A novel fluorophosphonate inhibitor of the biosynthesis of the endocannabinoid 2-arachidonoylglycerol with potential anti-obesity effects

Br J Pharmacol. 2013 Jun;169(4):784-93. doi: 10.1111/bph.12013.

Abstract

Background and purpose: 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.

Experimental approach: 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.

Key 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 and implications: 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.

Publication types

  • Comparative Study
  • Research Support, N.I.H., Extramural

MeSH terms

  • Animals
  • Anti-Obesity Agents / administration & dosage
  • Anti-Obesity Agents / pharmacology
  • Anti-Obesity Agents / therapeutic use*
  • Arachidonic Acids / antagonists & inhibitors*
  • Arachidonic Acids / metabolism
  • Behavior, Animal / drug effects
  • Cell Line
  • Chlorocebus aethiops
  • Dose-Response Relationship, Drug
  • Endocannabinoids / antagonists & inhibitors*
  • Endocannabinoids / metabolism
  • Energy Intake / drug effects
  • Enzyme Inhibitors / pharmacology
  • Enzyme Inhibitors / therapeutic use*
  • Glycerides / antagonists & inhibitors*
  • Glycerides / metabolism
  • Glycerophospholipids / administration & dosage
  • Glycerophospholipids / pharmacology
  • Glycerophospholipids / therapeutic use*
  • Humans
  • Hypothalamus / drug effects
  • Hypothalamus / enzymology
  • Hypothalamus / metabolism
  • Lipoprotein Lipase / antagonists & inhibitors*
  • Lipoprotein Lipase / genetics
  • Lipoprotein Lipase / metabolism
  • Liver / drug effects
  • Liver / enzymology
  • Liver / metabolism
  • Male
  • Mice
  • Mice, Inbred C57BL
  • Nerve Tissue Proteins / antagonists & inhibitors
  • Nerve Tissue Proteins / genetics
  • Nerve Tissue Proteins / metabolism
  • Neurons / drug effects
  • Neurons / enzymology
  • Neurons / metabolism
  • Obesity / drug therapy*
  • Obesity / enzymology
  • Obesity / metabolism
  • Oleic Acids / administration & dosage
  • Oleic Acids / pharmacology
  • Oleic Acids / therapeutic use*
  • Organophosphonates / administration & dosage
  • Organophosphonates / pharmacology
  • Organophosphonates / therapeutic use*
  • Rats
  • Recombinant Proteins / chemistry
  • Recombinant Proteins / metabolism
  • Sterol Esterase / antagonists & inhibitors
  • Sterol Esterase / metabolism

Substances

  • 1-((fluoro(methyl)phosphoryl)oxy)-3-isopropoxypropan-2-yl oleate
  • Anti-Obesity Agents
  • Arachidonic Acids
  • Endocannabinoids
  • Enzyme Inhibitors
  • Glycerides
  • Glycerophospholipids
  • Nerve Tissue Proteins
  • Oleic Acids
  • Organophosphonates
  • Recombinant Proteins
  • glyceryl 2-arachidonate
  • Nceh1 protein, mouse
  • Sterol Esterase
  • DAGLA protein, human
  • Lipoprotein Lipase