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Pharmaceutical Research

, Volume 23, Issue 7, pp 1543–1553 | Cite as

Development of an in Vitro Rat Intestine Segmental Perfusion Model to Investigate Permeability and Predict Oral Fraction Absorbed

  • Marc-Etienne Castella
  • Marianne Reist
  • Joachim M. Mayer
  • Jean-Jacques Turban
  • Bernard Testa
  • Claire Boursier-Neyret
  • Bernard Walther
  • Jean-Marie Delbos
  • Pierre-Alain Carrupt
Research Paper

Purpose

The aims of the study are to develop and evaluate an in vitro rat intestine segmental perfusion model for the prediction of the oral fraction absorbed of compounds and to assess the ability of the model to study intestinal metabolism.

Methods

The system consisted of a perfusion cell with a rat intestinal segment and three perfusion circulations (donor, receiver, and rinsing circulation). Lucifer yellow (LY) was applied as internal standard together with test compounds in the donor circulation. To validate the model, the permeability of eight noncongeneric passively absorbed drugs was determined. Intestinal N-demethylation of verapamil into norverapamil was followed in the donor and receiver circulations by high-performance liquid chromatography analysis.

Results

The in vitro model allowed ranking of the tested compounds according to their in vivo absorption potential. The Spearman's correlation coefficient between the oral fraction absorbed in humans and the ratio of permeation coefficient of test compound to the permeation coefficient of LY within the same experiment was 0.98 (P < 0.01). Moreover, intestinal N-demethylation of verapamil, its permeation, and the permeation of its metabolite norverapamil could be assessed in parallel.

Conclusions

Up to six permeation kinetics can be obtained per rat, and the method has shown to be a valuable tool to estimate human oral absorption.

Key Words

absorption model in vitro intestinal metabolism in vitro intestinal permeability in vitroin vivo correlation rat jejunal perfusion 

Abbreviations

Fa

oral fraction absorbed

HPLC

high-performance liquid chromatography

LC-MS/MS

liquid chromatography with tandem mass spectrometry

log Doct7.4

octanol water distribution coefficient at pH 7.4

log PN

octanol water partition coefficient of the uncharged species

LY

Lucifer yellow

MW

molecular weight

Papp

coefficient of apparent permeability

Peff

effective permeability coefficients

Notes

Acknowledgments

We thank Marie-Paule Le Bon, Josseline Le Gourrierec, and Jacky Pothier for the analyses of nadolol samples. Also, we are grateful to Catherine Canovi for her advices and skilful technical assistance.

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Copyright information

© Springer Science + Business Media, Inc. 2006

Authors and Affiliations

  • Marc-Etienne Castella
    • 1
  • Marianne Reist
    • 1
  • Joachim M. Mayer
    • 1
  • Jean-Jacques Turban
    • 1
  • Bernard Testa
    • 2
  • Claire Boursier-Neyret
    • 3
  • Bernard Walther
    • 3
  • Jean-Marie Delbos
    • 3
  • Pierre-Alain Carrupt
    • 1
  1. 1.LCT-Pharmacochemistry, School of Pharmaceutical Sciences, EPGLUniversity of Geneva, University of LausanneGeneva 4Switzerland
  2. 2.Département de PharmacieCentre Hospitalier Universitaire VaudoisLausanneSwitzerland
  3. 3.Technologie ServierOrléansFrance

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