Abstract
Purpose. To synthesize amino acid ester prodrugs of floxuridine (FUdR) and to investigate the effects of structure, stereochemistry, and site of esterification of promoiety on the rates of hydrolysis of these prodrugs in Caco-2 cell homogenates.
Methods. Amino acid ester prodrugs of FUdR were synthesized using established procedures. The kinetics of hydrolysis of prodrugs was evaluated in human adenocarcinoma cell line (Caco-2) homogenates and pH 7.4 phosphate buffer.
Results. 3′-Monoester, 5′-monoester, and 3′,5′-diester prodrugs of FUdR utilizing proline, L-valine, D-valine, L-phenylalanine, and D-phenylalanine as promoieties were synthesized and characterized. In Caco-2 cell homogenates, the L-amino acid ester prodrugs hydrolyzed 10 to 75 times faster than the corresponding D-amino acid ester prodrugs. Pro and Phe ester prodrugs hydrolyzed much faster (3- to 30-fold) than the corresponding Val ester prodrugs. Further, the 5′-monoester prodrugs hydrolyzed significantly faster (3-fold) than the 3′,5′-diester prodrugs.
Conclusions. Novel amino acid ester prodrugs of FUdR were successfully synthesized. The results presented here clearly demonstrate that the rate of FUdR prodrug activation in Caco-2 cell homogenates is affected by the structure, stereochemistry, and site of esterification of the promoiety. Finally, the 5′-Val and 5′-Phe monoesters exhibited desirable characteristics such as good solution stability and relatively fast enzymatic conversion rates.
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N. Kemeny, Y. Huang, A. M. Cohen, W. Shi, J. A. Conti, M. F. Brennan, J. R. Bertino, A. D. Turnbull, D. Sullivan, J. Stockman, L. H. Blumgart, and Y. Fong. Hepatic arterial infusion of chemotherapy after resection of hepatic metastases from colorectal cancer. N. Engl. J. Med. 341:2039–2048 (1999).
J. L. Grem. 5–Fluorouracil: Forty–plus and still ticking. A review of its preclinical and clinical development. Invest. New Drugs 18:299–313 (2000).
B. A. Chabner, D. P. Ryan, L. Paz–Area, R. Garcia–Carbonero, and P. Calabresi. Antineoplastic agents. In G. Hardman and L. E. Limbird (eds.), Goodman & Gilman's The Pharmacological Basis of Therapeutics, 10th ed., Mc–Graw Hill, New York, 2001, pp. 1389–1459.
J. A. Van Laar, Y. M. Rustum, S. P. Ackland, C. J. Van Groeningen, and G. J. Peters. Comparison of 5–fluoro–2′–deoxyuridine with 5–fluorouracil and their role in the treatment of colorectal cancer. Eur. J. Cancer 34:296–306 (1998).
H. K. Han and G. L. Amidon. Targeted prodrug design to optimize drug delivery. AAPS Pharmsci. 2:E6(2000).
W. A. Denny. Prodrug strategies in cancer therapy. Eur. J. Med. Chem. 36:577–595 (2001).
R. A. Schwendener, A. Supersaxo, W. Rubas, H. G. Weder, H. R. Hartmann, H. Schott, A. Ziegler, and H. Hengartner. 5′–O–Palmitoyl–and 3′,5′–O–dipalmitoyl–5–fluoro–2′–deoxyuridine—novel lipophilic analogues of 5′–fluoro–2′–deoxyuridine: Synthesis, incorporation into liposomes and preliminary biological results. Biochem. Biophys. Res. Commun. 126:660–666 (1985).
S. Fukushima, T. Kawaguchi, M. Nishida, K. Juni, Y. Yamashita, M. Takahashi, and M. Nakano. Selective anticancer effects of 3′,5′–dioctanoyl–5–fluoro–2′–deoxyuridine, a lipophilic prodrug of 5–fluoro–2′–deoxyuridine, dissolved in an oily lymphographic agent on hepatic cancer of rabbits bearing vx–2 tumor. Cancer Res. 47:1930–1934 (1987).
T. Kawaguchi, S. Fukushima, Y. Hayashi, and M. Nakano. Nonenzymatic enzymatic hydrolysis of 5–fluoro–2′–deoxyuridine (FUdR) esters. Pharm. Res. 5:741–744 (1988).
T. Halmos, P. Moroni, K. Antonakis, and J. Uriel. Fatty acid conjugates of 2′–deoxy–5–fluorouridine as prodrugs for the selective delivery of 5–fluorouracil to tumor cells. Biochem. Pharmacol. 44:149–155 (1992).
