Therapeutic modulation of cannabinoid lipid signaling: metabolic profiling of a novel antinociceptive cannabinoid-2 receptor agonist

Life Sci. 2013 Mar 19;92(8-9):482-91. doi: 10.1016/j.lfs.2012.06.019. Epub 2012 Jun 28.

Abstract

Aims: AM-1241, a novel, racemic cannabinoid-2 receptor (CB2) ligand, is the primary experimental agonist used to characterize the role of CB2-mediated lipid signaling in health and disease, including substance abuse disorders. In vivo pharmacological effects have been used as indirect proxies for AM-1241 biotransformation processes that could modulate CB2 activity. We report the initial pre-clinical characterization of AM-1241 biotransformation and in vivo distribution.

Main methods: AM-1241 metabolism was characterized in a variety of predictive in vitro systems (Caco-2 cells; mouse, rat and human microsomes) and in the mouse in vivo. Liquid chromatography and mass spectrometry techniques were used to quantify AM-1241 tissue distribution and metabolic conversion.

Key findings: AM-1241 bound extensively to plasma protein/albumin. A pharmacological AM-1241 dose (25mg/kg, i.v.) was administered to mice for direct determination of its plasma half-life (37 min), following which AM-1241 was quantified in brain, spleen, liver, and kidney. After p.o. administration, AM-1241 was detected in plasma, spleen, and kidney; its oral bioavailability was ~21%. From Caco-2 permeability studies and microsomal-based hepatic clearance estimates, in vivo AM-1241 absorption was moderate. Hepatic microsomal metabolism of AM-1241 in vitro generated hydroxylation and demethylation metabolites. Species-dependent differences were discovered in AM-1241's predicted hepatic clearance. Our data demonstrate that AM-1241 has the following characteristics: a) short plasma half-life; b) limited oral bioavailability; c) extensive plasma/albumin binding; d) metabolic substrate for hepatic hydroxylation and demethylation; e) moderate hepatic clearance.

Significance: These results should help inform the design, optimization, and pre-clinical profiling of CB2 ligands as pharmacological tools and medicines.

Publication types

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

MeSH terms

  • Analgesics / pharmacology*
  • Animals
  • Biotransformation
  • Blood Proteins / metabolism
  • Caco-2 Cells
  • Cannabinoids / chemistry
  • Cannabinoids / pharmacokinetics
  • Cannabinoids / pharmacology
  • Chromatography, High Pressure Liquid
  • Endocannabinoids / physiology*
  • Humans
  • Indicators and Reagents
  • Injections, Intraperitoneal
  • Injections, Subcutaneous
  • Lipids / physiology*
  • Liver / metabolism
  • Male
  • Mice
  • Protein Binding
  • Receptor, Cannabinoid, CB2 / agonists*
  • Signal Transduction / drug effects
  • Spectrum Analysis
  • Tissue Distribution

Substances

  • AM 1241
  • Analgesics
  • Blood Proteins
  • Cannabinoids
  • Endocannabinoids
  • Indicators and Reagents
  • Lipids
  • Receptor, Cannabinoid, CB2