Determination of cobimetinib in human plasma using protein precipitation extraction and high-performance liquid chromatography coupled to mass spectrometry

J Chromatogr B Analyt Technol Biomed Life Sci. 2014 Dec 1:972:117-23. doi: 10.1016/j.jchromb.2014.09.034. Epub 2014 Oct 7.

Abstract

Inhibition of MAP/ERK kinase (MEK) is a promising strategy to control the growth of tumors that are dependent on aberrant signaling in the MEK pathway. Cobimetinib (GDC-0973) (S)-[3,4-Difluoro-2-(2-fluoro-4-iodo-phenylamino)-phenyl]-((S)-3-hydroxy-3-piperidin-2-yl-azetidin-1-yl)-methanone) inhibits proliferation of a variety of human tumor cell lines by inhibiting MEK1 and MEK2. A specific high performance liquid chromatography-mass spectrometric assay was developed and validated for the determination of cobimetinib in human plasma. The overall mean recovery using protein precipitation extraction with acetonitrile was found to be 54.1%. The calibration curve was ranged from 0.20 to 100ng/mL. The LLOQ was sensitive enough to detect terminal phase concentrations of the drug. The intra- and inter-assay precision (%CV) was within 10.3% and 9.5% for cobimetinib. The assay accuracy (%RE) was within ±13.7% of the nominal concentration values for cobimetinib with the normal analytical QCs. The developed assay was successfully used to analyze the human plasma samples (for pharmacokinetic analysis) from clinical trials.

Keywords: Cobimetinib; GDC-0973; LC–MS/MS; Validation.

Publication types

  • Randomized Controlled Trial

MeSH terms

  • Azetidines / blood*
  • Azetidines / pharmacokinetics
  • Cell Line, Tumor
  • Chromatography, High Pressure Liquid / methods*
  • Cross-Over Studies
  • Humans
  • Limit of Detection
  • MAP Kinase Kinase 1 / antagonists & inhibitors*
  • Mass Spectrometry / methods*
  • Piperidines / blood*
  • Piperidines / pharmacokinetics
  • Protein Kinase Inhibitors / blood*
  • Protein Kinase Inhibitors / pharmacokinetics
  • Proteins / chemistry*
  • Reproducibility of Results

Substances

  • Azetidines
  • Piperidines
  • Protein Kinase Inhibitors
  • Proteins
  • MAP Kinase Kinase 1
  • cobimetinib