Development of the first potent and specific inhibitors of endocannabinoid biosynthesis

Tiziana Bisogno, Maria Grazia Cascio, Bijali Saha, Anu Mahadevan, Paolo Urbani, Alberto Minassi, Giovanni Appendino, Carmela Saturnino, Billy Martin, Raj Razdan, Vincenzo Di Marzo*

*Corresponding author for this work

Research output: Contribution to journalArticlepeer-review

118 Citations (Scopus)


Enzymes for the biosynthesis and degradation of the endocannabinoid 2-arachidonoyl glycerol (2-AG) have been cloned and are the sn-1-selective-diacylglycerol lipases α and β (DAGLα and β) and the monoacylglycerol lipase (MAGL), respectively. Here, we used membranes from COS cells over-expressing recombinant human DAGLα to screen new synthetic substances as DAGLα inhibitors, and cytosolic fractions from wild-type COS cells to look for MAGL inhibitors. DAGLα and MAGL activities were assessed by using sn-1-[14C]-oleoyl-2-arachidonoyl-glycerol and 2-[3H]-arachidonoylglycerol as substrates, respectively. We screened known compounds as well as new phosphonate derivatives of oleic acid and fluoro-phosphinoyl esters of different length. Apart from the general lipase inhibitor tetrahydrolipstatin (orlistat®) (IC50 ∼ 60 nM), the most potent inhibitors of DAGLα were O-3640 [octadec-9-enoic acid-1-(fluoro-methyl-phosphoryloxymethyl)-propylester] (IC50 = 500 nM), and O-3841 [octadec-9-enoic acid 1-methoxymethyl-2-(fluoro-methyl-phosphinoyloxy)-ethyl ester] (IC50 = 160 nM). Apart from being almost inactive on MAGL, these two compounds showed high selectivity over rat liver triacylglycerol lipase, rat N-acylphosphatidyl-ethanolamine-selective phospholipase D (involved in anandamide biosynthesis), rat fatty acid amide hydrolase and human recombinant cannabinoid CB1 and CB2 receptors. Methylarachidonoyl-fluorophosphonate and the novel compound UP-101 [O-ethyl-O-p-nitro-phenyl oleylphosphonate] inhibited both DAGLα and MAGL with similar potencies (IC50 = 0.8-0.1 and 3.7-3.2 μM, respectively). Thus, we report the first potent and specific inhibitors of the biosynthesis of 2-AG that may be used as pharmacological tools to investigate the biological role of this endocannabinoid.

Original languageEnglish
Pages (from-to)205-212
Number of pages8
JournalBiochimica et Biophysica Acta - Molecular and Cell Biology of Lipids
Issue number2
Early online date23 Jan 2006
Publication statusPublished - Feb 2006

Bibliographical note

Funding Information:
The authors acknowledge the National Institute of Drug Abuse (Grants DA-09789, DA-08904 and subaward 520479/PO P642467) for partly funding this work. We are grateful to Prof. N. Ueda, Kagawa University, Japan, for the generous gift of NAPE-PLD cDNA construct.


  • 2-Arachidonoylglycerol
  • Cannabinoid
  • Diacylglycerol
  • Inhibitor
  • Lipase


Dive into the research topics of 'Development of the first potent and specific inhibitors of endocannabinoid biosynthesis'. Together they form a unique fingerprint.

Cite this