TY - JOUR
T1 - Development of the first potent and specific inhibitors of endocannabinoid biosynthesis
AU - Bisogno, Tiziana
AU - Cascio, Maria Grazia
AU - Saha, Bijali
AU - Mahadevan, Anu
AU - Urbani, Paolo
AU - Minassi, Alberto
AU - Appendino, Giovanni
AU - Saturnino, Carmela
AU - Martin, Billy
AU - Razdan, Raj
AU - Di Marzo, Vincenzo
N1 - 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.
PY - 2006/2
Y1 - 2006/2
N2 - 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.
AB - 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.
KW - 2-Arachidonoylglycerol
KW - Cannabinoid
KW - Diacylglycerol
KW - Inhibitor
KW - Lipase
UR - http://www.scopus.com/inward/record.url?scp=33646472897&partnerID=8YFLogxK
U2 - 10.1016/j.bbalip.2005.12.009
DO - 10.1016/j.bbalip.2005.12.009
M3 - Article
SN - 1388-1981
VL - 1761
SP - 205
EP - 212
JO - Biochimica et Biophysica Acta - Molecular and Cell Biology of Lipids
JF - Biochimica et Biophysica Acta - Molecular and Cell Biology of Lipids
IS - 2
ER -