Saiz JC; Departamento de Biotecnología, Instituto Nacional de Investigación y Tecnología Agraria y Alimentaria, (INIA, CSIC), 289040, Madrid, Spain., Del Aguila C; Facultad de Farmacia, Universidad San Pablo-CEU, CEU Universities, Urbanización Montepríncipe, 28660, Boadilla del Monte, Spain., Martín-Acebes MA; Departamento de Biotecnología, Instituto Nacional de Investigación y Tecnología Agraria y Alimentaria, (INIA, CSIC), 289040, Madrid, Spain., Pérez-Pérez MJ; Instituto de Química Médica (IQM, CSIC), 28006, Madrid, Spain., Agudo R; Facultad de Farmacia, Universidad San Pablo-CEU, CEU Universities, Urbanización Montepríncipe, 28660, Boadilla del Monte, Spain. Electronic address: ruben.agudotorres@ceu.es.
Jazyk: angličtina
Zdroj: Antiviral research [Antiviral Res] 2023 Apr; Vol. 212, pp. 105568. Date of Electronic Publication: 2023 Feb 24.
DOI: 10.1016/j.antiviral.2023.105568
Abstrakt: West Nile virus (WNV) is a re-emergent mosquito-borne RNA virus that causes major outbreaks of encephalitis around the world. However, there is no therapeutic treatment to struggle against WNV, and the current treatment relies on alleviating symptoms. Therefore, due to the threat virus poses to animal and human health, there is an urgent need to come up with fast strategies to identify and assess effective antiviral compounds. A relevant target when developing drugs against RNA viruses is the viral RNA-dependent RNA polymerase (RdRp), responsible for the replication of the viral genome within a host cell. RdRps are key therapeutic targets based on their specificity for RNA and their essential role in the propagation of the infection. We have developed a fluorescence-based method to measure WNV RdRp activity in a fast and reliable real-time way. Interestingly, rilpivirine has shown in our assay inhibition of the WNV RdRp activity with an IC 50 value of 3.3 μM and its antiviral activity was confirmed in cell cultures. Furthermore, this method has been extended to build up a high-throughput screening platform to identify WNV polymerase inhibitors. By screening a small chemical library, novel RdRp inhibitors 1-4 have been identified. When their antiviral activity was tested against WNV in cell culture, 4 exhibited an EC 50 value of 2.5 μM and a selective index of 12.3. Thus, rilpivirine shows up as an interesting candidate for repurposing against flavivirus. Moreover, the here reported method allows the rapid identification of new WNV RdRp inhibitors.
Competing Interests: Declaration of competing interest The authors declare that they have no known competing financial interests or personal relationships that could have appeared to influence the work reported in this paper.
(Copyright © 2023 The Authors. Published by Elsevier B.V. All rights reserved.)
Databáze: MEDLINE
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