Wojnicz Aneta, Colom-Fernández Beatriz, Steegmann Juan L, Muñoz-Calleja Cecilia, Abad-Santos Francisco, Ruiz-Nuño Ana
*Instituto-Fundación Teófilo Hernando, Universidad Autónoma de Madrid, Madrid, Spain;†Departamento de Farmacología y Terapéutica, Facultad de Medicina, Universidad Autónoma de Madrid, Madrid, Spain;‡Servicio de Farmacología Clínica, Instituto de Investigación Sanitaria, Hospital Universitario de la Princesa, Universidad Autónoma de Madrid, Madrid, Spain;§Servicio de Inmunología, Instituto de Investigación Sanitaria, Hospital Universitario de la Princesa, Universidad Autónoma de Madrid, Madrid, Spain; and¶Servicio de Hematología, Instituto de Investigación Sanitaria, Hospital Universitario de la Princesa, Universidad Autónoma de Madrid, Madrid, Spain.
Ther Drug Monit. 2017 Jun;39(3):252-262. doi: 10.1097/FTD.0000000000000406.
Imatinib, dasatinib, and nilotinib are tyrosine kinase inhibitors (TKIs) used as first-line treatment of chronic myeloid leukemia. Therapeutic drug monitoring is important to achieve treatment efficacy in the case of imatinib and nilotinib, and to control toxicity in the case of dasatinib. New high-sensitivity methods to monitor those drugs are needed, especially for dasatinib. Thus, a simple method to determine plasma levels of imatinib, dasatinib, and nilotinib for application in clinical practice was developed.
TKIs were eluted with a Poroshell 120 EC-C18 column (2.1 × 75 mm, 2.7 μm) at 0.5 mL/min and 60°C, under gradient conditions through a mobile phase consisting of 4 mmol/L ammonium formate, pH 3.2 (65%), and acetonitrile (35%). TKIs were detected and quantified by liquid chromatography in tandem with mass spectrometry (LC/MS-MS) with positive electrospray ionization and analytes were extracted using solid phase extraction (Versaplate-SCX). Internal standards were isotope-labeled for each analyte.
The method was linear in the range of 2.5-5000 ng/mL for imatinib, 0.75-400 ng/mL for dasatinib, and 2-4000 ng/mL for nilotinib. The validation assays for accuracy and precision, matrix effect, extraction recovery, carryover, and stability of the samples for all the TKIs were appropriate according to regulatory agencies. Furthermore, imatinib plasma samples, stored for 4 years at -80°C were quite stable in approximately half of the samples.
The method enables rapid quantification of TKI concentrations and is being applied to therapeutic drug monitoring to adjust dose and to manage adverse reactions in clinical practice.
伊马替尼、达沙替尼和尼洛替尼是酪氨酸激酶抑制剂（TKIs），用于慢性髓性白血病的一线治疗。对于伊马替尼和尼洛替尼，治疗药物监测对于实现治疗效果很重要，而对于达沙替尼，治疗药物监测对于控制毒性很重要。需要新的高灵敏度方法来监测这些药物，尤其是对于达沙替尼。因此，开发了一种用于临床实践中测定伊马替尼、达沙替尼和尼洛替尼血浆水平的简单方法。
使用Poroshell 120 EC-C18柱（2.1×75 mm，2.7μm），以0.5 mL/min的流速和60°C的温度，在梯度条件下通过由4 mmol/L甲酸铵（pH 3.2，65%）和乙腈（35%）组成的流动相洗脱TKIs。通过液相色谱串联质谱（LC/MS-MS），采用正电喷雾电离检测和定量TKIs，并使用固相萃取（Versaplate-SCX）提取分析物。每种分析物的内标均为同位素标记。
该方法对伊马替尼的线性范围为2.5 - 5000 ng/mL，对达沙替尼为0.75 - 400 ng/mL，对尼洛替尼为2 - 4000 ng/mL。根据监管机构的要求，所有TKIs的准确性和精密度、基质效应、提取回收率、残留和样品稳定性的验证试验均符合要求。此外，在-80°C下储存4年的伊马替尼血浆样本中，约一半样本相当稳定。
该方法能够快速定量TKIs浓度，正在应用于治疗药物监测，以在临床实践中调整剂量和管理不良反应。
