Modeling metabolism of the synthetic cannabinoids Bay 59‐3074 and JWH‐015 by human liver microsomes and cytochrome p450
Authors: Vanderpuye Oluseyi Adewale , Lampkin Alexius , Dunn Christina , Bailey Alexis 
Published: June 2021

Synthetic cannabinoids (SCs) produce profound physiological effects by binding to the CB1 and CB2 cannabinoid receptors in the brain and peripheral tissues. A problem exists in detecting abuse of novel SCs that become introduced for drug abuse because the metabolites are initially unknown and must be identified and their analytical standards produced. It is hypothesized that incubation with human liver microsomes and cytochrome p450 will produce metabolites of SCs JWH‐015 and Bay 59‐3074 found in vivo. It is also postulated that oxidation of SCs with chemical reagents will produce some of the same compounds produced by cytochrome p450 enzymes and enable convenient production of metabolite standards.JWH‐015 and Bay 59‐3074 and controls eosin and coumarin were incubated with human liver microsomes (HLM) and recombinant cytochrome p450 2D6 (CYP 2D6) and were also subjected to chemical oxidation by sodium hypochlorite. Chromatography and spectroscopy were used to detect changes in the SCs. Millimolar concentrations of the synthetic cannabinoids JWH‐015 and Bay 59‐3074 and eosin and coumarin controls were detectable by thin layer chromatography and uv‐visible spectroscopy. On TLC plates, the uv fluorescence of the main component of eosin was unaffected but the fluorescence of a faster moving component was decreased after incubation with HLM. After incubation with HLM but not after incubation in the control buffer the coumarin samples contained a new higher Rf value blue component. The JWH‐015 and Bay 59‐3074 were not detected on TLC plates after the dilutions necessary for the incubations with HLM and CYP 2D6 or mock treatments. Uv‐vis spectroscopy detected increases in absorbance at 340nm, 337 nm and 310 nm in the JWH‐015 after incubation with HLM compared to the control. For the Bay 59‐3074, uv‐vis spectroscopy detected a decrease in absorbance at 337nm and 340nm and an increase at 310 nm after incubation with HLM. Sodium hypochlorite oxidation of JWH‐015 and Bay 59‐3074 was done to see if oxidation could be detected by TLC and for future comparison to changes produced by cytochrome p450 and HLM. The Rf value of JWH‐015 on TLC plates was decreased after incubation with sodium hypochlorite, but that of Bay 59‐3074 was unaffected. Eosin and coumarin incubated with sodium hypochlorite showed differences detected by both TLC and uv‐visible spectroscopy relative to controls.In conclusion, an issue with the incubations of the synthetic cannabinoids with HLM and CYP 2D6 is the necessity to work with high dilutions of these compounds to minimize protein‐denaturing concentrations of organic solvents in which they are soluble..Chemical oxidation and in vitro incubation with human liver microsomes show some promise for producing metabolites for chemical characterization for the synthetic cannabinoids Bay 59‐3074 and JWH‐015. This may help in detecting metabolites of these compounds in biological samples.Support or Funding InformationNSF REU Grant CHE‐146806 award to K. Buchmueller, Tim Hanks, Furman UniversityThis abstract is from the Experimental Biology 2018 Meeting. There is no full text article associated with this abstract published in The FASEB Journal.