Zuo Lihua, Liu Liwei, Yang Yantao, Yang Jie, Chen Min, Zhang Huafeng, Kang Jian, Zhang Xiaojian, Wang Jiabo, Sun Zhi
Department of Pharmacy, the First Affiliated Hospital of Zhengzhou University, Zhengzhou, China.
Henan Engineering Research Center of Clinical Mass Spectrometry for Precision Medicine, Zhengzhou, China.
Front Pharmacol. 2021 May 7;12:610226. doi: 10.3389/fphar.2021.610226. eCollection 2021.
Prucalopride was widely used for chronic constipation, which is difficult to be adequately relieved by laxatives in adult patients in clinic. Due to the difficulty in metabolite identification, metabolic process of prucalopride had not been investigated in vivo. In this study, an efficient strategy was proposed for comprehensive metabolite profiling of prucalopride after oral administration in rat plasma, urine, and feces samples. This strategy was composed of five steps. First, the samples at multiple time points after oral administration were collected to increase the representativeness of the samples. Second, different sample preparation methods were investigated to obtain superior extraction efficiency. Third, the raw data of test sample and blank sample were acquired using ultra-performance liquid chromatography with Q-Exactive hybrid quadrupole-orbitrap high-resolution accurate mass spectrometry under the positive and negative full-scan/dd MS mode. Fourth, combined mass defect filter with background subtraction model in soft of compound discovery, all peaks were constructed to filter potential metabolites after retention time alignment and ion filtration, which could remove large amounts of interference ions. Besides, it can predict potential biotransformation, promoting to understand how to metabolize the drug. This provides multiple possibilities and prevents us conjecturing the potential metabolites blindly. Finally, the verification procedure was implemented through exporting the structure and MS spectrum to the analytical tool of Mass Frontier. The proposed strategy significantly improved the targeted detection and identification for metabolites in vivo. A total of 47 metabolites were tentatively characterized in the plasma, urine, and feces samples after oral administration of prucalopride. This study could provide a valuable reference for systematic metabolite profile of drug in vivo.
普芦卡必利广泛用于治疗慢性便秘，临床上成年患者使用泻药难以充分缓解该症状。由于代谢物鉴定困难，尚未在体内研究普芦卡必利的代谢过程。本研究提出了一种高效策略，用于在大鼠血浆、尿液和粪便样本中对口服普芦卡必利后的代谢物进行全面分析。该策略包括五个步骤。首先，收集口服给药后多个时间点的样本，以提高样本的代表性。其次，研究不同的样品制备方法以获得更高的提取效率。第三，在正、负全扫描/dd MS模式下，使用超高效液相色谱与Q-Exactive混合型四极杆-轨道阱高分辨率精确质谱联用仪获取测试样品和空白样品的原始数据。第四，在化合物发现软件中结合质量缺陷过滤器和背景扣除模型，在保留时间对齐和离子过滤后构建所有峰以筛选潜在代谢物，这可以去除大量干扰离子。此外，它可以预测潜在的生物转化，有助于了解药物的代谢方式。这提供了多种可能性，避免我们盲目推测潜在代谢物。最后，通过将结构和质谱导出到Mass Frontier分析工具中实施验证程序。所提出的策略显著提高了体内代谢物的靶向检测和鉴定能力。口服普芦卡必利后，在血浆、尿液和粪便样本中初步鉴定出总共47种代谢物。本研究可为药物体内系统代谢物分析提供有价值的参考。
