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Floxuridine Amino Acid Ester Prodrugs: Enhancing Caco-2 Permeability and Resistance to Glycosidic Bond Metabolism

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Abstract

Purpose

The aim of this study was to synthesize amino acid ester prodrugs of 5-fluoro-2′-deoxyuridine (floxuridine) to enhance intestinal absorption and resistance to glycosidic bond metabolism.

Methods

Amino acid ester prodrugs were synthesized and examined for their hydrolytic stability in human plasma, in Caco-2 cell homogenates, and in the presence of thymidine phosphorylase. Glycyl-l-sarcosine uptake inhibition and direct uptake studies with HeLa/PEPT1 cells [HeLa cells overexpressing oligopeptide transporter (PEPT1)] were conducted to determine PEPT1-mediated transport and compared with permeability of the prodrugs across Caco-2 monolayers.

Results

Isoleucyl prodrugs exhibited the highest chemical and enzymatic stability. The prodrugs enhanced the stability of the glycosidic bond of floxuridine. Thymidine phosphorylase rapidly cleaved floxuridine to 5-fluorouracil, whereas with the prodrugs no detectable glycosidic bond cleavage was observed. The 5′-l-isoleucyl and 5′-l-valyl monoester prodrugs exhibited 8- and 19-fold PEPT1-mediated uptake enhancement in HeLa/PEPT1 cells, respectively. Uptake enhancement in HeLa/PEPT1 cells correlated highly with Caco-2 permeability for all prodrugs tested. Caco-2 permeability of 5′-l-isoleucyl and 5′-l-valyl prodrugs was 8- to 11-fold greater compared with floxuridine.

Conclusions

Amino acid ester prodrugs such as isoleucyl floxuridine that exhibit enhanced Caco-2 transport and slower rate of enzymatic activation to parent, and that are highly resistant to metabolism by thymidine phosphorylase may improve oral delivery and therapeutic index of floxuridine.

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Abbreviations

HeLa/PEPT1:

HeLa cells overexpressing PEPT1

PEPT1:

oligopeptide transporter

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Acknowledgment

This work was supported by grant NIGMD-1R01GM 37188.

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Authors and Affiliations

  1. Department of Pharmaceutical Sciences, College of Pharmacy, University of Michigan, Ann Arbor, Michigan, 48109-1065, USA

    Christopher P. Landowski, Xueqin Song, Philip L. Lorenzi & Gordon L. Amidon

  2. TSRL, Inc., 540 Avis Drive, Suite A, Ann Arbor, Michigan, 48108, USA

    John M. Hilfinger

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  1. Christopher P. Landowski
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  2. Xueqin Song
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  4. John M. Hilfinger
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  5. Gordon L. Amidon
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Correspondence to Gordon L. Amidon.

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Landowski, C.P., Song, X., Lorenzi, P.L. et al. Floxuridine Amino Acid Ester Prodrugs: Enhancing Caco-2 Permeability and Resistance to Glycosidic Bond Metabolism. Pharm Res 22, 1510–1518 (2005). https://doi.org/10.1007/s11095-005-6156-9

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