Research Institute for Doping Control, Shanghai University of Sport, 900 Jiangwancheng Road, Shanghai, 200438, China.
School of Exercise and Health, Shanghai University of Sport, 399 Changhai Road, Shanghai, 200438, China.
Anal Bioanal Chem. 2024 Nov;416(28):6575-6588. doi: 10.1007/s00216-024-05555-8. Epub 2024 Oct 4.
Voxelotor was approved for the treatment of sickle cell anemia as a potent hemoglobin S polymerization inhibitor. Owing to its ability to affect blood components and its potential to enhance athletic performance, voxelotor was included in the prohibited list issued by the World Anti-Doping Agency in 2023, banning its use both in and out of competition. This study aimed to comprehensively investigate the metabolic profile of voxelotor in human urine and identify suitable metabolites for long-term analytical retrospectivity in doping control. A novel strategy for metabolite identification was established by combining in vivo human administration with isotope labeling-based in vitro metabolism analysis. A single microdose of voxelotor was administered orally to five volunteers, and urine samples were collected for up to 28 days post-administration. Concurrently, in vitro incubation of human liver microsomes with voxelotor and D3-voxelotor was conducted, and the microsomal incubates were analyzed via liquid chromatography-high-resolution mass spectrometry. Targeted metabolite searches in human urine samples and automated nontargeted screening of isotope metabolite ion pairs in incubation samples led to the discovery of 9 phase I metabolites and 23 phase II metabolites. Analysis of the urine excretion curves revealed that 4 metabolites, along with voxelotor, were suitable for long-term anti-doping monitoring, with a detection window exceeding 20 days. Using both in vivo and in vitro metabolic models, this study provides comprehensive insight into the metabolic profile of voxelotor in human urine for the first time, enhancing the capacity for doping screening and extending the retrospectivity of voxelotor detection.
沃瑟罗特被批准用于治疗镰状细胞贫血，作为一种有效的血红蛋白 S 聚合抑制剂。由于其影响血液成分的能力及其增强运动表现的潜力，沃瑟罗特被列入世界反兴奋剂机构在 2023 年发布的禁用清单中，禁止在比赛内外使用。本研究旨在全面研究沃瑟罗特在人尿液中的代谢特征，并确定适合长期兴奋剂控制分析回溯的合适代谢物。通过将体内人体给药与基于同位素标记的体外代谢分析相结合，建立了一种新的代谢物鉴定策略。给五名志愿者口服单微剂量的沃瑟罗特，并在给药后长达 28 天内收集尿液样本。同时，用人肝微粒体与沃瑟罗特和 D3-沃瑟罗特进行体外孵育，并通过液相色谱-高分辨率质谱对微粒体孵育物进行分析。对人尿液样本中的靶向代谢物进行搜索和对孵育样本中同位素代谢物离子对的自动非靶向筛选，发现了 9 种 I 相代谢物和 23 种 II 相代谢物。对尿液排泄曲线的分析表明，有 4 种代谢物与沃瑟罗特一起，适合长期兴奋剂监测，检测窗口超过 20 天。本研究通过体内和体外代谢模型，首次全面了解了沃瑟罗特在人尿液中的代谢特征，增强了兴奋剂筛选的能力，并延长了沃瑟罗特检测的回溯性。
