Panyain Nattawadee, Godinat Aurélien, Thawani Aditya Raymond, Lachiondo-Ortega Sofía, Mason Katie, Elkhalifa Sarah, Smith Lisa M, Harrigan Jeanine A, Tate Edward W
Department of Chemistry, Molecular Sciences Research Hub, Imperial College London London W12 0BZ UK
Mission Therapeutics Ltd, The Glenn Berge Building, Babraham Research Campus Babraham Cambridge CB22 3FH UK.
RSC Med Chem. 2021 Aug 16;12(11):1935-1943. doi: 10.1039/d1md00218j. eCollection 2021 Nov 17.
Ubiquitin carboxy-terminal hydrolase L1 (UCHL1), a deubiquitinating enzyme (DUB), is a potential drug target in various cancers, and liver and lung fibrosis. However, functions and substrates of UCHL1 remain poorly understood. Herein, we report the characterization of UCHL1 covalent inhibitor based on a thiazole cyanopyrrolidine scaffold. In combination with chemical proteomics, a closely related activity-based probe () was used to generate a comprehensive quantitative profile for on- and off-targets at endogenous cellular abundance. Both compounds are selective for UCHL1 over other DUBs in intact cells but also engage a range of other targets with good selectivity over the wider proteome, including aldehyde dehydrogenases, redox-sensitive Parkinson's disease related protein PARK7, and glutamine amidotransferase. Taken together, these results underline the importance of robust profiling of activity-based probes as chemical tools and highlight the cyanopyrrolidine warhead as a versatile platform for liganding diverse classes of protein with reactive cysteine residues which can be used for further inhibitor screening, and as a starting point for inhibitor development.
泛素羧基末端水解酶L1（UCHL1）是一种去泛素化酶（DUB），是多种癌症以及肝肺纤维化中的潜在药物靶点。然而，UCHL1的功能和底物仍知之甚少。在此，我们报道了基于噻唑氰基吡咯烷支架的UCHL1共价抑制剂的特性。结合化学蛋白质组学，使用一种密切相关的基于活性的探针（）来生成内源性细胞丰度下的靶标和脱靶标的全面定量图谱。这两种化合物在完整细胞中对UCHL1的选择性高于其他DUB，但对更广泛的蛋白质组也具有良好的选择性，可作用于一系列其他靶点，包括醛脱氢酶、氧化还原敏感的帕金森病相关蛋白PARK7和谷氨酰胺酰胺转移酶。综上所述，这些结果强调了将基于活性的探针作为化学工具进行全面分析的重要性，并突出了氰基吡咯烷弹头作为一个通用平台的作用，该平台可用于连接具有反应性半胱氨酸残基的不同类型蛋白质，可用于进一步的抑制剂筛选，并作为抑制剂开发的起点。
