开发和验证一种用于检测人血浆中 CDK4/6 抑制剂帕博西利、瑞博西利、阿贝西利和阿贝西利 M2 的定量 LC-MS/MS 方法。
Development and Validation of a Quantitative LC-MS/MS Method for CDK4/6 Inhibitors Palbociclib, Ribociclib, Abemaciclib, and Abemaciclib-M2 in Human Plasma.
机构信息
Bioanalytics, Metabolomics, and Pharmacokinetics Shared Resource, Roswell Park Comprehensive Cancer Center, Buffalo, New York.
Department of Pharmacology and Therapeutics, Roswell Park Comprehensive Cancer Center, Buffalo, New York.
出版信息
Ther Drug Monit. 2023 Jun 1;45(3):327-336. doi: 10.1097/FTD.0000000000001063. Epub 2023 Jan 10.
BACKGROUND
The cyclin-dependent kinase 4 and 6 (CDK4/6) inhibitors, palbociclib, ribociclib, and abemaciclib, are standard-of-care agents for patients with hormone receptor-positive human epidermal growth factor receptor 2-negative metastatic breast cancer. In support of therapeutic drug monitoring and clinical pharmacokinetic studies, a liquid chromatography coupled with tandem mass spectrometry assay for the simultaneous quantitation of CDK4/6 inhibitors and the major active metabolite M2 of abemaciclib in human plasma has been developed.
METHODS
Analytes were extracted from 50 μL of human plasma by precipitating proteins with methanol and then collecting the supernatant. Reversed-phase high-performance liquid chromatography was performed for analyte separation using a biphasic gradient at a flow rate of 0.25-0.5 mL/min. The total run time was 9.5 minutes. The analytes were detected using MS/MS with electrospray ionization operating in positive ion mode.
RESULTS
Validation according to the US Food and Drug Administration's guidance showed that the new assay produced accurate (94.7%-107%) and precise (within-run: 1.2%-8.2%; between-run: 0.6%-7.5%) measurements of all analytes over a concentration range of 5-2000 ng/mL. Overall, analyte recoveries were consistent (mean values: 110%-129%). The analytes were also stable in human plasma and the final extract under various storage conditions. Finally, the clinical applicability of the assay was confirmed by quantitation of all analytes in plasma samples obtained from patients treated with CDK4/6 inhibitors. Reproducibility of the measured analyte concentrations in study samples was confirmed successfully by incurred sample reanalysis.
CONCLUSIONS
A sensitive liquid chromatography coupled with tandem mass spectrometry method to measure CDK4/6 inhibitors was developed and validated according to the Food and Drug Administration criteria. Quantitation of all analytes in clinical plasma samples confirmed that the assay is suitable for therapeutic drug monitoring and clinical pharmacokinetic studies of CDK4/6 inhibitors.
背景
细胞周期蛋白依赖性激酶 4 和 6（CDK4/6）抑制剂，如哌柏西利、瑞博西林和阿贝西利，是激素受体阳性人表皮生长因子受体 2 阴性转移性乳腺癌患者的标准治疗药物。为了支持治疗药物监测和临床药代动力学研究，已经开发了一种液相色谱-串联质谱法，用于同时定量测定人血浆中的 CDK4/6 抑制剂和阿贝西利的主要活性代谢物 M2。
方法
用甲醇沉淀蛋白质提取 50μL 人血浆中的分析物，然后收集上清液。采用双相梯度以 0.25-0.5mL/min 的流速进行反相高效液相色谱分离分析物。总运行时间为 9.5 分钟。采用电喷雾正离子模式的 MS/MS 进行分析物检测。
结果
根据美国食品和药物管理局的指导进行验证表明，新的测定方法在 5-2000ng/mL 的浓度范围内产生了准确（94.7%-107%）和精确（批内：1.2%-8.2%；批间：0.6%-7.5%）的测量结果。总体而言，分析物回收率一致（平均值：110%-129%）。在各种储存条件下，分析物在人血浆和最终提取物中也稳定。最后，通过测定接受 CDK4/6 抑制剂治疗的患者的血浆样本中的所有分析物，证实了该测定方法的临床适用性。通过对研究样本中测定的分析物浓度进行重复样品分析，成功地证实了测量结果的重现性。
结论
根据食品和药物管理局的标准，开发并验证了一种灵敏的液相色谱-串联质谱法来测定 CDK4/6 抑制剂。对临床血浆样本中所有分析物的定量证实了该测定方法适用于 CDK4/6 抑制剂的治疗药物监测和临床药代动力学研究。
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