Romański Michał, Kasprzyk Anna, Teżyk Artur, Widerowska Agnieszka, Żaba Czesław, Główka Franciszek
Department of Physical Pharmacy and Pharmacokinetics, Poznan University of Medical Sciences, 6 Święcickiego Street, 60-781 Poznan, Poland.
Department of Physical Pharmacy and Pharmacokinetics, Poznan University of Medical Sciences, 6 Święcickiego Street, 60-781 Poznan, Poland.
J Pharm Biomed Anal. 2017 Jun 5;140:122-129. doi: 10.1016/j.jpba.2017.03.023. Epub 2017 Mar 18.
A prodrug treosulfan (TREO) is currently investigated in clinical trials for conditioning prior to hematopoietic stem cell transplantation. Bioanalysis of TREO and its active derivatives, monoepoxide (S,S-EBDM) and diepoxide, in plasma and urine underlay the pharmacokinetic studies of these compounds but cannot explain an organ pharmacological action or toxicity. Recently, distribution of TREO and S,S-EBDM into brain, cerebrospinal fluid, and aqueous humor of the eye has been investigated in animal models and the obtained results presented clinical relevance. In this paper, a selective and rapid HPLC-ESI-MS/MS method was elaborated and validated for the studies of disposition of TREO and S,S-EBDM in rat plasma, liver, lungs, kidneys, muscle, and brain. The two analytes and codeine, internal standard (IS), were isolated from 50μL of plasma and 100μL of supernatants of the tissues homogenates using ultrafiltration Amicon vials. Chromatographic resolution was accomplished on C18 column with isocratic elution. The limits of quantitation of TREO and S,S-EBDM in the studied matrices ranged from 0.11 to 0.93μM. The HPLC-MS/MS method was adequately precise and accurate within and between runs. The IS-normalized matrix effect differed among the tissues and was the most pronounced in a liver homogenate supernatant (approximately 0.55 for TREO and 0.35 for S,S-EBDM). Stability of the analytes in experimental samples was also established. The validated method for the first time enabled determination of TREO and S,S-EBDM in the six life-important tissues in rats following administration of the prodrug.
一种前体药物苏消安（TREO）目前正在进行临床试验，用于造血干细胞移植前的预处理。对TREO及其活性衍生物单环氧化物（S,S-EBDM）和双环氧化物在血浆和尿液中的生物分析是这些化合物药代动力学研究的基础，但无法解释其器官药理作用或毒性。最近，在动物模型中研究了TREO和S,S-EBDM在脑、脑脊液和眼房水中的分布情况，所得结果具有临床相关性。本文阐述并验证了一种选择性快速的HPLC-ESI-MS/MS方法，用于研究TREO和S,S-EBDM在大鼠血浆、肝脏、肺、肾脏、肌肉和脑中的处置情况。使用超滤Amicon小瓶从50μL血浆和100μL组织匀浆上清液中分离出两种分析物和内标（IS）可待因。在C18柱上采用等度洗脱实现色谱分离。在所研究的基质中，TREO和S,S-EBDM的定量限范围为0.11至0.93μM。HPLC-MS/MS方法在批内和批间都具有足够的精密度和准确度。经内标归一化后的基质效应在不同组织中有所不同，在肝脏匀浆上清液中最为明显（TREO约为0.55，S,S-EBDM约为0.35）。还确定了分析物在实验样品中的稳定性。该经过验证的方法首次能够在给予前体药物后测定大鼠六种重要生命组织中的TREO和S,S-EBDM。
