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Characterization of the Efflux Transporter(s) Responsible for Restricting Intestinal Mucosa Permeation of an Acyloxyalkoxy-Based Cyclic Prodrug of the Opioid Peptide DADLE

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Abstract

Purpose. To elucidate the efflux transporter(s) responsible for restricting the permeation of an acyloxyalkoxy-based cyclic prodrug of the opioid peptide DADLE (AD) through Caco-2 cell monolayers.

Methods. The cellular permeation characteristics of AD were investigated using Caco-2 cells, Madin-Darby canine kidney wild-type II cells (MDCK-WT), MDCK cells transfected with the human MDR1 gene (MDCK-MDR1), and MDCK cells transfected with the human MRP2 gene (MDCK-MRP2). These cells were grown as monolayers onto microporous membranes. The disappearance of AD from the donor side and its appearance on the receiver side were monitored by high-performance liquid chromatography. The substrate activity of AD for P-glycoprotein (P-gp) was determined using GF120918, a known P-gp specific inhibitor. The substrate activity of AD for MRP2 was determined by using cyclosporin A, a known MRP2 and P-gp inhibitor.

Results. In Caco-2 cells, the ratio of the apparent permeability coefficients (Papp) of AD flux measured in the basolateral (BL) to apical (AP) direction vs. the flux in the AP-to-BL direction (Papp BL-to-AP/

Papp AP-to-BL) was 99. In the presence of 2 μM GF120918 or 25 μM cyclosporin A, the Papp BL-to-AP/Papp AP-to-BL ratio was decreased to 11. In MDCK-WT, MDCK-MDR1, and MDCK-MRP2 cells, the Papp BL-to-AP/Papp AP-to-BL ratios of AD were 4.7, 10, and 5.8, respectively. A mixture of GF120918 (2 μM) and cyclosporin A (25 μM) decreased the Papp BL-to-AP/Papp AP-to-BL ratios of AD in MDCK-WT, MDCK-MDR1, and MDCK-MRP2 cells to 1.2, 1.8, and 2.3, respectively.

Conclusions. These data suggest that AD is a much better substrate for P-gp than MRP2 and that the restricted permeation of this cyclic prodrug in Caco-2 cells and in the intestinal mucosa probably is due primarily to its substrate activity for P-gp.

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  1. Fuxing Tang

    Present address: Forest Laboratories, Inc., Farmingdale, New York, 11735

Authors and Affiliations

  1. Department of Pharmaceutical Chemistry, The University of Kansas, Lawrence, Kansas, 66047

    Fuxing Tang

  2. Forest Laboratories, Inc., Farmingdale, New York, 11735

    Ronald T. Borchardt

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Tang, F., Borchardt, R.T. Characterization of the Efflux Transporter(s) Responsible for Restricting Intestinal Mucosa Permeation of an Acyloxyalkoxy-Based Cyclic Prodrug of the Opioid Peptide DADLE. Pharm Res 19, 780–786 (2002). https://doi.org/10.1023/A:1016144530146

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