Drug Administration Routes Impact the Metabolism of a Synthetic Cannabinoid in the Zebrafish Larvae Model

Abstract

Zebrafish (Danio rerio) larvae have gained attention as a valid model to study in vivo drug metabolism and to predict human metabolism. The microinjection of compounds, oligonucleotides, or pathogens into zebrafish embryos at an early developmental stage is a well-established technique. Here, we investigated the metabolism of zebrafish larvae after microinjection of methyl 2-(1-(5-fluoropentyl)-1H-pyrrolo[2,3-b]pyridine-3-carboxamido)-3,3-dimethylbutanoate (7′N-5F-ADB) as a representative of recently introduced synthetic cannabinoids. Results were compared to human urine data and data from the in vitro HepaRG model and the metabolic pathway of 7′N-5F-ADB were reconstructed. Out of 27 metabolites detected in human urine samples, 19 and 15 metabolites were present in zebrafish larvae and HepaRG cells, respectively. The route of administration to zebrafish larvae had a major impact and we found a high number of metabolites when 7′N-5F-ADB was microinjected into the caudal vein, heart ventricle, or hindbrain. We further studied the spatial distribution of the parent compound and its metabolites by mass spectrometry imaging (MSI) of treated zebrafish larvae to demonstrate the discrepancy in metabolite profiles among larvae exposed through different administration routes. In conclusion, zebrafish larvae represent a superb model for studying drug metabolism, and when combined with MSI, the optimal administration route can be determined based on in vivo drug distribution.

Recent trends in drugs of abuse metabolism studies for mass spectrometry–based analytical screening procedures

“…Zebrafish larvae were found to produce the highest number of phase I but also phase II metabolites (79 metabolites in total), followed by HepaRG cells (66 metabolites), and pooled HLS9 (57 metabolites). However, the findings by Richter et al [27] indicated that the route of application influences the experimental outcome and Park et al recently confirmed this assumption and detected a high number of 5F-MDMB-P7AICA metabolites in zebrafish larvae after microinjection into the caudal vein, heart ventricle, or hindbrain [28].…”

Induction of Liver Size Reduction in Zebrafish Larvae by the Emerging Synthetic Cannabinoid 4F-MDMB-BINACA and Its Impact on Drug Metabolism

“…Over the past few years, the ZF model has been increasingly used not only in functional and safety studies but also in drug metabolism studies, and its reproducibility and high coverage of human metabolites have been demonstrated [19]. Numerous studies have been published on ZF xenobiotic metabolites [19][20][21][22][23][24], and importantly, several new psychoactive substances (NPSs) and their phase I and phase II metabolites in ZF have been evaluated and were found to well correlate with human metabolism [25][26][27][28].…”

“…Numerous studies have been published on ZF xenobiotic metabolites [19][20][21][22][23][24], and importantly, several new psychoactive substances (NPSs) and their phase I and phase II metabolites in ZF have been evaluated and were found to well correlate with human metabolism [25][26][27][28].…”

“…In our previous study [28], we assessed various administration routes into ZF larvae, and an authentic spectrum of metabolites of the synthetic cannabinoid (SC) methyl 2-(1-(5-fluoropentyl)-1H-pyrrolo [2,3-b]pyridine-3-carboxamido)-3,3-dimethylbutanoate (7′N-5F-ADB; Figure 1a) was obtained in ZF larvae. Using mass spectrometry imaging (MSI), the spatial distribution pattern in ZF larvae of 7′N-5F-ADB and its metabolites was studied and revealed the impact of different administration routes on in vivo distribution and drug metabolism.…”

Zebrafish: An Attractive Model to Study Staphylococcus aureus Infection and Its Use as a Drug Discovery Tool

“…They also assessed the spatial drug distribution of the compound and its metabolites by mass spectrometry imaging (MSI) in treated zebrafish larvae. Their findings presented an additional insight related to the study of metabolites in zebrafish larvae upon drug injection in different sites [ 70 ].…”