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Evidence for Overlapping Substrate Specificity Between Large Neutral Amino Acid (LNAA) and Dipeptide (hPEPTl) Transporters for PD 158473, an NMDA Antagonist

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Abstract

Purpose. The objective of this research was to investigate the substrate specificity of large neutral amino acid carrier (LNAA) and di/tripeptide (hPEPTl) transporters with respect to PD 158473, an NMDA antagonist.

Methods. Cellular uptake studies were carried out using two types of Chinese Hamster Ovary (CHO). CHO-K1 cells represent the wild type with inherent large neutral amino acid (LNAA) activity. CHO-PEPT1 cells were generated by stable transfection of hPEPTl gene into CHO cells. Therefore, these cells possess both LNAA activity and di/tripeptide transporter activities as a result of the transfection. Cellular uptake of PD 158473 was quantified using a HPLC method previously developed in our laboratory.

Results. The utility of the CHO-PEPT1 cell model was demonstrated by determining the uptake kinetics of Gly-Sar, a prototypical dipeptide transporter substrate. Uptake kinetics of PD 158473 displayed two carrier-mediated transport components in CHO-PEPT1 cells, while in CHO-K1 cells the relationship was consistent with classic one component Michaelis-Menten kinetics. These results confirmed the affinity of PD 158473 for both LNAA and di/tripeptide transporters. Further, results from inhibition experiments using these two cell types indicate that the high affinity-low capacity system was the LNAA carrier and the low affinity-high capacity carrier was the di/tripeptide transporter.

Conclusions. This study demonstrates overlapping substrate specificity between LNAA carrier and di/tripeptide transporter (hPEPTl) for PD 158473, an amino acid analog. Establishing Structure Transport Relationship (STR) for this overlap will aid in a design strategy for increasing oral absorption or targeting specific drugs to selected tissues.

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Authors and Affiliations

  1. Department of Pharmacokinetics, Dynamics and Metabolism, Parke-Davis Pharmaceutical Research, Division of Warner-Lambert Company, Ann Arbor, Michigan, 48105

    Narayanan Surendran, Christopher F. Bigge & Barbra H. Stewart

  2. Department of Biopharmaceutical Sciences and Pharmaceutical Chemistry, University of California, San Francisco, California

    Kuang-Ming Y. Covitz & Wolfgang Sadee

  3. College of Pharmacy, The University of Michigan, Ann Arbor, Michigan

    Hyo-kyung Han & Doo-Man Oh

  4. Department of Chemistry, Parke-Davis Pharmaceutical Research, Division of Warner-Lambert Company, Ann Arbor, Michigan

    Gordon L. Amidon

  5. Department of Molecular Biology, Parke-Davis Pharmaceutical Research, Division of Warner-Lambert Company, Ann Arbor, Michigan, 48105

    Rufus M. Williamson

Authors
  1. Narayanan Surendran
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  2. Kuang-Ming Y. Covitz
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  3. Hyo-kyung Han
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  4. Wolfgang Sadee
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  5. Doo-Man Oh
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  6. Gordon L. Amidon
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  7. Rufus M. Williamson
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  8. Christopher F. Bigge
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  9. Barbra H. Stewart
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Correspondence to Narayanan Surendran.

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Surendran, N., Covitz, KM.Y., Han, Hk. et al. Evidence for Overlapping Substrate Specificity Between Large Neutral Amino Acid (LNAA) and Dipeptide (hPEPTl) Transporters for PD 158473, an NMDA Antagonist. Pharm Res 16, 391–395 (1999). https://doi.org/10.1023/A:1018821718340

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