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Hepatic Disposition of Ximelagatran and Its Metabolites in Pig; Prediction of the Impact of Membrane Transporters Through a Simple Disposition Model

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Abstract

Purpose

The double prodrug, ximelagatran, is bioconverted, via the intermediates ethylmelagatran and N-hydroxymelagatran, to the direct thrombin inhibitor, melagatran. The primary aim of this study was to investigate the hepatic metabolism and disposition of ximelagatran and the intermediates in pig. A secondary aim was to explore a simple in vitro methodology for quantitative investigations of the impact of membrane transporters on the disposition of metabolized drugs.

Methods

Porcine S1 (supernatant fraction obtained by centrifuging at 1,000 g for 10 min) liver fractions and hepatocytes were incubated in the absence and presence of known membrane transporter inhibitors. The in vitro kinetics and disposition were determined by simultaneously fitting the disappearance of ximelagatran and the appearance of the metabolites.

Results

In S1 liver fractions, the metabolism was significantly inhibited by co-incubation of verapamil or ketoconazole, but not by erythromycin, quinine or quinidine. The disposition of ximelagatran and the intermediate metabolites in hepatocytes were influenced, to various degrees, by carrier-mediated transport processes.

Conclusion

This work demonstrates that it is possible to obtain profound information on the general mechanisms that are important in the drug liver disposition using the combination of common in vitro systems and the simple disposition model proposed in this study.

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Abbreviations

ACTB:

β actin

BCRP:

breast cancer resistance protein

Cu :

unbound concentration

CYP:

cytochrome P450

ETM:

ethylmelagatran

fu :

fraction unbound

GAPDH:

glyceraldehyde-3-phosphate dehydrogenase

HBSS:

Hank’s buffer salt solution

HPRT:

hypoxanthine phosphoribosyltransferase

HSB:

Hepatocyte suspension buffer

Km :

Michaelis constant

MEL:

melagatran

MRP2:

multi-drug resistance protein 2

NADH:

reduced β-nicotinamide adenine dinucleotide

NADPH:

reduced β-nicotinamide adenine dinucleotide phosphate

OHM:

N-hydroxymelagatran

P-gp:

P-glycoprotein

S1:

supernatant fraction obtained by centrifuging at 1,000 g for 10 min

Vmax :

theoretical maximum reaction rate

XIM:

ximelagatran

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ACKNOWLEDGEMENTS

This work was supported by AstraZeneca R&D Mölndal, Mölndal, Sweden.

Author information

Author notes

  1. Björn Arvidsson

    Present address: FOI, Swedish Defence Research Agency, CBRN Defence and Security, Umeå, Sweden

Authors and Affiliations

  1. Department of Pharmacy, Uppsala University, Box 580, 751 23, Uppsala, Sweden

    Erik Sjögren & Hans Lennernäs

  2. Discovery DMPK & Bioanalytical Chemistry, AstraZeneca R&D, Mölndal, Sweden

    Ulf Bredberg

  3. Analytical Chemistry, Department of Physical and Analytical Chemistry, Uppsala University, Uppsala, Sweden

    Erik Allard, Björn Arvidsson & Jonas Bergquist

  4. Clinical Pharmacology and DMPK, AstraZeneca R&D, Mölndal, Sweden

    Tommy B. Andersson

  5. Section of Pharmacogenetics, Department of Physiology and Pharmacology, Karolinska Institutet, Stockholm, Sweden

    Tommy B. Andersson

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  1. Erik Sjögren
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Correspondence to Hans Lennernäs.

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Sjögren, E., Bredberg, U., Allard, E. et al. Hepatic Disposition of Ximelagatran and Its Metabolites in Pig; Prediction of the Impact of Membrane Transporters Through a Simple Disposition Model. Pharm Res 27, 597–607 (2010). https://doi.org/10.1007/s11095-009-0016-y

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