Development of an in Vitro Rat Intestine Segmental Perfusion Model to Investigate Permeability and Predict Oral Fraction Absorbed
- 220 Downloads
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.
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.
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.
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 Wordsabsorption model in vitro intestinal metabolism in vitro intestinal permeability in vitro–in vivo correlation rat jejunal perfusion
oral fraction absorbed
high-performance liquid chromatography
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
coefficient of apparent permeability
effective permeability coefficients
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.
- 1.Tukker, J. J. 1996In vitro methods for the assessment of permeabilityDressman, J. B.Lennernäs, H. eds. Oral Drug Absorption—Prediction and Assessment Vol. 106Marcel Dekker Inc.New York5172Google Scholar
- 10.Smith, P. L. 1996Methods for evaluating intestinal permeability and metabolism in vitroBorchardt, R. T.Smith, P. L.Wilson, G. eds. Models for Assessing Drug Absorption and Metabolism Vol. 8PlenumNew York1334Google Scholar
- 13.Palm, K., Artursson, P., Luthman, K. 1997Experimental and theoretical predictions of intestinal drug absorptionWaterbeemd, H.Testa, B.Folkers, G. eds. Computer-Assisted Lead Finding and OptimizationWiley-VCHWeinheim277289Google Scholar
- 18.Johnson, B. M., Charman, W. N., Porter, C. J. 2002An in vitro examination of the impact of polyethylene glycol 400, pluronic P85 and vitamin E d-a-tocopheryl polyethylene glycol 1000 succinate on p-glycoprotein efflux and enterocyte-based metabolism in excised rat intestineAAPS Pharm. Sci.4E40CrossRefGoogle Scholar
- 30.Polentarutti, B. I., Peterson, A. L., Sjoberg, A. K., Anderberg, E. K., Utter, L. M., Ungell, A. L. 1999Evaluation of viability of excised rat intestinal segments in the Ussing chamber: investigation of morphology, electrical parameters, and permeability characteristicsPharm. Res.16446454PubMedCrossRefGoogle Scholar
- 42.Eichelbaum, M., Ochs, H. R., Roberts, G., Somogyi, A. 1982Pharmacokinetics and metabolism of antipyrine (phenazone) after intravenous and oral administrationArzneim.-Forsch.32575578Google Scholar
- 44.Caron, G., Steyaert, G., Pagliara, A., Reymond, F., Crivori, P., Gaillard, P., Carrupt, P. A., Avdeef, A., Comer, J., Box, K. J., Girault, H. H., Testa, B. 1999Structure–lipophilicity relationships of neutral and protonated β-blockers. Part I. Intra- and intermolecular effects in isotropic systemsHelv. Chim. Acta8212111222CrossRefGoogle Scholar