Assay and inhibition of diacylglycerol lipase activity

作者：Johnston Meghan; Bhatt Shachi R; Sikka Surina; Mercier Richard W; West Jay M; Makriyannis Alexandros; Gatley S John; Duclos Richard I Jr^*

来源：Bioorganic & Medicinal Chemistry Letters, 2012, 22(14): 4585-4592.

DOI：10.1016/j.bmcl.2012.05.101

• Diacylglycerol lipase
• 2-Arachidonoylglycerol
• Endocannabinoid
• Labeled
• Assay
• Fluorescence resonance energy transfer

A series of N-formyl-alpha-amino acid esters of beta-lactone derivatives structurally related to tetrahydrolipstatin (THL) and O-3841 were synthesized that inhibit human and murine diacylglycerol lipase (DAGL) activities. New ether lipid reporter compounds were developed for an in vitro assay to efficiently screen inhibitors of 1,2-diacyl-sn-glycerol hydrolysis and related lipase activities using fluorescence resonance energy transfer (FRET). A standardized thin layer chromatography (TLC) radioassay of diacylglycerol lipase activity utilizing the labeled endogenous substrate [1 ''-C-14]1-stearoyl-2-arachidonoyl-sn-glycerol with phosphorimaging detection was used to quantify inhibition by following formation of the initial product [1 ''-C-14]2-arachidonoylglycerol and further hydrolysis under the assay conditions to [1-C-14]arachidonic acid.

• 出版日期2012-7-15
• 单位东北大学