Discovery of thiosemicarbazone derivatives as promising SARS-CoV-2 Mpro inhibitors by spectroscopy and microscale thermophoresis

IF 3.5 3区医学 Q2 BIOCHEMISTRY & MOLECULAR BIOLOGY

Bioorganic & Medicinal Chemistry Pub Date : 2026-02-01 Epub Date: 2025-11-29 DOI:10.1016/j.bmc.2025.118501

Xing Xue , Leyao Chen , Miao Zhang , Xiaoyu Chang , Ruiyong Wang

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Abstract

Since the outbreak of COVID-19, one key strategy has been to screen inhibitors of the SARS-CoV-2 main protease (Mpro). Mpro is critical for viral replication and a key target for therapeutic development. In this work, we report the design and synthesis of 29 thiosemicarbazone derivatives. Enzymatic inhibition assays showed that 27 compounds exhibited inhibitory activity against Mpro, among which 4 compounds (1e, 2c, 2f, 3a) demonstrated significant inhibitory potency, with IC₅₀ values of 5.97 ± 0.98 μM, 4.43 ± 1.67 μM, 10.99 ± 2.39 μM, and 28.88 ± 3.57 μM, respectively. Fluorescence spectroscopy, molecular docking, microscale thermophoresis and molecular dynamics simulations were used to analyze the interaction mechanisms between thiosemicarbazone derivatives and Mpro. Fluorescence results indicated that the fluorescence quenching mechanism of the compounds on Mpro was predominantly static quenching. Hydrogen bonding and hydrophobic interactions were observed in the binding process. Microscale thermophoresis results confirmed that compounds 1e, 2c and 2f exhibit significant binding affinity for Mpro, with K_d values of 4.21 ± 0.35 μM, 3.24 ± 0.28 μM, and 24.26 ± 1.82 μM, respectively. This study provides valuable information for understanding the interaction mechanism between thiosemicarbazone derivatives and Mpro and offers insights into novel Mpro inhibitors.

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通过光谱和微尺度热电泳发现硫代氨基脲衍生物作为有前途的SARS-CoV-2 Mpro抑制剂。

自COVID-19爆发以来，一项关键策略是筛选SARS-CoV-2主要蛋白酶（Mpro）的抑制剂。Mpro对病毒复制至关重要，也是治疗开发的关键靶点。在这项工作中，我们报道了29个硫代氨基脲衍生物的设计和合成。酶促抑制实验表明，27个化合物对Mpro具有抑制活性，其中4个化合物（1e、2c、2f、3a）表现出显著的抑制作用，IC50值分别为5.97±0.98 μM、4.43±1.67 μM、10.99±2.39 μM和28.88±3.57 μM。采用荧光光谱、分子对接、微尺度热电泳和分子动力学模拟等方法分析了硫代氨基脲衍生物与Mpro的相互作用机理。荧光结果表明，化合物在Mpro上的荧光猝灭机制以静态猝灭为主。在结合过程中观察到氢键和疏水相互作用。微尺度热泳实验结果表明，化合物1e、2c和2f对Mpro具有明显的结合亲和力，Kd值分别为4.21±0.35 μM、3.24±0.28 μM和24.26±1.82 μM。本研究为了解硫代氨基脲衍生物与Mpro之间的相互作用机制提供了有价值的信息，并为开发新型Mpro抑制剂提供了见解。

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来源期刊

Bioorganic & Medicinal Chemistry 医学-生化与分子生物学

CiteScore

6.80

自引率

2.90%

发文量

413

审稿时长

17 days

期刊介绍： Bioorganic & Medicinal Chemistry provides an international forum for the publication of full original research papers and critical reviews on molecular interactions in key biological targets such as receptors, channels, enzymes, nucleotides, lipids and saccharides. The aim of the journal is to promote a better understanding at the molecular level of life processes, and living organisms, as well as the interaction of these with chemical agents. A special feature will be that colour illustrations will be reproduced at no charge to the author, provided that the Editor agrees that colour is essential to the information content of the illustration in question.