Validation of a robust and rapid liquid chromatography tandem mass spectrometric method for the quantitative analysis of VK-2019, a selective EBNA1 inhibitor

  • Biomed Chromatogr. 2024 Feb;38(2):e5775. doi: 10.1002/bmc.5775.
Michael T Davis  1 ,  Nicole M Anders  1  2 ,  A Dimitrios Colevas  3 ,  Troy E Messick  4 ,  Michelle A Rudek  1  5  6
Affiliations
  • 1. The Sidney Kimmel Comprehensive Cancer Center at Johns Hopkins University, Baltimore, Maryland, USA.
  • 2. Takeda Pharmaceutical Company, San Diego, California, USA.
  • 3. Division of Medical Oncology, Stanford Cancer Center, Stanford University, Stanford, California, USA.
  • 4. The Wistar Institute, Philadelphia, Pennsylvania, USA.
  • 5. Department of Oncology, School of Medicine, Johns Hopkins University, Baltimore, Maryland, USA.
  • 6. Division of Clinical Pharmacology, Department of Medicine, School of Medicine, Johns Hopkins University, Baltimore, Maryland, USA.
PMID: 37942577 DOI: 10.1002/bmc.5775
Abstract

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.

Keywords
VK-2019 quantitative analysis; tandem mass spectrometry; validation.
Products
  • Cat. No.
    Product Name
    Description
    Target
    Research Area
  • HY-W725179
    VK-2019
    99.97%, EBNA1 Inhibitor
    target: EBV
    Research Areas: Cancer