Diacylglycerol lipases (DAGL) are responsible for the biosynthesis of the endocannabinoid 2‐arachidonoylglycerol. The fluorescent activity‐based probes DH379 and HT‐01 have been previously shown to label DAGLs and to cross‐react with the serine hydrolase ABHD6. Here, we report the synthesis and characterization of two new quenched activity‐based probes 1 and 2, the design of which was based on the structures of DH379 and HT‐01, respectively. Probe 1 contains a BODIPY‐FL and a 2,4‐dinitroaniline moiety as a fluorophore–quencher pair, whereas probe 2 employs a Cy5‐fluorophore and a cAB40‐quencher. The fluorescence of both probes was quenched with relative quantum yields of 0.34 and 0.0081, respectively. The probes showed target inhibition as characterized in activity‐based protein profiling assays using human cell‐ and mouse brain lysates, but were unfortunately not active in living cells, presumably due to limited cell permeability.

中文翻译：

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基于淬灭活性的二酰基甘油脂肪酶和α，β-水合酶域蛋白6探针的设计与合成

二酰基甘油脂酶（DAGL）负责内源性大麻素2-花生四烯酸甘油酯的生物合成。以前已经证明基于荧光活性的探针DH379和HT-01可以标记DAGL并与丝氨酸水解酶ABHD6交叉反应。在这里，我们报告了两种新的基于淬灭活性的探针1和2的合成和表征 ，它们的设计分别基于DH379和HT-01的结构。探针 1包含一个BODIPY-FL和一个2,4-二硝基苯胺部分作为荧光团-猝灭剂对，而探针 2使用Cy5荧光团和cAB40猝灭剂。两种探针的荧光均以相对量子产率分别为0.34和0.0081淬灭。如使用人类细胞和小鼠脑裂解物进行的基于活动的蛋白质谱分析所描述，这些探针显示出靶标抑制作用，但不幸的是，在活细胞中没有活性，可能是由于细胞通透性有限。