Utrecht University, Faculty of Science, Department of Pharmaceutical Sciences, Division of Pharmacoepidemiology & Clinical Pharmacology, Universiteitsweg 99, 3584 CG, Utrecht, The Netherlands; Utrecht University, Faculty of Science, Department of Pharmaceutical Sciences, Division of Chemical Biology & Drug Development, Universiteitsweg 99, 3584 CG, Utrecht, The Netherlands.
The Netherlands Cancer Institute, Division of Pharmacology, Plesmanlaan 121, 1066 CX Amsterdam, The Netherlands.
J Pharm Biomed Anal. 2018 Nov 30;161:136-143. doi: 10.1016/j.jpba.2018.08.038. Epub 2018 Aug 19.
Several second and third generation ALK inhibitors have been introduced in recent years. A bioanalytical assay for simultaneous quantification of alectinib, brigatinib, and lorlatinib was developed and validated for human plasma. The method was also partially validated for diluted mouse plasma and tissue homogenates of brain, liver, kidney, and spleen. Samples (40 μl) were pretreated in a 96-well plate by protein precipitation with acetonitrile containing the internal standard [H]-alectinib. After chromatographic separation on an ethylene bridged octadecyl silica column by gradient elution at 600 μl/min using 1% (v/v) formic acid (in water) and acetonitrile, compounds were ionized by a turbo electrospray and monitored by selected reaction monitoring on a triple quadrupole mass spectrometer. Validation was performed in a 2-2000 ng/ml concentration range for alectinib and lorlatinib and a 4-4000 ng/ml range for brigatinib. Precisions (within-day and between-day) were in the range 2.2-15.0% and accuracies were in between 87.2 and 110.2% for all matrices and levels. Compounds were sufficiently stable under most investigated conditions. Results of a pilot pharmacokinetic and tissue distribution study for brigatinib in mice are reported. Finally, successful incurred samples reanalysis of tissue homogenate samples containing brigatinib and lorlatinib is presented. Lorlatinib homogenate samples were also successfully reanalyzed using a second independent assay (cross-validation).
近年来已经引入了几种第二代和第三代 ALK 抑制剂。本文建立并验证了一种用于同时定量检测阿来替尼、布加替尼和劳拉替尼的人血浆的生物分析测定法。该方法还部分验证了经稀释的小鼠血浆和脑、肝、肾和脾组织匀浆。通过用含有内标[H]-阿来替尼的乙腈进行 96 孔板中的蛋白沉淀预处理样品（40 μl）。在以 600 μl/min 的流速通过梯度洗脱在乙烯桥联十八烷基硅烷柱上进行色谱分离，使用 1%（v/v）甲酸（在水中）和乙腈，通过涡轮电喷雾使化合物离子化，并在三重四极杆质谱仪上通过选择反应监测进行监测。在 2-2000 ng/ml 浓度范围内对阿来替尼和劳拉替尼以及 4-4000 ng/ml 浓度范围内对布加替尼进行验证。在所有基质和水平下，精密度（日内和日间）在 2.2-15.0%范围内，准确度在 87.2-110.2%之间。在大多数研究条件下，化合物均具有足够的稳定性。本文还报告了布加替尼在小鼠中的初步药代动力学和组织分布研究的结果。最后，成功地对含有布加替尼和劳拉替尼的组织匀浆样品进行了已分析样品的再分析。劳拉替尼的匀浆样品也使用第二个独立的测定法（交叉验证）成功地进行了再分析。
