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Carrier Mechanisms Involved in the Transepithelial Transport of bis(POM)-PMEA and Its Metabolites Across Caco-2 Monolayers

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Abstract

Purpose. To investigate the role of carrier mechanisms in: [1] the polarized transport of the bis(pivaloyloxymethyl)- [bis(POM)-] ester prodrug of the antiviral agent 9-(2-phosphonylmethoxyethyl)adenine [PMEA] and [2] the directional secretion of its metabolites.

Methods. Caco-2 monolayers were used to study the modulating effect of carriers on the transport of bis(POM)-PMEA and the efflux of intracellularly formed metabolites mono(POM)-PMEA and PMEA from the cells. The interaction of bis(POM)-PMEA and its metabolites with the efflux mechanisms present in Caco-2 monolayers was investigated by testing the effect of various concentrations of verapamil (30, 100, 300 μM) or indomethacin (10-500 μM) on transport and efflux.

Results. Polarity in transport of bis(POM)-PMEA (50 μM) across Caco-2 monolayers was noted: transport of total PMEA [=bis(POM)-PMEA, mono(POM)-PMEA and PMEA] was significantly higher in basolateral (BL) to apical (AP) direction (14.5 ± 0.4%) than transport in the opposite (AP to BL) direction (1.7 ± 0.2%). This difference was reduced in a concentration dependent way when verapamil (0−100 μM) was included in both AP and BL incubation media. After loading the cells with bis(POM)-PMEA (100 μM) for 1 hr, studies on efflux of PMEA and mono(POM)-PMEA from the Caco-2 monolayers over a 3 hr period, revealed that both metabolites were preferentially secreted towards the AP compartment. Efflux of PMEA towards AP and BL compartments amounted to 14.6 ± 1.1 % and 5.3 ± 0.4%, respectively, of the initial intracellular amount of total PMEA, while efflux of mono(POM)-PMEA towards AP and BL compartments was limited to 2.3 ± 0.1 % and 0.5 ± 0.1 %, respectively. When 10 μM indomethacin was included in the AP incubation medium, efflux of PMEA was decreased to 7.8 ± 0.3% and 3.3 ± 0.3% towards the AP and BL compartments, respectively. The decrease in efflux by indomethacin was concentration-dependent up to 100 μM. Transepithelial transport of total PMEA was also reduced in the presence of 30 μM indomethacin, as reflected in smaller concentrations of PMEA and mono(POM)-PMEA in the acceptor compartment, irrespective of the transport direction.

Conclusions. The data obtained in this study suggest that bis(POM)-PMEA is substrate for a P-glycoprotein-like carrier mechanism in Caco-2 monolayers, while its metabolites mono(POM)-PMEA and PMEA are transported by a non-P-glycoprotein efflux protein.

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Authors and Affiliations

  1. Laboratorium voor Farmacotechnologie en Biofarmacie, KULeuven, Campus Gasthuisberg, O + N, Herestraat 49, B-3000, Leuven, Belgium

    P. Annaert, J. Van Gelder, G. Van den Mooter, R. Kinget & P. Augustijns

  2. Rega Institute for Medical Research, KULeuven, Minderbroedersstraat 10, B-3000, Leuven, Belgium

    L. Naesens & E. De Clercq

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  1. P. Annaert
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  2. J. Van Gelder
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  6. R. Kinget
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  7. P. Augustijns
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Annaert, P., Van Gelder, J., Naesens, L. et al. Carrier Mechanisms Involved in the Transepithelial Transport of bis(POM)-PMEA and Its Metabolites Across Caco-2 Monolayers. Pharm Res 15, 1168–1173 (1998). https://doi.org/10.1023/A:1011923420719

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