A novel fluorophosphonate inhibitor of the biosynthesis of the endocannabinoid 2-arachidonoylglycerol with potential anti-obesity effects.
OBJECTIVE
The development of potent and selective inhibitors of the biosynthesis of the endocannabinoid 2-arachidonoylglycerol (2-AG) via DAG lipases (DAGL) α and β is just starting to be considered as a novel and promising source of pharmaceuticals for the treatment of disorders that might benefit from a reduction in endocannabinoid tone, such as hyperphagia in obese subjects.


METHODS
Three new fluorophosphonate compounds O-7458, O-7459 and O-7460 were synthesized and characterized in various enzymatic assays. The effects of O-7460 on high-fat diet intake were tested in mice.


RESULTS
Of the new compounds, O-7460 exhibited the highest potency (IC₅₀ = 690 nM) against the human recombinant DAGLα, and selectivity (IC₅₀ > 10 μM) towards COS-7 cell and human monoacylglycerol lipase (MAGL), and rat brain fatty acid amide hydrolase. Competitive activity-based protein profiling confirmed that O-7460 inhibits mouse brain MAGL only at concentrations ≥ 10 μM, and showed that this compound has only one major 'off-target', that is, the serine hydrolase KIAA1363. O-7460 did not exhibit measurable affinity for human recombinant CB₁ or CB₂ cannabinoid receptors (Ki > 10 μM). In mouse neuroblastoma N18TG2 cells stimulated with ionomycin, O-7460 (10 μM) reduced 2-AG levels. When administered to mice, O-7460 dose-dependently (0-12 mg·kg⁻¹, i.p.) inhibited the intake of a high-fat diet over a 14 h observation period, and, subsequently, slightly but significantly reduced body weight.


CONCLUSIONS
O-7460 might be considered a useful pharmacological tool to investigate further the role played by 2-AG both in vitro and in vivo under physiological as well as pathological conditions.
Endocannabinoid Research Group, Institute of Biomolecular Chemistry, C.N.R., Pozzuoli, Italy. tbisogno@icb.cnr.it
Tiziana
T
Bisogno
Anu
A
Mahadevan
Roberto
R
Coccurello
Jae Won
JW
Chang
Marco
M
Allarà
Yugang
Y
Chen
Giacomo
G
Giacovazzo
Aron
A
Lichtman
Benjamin
B
Cravatt
Anna
A
Moles
Vincenzo
V
Di Marzo
D000818:Animals; D019440:Anti-Obesity Agents / Q000008:administration & dosage / Q000494:pharmacology / Q000627:therapeutic use*; D001095:Arachidonic Acids / Q000037:antagonists & inhibitors* / Q000378:metabolism; D001522:Behavior, Animal / Q000187:drug effects; D002460:Cell Line; D002522:Chlorocebus aethiops; D004305:Dose-Response Relationship, Drug; D063388:Endocannabinoids / Q000037:antagonists & inhibitors* / Q000378:metabolism; D002149:Energy Intake / Q000187:drug effects; D004791:Enzyme Inhibitors / Q000494:pharmacology / Q000627:therapeutic use*; D005989:Glycerides / Q000037:antagonists & inhibitors* / Q000378:metabolism; D020404:Glycerophospholipids / Q000008:administration & dosage / Q000494:pharmacology / Q000627:therapeutic use*; D006801:Humans; D007031:Hypothalamus / Q000187:drug effects / Q000201:enzymology / Q000378:metabolism; D008071:Lipoprotein Lipase / Q000037:antagonists & inhibitors* / Q000235:genetics / Q000378:metabolism; D008099:Liver / Q000187:drug effects / Q000201:enzymology / Q000378:metabolism; D008297:Male; D051379:Mice; D008810:Mice, Inbred C57BL; D009419:Nerve Tissue Proteins / Q000037:antagonists & inhibitors / Q000235:genetics / Q000378:metabolism; D009474:Neurons / Q000187:drug effects / Q000201:enzymology / Q000378:metabolism; D009765:Obesity / Q000188:drug therapy* / Q000201:enzymology / Q000378:metabolism; D009829:Oleic Acids / Q000008:administration & dosage / Q000494:pharmacology / Q000627:therapeutic use*; D063065:Organophosphonates / Q000008:administration & dosage / Q000494:pharmacology / Q000627:therapeutic use*; D051381:Rats; D011994:Recombinant Proteins / Q000737:chemistry / Q000378:metabolism; D002787:Sterol Esterase / Q000037:antagonists & inhibitors / Q000378:metabolism
C000591836:1-((fluoro(methyl)phosphoryl)oxy)-3-isopropoxypropan-2-yl oleate; D019440:Anti-Obesity Agents; D001095:Arachidonic Acids; D063388:Endocannabinoids; D004791:Enzyme Inhibitors; D005989:Glycerides; D020404:Glycerophospholipids; D009419:Nerve Tissue Proteins; D009829:Oleic Acids; D063065:Organophosphonates; D011994:Recombinant Proteins; C094503:glyceryl 2-arachidonate; C514712:Nceh1 protein, mouse; D002787:Sterol Esterase; C480280:DAGLA protein, human; D008071:Lipoprotein Lipase