Evaluation of Acute Cell Toxicity of Pyrolytic Products of Synthetic Cannabinoids

As submitted by the applicant: Synthetic cannabinoids have become a ubiquitous challenge in forensic toxicology and seized drug analysis. Aside from various degrees of impairment, acute toxic effects associated with these drugs include tachycardia, seizures, depression, possible suicidal tendencies, and the onset of psychotic episodes. To date, these effects are poorly understood. These drugs are ingested primarily by smoking, which introduces additional complexity to the problem of understanding and characterizing acute and chronic toxic effects as well as degrees and types of impairment that occur after ingestion.

This objective of this proposal is to evaluate the toxicity of confirmed pyrolytic products produced from the smoking process of selected representative synthetic cannabinoids. Recent work in our laboratory has shown that some of the thermal degradation products would be expected to bind with the cannabinoid receptors, an unexpected finding. Additionally, recent literature reports describe the detection of thermal degradation of the synthetic cannabinoids in traditional toxicological matrices of blood and urine so the route of ingestion for these drugs is clearly an important contributor to their acute and potentially chronic toxic effects.

Toxicity of thermal degradation products (confirmed through recent literature and exact mass characterization in our laboratory) will be evaluated and compared to parent and metabolic compounds to assess the overall effects of abuse. The data will be used to develop detailed study of the mechanism of toxicity in various organs including the brain, lung, liver, buccal, and heart. This information is essential for understanding toxicological responses, degrees of incapacitation, prediction of acute and chronic toxic responses, and for developing assays for the appropriate body fluids and organs. The analytical instruments and protocols used in the study (GC/MS, LC/MS/MS, LTQ-FTICR/MS, TECAN Infinite M1000 plate reader with fluorescence, and BioPlex 200 Flow Cytometer) are all in place and no new equipment or instrumentation is needed for this project. All analytical protocols will be validated as per the Standard Practices for Method Validation in Forensic Toxicology. All of the expertise necessary for completion of the project is available through the student’s established research committee and the PI’s research group has many years of experience in forensic chemical analysis including the analysis of smoked drugs of abuse, working with liver microsomes, method development, and toxicological analyses.

Note: This project contains a research and/or development component, as defined in applicable law.

nca/ncf.

The objective of this proposal is to evaluate the toxicity of confirmed pyrolytic products produced from the smoking process of selected representative synthetic cannabinoids. This is a timely project given that current literature reports describe the detection of thermal degradation of the synthetic cannabinoids in traditional toxicological matrices of blood and urine.

Fundamental understanding of the toxic effects of thermal degradation products would assist forensic toxicologists assess intoxication and direct development of new assays.

This knowledge could also be of use to the field of medicolegal death investigation and finally in the broader context of public health and safety. Toxicity of thermal degradation products (confirmed through recent literature and exact mass characterization in our laboratory) will be evaluated and compared to parent and metabolic compounds to assess the overall effects of abuse.

The data will be used to develop the mechanism of toxicity caused by the pyrolytic products in various organs including the brain, lung, liver, buccal and heart. The analytical methods used in the study (GC/MS, LC/MS/MS, LTQ-FTICR/MS, TECAN Infinite M1000 plate reader with fluorescence, and BioPlex 200 Flow Cytometer) will be validated according to the guidelines set forth by the Standard Practices for Method Validation in Forensic Toxicology, and the project will aid in achieving research goals set out by SWGTOX in August 2014.

All of the equipment, instrumentation, and expertise necessary for completion of the proposed project are in place and the PI’s research group has many years of experience in forensic chemical analysis including the analysis of smoked drugs of abuse, working with liver microsomes, method development, and toxicological analyses. There will also be a collaborative effort with colleagues in the medical examiner’s community to ensure connection of the research to findings observed in case work.

This project contains a research and/or development component, as defined in the applicable law.

nca/ncf