Defense Date
2024
Document Type
Directed Research Project
First Advisor
Emanuele Alves
Abstract
In 2021, opioids were involved in 80,411 overdose-related deaths in the United States accounting for 74.5 % of all drug overdose deaths. In 2020, synthetic opioid-related deaths accounted for over 82% of opioid deaths. Bucinnazine (AP-237) and its derivatives (2-methyl-AP-237, para-methyl-AP-237, and AP-238) have gained popularity due to its online easy availability. Forensic and clinical identification of these substances are challenging and usually relies on the identification of their metabolites and the parent drug in biological samples. This study aimed to identify the metabolites for bucinnazine and its derivatives using in silico predictions and experimental data using Liquid Chromatography – Quadrupole Time-of-Flight – Mass Spectrometry (LC-QTOF-MS). In silico prediction of the metabolites was applied by combining four different software. Way2Drug was used to predict the sites of metabolism (SOM), SwissADME and ADMETlab for pharmacokinetic information, and GLORYx to predict phase I and phase II metabolites for all 4 drugs. In vitro experiments were performed for each drug using human, mouse, rat, and rabbit microsomes. The supernatants were collected and analyzed using an LC-QTOF-MS and Molecule Profiler. Specific metabolites with possible biomarker application were detected for each specie. Biomarkers for previous use of AP-237, 2-methyl-AP-237, para-methyl-AP-237, and AP-238 were identified in humans. Across species, it was not possible to identify the target drug associated to a specie in all cases. Among the species analyzed, mice, rats, and rabbits presented high metabolic similarity with humans with metabolites M2-M4, M6, M9, M12, M35, and M44-M53 in common. Therefore, studies aiming to evaluate the metabolism of cinnamylpiperazines for drug development or other applications could be performed in mice. The forensic study of metabolites of synthetic opioids is a powerful tool to improve early detection of these derivatives improving the response in cases of intoxications.
Rights
© The Author(s)
Is Part Of
VCU Master of Science in Forensic Science Directed Research Projects
Date of Submission
4-29-2024