Discovery of a NAPE-PLD inhibitor that modulates emotional behavior in mice

Nat Chem Biol. 2020 Jun;16(6):667-675. doi: 10.1038/s41589-020-0528-7. Epub 2020 May 11.


N-acylethanolamines (NAEs), which include the endocannabinoid anandamide, represent an important family of signaling lipids in the brain. The lack of chemical probes that modulate NAE biosynthesis in living systems hamper the understanding of the biological role of these lipids. Using a high-throughput screen, chemical proteomics and targeted lipidomics, we report here the discovery and characterization of LEI-401 as a CNS-active N-acylphosphatidylethanolamine phospholipase D (NAPE-PLD) inhibitor. LEI-401 reduced NAE levels in neuroblastoma cells and in the brain of freely moving mice, but not in NAPE-PLD KO cells and mice, respectively. LEI-401 activated the hypothalamus-pituitary-adrenal axis and impaired fear extinction, thereby emulating the effect of a cannabinoid CB1 receptor antagonist, which could be reversed by a fatty acid amide hydrolase inhibitor. Our findings highlight the distinctive role of NAPE-PLD in NAE biosynthesis in the brain and suggest the presence of an endogenous NAE tone controlling emotional behavior.

Publication types

  • Research Support, N.I.H., Extramural
  • Research Support, N.I.H., Intramural
  • Research Support, Non-U.S. Gov't

MeSH terms

  • Amidohydrolases / metabolism
  • Animals
  • Behavior, Animal / drug effects*
  • Blood Proteins / metabolism
  • Brain / metabolism
  • Cannabinoid Receptor Antagonists / metabolism
  • Cell Line, Tumor
  • Drug Evaluation, Preclinical
  • Enzyme Inhibitors / chemistry*
  • Enzyme Inhibitors / metabolism
  • Enzyme Inhibitors / pharmacokinetics
  • Fear / drug effects
  • Humans
  • Lipid Metabolism / drug effects*
  • Male
  • Mice
  • Mice, Inbred C57BL
  • Molecular Structure
  • Phosphatidylethanolamines / metabolism*
  • Phospholipase D / antagonists & inhibitors*
  • Receptors, Cannabinoid / metabolism
  • Signal Transduction


  • Blood Proteins
  • Cannabinoid Receptor Antagonists
  • Enzyme Inhibitors
  • N-acylphosphatidylethanolamine
  • Phosphatidylethanolamines
  • Receptors, Cannabinoid
  • Phospholipase D
  • Amidohydrolases
  • fatty-acid amide hydrolase