Yang Mingxue, Cao Xu, Wu Zhaobo, Huang Xiaochen, Ma Aoxin, Li Yunda, Li Guojun, Lian Kaoqi
School of Public Health, Hebei Medical University, Shijiazhuang, 050017, China.
Hebei Research Institute of Sports Science, Shijiazhuang, 050017, China.
Anal Bioanal Chem. 2025 Sep;417(22):5087-5104. doi: 10.1007/s00216-025-06031-7. Epub 2025 Jul 30.
Troponin activators, including Reldesemtiv and Tirasemtiv, represent a class of drugs that enhance the contractility of cardiac muscle and delay the onset of muscle fatigue. Consequently, they were classified as prohibited substances by the World Anti-Doping Agency (WADA) in 2024. This study aimed to develop an ultra-high-performance liquid chromatography-mass spectrometry (UHPLC-MS)-based method to quantify Reldesemtiv and Tirasemtiv levels in rat urine, blood, and feces over a 72-h period following transgastric administration. Additionally, Compound Discoverer 3.3 was utilized to investigate the metabolites and metabolic pathways of these two drugs in vivo. Twelve metabolites of Reldesemtiv and seven metabolites of Tirasemtiv were identified. Notably, ten of the twelve Reldesemtiv metabolites had not been previously reported in the literature. Using UHPLC-MS, the concentration of each metabolite in urine and blood was measured at various time points, enabling the generation of a metabolic profile. Six metabolites of Reldesemtiv are detectable 72 h after administration, while Tirasemtiv and its metabolite T-M1a were also detectable at the same time point. This analytical method can characterize the metabolic profiles of the drugs and their metabolites, as well as identify unknown and long-lived metabolites in drug testing. These capabilities are critical for the quantitative analysis of troponin activators within complex biological matrices, supporting their study in physiological and doping-control contexts.
肌钙蛋白激活剂，包括雷地西明和替拉西明，是一类可增强心肌收缩力并延缓肌肉疲劳发作的药物。因此，它们在2024年被世界反兴奋剂机构（WADA）列为违禁物质。本研究旨在开发一种基于超高效液相色谱-质谱联用（UHPLC-MS）的方法，用于定量大鼠经胃给药后72小时内尿液、血液和粪便中雷地西明和替拉西明的含量。此外，还利用Compound Discoverer 3.3研究了这两种药物在体内的代谢产物和代谢途径。鉴定出了雷地西明的12种代谢产物和替拉西明的7种代谢产物。值得注意的是，雷地西明的12种代谢产物中有10种此前未见文献报道。使用UHPLC-MS在不同时间点测量尿液和血液中各代谢产物的浓度，从而生成代谢图谱。给药72小时后可检测到雷地西明的6种代谢产物，同时也可检测到替拉西明及其代谢产物T-M1a。这种分析方法可以表征药物及其代谢产物的代谢图谱，以及在药物检测中识别未知和长效代谢产物。这些能力对于在复杂生物基质中定量分析肌钙蛋白激活剂至关重要，有助于在生理和兴奋剂控制背景下对其进行研究。
