LC–MS/MS method for the quantitation of a dual PI3K/BRD4 inhibitor SF2523 in mouse plasma: Application to plasma protein binding and metabolism studies

Veenu Bala, Yashpal S. Chhonker, Guillermo A. Morales, Krishnaiah Maddeboina, Dhananjaya Pal, Donald L. Durden, Daryl J. Murry

Research output: Contribution to journalArticlepeer-review

1 Scopus citations

Abstract

A sensitive and selective liquid chromatography coupled with tandem mass spectrometry (LC–MS/MS) method was developed and validated for the quantitation of dual PI3K/BRD4 inhibitor SF2523 in mouse plasma. The analysis was performed on a UPLC system connected to a Shimadzu 8060 mass spectrometer by electrospray ionization in positive multiple reaction monitoring mode. Chromatographic separation was carried out on an ACE Excel C18 column with a gradient elution containing 0.1% formic acid and methanol as the mobile phase. The linearity was conducted in the concentration range 0.1–500 ng/ml for SF2523 in 100 μl of plasma. The inter- and intra-batch precision (RSD) were both lower than 13.5%, with the accuracy (percentage bias) ranging from −10.03 to 11.56%. The validated method was successfully applied to plasma protein binding and in vitro metabolism studies. SF2523 was highly bound to mouse plasma proteins (>95% bound). Utilizing mouse S9 fractions, a total of seven phase I and II metabolites were identified with hydroxylation found to be the major metabolic pathway. Metabolite identification included analysis of retention behaviors, molecular weight changes and MS/MS fragment patterns of SF2523 and the metabolites. This newly developed and validated method allows the rapid and easy determination of the SF2523 concentration with high sensitivity in a low sample volume and can be applied to future pre-clinical studies.

Original languageEnglish (US)
Article numbere5643
JournalBiomedical Chromatography
Volume37
Issue number8
DOIs
StatePublished - Aug 2023

Keywords

  • LC–MS/MS
  • SF2523
  • dual PI3K/BRD4 inhibitor

ASJC Scopus subject areas

  • Analytical Chemistry
  • Biochemistry
  • Molecular Biology
  • Pharmacology
  • Drug Discovery
  • Clinical Biochemistry

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