bcj-2021-0244 (1).pdf (2.91 MB)
Identifying SARS-CoV-2 antiviral compounds by screening for small molecule inhibitors of Nsp3 papain-like protease.
journal contributionposted on 2021-07-09, 08:42 authored by Chew Theng Lim, Kang Wei Tan, Mary Wu, Rachel Ulferts, Lee A Armstrong, Eiko Ozono, Lucy S Drury, Jennifer C Milligan, Theresa U Zeisner, Jingkun Zeng, Florian Weissmann, Berta Canal, Ganka Bineva-Todd, Michael Howell, Nicola O'Reilly, Rupert Beale, Yogesh Kulathu, Karim Labib, John FX Diffley
The COVID-19 pandemic has emerged as the biggest life-threatening disease of this century. Whilst vaccination should provide a long-term solution, this is pitted against the constant threat of mutations in the virus rendering the current vaccines less effective. Consequently, small molecule antiviral agents would be extremely useful to complement the vaccination program. The causative agent of COVID-19 is a novel coronavirus, SARS-CoV-2, which encodes at least nine enzymatic activities that all have drug targeting potential. The papain-like protease (PLpro) contained in the nsp3 protein generates viral non-structural proteins from a polyprotein precursor, and cleaves ubiquitin and ISG protein conjugates. Here we describe the expression and purification of PLpro. We developed a protease assay that was used to screen a custom compound library from which we identified dihydrotanshinone I and Ro 08-2750 as compounds that inhibit PLpro in protease and isopeptidase assays and also inhibit viral replication in cell culture-based assays.
Crick (Grant ID: 10827, Grant title: Beale FC001827) Crick (Grant ID: 10066, Grant title: Diffley FC001066) Crick (Grant ID: 10008, Grant title: STP High Throughput Screening) Crick (Grant ID: 10013, Grant title: STP Peptide Chemistry) Wellcome Trust (Grant ID: 106252/Z/14/Z, Grant title: WT 106252/Z/14/Z) European Commission (Grant ID: 844211 - HUMAN REPL MECH, Grant title: EC 844211 - HUMAN REPL MECH)
coronavirusCOVID-19proteaseubiquitinAdenosine MonophosphateAlanineAniline CompoundsAnimalsAntiviral AgentsBenzamidesChlorocebus aethiopsCoronavirus Papain-Like ProteasesDrug Evaluation, PreclinicalDrug SynergismEnzyme AssaysFlavinsFluorescence Resonance Energy TransferFuransHigh-Throughput Screening AssaysInhibitory Concentration 50NaphthalenesPhenanthrenesQuinonesReproducibility of ResultsSARS-CoV-2Small Molecule LibrariesVero CellsVirus ReplicationDiffley FC001066Beale FC001827HTSPC03 Chemical Sciences06 Biological Sciences11 Medical and Health SciencesBiochemistry & Molecular Biology