A practical strategy for characterization of the metabolic profile of chiral drugs using combinatory liquid chromatography-mass spectrometric techniques: application to tetrahydropalmatine enantiomers and their metabolites in rat urine.
A practical strategy for characterization of the metabolic profile of chiral drugs using combinatory liquid chromatography-mass spectrometric techniques: application to tetrahydropalmatine enantiomers and their metabolites in rat urine.
复制标题
DOI：
10.1016/j.jpba.2014.01.045
复制
发表时间：
2014-06
影响因子：
3.4
通讯作者：
Yinying Zhang;Xin Dong;Jian Le;J. Wen;Z. Lin;Yinli Liu;Z. Lou;Y. Chai;Z. Hong
中科院分区：
文献类型：
--
作者：
Yinying Zhang;Xin Dong;Jian Le;J. Wen;Z. Lin;Yinli Liu;Z. Lou;Y. Chai;Z. Hong
由于代谢物的复杂性并且大多数手性药物代谢物无法商业化，因此手性药物代谢物的表征和定量仍然是一个巨大的挑战。在这项研究中，提出了一种基于组合液相色谱-质谱技术的实用方法，用于评估手性药物及其代谢物的代谢特征和尿排泄动力学。外消旋延胡索乙素（rac-THP）是从传统中药延胡索中分离出来的生物活性成分，被选为模型手性药物。采用超高效液相色谱-四极杆飞行时间质谱（UPLC-QTOF/MS）对（+）-THP 或（−）-THP 给药后大鼠尿液中 THP 对映体的代谢物进行表征。精确的质量测量用于确定代谢物的元素组成，从而确认这些代谢物的拟议结构。在大鼠尿液中发现了30多种潜在代谢物，其中大部分为首次鉴定，其体内代谢途径涉及去甲基化、氧化、葡萄糖醛酸结合和硫酸化等。并且仅在(+)-THP处理的大鼠中发现了三去甲基化代谢物和二去甲基化与氧化代谢物。随后，开发并验证了液相色谱串联质谱 (LC-QqQ/MS) 测定法，用于测定尿液中 THP 对映体及其代谢物的水平。对三个 I 相代谢物和两个 II 相代谢物进行半定量。获得了THP及其五种代谢物的对映体(-/+)累积尿排泄比，这表明THP对映体代谢物在体内的立体选择性。该研究证明了该策略在手性药物及其代谢物的定性表征、定量测定和立体选择性方面的巨大潜力。
The characterization and quantification of the metabolites of chiral drugs still remain a great challenge due to the complexity of the metabolites and most of them are not commercially available. In this study, a practical approach based on the combinatory liquid chromatography–mass spectrometric techniques has been proposed for the evaluation of metabolism profiles and urinary excretion kinetics of chiral drugs and their metabolites. Racemic tetrahydropalmatine (rac-THP), a biologically active ingredient isolated from a traditional Chinese herbRhizoma Corydalis, was chosen as the model chiral drug. Ultra-high performance liquid chromatography–quadrupole time-of-flight mass spectrometry (UPLC–QTOF/MS) was applied to characterize the metabolites of THP enantiomers in rat urine after administration of (+)-THP or (−)-THP. Accurate mass measurement was used to determine the elemental composition of metabolites and thus to confirm the proposed structures of these metabolites. More than 30 potential metabolites were found in rat urine, most of which were identified for the first time, and the metabolic pathwaysin vivowere involved in demethylation, oxidation, glucuronide conjugation and sulfation,etc. And the tridesmethlyzed metabolite and didesmethlyzed coupled with oxidation metabolite were found only in (+)-THP treated rats. Afterwards, a liquid chromatography tandem mass spectrometry (LC–QqQ/MS) assay was developed and validated for the determination of the urine level of THP enantiomers and their metabolites. Semi-quantification of three phase I metabolites and two phase II metabolites were performed. Enantiomeric (−/+) cumulative urinary excretion ratios of THP and its five metabolites were obtained, which indicated the stereoselective aspects of metabolites of THP enantiomersin vivo. The study demonstrated the enormous potential of this strategy for the qualitative characterization, quantitative assay and the stereoselectivity of chiral drugs and their metabolites.