Kim Kyu-Pyo, Parise Robert A, Holleran Julianne L, Lewis Lionel D, Appleman Leonard, van Erp Nielka, Morris Michael J, Beumer Jan H
Cancer Therapeutics Program, University of Pittsburgh Cancer Institute, Pittsburgh, PA, USA; Department of Oncology, Asan Medical Center, University of Ulsan College of Medicine, Seoul, South Korea; Division of Hematology-Oncology, Department of Medicine, University of Pittsburgh School of Medicine, Pittsburgh, PA, USA.
Cancer Therapeutics Program, University of Pittsburgh Cancer Institute, Pittsburgh, PA, USA; Department of Pharmaceutical Sciences, School of Pharmacy, University of Pittsburgh, Pittsburgh, PA, USA.
J Pharm Biomed Anal. 2017 May 10;138:197-205. doi: 10.1016/j.jpba.2017.02.018. Epub 2017 Feb 13.
Inhibiting the androgen receptor (AR) pathway is an important clinical strategy in metastatic prostate cancer. Novel agents including abiraterone acetate and enzalutamide have been shown to prolong life in men with metastatic, castration-resistant prostate cancer (mCRPC). To evaluate the pharmacokinetics of AR-targeted agents, we developed and validated an LC-MS/MS assay for the quantitation of enzalutamide, N-desmethyl enzalutamide, abiraterone and bicalutamide in 0.05mL human plasma. After protein precipitation, chromatographic separation was achieved with a Phenomenex Synergi Polar-RP column and a linear gradient of 0.1% formic acid in methanol and water. Detection with an ABI 4000Q mass spectrometer utilized electrospray ionization in positive multiple reaction monitoring mode. The assay was linear over the ranges of 1-1000ng/mL for abiraterone and bicalutamide and 100-30,000ng/mL for N-desmethyl enzalutamide and enzalutamide and proved to be accurate (92.8-107.7%) and precise (largest was 15.3% CV at LLOQ for bicalutamide), and fulfilled FDA criteria for bioanalytical method validation. We demonstrated the suitability of this assay in plasma from patients who were administered enzalutamide 160mg, abiraterone 1000mg and bicalutamide 50mg once a day as monotherapy or in combination. The LC-MS/MS assay that has been developed will be an essential tool that further defines the pharmacology of the combinations of androgen synthesis or AR-receptor targeted agents.
抑制雄激素受体（AR）途径是转移性前列腺癌的一项重要临床策略。包括醋酸阿比特龙和恩杂鲁胺在内的新型药物已被证明可延长转移性去势抵抗性前列腺癌（mCRPC）男性患者的生命。为了评估AR靶向药物的药代动力学，我们开发并验证了一种液相色谱-串联质谱（LC-MS/MS）测定法，用于定量0.05mL人血浆中的恩杂鲁胺、N-去甲基恩杂鲁胺、阿比特龙和比卡鲁胺。蛋白质沉淀后，使用Phenomenex Synergi Polar-RP柱以及甲醇和水中含0.1%甲酸的线性梯度进行色谱分离。使用ABI 4000Q质谱仪在正离子多反应监测模式下利用电喷雾电离进行检测。该测定法在阿比特龙和比卡鲁胺1-1000ng/mL以及N-去甲基恩杂鲁胺和恩杂鲁胺100-30,000ng/mL的范围内呈线性，并且被证明是准确的（92.8-107.7%）和精密的（比卡鲁胺在LLOQ时最大变异系数为15.3%），并符合美国食品药品监督管理局（FDA）生物分析方法验证标准。我们证明了该测定法适用于接受恩杂鲁胺160mg、阿比特龙1000mg和比卡鲁胺50mg每日一次单药治疗或联合治疗患者的血浆。已开发的LC-MS/MS测定法将成为进一步明确雄激素合成或AR受体靶向药物组合药理学的重要工具。
