Identifying SARS-CoV-2 Antiviral Compounds by Screening for Small Molecule Inhibitors of Nsp15 Endoribonuclease
Identifying SARS-CoV-2 Antiviral Compounds by Screening for Small Molecule Inhibitors of Nsp15 Endoribonuclease
Summary SARS-CoV-2 is responsible for COVID-19, a human disease that has caused over 2 million deaths, stretched health systems to near-breaking point and endangered the economies of countries and families around the world. Antiviral treatments to combat COVID-19 are currently lacking. Remdesivir, the only antiviral drug approved for the treatment of COVID-19, can affect disease severity, but better treatments are needed. SARS-CoV-2 encodes 16 non-structural proteins (nsp) that possess different enzymatic activities with important roles in viral genome replication, transcription and host immune evasion. One key aspect of host immune evasion is performed by the uridine-directed endoribonuclease activity of nsp15. Here we describe the expression and purification of nsp15 recombinant protein. We have developed biochemical assays to follow its activity, and we have found evidence for allosteric behaviour. We screened a custom chemical library of over 5000 compounds to identify nsp15 endoribonuclease inhibitors, and we identified and validated NSC95397 as an inhibitor of nsp15 endoribonuclease in vitro. Although NSC95397 did not inhibit SARS-CoV-2 growth in VERO E6 cells, further studies will be required to determine the effect of nsp15 inhibition on host immune evasion.
Fujisawa Ryo、Canal Berta、McClure Allison W.、Deegan Tom、Wu Mary、Drury Lucy S.、Howell Michael、Labib Karim、Zeng Jingkun、Ulferts Rachel、Weissmann Florian、Diffley John F.X、Bertolin Agustina P.、Beale Rupert
The MRC Protein Phosphorylation and Ubiquitylation Unit, School of Life Sciences, University of DundeeChromosome Replication Laboratory, The Francis Crick InstituteChromosome Replication Laboratory, The Francis Crick InstituteThe MRC Protein Phosphorylation and Ubiquitylation Unit, School of Life Sciences, University of DundeeHigh Throughput Screening, The Francis Crick InstituteChromosome Replication Laboratory, The Francis Crick InstituteHigh Throughput Screening, The Francis Crick InstituteThe MRC Protein Phosphorylation and Ubiquitylation Unit, School of Life Sciences, University of DundeeChromosome Replication Laboratory, The Francis Crick InstituteCell Biology of Infection Laboratory, the Francis Crick InstituteChromosome Replication Laboratory, The Francis Crick InstituteChromosome Replication Laboratory, The Francis Crick InstituteChromosome Replication Laboratory, The Francis Crick InstituteCell Biology of Infection Laboratory, the Francis Crick Institute
医学研究方法基础医学生物科学研究方法、生物科学研究技术
Fujisawa Ryo,Canal Berta,McClure Allison W.,Deegan Tom,Wu Mary,Drury Lucy S.,Howell Michael,Labib Karim,Zeng Jingkun,Ulferts Rachel,Weissmann Florian,Diffley John F.X,Bertolin Agustina P.,Beale Rupert.Identifying SARS-CoV-2 Antiviral Compounds by Screening for Small Molecule Inhibitors of Nsp15 Endoribonuclease[EB/OL].(2025-03-28)[2025-04-28].https://www.biorxiv.org/content/10.1101/2021.04.07.438811.点此复制
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