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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

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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.

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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).

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Authors and Affiliations

  1. Department of Pharmacy, National University of Singapore, 10 Kent Ridge Crescent, Singapore, 119260, Singapore

    Samuel Chao Ming Yeo, Meng Huang & Hai-Shu Lin

  2. Department of Molecular and Cellular Biochemistry, University of Kentucky, Lexington, KY, 40506-0509, USA

    Vitaliy M. Sviripa, Liliia Kril, David S. Watt & Chunming Liu

  3. Center for Pharmaceutical Research and Innovation, College of Pharmacy, University of Kentucky, Lexington, KY, 40506-0509, USA

    Vitaliy M. Sviripa, Liliia Kril & David S. Watt

  4. Lucille Parker Markey Cancer Center, University of Kentucky, Lexington, KY, 40536-0093, USA

    David S. Watt & Chunming Liu

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  1. Samuel Chao Ming Yeo
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Correspondence to Hai-Shu Lin.

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Yeo, S.C.M., Sviripa, V.M., Huang, M. et al. 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. Anal Bioanal Chem 407, 7319–7332 (2015). https://doi.org/10.1007/s00216-015-8893-x

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