Bilel S, Murari M, Pesavento S, Arfè R, Tirri M, Torroni L, Marti M, Tagliaro F, Gottardo R
Department of Translational Medicine, Section of Legal Medicine and LTTA Centre, University of Ferrara, Italy.
Unit of Forensic Medicine, Department of Diagnostics and Public Health, University of Verona, Verona, Italy.
Neurotoxicology. 2023 Mar;95:83-93. doi: 10.1016/j.neuro.2023.01.003. Epub 2023 Jan 10.
The introduction of the so-called New Psychoactive Substances represents a problem of global concern due to several factors, including multiplicity of structures, poorly known activity, short half-life in the market, lack of pure standards etc. Among these problems, of the highest relevance is also the lack of information about metabolism and adverse effects, which must be faced using simple and low-cost animal models. On these grounds, the present work has been carried out on 5 days post fertilization zebrafish (Danio rerio) larvae in comparison with adult mice (Mus musculus). Ocfentanil and 2-furanylfentanyl were administered at different concentrations to zebrafish larvae (1, 10 µM) and mice (0.1, 1, 6, 15 mg/kg). The behavioural assay showed a decrease in basal locomotor activity in zebrafish, whereas in mice this effect was evident only after the mechanical stimulus. Larva extracts and mice urine were analysed by using liquid chromatography coupled to high resolution mass spectrometry to identify the metabolic pathways of the fentanyl analogs. For 2-furanylfentanyl, the most common biotransformations observed were hydroxylation, hydration and oxidation in zebrafish larvae, whereas mice produced mainly the dihydrodiol metabolite. Hydroxylation was the major route of metabolism for ocfentanil in zebrafish larvae, while in mice the O-demethylated derivative was the main metabolite. In addition, a study was conducted to evaluate morphological effects of the two drugs on zebrafish larvae. Malformations were noticeable only at the highest concentration of 2-furanylfentanyl, whereas no significant damage was observed with ocfentanil. In conclusion, the two animal models show similarities in behavioral response and in metabolism, considering the different biological investigated.
所谓新型精神活性物质的出现引发了一个全球关注的问题，原因有多个，包括结构多样、活性了解不足、在市场上的半衰期短、缺乏纯标准品等。在这些问题中，与代谢和不良反应相关的信息匮乏也是最为关键的，必须通过简单且低成本的动物模型来解决。基于这些原因，本研究以受精后5天的斑马鱼（Danio rerio）幼体为对象，并与成年小鼠（Mus musculus）进行比较。将奥芬太尼和2-呋喃基芬太尼以不同浓度分别给予斑马鱼幼体（1、10 μM）和小鼠（0.1、1、6、15 mg/kg）。行为学检测显示，斑马鱼的基础运动活性降低，而在小鼠中，这种效应仅在机械刺激后才明显。利用液相色谱-高分辨率质谱联用技术分析幼体提取物和小鼠尿液，以确定芬太尼类似物的代谢途径。对于2-呋喃基芬太尼，在斑马鱼幼体中观察到的最常见生物转化是羟基化、水合作用和氧化，而小鼠主要产生二氢二醇代谢物。羟基化是斑马鱼幼体中奥芬太尼的主要代谢途径，而在小鼠中，O-去甲基化衍生物是主要代谢物。此外，还进行了一项研究，以评估这两种药物对斑马鱼幼体的形态学影响。仅在2-呋喃基芬太尼的最高浓度下才观察到畸形，而奥芬太尼未观察到明显损害。总之，考虑到所研究的不同生物学特性，这两种动物模型在行为反应和代谢方面表现出相似性。
