Serine hydrolases comprise a large family of enzymes that have diverse roles in key cellular processes, such as lipid metabolism, cell signaling, and regulation of post-translation modifications of proteins. They are also therapeutic targets for multiple human pathologies, including viral infection, diabetes, hypertension, and Alzheimer disease; however, few have well-defined substrates and biological functions. Activity-based probes (ABPs) have been used as effective tools to both profile activity and screen for selective inhibitors of serine hydrolases. One broad-spectrum ABP containing a fluorophosphonate electrophile has been used extensively to advance our understanding of diverse serine hydrolases. Due to the success of this single reagent, several robust chemistries have been developed to further diversify and tune the selectivity of ABPs used to target serine hydrolases. In this review, we highlight approaches to identify selective serine hydrolase ABPs and suggest new synthetic methodologies that could be applied to further advance probe development.

丝氨酸水解酶包含一个大家族的酶，它们在关键细胞过程中发挥不同的作用，例如脂质代谢、细胞信号传导和蛋白质翻译后修饰的调节。它们也是多种人类疾病的治疗靶点，包括病毒感染、糖尿病、高血压和阿尔茨海默病；然而，很少有明确的底物和生物功能。基于活性的探针（ABP）已被用作分析丝氨酸水解酶活性和筛选选择性抑制剂的有效工具。一种含有氟膦酸亲电子试剂的广谱 ABP 已被广泛用于增进我们对各种丝氨酸水解酶的理解。由于这种单一试剂的成功，几种强大的化学物质已经被开发出来，以进一步多样化和调整用于靶向丝氨酸水解酶的 ABP 的选择性。在这篇综述中，我们重点介绍了识别选择性丝氨酸水解酶 ABP 的方法，并提出了可用于进一步推进探针开发的新合成方法。