A cell-based assay to discover inhibitors of Zika virus RNA-dependent RNA polymerase

Lidan Wang; Rui Zhou; Yitong Liu; Saisai Guo; Dongrong Yi; Jianyuan Zhao; Quanjie Li; Yongxin Zhang; Chen Liang; Jing Wang; Guangzhi Shan; Shan Cen

doi:10.1016/j.virol.2023.109939

A cell-based assay to discover inhibitors of Zika virus RNA-dependent RNA polymerase

Virology. 2024 Jan:589:109939. doi: 10.1016/j.virol.2023.109939. Epub 2023 Nov 11.

Authors

Lidan Wang¹, Rui Zhou¹, Yitong Liu¹, Saisai Guo¹, Dongrong Yi¹, Jianyuan Zhao¹, Quanjie Li¹, Yongxin Zhang¹, Chen Liang², Jing Wang³, Guangzhi Shan⁴, Shan Cen⁵

Affiliations

¹ CAMS Key Laboratory of Antiviral Drug Research, Institute of Medicinal Biotechnology, Chinese Academy of Medical Sciences and Peking Union Medical College, 1 Tiantanxili, Beijing, 100050, China.
² Lady Davis Institute for Medical Research, Jewish General Hospital, McGill University, Montreal, H3T 1E2, Canada.
³ CAMS Key Laboratory of Antiviral Drug Research, Institute of Medicinal Biotechnology, Chinese Academy of Medical Sciences and Peking Union Medical College, 1 Tiantanxili, Beijing, 100050, China. Electronic address: jingwang@imb.pumc.edu.cn.
⁴ CAMS Key Laboratory of Antiviral Drug Research, Institute of Medicinal Biotechnology, Chinese Academy of Medical Sciences and Peking Union Medical College, 1 Tiantanxili, Beijing, 100050, China. Electronic address: shanguangzhi@imb.pumc.edu.cn.
⁵ CAMS Key Laboratory of Antiviral Drug Research, Institute of Medicinal Biotechnology, Chinese Academy of Medical Sciences and Peking Union Medical College, 1 Tiantanxili, Beijing, 100050, China. Electronic address: shancen@imb.pumc.edu.cn.

PMID: 37979208
DOI: 10.1016/j.virol.2023.109939

Abstract

Zika virus (ZIKV) belongs to Flaviviridae, the Flavivirus genus. Its infection causes congenital brain abnormalities and Guillain-Barré syndrome. However, there are no effective vaccines, no FDA-approved drugs to manage ZIKV infection. The non-structural protein NS5 of ZIKV has been recognized as a valuable target of antivirals because of its RNA-dependent RNA polymerase (RdRp) and methyltransferase (MTase) activities essential for viral RNA synthesis. Here, we report a cell-based assay for discovering inhibitors of ZIKV NS5 and found that 5-Azacytidine potently inhibits ZIKV NS5, with EC₅₀ of 4.9 μM. Furthermore, 5-Azacytidine suppresses ZIKV replication by inhibiting NS5-mediated viral RNA transcription. Therefore, we have developed a cell-based ZIKV NS5 assay which can be deployed to discover ZIKV NS5 inhibitors and demonstrated the potential of 5-Azacytidine for further development as a ZIKV NS5 inhibitor.

Keywords: 5-Azacytidine; Antiviral; NS5; Nucleotide analog inhibitor; ZIKV.

MeSH terms

Antiviral Agents / chemistry
Azacitidine / metabolism
Azacitidine / pharmacology
Azacitidine / therapeutic use
Humans
RNA, Viral / genetics
RNA, Viral / metabolism
RNA-Dependent RNA Polymerase / metabolism
Viral Nonstructural Proteins / metabolism
Virus Replication
Zika Virus Infection* / drug therapy
Zika Virus* / genetics

Substances

Antiviral Agents
RNA-Dependent RNA Polymerase
Viral Nonstructural Proteins
RNA, Viral
Azacitidine