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

Yu Mi Park,Markus R Meyer,Jennifer Herrmann,Rolf Müller

doi:10.3390/molecules25194474

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.

Highlights

Zebrafish (Danio rerio; ZF) has become a very prominent in vivo model organism in various research fields, such as toxicology, drug discovery, disease models, and neurobiology [1,2,3,4,5]
After addition of the compound compound to larvae either through the larvae-surrounding water or through microinjection into sac to ZF larvae either through the larvae-surrounding water or through microinjection into yolk yolk sac resulting in a reduction from 18 to only one detected metabolite when the yolk sac resulting in a reduction from 18 to only one detected metabolite when the yolk sac administration administration route was chosen
ZF larvae were exposed to 70 N-5F-ADB by different administration routes and the total number of metabolites observed in these microinjected samples was higher than the number of metabolites detected after conventional waterborne exposure (Table 1)

Summary

Introduction

Zebrafish (Danio rerio; ZF) has become a very prominent in vivo model organism in various research fields, such as toxicology, drug discovery, disease models, and neurobiology [1,2,3,4,5]. The use of self-feeding ZF embryos and larvae that are younger than 120 h post-fertilization (hpf) is popular because such experiments are not considered as animal experiments according to European legislation (EU directive 2010/63/EU). Experiments with larvae at ≤120 hpf are in compliance with the 3R principle (Replacement, Reduction, Refinement) as they contribute to the reduction of animal experiments. ZF embryos and larvae have been widely applied in studies of e.g., human disease [9,10], infection [11,12], antibiotics [13,14], and human metabolism [15,16,17], to name just a few

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Results

Conclusion