Division of Clinical Pharmacology, Department of Medicine, School of Medicine, Johns Hopkins University, Baltimore, MD, USA.
The Sidney Kimmel Comprehensive Cancer Center at Johns Hopkins University, Baltimore, MD, USA.
Biomed Chromatogr. 2022 Nov;36(11):e5467. doi: 10.1002/bmc.5467. Epub 2022 Aug 31.
Navtemadlin is an orally bioavailable small molecule that blocks the protein-protein interaction between murine double minute 2 protein (MDM2) and the tumor suppressor protein p53, leading to p53-mediated cell cycle arrest and apoptosis. It is being evaluated in clinical trials for a variety of malignancies, both as a single agent and in combination regimens. A sensitive, robust LC-tandem mass spectrometry (LC-MS/MS) method was developed to quantitate navtemadlin in plasma, and this method was also validated using brain tissue homogenate. Sample preparation involved protein precipitation of plasma or brain tissue homogenate using acetonitrile. Navtemadlin, navtemadlin glucuronide, and the internal standard, D -navtemadlin, were separated from microsomal incubation extracts using gradient elution and a ZORBAX XDB C column. Analytes were detected using a SCIEX 5500 triple quadrupole mass spectrometer in positive electrospray ionization mode. The assay range of 1-1000 ng/mL was shown to be accurate (96.1-102.0% and 95.7-104%) and precise (coefficient of variation ≤ 10.6% and ≤ 6.6%) in plasma and brain tissue homogenate, respectively. An 8000 ng/mL navtemadlin sample diluted 1:10 (v/v) with plasma was also accurately quantitated. Navtemadlin has been stable in frozen plasma at -70°C for at least 20 months. This validated LC-MS/MS method was applied to determine navtemadlin concentrations in plasma and brain tissue samples from two separate patients receiving 120 mg/day navtemadlin on protocol ABTC1604.
纳维他林是一种口服生物可利用的小分子，可阻断鼠双微体 2 蛋白（MDM2）和肿瘤抑制蛋白 p53 之间的蛋白-蛋白相互作用，导致 p53 介导的细胞周期停滞和细胞凋亡。它正在临床试验中评估用于各种恶性肿瘤，无论是作为单一药物还是联合治疗方案。开发了一种灵敏、稳健的液相色谱-串联质谱（LC-MS/MS）方法来定量血浆中的纳维他林，并且还使用脑组织匀浆验证了该方法。样品制备涉及使用乙腈沉淀血浆或脑组织匀浆中的蛋白质。纳维他林、纳维他林葡萄糖醛酸苷和内标 D-纳维他林使用梯度洗脱和 ZORBAX XDB C 柱从微粒体孵育提取物中分离。使用 SCIEX 5500 三重四极杆质谱仪在正电喷雾电离模式下检测分析物。在血浆和脑组织匀浆中，1-1000ng/mL 的测定范围分别显示出准确性（96.1-102.0%和 95.7-104%）和精密度（变异系数≤10.6%和≤6.6%）。用血浆稀释 1:10（v/v）的 8000ng/mL 纳维他林样品也被准确定量。纳维他林在 -70°C 下冷冻血浆中至少稳定 20 个月。该经过验证的 LC-MS/MS 方法应用于确定接受 ABTC1604 方案每天 120mg 纳维他林的两名患者的血浆和脑组织样本中的纳维他林浓度。
