Abstract
The 3-chymotrypsin-like cysteine protease (3CLpro) of severe acute respiratory syndrome conoravirus 2 (SARS-CoV-2) remains a promising therapeutic target to combat COVID-19. Our group recently described a novel duplexed biochemical assay that combines self-assembled monolayers of alkanethiolates on gold with matrix assisted laser desorption ionization (MALDI) time of flight (TOF) mass spectrometry (MS) to simultaneously measure 3CLpro and human rhinovirus 3C protease activities. This study describes applying the assay for the completion of a high-throughput duplexed screen of 300,000 diverse, drug-like small molecules in 3 days. The hits were confirmed and evaluated in dose response analyses against recombinant 3CLpro, HRV3C, and the human Cathepsin L proteases. The 3CLpro specific inhibitors were further assessed for activity in cellular cytotoxicity and anti-viral assays. Structure activity relationship studies informed on structural features required for activity and selectivity to 3CLpro over HRV3C. These results will guide the optimization of 3CLpro selective inhibitors to combat COVID-19 along with antiviral compounds against coronaviruses and rhinoviruses.
Keywords: COVID-19; Coronavirus; High-throughput screening; Mass spectrometry; Protease inhibitor; Rhinovirus.
【저자키워드】 COVID-19, coronavirus, mass spectrometry, High-throughput screening, Protease inhibitor, Rhinovirus., 【초록키워드】 SARS-CoV-2, mass spectrometry, Antiviral compounds, 3CLpro, protease, Anti-viral, Protease inhibitor, Severe acute respiratory syndrome, rhinovirus, gold, small molecule, Proteases, selectivity, inhibitor, optimization, cathepsin, therapeutic target, cysteine, cathepsin L, structure activity relationship, specific inhibitor, matrix, acute respiratory syndrome, mass, cysteine protease, activities, completion, dose response, chymotrypsin, selective, rhinoviruses, monolayer, Human rhinovirus, Antiviral compound, feature, cellular cytotoxicity, biochemical assay, described, assays, evaluated, required, coronavirus, analysis, 【제목키워드】 SARS-CoV-2, inhibitor, reveal,