Design and Synthesis of Quenched Activity‐based Probes for Diacylglycerol Lipase and α,β‐Hydrolase Domain Containing Protein 6

다국어 초록 (Multilingual Abstract)

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.
Pairing up: The synthesis and characterization of two new quenched activity‐based probes for diacylglycerol lipase and α,β‐hydrolase domain containing protein 6 with BODIPY‐FL and 2,4‐dinitroaniline or Cy5‐fluorophore and a cAB40‐quencher as a fluorophore–quencher pair is reported. The probes showed target inhibition as characterized in activity‐based protein profiling assays using human cell and mouse brain lysates, but not in living cells.