Development and validation of a sensitive UHPLC-MS/MS analytical method for venetoclax in mouse plasma, and its application to pharmacokinetic studies

J Chromatogr B Analyt Technol Biomed Life Sci. 2020 Sep 1:1152:122176. doi: 10.1016/j.jchromb.2020.122176. Epub 2020 May 20.

Abstract

A rapid and sensitive analytical method was developed to quantify venetoclax, an oral BH3-mimetic that blocks the anti-apoptotic protein BCL-2, in mouse plasma using ultra-high-performance liquid chromatography with electrospray ionization tandem mass spectrometric detection. Plasma protein precipitation was performed on 5 µL samples, and separation of the analytes was accomplished on an Accucore aQ column using gradient elution at a flow rate of 0.4 mL/min. The calibration curve was linear (R2 ≥ 0.99) over the concentration range of 5-1,000 ng/mL, and the lower limit of quantitation was 5 ng/mL. The intra-day and inter-day precisions (RSD%) were < 10.5%, and accuracies ranged from 94.4 to 106%. The developed method was successfully applied to pharmacokinetic studies involving serial 30 µL blood sampling from male and female mice after oral administration of venetoclax (10 mg/kg) alone or 30 min after oral administration of ketoconazole (50 mg/kg) or vehicle (PEG400). The observed pharmacokinetic profiles suggest venetoclax undergoes sexually dimorphic disposition in mice. However, regardless of sex, pharmacokinetic studies demonstrated that venetoclax AUC(0-6h) was increased greater than 2-fold with prior administration of ketoconazole. Overall, our pharmacokinetic studies suggest that mice could be a translationally relevant model for the characterization of venetoclax pharmacokinetics. We have developed an analytical method suitable for such murine pharmacokinetic studies.

Keywords: Mouse plasma; Pharmacokinetics; UHPLC-MS/MS; Venetoclax.

MeSH terms

  • Administration, Oral
  • Animals
  • Bridged Bicyclo Compounds, Heterocyclic / administration & dosage
  • Bridged Bicyclo Compounds, Heterocyclic / blood*
  • Bridged Bicyclo Compounds, Heterocyclic / chemistry
  • Bridged Bicyclo Compounds, Heterocyclic / pharmacokinetics*
  • Chromatography, High Pressure Liquid / methods*
  • Drug Interactions
  • Female
  • Ketoconazole / administration & dosage
  • Ketoconazole / pharmacokinetics
  • Linear Models
  • Male
  • Mice
  • Reproducibility of Results
  • Sensitivity and Specificity
  • Sulfonamides / administration & dosage
  • Sulfonamides / blood*
  • Sulfonamides / chemistry
  • Sulfonamides / pharmacokinetics*
  • Tandem Mass Spectrometry / methods*

Substances

  • Bridged Bicyclo Compounds, Heterocyclic
  • Sulfonamides
  • venetoclax
  • Ketoconazole