Kiesel Brian F, Parise Robert A, Wong Alvin, Keyvanjah Kiana, Jacobs Samuel, Beumer Jan H
Cancer Therapeutics Program, University of Pittsburgh Cancer Institute, Pittsburgh, PA, USA; Department of Pharmaceutical Sciences, School of Pharmacy, University of Pittsburgh, Pittsburgh, PA, USA.
Cancer Therapeutics Program, University of Pittsburgh Cancer Institute, Pittsburgh, PA, USA.
J Pharm Biomed Anal. 2017 Feb 5;134:130-136. doi: 10.1016/j.jpba.2016.11.035. Epub 2016 Nov 22.
Neratinib is an orally available tyrosine kinase inhibitor targeting HER2 (ERBB2) and EGFR (ERBB). It is being clinically evaluated for the treatment of breast and other solid tumors types as a single agent or in combination with other chemotherapies. In support of several phase I/II clinical trials investigating neratinib combinations, we developed and validated a novel LC-MS/MS assay for the quantification of neratinib in 100μL of human plasma with a stable isotopic internal standard. Analytes were extracted from plasma using protein precipitation and evaporation of the resulting supernatant followed by resuspension. Chromatographic separation was achieved using an Acquity UPLC BEH Shield RP18 column and a gradient methanol-water mobile phase containing 10% ammonium acetate. An ABI 4000 mass spectrometer and electrospray positive mode ionization were used for detection. The assay was linear from 2 to 1,000ng/mL and proved to be accurate (98.9-106.5%) and precise (<6.2%CV), and met the FDA guidance for bioanalytical method validation. This LC-MS/MS assay will be an essential tool to further define the pharmacokinetics of neratinib.
奈拉替尼是一种口服可用的酪氨酸激酶抑制剂，靶向HER2（ERBB2）和EGFR（ERBB）。它正在作为单一药物或与其他化疗药物联合用于治疗乳腺癌和其他实体瘤类型的临床评估中。为支持多项研究奈拉替尼联合用药的I/II期临床试验，我们开发并验证了一种新型液相色谱-串联质谱法（LC-MS/MS），用于定量100μL人血浆中的奈拉替尼，并使用稳定同位素内标。通过蛋白质沉淀从血浆中提取分析物，蒸发所得上清液，然后再悬浮。使用Acquity UPLC BEH Shield RP18柱和含有10%乙酸铵的甲醇-水梯度流动相实现色谱分离。使用ABI 4000质谱仪和电喷雾正模式电离进行检测。该方法在2至1000ng/mL范围内呈线性，证明准确（98.9-106.5%）且精密（<6.2%CV），并符合美国食品药品监督管理局（FDA）生物分析方法验证指南。这种LC-MS/MS方法将是进一步确定奈拉替尼药代动力学的重要工具。
