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Eur J Med Chem . Discovery, synthesis and mechanism study of 2,3,5-substituted [1,2,4]-thiadiazoles as covalent inhibitors targeting 3C-Like protea

tetano

Editor, Senior Moderator
Eur J Med Chem


. 2023 Jan 18;249:115129.
doi: 10.1016/j.ejmech.2023.115129. Online ahead of print.
Discovery, synthesis and mechanism study of 2,3,5-substituted [1,2,4]-thiadiazoles as covalent inhibitors targeting 3C-Like protease of SARS-CoV-2


Pengxuan Ren[SUP] 1 [/SUP], Changyue Yu[SUP] 2 [/SUP], Ruxue Zhang[SUP] 3 [/SUP], Tianqing Nie[SUP] 4 [/SUP], Qiaoyu Hu[SUP] 5 [/SUP], Hui Li[SUP] 4 [/SUP], Xianglei Zhang[SUP] 1 [/SUP], Xueyuan Zhang[SUP] 1 [/SUP], Shiwei Li[SUP] 1 [/SUP], Lu Liu[SUP] 1 [/SUP], Wenhao Dai[SUP] 2 [/SUP], Jian Li[SUP] 2 [/SUP], Yechun Xu[SUP] 6 [/SUP], Haixia Su[SUP] 7 [/SUP], Leike Zhang[SUP] 8 [/SUP], Hong Liu[SUP] 9 [/SUP], Fang Bai[SUP] 10 [/SUP]



Affiliations

Abstract

The 3C-like protease (3CL[SUP]pro[/SUP]) is essential for the replication and transcription of severe acute respiratory syndrome coronavirus 2 (SARS-CoV-2), making it a promising target for the treatment of corona virus disease 2019 (COVID-19). In this study, a series of 2,3,5-substituted [1,2,4]-thiadiazole analogs were discovered to be able to inhibit 3CL[SUP]pro[/SUP] as non-peptidomimetic covalent binders at submicromolar levels, with IC[SUB]50[/SUB] values ranging from 0.118 to 0.582 μM. Interestingly, these compounds were also shown to inhibit PL[SUP]pro[/SUP] with the same level of IC[SUB]50[/SUB] values, but had negligible effect on proteases such as chymotrypsin, cathepsin B, and cathepsin L. Subsequently, the antiviral abilities of these compounds were evaluated in cell-based assays, and compound 6g showed potent antiviral activity with an EC[SUB]50[/SUB] value of 7.249 μM. It was proposed that these compounds covalently bind to the catalytic cysteine 145 via a ring-opening metathesis reaction mechanism. To understand this covalent-binding reaction, we chose compound 6a, one of the identified hit compounds, as a representative to investigate the reaction mechanism in detail by combing several computational predictions and experimental validation. The process of ring-opening metathesis was theoretically studied using quantum chemistry calculations according to the transition state theory. Our study revealed that the 2,3,5-substituted [1,2,4]-thiadiazole group could covalently modify the catalytic cysteine in the binding pocket of 3CL[SUP]pro[/SUP] as a potential warhead. Moreover, 6a was a known GPCR modulator, and our study is also a successful computational method-based drug-repurposing study.

Keywords: 2,3,5-substituted [1,2,4]-thiadiazole; 3CL(pro); Covalent inhibitors; Ring-opening metathesis; SARS-CoV-2; Warhead.
 
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