Breusova Kateryna, Ernstsen Kristian Goldeman, Palner Mikael, Linnet Kristian, Kristensen Jesper Langgaard, Kretschmann Andreas Christopher
Department of Drug Design and Pharmacology, Faculty of Health and Medical Sciences, University of Copenhagen, Universitetsparken 2, 2100, Copenhagen, Denmark.
Department of Pharmacy, Faculty of Health and Medical Sciences, University of Copenhagen, Universitetsparken 2, 2100, Copenhagen, Denmark.
J Pharm Biomed Anal. 2021 May 30;199:114016. doi: 10.1016/j.jpba.2021.114016. Epub 2021 Mar 8.
In recent years, agonists of the 5-HT receptor have gained increasing attention for their potential therapeutic use to treat psychological disorders such as anxiety and depression. Here, we report the development and validation of an LC-MSMS based analytical method for the quantification of the novel selective 5-HT agonist 25CN-NBOH in rat plasma and brain. As simple and efficient sample clean-up we applied the Phree Phospholipid Removal approach from Phenomenex, which is particularly novel for brain samples. In order to investigate the metabolic stability of 25CN-NBOH in vitro biotransformation studies with recombinant enzymes and human liver microsomes were conducted. Several biotransformation products and pathways could be identified. Based on the in vitro study one of the putative metabolites (2C-CN) was included in the analytical method development. To test the methods applicability 25CN-NBOH was quantified in plasma and brain samples from a pharmacokinetic in vivo study with Wildtype Long Evans rats. Both the in vitro metabolism data as well as the in vivo PK data suggest that 25CN-NBOH is susceptible to metabolism, but is degraded slower and is more stable compared to other NBOMe's investigated to date. The developed analytical method might serve as basis to include further 25CN-NBOH metabolites. It is expected to facilitate further preclinical and clinical investigations of 25CN-NBOH in biological matrices.
近年来，5-羟色胺（5-HT）受体激动剂因其在治疗焦虑和抑郁等心理障碍方面的潜在治疗用途而受到越来越多的关注。在此，我们报告了一种基于液相色谱-串联质谱（LC-MSMS）的分析方法的开发与验证，该方法用于定量大鼠血浆和脑中新型选择性5-HT激动剂25CN-NBOH。作为简单有效的样品净化方法，我们应用了菲罗门公司的Phree磷脂去除方法，这对于脑样品来说尤其新颖。为了研究25CN-NBOH的代谢稳定性，我们用重组酶和人肝微粒体进行了体外生物转化研究。可以鉴定出几种生物转化产物和途径。基于体外研究，一种推定的代谢物（2C-CN）被纳入分析方法的开发中。为了测试该方法的适用性，在对野生型长爪沙鼠进行的体内药代动力学研究的血浆和脑样品中对25CN-NBOH进行了定量。体外代谢数据以及体内药代动力学数据均表明，25CN-NBOH易于代谢，但与迄今为止研究的其他NBOMe相比，其降解速度较慢且更稳定。所开发的分析方法可能作为纳入更多25CN-NBOH代谢物的基础。预计这将有助于在生物基质中对25CN-NBOH进行进一步的临床前和临床研究。
