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Biochemistry . BRD4354 Is a Potent Covalent Inhibitor against the SARS-CoV-2 Main Protease

tetano

Editor, Senior Moderator
Biochemistry


. 2024 Feb 8.
doi: 10.1021/acs.biochem.3c00685. Online ahead of print. BRD4354 Is a Potent Covalent Inhibitor against the SARS-CoV-2 Main Protease

Yan J Sheng[SUP] 1 [/SUP], Syuan-Ting A Kuo[SUP] 1 [/SUP], Tingyuan Yang[SUP] 1 [/SUP], Hanyuan E Zhang[SUP] 1 [/SUP], David H Russell[SUP] 1 [/SUP], Xin Yan[SUP] 1 [/SUP], Shiqing Xu[SUP] 1 2 [/SUP], Wenshe R Liu[SUP] 1 3 4 5 [/SUP], Carol A Fierke[SUP] 6 [/SUP]



Affiliations
Abstract

Numerous organic molecules are known to inhibit the main protease (M[SUP]Pro[/SUP]) of SARS-CoV-2, the pathogen of Coronavirus Disease 2019 (COVID-19). Guided by previous research on zinc-ligand inhibitors of M[SUP]Pro[/SUP] and zinc-dependent histone deacetylases (HDACs), we identified BRD4354 as a potent inhibitor of M[SUP]Pro[/SUP]. The in vitro protease activity assays show that BRD4354 displays time-dependent inhibition against M[SUP]Pro[/SUP] with an IC[SUB]50[/SUB] (concentration that inhibits activity by 50%) of 0.72 ± 0.04 μM after 60 min of incubation. Inactivation follows a two-step process with an initial rapid binding step with a K[SUB]I[/SUB] of 1.9 ± 0.5 μM followed by a second slow inactivation step, k[SUB]inact,max[/SUB] of 0.040 ± 0.002 min[SUP]-1[/SUP]. Native mass spectrometry studies indicate that a covalent intermediate is formed where the ortho-quinone methide fragment of BRD4354 forms a covalent bond with the catalytic cysteine C145 of M[SUP]Pro[/SUP]. Based on these data, a Michael-addition reaction mechanism between M[SUP]Pro[/SUP] C145 and BRD4354 was proposed. These results suggest that both preclinical testing of BRD4354 and structure-activity relationship studies based on BRD4354 are warranted to develop more effective anti-COVID therapeutics.



Yan J Sheng[SUP] 1 [/SUP], Syuan-Ting A Kuo[SUP] 1 [/SUP], Tingyuan Yang[SUP] 1 [/SUP], Hanyuan E Zhang[SUP] 1 [/SUP], David H Russell[SUP] 1 [/SUP], Xin Yan[SUP] 1 [/SUP], Shiqing Xu[SUP] 1 2 [/SUP], Wenshe R Liu[SUP] 1 3 4 5 [/SUP], Carol A Fierke[SUP] 6 [/SUP]



Affiliations
Abstract

Numerous organic molecules are known to inhibit the main protease (M[SUP]Pro[/SUP]) of SARS-CoV-2, the pathogen of Coronavirus Disease 2019 (COVID-19). Guided by previous research on zinc-ligand inhibitors of M[SUP]Pro[/SUP] and zinc-dependent histone deacetylases (HDACs), we identified BRD4354 as a potent inhibitor of M[SUP]Pro[/SUP]. The in vitro protease activity assays show that BRD4354 displays time-dependent inhibition against M[SUP]Pro[/SUP] with an IC[SUB]50[/SUB] (concentration that inhibits activity by 50%) of 0.72 ± 0.04 μM after 60 min of incubation. Inactivation follows a two-step process with an initial rapid binding step with a K[SUB]I[/SUB] of 1.9 ± 0.5 μM followed by a second slow inactivation step, k[SUB]inact,max[/SUB] of 0.040 ± 0.002 min[SUP]-1[/SUP]. Native mass spectrometry studies indicate that a covalent intermediate is formed where the ortho-quinone methide fragment of BRD4354 forms a covalent bond with the catalytic cysteine C145 of M[SUP]Pro[/SUP]. Based on these data, a Michael-addition reaction mechanism between M[SUP]Pro[/SUP] C145 and BRD4354 was proposed. These results suggest that both preclinical testing of BRD4354 and structure-activity relationship studies based on BRD4354 are warranted to develop more effective anti-COVID therapeutics.


 
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