Comprehensive investigation on synthetic cannabinoids: Metabolic behavior and potency testing, using 5F-APP-PICA and...

Area of Science:

• Forensic Toxicology
• Pharmacology
• Organic Chemistry

Background:

• Synthetic cannabinoids (SCs) are a growing concern, comprising 45% of new psychoactive substance seizures in Europe.
• Their unknown toxicity, diverse structures, and rapid modification pose significant challenges for detection and regulation.
• SCs are extensively metabolized, necessitating metabolite analysis for accurate consumption monitoring in forensic samples.

Purpose of the Study:

• To develop and apply a comprehensive workflow for studying SC consumption.
• To elucidate metabolites of 5F-APP-PICA and AMB-FUBINACA for use as consumption markers.
• To assess the functional activity of these SCs at CB1 and CB2 receptors.

Main Methods:

• High-resolution mass spectrometry was used to identify metabolites in human hepatocytes.
• A workflow integrating metabolite identification and functional assays (NanoLuc approach) was employed.
• Bioassays measured the half maximal effective concentration (EC50) and maximum response (Emax) at CB1 and CB2 receptors.

Main Results:

• 12 and 8 metabolites were elucidated for 5F-APP-PICA and AMB-FUBINACA, respectively, confirming complete metabolism.
• Suitable consumption markers were proposed based on identified metabolites.
• 5F-APP-PICA exhibited lower activity at CB1 and CB2 receptors compared to AMB-FUBINACA.

Conclusions:

• The proposed workflow effectively identifies SC metabolites for forensic analysis.
• Metabolites of 5F-APP-PICA and AMB-FUBINACA are suitable markers for consumption monitoring.
• Observed differences in receptor activity align with reported clinical intoxication cases, particularly for AMB-FUBINACA.