tetano
Editor, Senior Moderator
Antiviral Res
. 2020 Sep 4;104924.
doi: 10.1016/j.antiviral.2020.104924. Online ahead of print.
Evaluation of SARS-CoV-2 3C-like protease inhibitors using self-assembled monolayer desorption ionization mass spectrometry
Zachary A Gurard-Levin[SUP] 1 [/SUP], Cheng Liu[SUP] 2 [/SUP], Andreas Jekle[SUP] 2 [/SUP], Ruchika Jaisinghani[SUP] 2 [/SUP], Suping Ren[SUP] 2 [/SUP], Koen Vandyck[SUP] 3 [/SUP], Dirk Jochmans[SUP] 4 [/SUP], Pieter Leyssen[SUP] 4 [/SUP], Johan Neyts[SUP] 4 [/SUP], Lawrence M Blatt[SUP] 2 [/SUP], Leonid Beigelman[SUP] 2 [/SUP], Julian A Symons[SUP] 2 [/SUP], Pierre Raboisson[SUP] 3 [/SUP], Michael D Scholle[SUP] 1 [/SUP], Jerome Deval[SUP] 5 [/SUP]
Affiliations
Abstract
Severe acute respiratory syndrome coronavirus 2 (SARS-CoV-2) is the cause of the COVID-19 pandemic that began in 2019. The coronavirus 3-chymotrypsin-like cysteine protease (3CLpro) controls replication and is therefore considered a major target for antiviral discovery. This study describes the evaluation of SARS-CoV-2 3CLpro inhibitors in a novel self-assembled monolayer desorption ionization mass spectrometry (SAMDI-MS) enzymatic assay. Compared with a traditional FRET readout, the label-free SAMDI-MS assay offers greater sensitivity and eliminates false positive inhibition from compound interference with the optical signal. The SAMDI-MS assay was optimized and validated with known inhibitors of coronavirus 3CLpro such as GC376 (IC[SUB]50[/SUB]= 0.060 μM), calpain inhibitors II and XII (IC[SUB]50[/SUB] ∼20-25 μM). The FDA-approved drugs shikonin, disulfiram, and ebselen did not inhibit SARS-CoV-2 3CLpro activity in the SAMDI-MS assay under physiologically relevant reducing conditions. The three drugs did not directly inhibit human β-coronavirus OC-43 or SARS-CoV-2 in vitro, but instead induced cell death. In conclusion, the SAMDI-MS 3CLpro assay, combined with antiviral and cytotoxic assessment, provides a robust platform to evaluate antiviral agents directed against SARS-CoV-2.
Keywords: 3CLpro; COVID-19; SAMDI-MS; coronavirus; mass spectrometry; protease inhibitor.
. 2020 Sep 4;104924.
doi: 10.1016/j.antiviral.2020.104924. Online ahead of print.
Evaluation of SARS-CoV-2 3C-like protease inhibitors using self-assembled monolayer desorption ionization mass spectrometry
Zachary A Gurard-Levin[SUP] 1 [/SUP], Cheng Liu[SUP] 2 [/SUP], Andreas Jekle[SUP] 2 [/SUP], Ruchika Jaisinghani[SUP] 2 [/SUP], Suping Ren[SUP] 2 [/SUP], Koen Vandyck[SUP] 3 [/SUP], Dirk Jochmans[SUP] 4 [/SUP], Pieter Leyssen[SUP] 4 [/SUP], Johan Neyts[SUP] 4 [/SUP], Lawrence M Blatt[SUP] 2 [/SUP], Leonid Beigelman[SUP] 2 [/SUP], Julian A Symons[SUP] 2 [/SUP], Pierre Raboisson[SUP] 3 [/SUP], Michael D Scholle[SUP] 1 [/SUP], Jerome Deval[SUP] 5 [/SUP]
Affiliations
- PMID: 32896566
- DOI: 10.1016/j.antiviral.2020.104924
Abstract
Severe acute respiratory syndrome coronavirus 2 (SARS-CoV-2) is the cause of the COVID-19 pandemic that began in 2019. The coronavirus 3-chymotrypsin-like cysteine protease (3CLpro) controls replication and is therefore considered a major target for antiviral discovery. This study describes the evaluation of SARS-CoV-2 3CLpro inhibitors in a novel self-assembled monolayer desorption ionization mass spectrometry (SAMDI-MS) enzymatic assay. Compared with a traditional FRET readout, the label-free SAMDI-MS assay offers greater sensitivity and eliminates false positive inhibition from compound interference with the optical signal. The SAMDI-MS assay was optimized and validated with known inhibitors of coronavirus 3CLpro such as GC376 (IC[SUB]50[/SUB]= 0.060 μM), calpain inhibitors II and XII (IC[SUB]50[/SUB] ∼20-25 μM). The FDA-approved drugs shikonin, disulfiram, and ebselen did not inhibit SARS-CoV-2 3CLpro activity in the SAMDI-MS assay under physiologically relevant reducing conditions. The three drugs did not directly inhibit human β-coronavirus OC-43 or SARS-CoV-2 in vitro, but instead induced cell death. In conclusion, the SAMDI-MS 3CLpro assay, combined with antiviral and cytotoxic assessment, provides a robust platform to evaluate antiviral agents directed against SARS-CoV-2.
Keywords: 3CLpro; COVID-19; SAMDI-MS; coronavirus; mass spectrometry; protease inhibitor.