A Methodology for Radiolabeling of the Endocannabinoid 2-Arachidonoylglycerol (2-AG)

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• 作者：Richard I. Duclos ; Jr. ; Meghan Johnston ; Subramanian K. Vadivel ; Alexandros Makriyannis ; Sherrye T. Glaser ; S. John Gatley
• 刊名：Journal of Organic Chemistry
• 出版年：2011
• 出版时间：April 1, 2011
• 年：2011
• 卷：76
• 期：7
• 页码：2049-2055
• 全文大小：905K
• 年卷期：v.76,no.7(April 1, 2011)
• ISSN：1520-6904

文摘

The metabolic intermediate and endocannabinoid signaling lipid 2-arachidonoylglycerol (2-AG) has not been readily labeled, primarily because of its instability toward rearrangement. We now detail a synthetic method that easily gives tritiated 2-AG from [5,6,8,9,11,12,14,15-³H(N)]arachidonic acid in two steps. We utilized a short chain 1,3-diacylglycerol and proceeded through the 鈥渟tructured lipid鈥?[5鈥测€?6鈥测€?8鈥测€?9鈥测€?11鈥测€?12鈥测€?14鈥测€?15鈥测€?³H(N)]2-arachidonoyl-1,3-dibutyrylglycerol, a triacylglycerol that was conveniently deprotected in ethanol with acrylic beads containing Candida antarctica lipase B to give [5鈥测€?6鈥测€?8鈥测€?9鈥测€?11鈥测€?12鈥测€?14鈥测€?15鈥测€?³H(N)]2-arachidonoylglycerol ([³H]2-AG). The flash chromatographic separation necessary to isolate the labeled 2-acylglycerol [³H]2-AG resulted in only 4% of the rearrangement byproducts that have been a particular problem with previous methodologies. This reliable 鈥渒it鈥?method to prepare the radiolabeled endocannabinoid as needed gave tritiated 2-arachidonoylglycerol [³H]2-AG with a specific activity of 200 Ci/mmol for enzyme assays, metabolic studies, and tissue imaging. It has been run on unlabeled materials on over 10 mg scales and should be generally applicable to other 2-acylglycerols.