S. C. Tobias and R. F. Borch. Synthesis and biological studies of novel nucleoside phosphoramidate prodrugs. J. Med. Chem. 44:4475–4480 (2001).
Y. Wei, Y. Yan, D. Pei, and B. Gong. A photoactivated prodrug. Bioorg. Med. Chem. Lett. 8:2419–2422 (1998).
Z. Xia, L. I. Wiebe, G. G. Miller, E. E. Knaus. Synthesis and biological evaluation of butanoate, retinoate, bis(2,2,2–trichloroethyl)phosphate derivatives of 5–fluoro–2′–deoxyuridine 2′,5–difluoro–2′–deoxyuridine as potential dual action anti–cancer prodrugs. Arch. Pharm. (Weinheim) 332:286–294 (1999).
H. K. Han, D. M. Oh, and G. L. Amidon. Cellular uptake mechanism of amino acid ester prodrugs in caco–2/hpept1 cells overexpressing a human peptide transporter. Pharm. Res. 15:1382–1386 (1998).
I. Rubio–Aliaga and H. Daniel. Mammalian peptide transporters as targets for drug delivery. Trends Pharmacol. Sci. 23:434–440 (2002).
D. M. Oh, H. Han, and G. L. Amidon. Drug transport and targeting. Intestinal transport. Pharm. Biotechnol. 12:59–88 (1999).
M. Sugawara, W. Huang, Y. L. Fei, F. H. Leibach, V. Ganapathy, and M. E. Ganapathy. Transport of valganciclovir, a ganciclovir prodrug, via peptide transporters pept1 and pept2. J. Pharm. Sci. 89:781–789 (2000).
T. Nakanishi, I. Tamai, A. Takaki, A. Tsuji. Cancer cell–targeted drug delivery utilizing oligopeptide transport activity. Int. J. Cancer 88:274–280 (2000).
D. E. Gonzalez, K. M. Covitz, W. Sadee, and R. J. Mrsny. An oligopeptide transporter is expressed at high levels in the pancreatic carcinoma cell lines aspc–1 and capan–2. Cancer Res. 58:519–525 (1998).
R. J. Mrsny. Oligopeptide transporters as putative therapeutic targets for cancer cells. Pharm. Res. 15:816–818 (1998).
V. H. Lee. Mucosal drug delivery. J. Natl. Cancer Inst. Monogr. 41:4(2001).
F. M. Williams. Clinical significance of esterases in man. Clinical Pharmacokin. 10:392–403 (1985).
P. K. Banerjee and G. L. Amidon. Design of prodrugs based on enzyme–substrate specificity. In H. Bundgaard (ed.), Design of Prodrugs, Elsevier, Sciences Publisher, New York, 1985, pp. 93–133.
T. Satoh, P. Taylor, W. F. Bosron, S. P. Sanghani, M. Hosokawa, and B. N. La Du. Current progress on esterases: From molecular structure to function. Drug Metab. Dispos. 30:488–493 (2002).
S. A. Varia, S. Schuller, and V. J. Stella. Phenytoin prodrugs iv: Hydrolysis of various 3–(hydroxymethyl)phenytoin esters. J. Pharm. Sci. 73:1074–1080 (1984).
T. C. Bruice and S. Benkovic. Bioorganic mechanisms, W. A. Benjamin, Inc., New York (1966).
R. Wolfenden. The mechanism of hydrolysis of amino acyl RNA. Biochemistry 2:1090–1092 (1963).
I. J. Hidalgo, T. J. Raub, and R. T. Borchardt. Characterization of the human colon carcinoma cell line (caco–2) as a model system for intestinal epithelial permeability. Gastroenterology 96:736–749 (1989).
K. Tamura, C. P. Lee, P. L. Smith, and R. T. Borchardt. Metabolism, uptake, transepithelial transport of the stereoisomers of val–val–val in the human intestinal cell line, caco–2. Pharm. Res. 13:1663–1667 (1996).
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Vig, B.S., Lorenzi, P.J., Mittal, S. et al. Amino Acid Ester Prodrugs of Floxuridine: Synthesis and Effects of Structure, Stereochemistry, and Site of Esterification on the Rate of Hydrolysis. Pharm Res 20, 1381–1388 (2003). https://doi.org/10.1023/A:1025745824632
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DOI: https://doi.org/10.1023/A:1025745824632