EBNA1 是一种 Epstein Barr 病毒 (EBV) 蛋白，在所有 EBV 相关癌症中表达。 EBNA1 在潜伏感染细胞中 EBV 附加体的复制和维持中发挥着关键作用。 VK-2019 被开发为 EBNA1 DNA 结合活性的高度特异性抑制剂，目前正处于 1 期开发，用于治疗 EBV 相关癌症。开发了一种灵敏可靠的方法，使用液相色谱法和串联质谱法定量人血浆中的 VK-2019，以进行详细的药代动力学研究。使用乙腈进行蛋白质沉淀，从血浆中提取 VK-2019。使用 Zorbax XDB C18 色谱柱，使用梯度流，在 6 分钟内实现 VK-2019、两种据称代谢物和内标 VK-2019-d6 的分离。使用以正电喷雾电离模式运行的 SCIEX 4500 三重四极杆质谱仪检测 VK-2019。测定范围为 0.5-500ng/mL，并被证明是准确和精确的。 1:10 的稀释度可准确定量。 VK-2019 在 -70°C 的血浆中稳定约 18 个月。该方法用于评估每日单次和多次口服剂量 120mg 的患者中 VK-2019 的总血浆浓度。© 2023 John Wiley

EBNA1 is an Epstein Barr virus (EBV) protein expressed in all EBV-associated cancers. EBNA1 plays a critical role in the replication and maintenance of EBV episomes in latently infected cells. VK-2019 was developed as a highly specific inhibitor of EBNA1 DNA binding activity and is currently in phase 1 development as a treatment for EBV-associated carcinomas. A sensitive and reliable method was developed to quantify VK-2019 in human plasma using liquid chromatography with tandem mass spectrometry to perform detailed pharmacokinetic studies. VK-2019 was extracted from plasma using protein precipitation with acetonitrile. Separation of VK-2019, two purported metabolites, and the internal standard, VK-2019-d6, was achieved with a Zorbax XDB C18 column using a gradient flow over 6 min. VK-2019 was detected using a SCIEX 4500 triple quadrupole mass spectrometer operating in positive electrospray ionization mode. The assay range was 0.5-500 ng/mL and proved to be accurate and precise. Dilutions of 1:10 were accurately quantified. VK-2019 was stable in plasma at -70°C for approximately 18 months. The method was applied to assess the total plasma concentrations of VK-2019 in a patient who received a single and multiple oral daily doses of 120 mg.© 2023 John Wiley & Sons Ltd.