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Secreted protein profile from HepG2 cells incubated by S(−) and R(+) enantiomers of chiral drug warfarin – An analysis in cell-based system and clinical samples

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Abstract

Purpose: Warfarin is a commonly prescribed oral anticoagulant with narrow therapeutic index. It interferes with the vitamin K cycle to achieve anti-coagulating effects. Warfarin has two enantiomers, S(−) and R(+) and undergoes stereoselective metabolism, with the S(−) enantiomer being more effective. Our target is to discover the biological differences of the two enantiomers for better warfarin therapy.

Experimental design: We reported the extracellular protein profile in HepG2 cells incubated with S(−) and R(+) warfarin, using iTRAQ-coupled 2-D LC-MS/MS. In addition, clinical sera from 30 patients taken warfarin were also analyzed by the same method as a long-term batch.

Results: In cell line batch in samples incubated with S(−) and R(+) warfarin alone, inter-α-trypsin inhibitor heavy chain H4, apolipoprotein A-I and α-2-HS-glycoprotein showed variations in cells incubated with S(−) warfarin and R(+) warfarin. For other proteins like α-2-macroglobulin and Fibrinogen γ chain, the expressions each were found to be the same in cells incubated with either S(−) or R(+) warfarin. Clinical results showed the same trends for protein ratio changes.

Conclusion and clinical relevance: Our results indicated that those proteins may interfere with blood coagulation process, as well as contribute to the warfarin's side-effect response. Taken together, our findings provided molecular evidence on a comprehensive protein profile on warfarin–cell interaction which may shed new lights on future improvement of warfarin therapy.

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