Thomann Jan, Rudin Deborah, Kraus Selina, Arikci Denis, Holze Friederike, Liechti Matthias E, Luethi Dino
Division of Clinical Pharmacology and Toxicology, Department of Biomedicine, University Hospital Basel, Basel, Switzerland; Division of Clinical Pharmacology and Toxicology, Department of Pharmaceutical Sciences, University of Basel, Basel, Switzerland.
Division of Clinical Pharmacology and Toxicology, Department of Pharmaceutical Sciences, University of Basel, Basel, Switzerland; Division of Clinical Pharmacology and Toxicology, Department of Clinical Research, University Hospital Basel, Basel, Switzerland.
Drug Metab Dispos. 2025 Jun;53(6):100086. doi: 10.1016/j.dmd.2025.100086. Epub 2025 Apr 28.
4-Bromo-2,5-dimethoxyphenethylamine (2C-B) is widely used recreationally and has recently gained interest as a treatment for mental health disorders. In this study, a liquid chromatography-tandem mass spectrometry method to quantify 2C-B and its metabolites 4-bromo-2,5-dimethoxyphenylacetic acid (BDMPAA) and 4-bromo-2-hydroxy-5-methoxyphenylacetic acid (B-2-HMPAA) in human plasma was developed and validated. Moreover, pharmacokinetic analysis was performed on samples from clinical study participants who received 30 mg of 2C-B. The metabolic degradation of 2C-B and its metabolites via monoamine oxidases (MAOs), cytosolic enzymes, and cytochrome P450 enzymes was assessed and their activation potencies at the serotonin 2A receptor were investigated. Optimal chromatographic separation was achieved using a Kinetex 2.6 μm XB-C18 analytical column and a mobile phase gradient of water and acetonitrile supplemented with 0.1% formic acid. Using electrospray ionization, a linear range of 0.5-100 ng/mL for 2C-B, 2.5-1000 ng/mL for BDMPAA, and 0.5-1000 ng/mL for B-2-HMPAA was achieved. The method demonstrated high accuracy, precision, and extraction recovery with minimal matrix effects. MAO-A, MAO-B, cytosolic enzymes, and CYP2D6 were identified as key enzymes involved in the metabolic degradation of 2C-B. Unlike 2C-B, BDMPAA and B-2-HMPAA did not activate the human serotonin 2A receptor, suggesting that these metabolites do not contribute to the psychedelic effect. This study provides insights into the pharmacokinetics, metabolism, and pharmacological activity of 2C-B and its metabolites. The validated liquid chromatography-tandem mass spectrometry method offers a reliable tool for future clinical studies investigating the therapeutic potential and metabolism of 2C-B. SIGNIFICANCE STATEMENT: A rapid and nonlaborious liquid chromatography-tandem mass spectrometry method was developed and validated for pharmacokinetic analysis of 4-bromo-2,5-dimethoxyphenethylamine (2C-B) and its metabolites 4-bromo-2,5-dimethoxyphenylacetic acid and 4-bromo-2-hydroxy-5-methoxyphenylacetic acid in human plasma. To assess the metabolites' relevance in psychedelic drug action, serotonin 2A receptor activity was studied. Unlike 2C-B, the metabolites failed to activate the receptor. Monoamine oxidase A and B, and cytosolic enzymes were confirmed in 4-bromo-2,5-dimethoxyphenylacetic acid formation, whereas CYP2D6 was found to metabolize 2C-B through a minor pathway.
4-溴-2,5-二甲氧基苯乙胺（2C-B）在娱乐场合被广泛使用，最近作为一种治疗精神健康障碍的药物引起了关注。在本研究中，开发并验证了一种液相色谱-串联质谱法，用于定量人血浆中的2C-B及其代谢物4-溴-2,5-二甲氧基苯乙酸（BDMPAA）和4-溴-2-羟基-5-甲氧基苯乙酸（B-2-HMPAA）。此外，对接受30 mg 2C-B的临床研究参与者的样本进行了药代动力学分析。评估了2C-B及其代谢物通过单胺氧化酶（MAO）、胞质酶和细胞色素P450酶的代谢降解情况，并研究了它们对5-羟色胺2A受体的激活能力。使用Kinetex 2.6 μm XB-C18分析柱以及水和乙腈组成的流动相梯度（添加0.1%甲酸）实现了最佳色谱分离。采用电喷雾电离，2C-B的线性范围为0.5 - 100 ng/mL，BDMPAA为2.5 - 1000 ng/mL，B-2-HMPAA为0.5 - 1000 ng/mL。该方法具有高准确性、精密度和提取回收率，基质效应最小。MAO-A、MAO-B、胞质酶和CYP2D6被确定为参与2C-B代谢降解的关键酶。与2C-B不同，BDMPAA和B-2-HMPAA不会激活人5-羟色胺2A受体，表明这些代谢物不会产生致幻效果。本研究为2C-B及其代谢物的药代动力学、代谢和药理活性提供了见解。经过验证的液相色谱-串联质谱法为未来研究2C-B治疗潜力和代谢的临床研究提供了可靠工具。重要声明：开发并验证了一种快速且简便的液相色谱-串联质谱法，用于分析人血浆中4-溴-2,5-二甲氧基苯乙胺（2C-B）及其代谢物4-溴-2,5-二甲氧基苯乙酸和4-溴-2-羟基-5-甲氧基苯乙酸的药代动力学。为评估代谢物在致幻药物作用中的相关性，研究了5-羟色胺2A受体活性。与2C-B不同，这些代谢物未能激活该受体。已证实MAO-A和MAO-B以及胞质酶参与4-溴-2,5-二甲氧基苯乙酸的形成，而CYP2D6通过次要途径代谢2C-B。
