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Validation and Invalidation of SARS-CoV-2 Papain-like Protease Inhibitors

[Image: see text] SARS-CoV-2 encodes two viral cysteine proteases, the main protease (M(pro)) and the papain-like protease (PL(pro)), both of which are validated antiviral drug targets. PL(pro) is involved in the cleavage of viral polyproteins as well as immune modulation by removing ubiquitin and i...

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Autores principales: Ma, Chunlong, Wang, Jun
Formato: Online Artículo Texto
Lenguaje:English
Publicado: American Chemical Society 2022
Acceso en línea:https://www.ncbi.nlm.nih.gov/pmc/articles/PMC8806001/
https://www.ncbi.nlm.nih.gov/pubmed/35178512
http://dx.doi.org/10.1021/acsptsci.1c00240
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author Ma, Chunlong
Wang, Jun
author_facet Ma, Chunlong
Wang, Jun
author_sort Ma, Chunlong
collection PubMed
description [Image: see text] SARS-CoV-2 encodes two viral cysteine proteases, the main protease (M(pro)) and the papain-like protease (PL(pro)), both of which are validated antiviral drug targets. PL(pro) is involved in the cleavage of viral polyproteins as well as immune modulation by removing ubiquitin and interferon-stimulated gene product 15 (ISG15) from host proteins. Therefore, targeting PL(pro) might be a two-pronged approach. Several compounds including YM155, cryptotanshinone, tanshinone I, dihydrotanshinone I, tanshinone IIA, SJB2-043, 6-thioguanine, and 6-mercaptopurine were recently identified as SARS-CoV-2 PL(pro) inhibitors through high-throughput screenings. In this study, we aim to validate/invalidate the reported PL(pro) inhibitors using a combination of PL(pro) target-specific assays including enzymatic FRET assay, thermal shift binding assay (TSA), and cell-based FlipGFP assay. Collectively, our results showed that all compounds tested either did not show binding or led to denaturation of PL(pro) in the TSA binding assay, which might explain their weak enzymatic inhibition in the FRET assay. In addition, none of the compounds showed cellular PL(pro) inhibition as revealed by the FlipGFP assay. Therefore, more efforts are needed to search for potent and specific SARS-CoV-2 PL(pro) inhibitors.
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spelling pubmed-88060012022-02-01 Validation and Invalidation of SARS-CoV-2 Papain-like Protease Inhibitors Ma, Chunlong Wang, Jun ACS Pharmacol Transl Sci [Image: see text] SARS-CoV-2 encodes two viral cysteine proteases, the main protease (M(pro)) and the papain-like protease (PL(pro)), both of which are validated antiviral drug targets. PL(pro) is involved in the cleavage of viral polyproteins as well as immune modulation by removing ubiquitin and interferon-stimulated gene product 15 (ISG15) from host proteins. Therefore, targeting PL(pro) might be a two-pronged approach. Several compounds including YM155, cryptotanshinone, tanshinone I, dihydrotanshinone I, tanshinone IIA, SJB2-043, 6-thioguanine, and 6-mercaptopurine were recently identified as SARS-CoV-2 PL(pro) inhibitors through high-throughput screenings. In this study, we aim to validate/invalidate the reported PL(pro) inhibitors using a combination of PL(pro) target-specific assays including enzymatic FRET assay, thermal shift binding assay (TSA), and cell-based FlipGFP assay. Collectively, our results showed that all compounds tested either did not show binding or led to denaturation of PL(pro) in the TSA binding assay, which might explain their weak enzymatic inhibition in the FRET assay. In addition, none of the compounds showed cellular PL(pro) inhibition as revealed by the FlipGFP assay. Therefore, more efforts are needed to search for potent and specific SARS-CoV-2 PL(pro) inhibitors. American Chemical Society 2022-01-24 /pmc/articles/PMC8806001/ /pubmed/35178512 http://dx.doi.org/10.1021/acsptsci.1c00240 Text en © 2022 American Chemical Society https://pubs.acs.org/page/vi/chemistry_coronavirus_researchThis article is made available via the ACS COVID-19 subset (https://pubs.acs.org/page/vi/chemistry_coronavirus_research) for unrestricted RESEARCH re-use and analyses in any form or by any means with acknowledgement of the original source. These permissions are granted for the duration of the World Health Organization (WHO) declaration of COVID-19 as a global pandemic.
spellingShingle Ma, Chunlong
Wang, Jun
Validation and Invalidation of SARS-CoV-2 Papain-like Protease Inhibitors
title Validation and Invalidation of SARS-CoV-2 Papain-like Protease Inhibitors
title_full Validation and Invalidation of SARS-CoV-2 Papain-like Protease Inhibitors
title_fullStr Validation and Invalidation of SARS-CoV-2 Papain-like Protease Inhibitors
title_full_unstemmed Validation and Invalidation of SARS-CoV-2 Papain-like Protease Inhibitors
title_short Validation and Invalidation of SARS-CoV-2 Papain-like Protease Inhibitors
title_sort validation and invalidation of sars-cov-2 papain-like protease inhibitors
url https://www.ncbi.nlm.nih.gov/pmc/articles/PMC8806001/
https://www.ncbi.nlm.nih.gov/pubmed/35178512
http://dx.doi.org/10.1021/acsptsci.1c00240
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