A multi-pronged evaluation of aldehyde-based tripeptidyl main protease inhibitors as SARS-CoV-2 antivirals.
A multi-pronged evaluation of aldehyde-based tripeptidyl main protease inhibitors as SARS-CoV-2 antivirals.
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DOI：
10.1016/j.ejmech.2022.114570
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发表时间：
2022-10-05
影响因子：
6.7
通讯作者：
Liu, Wenshe Ray
中科院分区：
文献类型：
--
作者：
Ma, Yuying;Yang, Kai S.;Geng, Zhi Zachary;Alugubelli, Yugendar R.;Shaabani, Namir;Vatansever, Erol C.;Ma, Xinyu R.;Cho, Chia-Chuan;Khatua, Kaustav;Xiao, Jing;Blankenship, Lauren R.;Yu, Ge;Sankaran, Banumathi;Li, Pingwei;Allen, Robert;Ji, Henry;Xu, Shiqing;Liu, Wenshe Ray
作为 COVID-19 病原体 SARS-CoV-2 的必需酶，主蛋白酶 (MPro) 是开发治疗 COVID-19 的抗病毒药物的可行靶点。通过改变 P2 和 P3 位点以及 N 端保护基团的化学组成，我们合成了 18 种三肽基 MPro 抑制剂，其中还包含醛弹头和 P1 位点的 β-(S-2-氧代吡咯烷-3-基)-丙醛醛。对这些抑制剂进行了系统表征，包括它们的体外酶抑制效力、它们与 MPro 复合物的 X 射线晶体结构、它们对 293T 细胞中瞬时表达的 MPro 的抑制，以及所选抑制剂的细胞毒性和 SARS-CoV-2 抗病毒效力。这些抑制剂的体外酶抑制 IC50 值变化很大，范围为 4.8 至 650 nM。测定的体外酶抑制IC50值表明，P2和P3位置上相对较小的侧链有利于获得高的体外MPro抑制效力，P3位置对α-碳上有两个烷基取代基的非天然氨基酸具有耐受性，并且抑制效力对N端保护基团敏感。测定了与 16 种抑制剂结合的 MPro 的 X 射线晶体结构。在所有结构中，MPro 活性位点半胱氨酸与结合抑制剂的醛弹头共价相互作用，形成 S 构型的半硫缩醛。对于所有抑制剂，N 端保护基团周围的选举密度较弱，表明该基团可能与 MPro 灵活结合。在 MPro 中，在残基 N142 和 Q189 上观察到较大的结构变化。与体外高酶抑制效力不同，大多数抑制剂对 293T 细胞中瞬时表达的 MPro 的抑制效力较低。能够高效抑制 293T 细胞中瞬时表达的 MPro 的抑制剂均在 P3 位点含有 O-叔丁基苏氨酸。这些抑制剂还表现出相对较低的细胞毒性和较高的抗病毒效力。总体而言，我们当前和之前的研究表明，P3 位点的 O-叔丁基苏氨酸是 MPro 三肽醛抑制剂实现高细胞和抗病毒效力的关键成分。
As an essential enzyme of SARS-CoV-2, the COVID-19 pathogen, main protease (MPro) is a viable target to develop antivirals for the treatment of COVID-19. By varying chemical compositions at both P2 and P3 positions and the N-terminal protection group, we synthesized 18 tripeptidyl MPro inhibitors that contained also an aldehyde warhead and β-(S-2-oxopyrrolidin-3-yl)-alaninal at the P1 position. Systematic characterizations of these inhibitors were conducted, including their in vitro enzymatic inhibition potency, X-ray crystal structures of their complexes with MPro, their inhibition of MPro transiently expressed in 293T cells, and cellular toxicity and SARS-CoV-2 antiviral potency of selected inhibitors. These inhibitors have a large variation of determined in vitro enzymatic inhibition IC50 values that range from 4.8 to 650 nM. The determined in vitro enzymatic inhibition IC50 values reveal that relatively small side chains at both P2 and P3 positions are favorable for achieving high in vitro MPro inhibition potency, the P3 position is tolerable toward unnatural amino acids with two alkyl substituents on the α-carbon, and the inhibition potency is sensitive toward the N-terminal protection group. X-ray crystal structures of MPro bound with 16 inhibitors were determined. In all structures, the MPro active site cysteine interacts covalently with the aldehyde warhead of the bound inhibitor to form a hemithioacetal that takes an S configuration. For all inhibitors, election density around the N-terminal protection group is weak indicating possible flexible binding of this group to MPro. In MPro, large structural variations were observed on residues N142 and Q189. Unlike their high in vitro enzymatic inhibition potency, most inhibitors showed low potency to inhibit MPro that was transiently expressed in 293T cells. Inhibitors that showed high potency to inhibit MPro transiently expressed in 293T cells all contain O-tert-butyl-threonine at the P3 position. These inhibitors also exhibited relatively low cytotoxicity and high antiviral potency. Overall, our current and previous studies indicate that O-tert-butyl-threonine at the P3 site is a key component to achieve high cellular and antiviral potency for tripeptidyl aldehyde inhibitors of MPro.
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影响因子：
16.8
作者：
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作者：
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