Analytical and Bioanalytical Chemistry

, Volume 407, Issue 24, pp 7319–7332 | Cite as

Analysis of trans-2,6-difluoro-4′-(N,N-dimethylamino)stilbene (DFS) in biological samples by liquid chromatography-tandem mass spectrometry: metabolite identification and pharmacokinetics

  • Samuel Chao Ming Yeo
  • Vitaliy M. Sviripa
  • Meng Huang
  • Liliia Kril
  • David S. Watt
  • Chunming Liu
  • Hai-Shu Lin
Research Paper

Abstract

The metabolism of a promising antineoplastic agent, trans-2,6-difluoro-4′-(N,N-dimethylamino)stilbene (DFS), was studied in mouse, rat, and human liver microsomes using liquid chromatography-tandem mass spectrometry (LC-MS/MS) with the multiple reaction monitoring-information-dependent acquisition-enhanced product ion scan (MRM-IDA-EPI) method. Ten putative metabolites were identified and the structures of four metabolites were confirmed using authentic standards. Since trans-2,6-difluoro-4′-(N-methylamino)stilbene (DMDFS, M1) was present in all species as metabolite and displayed in vitro growth inhibition superior to DFS, its pharmacokinetic profiles were examined in Sprague–Dawley rats using DFS as a comparator. A reliable LC-MS/MS multiple reaction monitoring (MRM) method was subsequently developed and validated for the simultaneous quantification of both DFS and DMDFS in rat plasma for this purpose. Upon intravenous administration (4 mg/kg), DFS had a moderate clearance (Cl = 62.7 ± 23.2 mL/min/kg), terminal elimination half-life (t 1/2 λZ  = 299 ± 73 min), and mean transit time (MTT = 123 ± 14 min) with demethylation metabolism accounting for about 10 % of its total clearance. DMDFS possessed an intravenous pharmacokinetic profile similar to DFS. During oral dosing (10 mg/kg) where both DFS and DMDFS were absorbed rapidly, the oral bioavailability of DFS was approximately 2-fold greater than that of DMDFS (DFS: F = 42.1 ± 12.8 %; DMDFS: F = 18.7 ± 3.9 %). Interestingly, the DMDFS exposure after oral dosing of DFS (10 mg/kg) was comparable to that after oral administration of DMDFS (10 mg/kg) alone. As DFS displayed potent anticancer activities and excellent pharmacokinetic profiles, it appears to be a favorable candidate for further pharmaceutical development.

Keywords

Liquid chromatography-tandem mass spectrometry Metabolite identification Pharmacokinetics trans-2,6-Difluoro-4′-(N-methylamino)stilbene trans-2,6-Difluoro-4′-(N,N-dimethylamino)stilbene 

Notes

Acknowledgments

HSL was supported by a start-up grant from the National University of Singapore (R-148-000-174-133). SCMY is a recipient of the President’s Graduate Fellowship of the National University of Singapore. CL and DSW were supported by R21 CA139359 and CA172379 from the NIH. DSW was supported by the Office of the Dean of the College of Medicine and by NIH Grant Number P20 RR020171 from the National Institute of General Medical Sciences to L. Hersh, PI. Its contents are solely the responsibility of the authors and do not necessarily represent the official views of the NIH or the NIGMS.

Conflict of interest

Samuel Chao Ming Yeo, Meng Huang, and Hai-Shu Lin declare that they have no conflict of interest. Vitaliy M. Sviripa, David S. Watt, and Chunming Liu declare that they are the inventors of “Stilbene analogs and methods of treating cancer” (US Patent 20120196874).

Supplementary material

216_2015_8893_MOESM1_ESM.pdf (36 kb)
ESM 1 (PDF 35.5 kb)

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Copyright information

© Springer-Verlag Berlin Heidelberg 2015

Authors and Affiliations

  • Samuel Chao Ming Yeo
    • 1
  • Vitaliy M. Sviripa
    • 2
    • 3
  • Meng Huang
    • 1
  • Liliia Kril
    • 2
    • 3
  • David S. Watt
    • 2
    • 3
    • 4
  • Chunming Liu
    • 2
    • 4
  • Hai-Shu Lin
    • 1
  1. 1.Department of PharmacyNational University of SingaporeSingaporeSingapore
  2. 2.Department of Molecular and Cellular BiochemistryUniversity of KentuckyLexingtonUSA
  3. 3.Center for Pharmaceutical Research and Innovation, College of PharmacyUniversity of KentuckyLexingtonUSA
  4. 4.Lucille Parker Markey Cancer CenterUniversity of KentuckyLexingtonUSA

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