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Competitive Inhibition of Glycylsarcosine Transport by Enalapril in Rabbit Renal Brush Border Membrane Vesicles: Interaction of ACE Inhibitors with High-Affinity H+/Peptide Symporter

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Abstract

Purpose. To examine the inhibitory potential of enalapril [and other angiotensin converting enzyme (ACE) inhibitors] on glycylsarcosine (GlySar) transport by the high-affinity renal peptide transporter.

Methods. Studies were performed in rabbit renal brush border membrane vesicles in which the uptake of radiolabeled GlySar was examined in the absence and presence of captopril, enalapril, enalaprilat, fosinopril, lisinopril, quinapril, quinaprilat, ramipril and zofenopril.

Results. Kinetic analyses demonstrated that enalapril inhibited the uptake of GlySar in a competitive manner (Ki ≈ 6 mM). Fosinopril and zofenopril had the greatest inhibitory potency (IC50 values of 55 and 81 μM, respectively) while the other ACE inhibitors exhibited low-affinity interactions with the renal peptide transporter. With respect to structure-function, ACE inhibitor affinity was strongly correlated with drug lipophilicity (r = 0.944, p < 0.001 for all ACE inhibitors; r = 0.983, p < 0.001 without enalaprilat, quinaprilat and quinapril).

Conclusions. The data suggest that enalapril and GlySar compete for the same substrate-binding site on the high-affinity peptide transporter in kidney, and that ACE inhibitors can interact with the renal carrier and inhibit dipeptide transport.

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REFERENCES

  1. H. Daniel. Function and molecular structure of brush border membrane peptide/H+ symporters. J. Memb. Biol. 154:197-203 (1996).

    Google Scholar 

  2. V. Ganapathy and F. H. Leibach. Peptide transporters. Curr. Opin. Nephrol. Hypertens. 5:395-400(1996).

    PubMed  Google Scholar 

  3. F. H. Leibach and V. Ganapathy. Peptide transporters in the intestine and the kidney. Annu. Rev. Nutr. 16:99-119 (1996).

    PubMed  Google Scholar 

  4. H. Daniel and M. Herget. Cellular and molecular mechanisms of renal peptide transport. Am. J. Physiol. 273:F1-F8 (1997).

    PubMed  Google Scholar 

  5. M. Boll, M. Herget, M. Wagener, W. M. Weber, D. Markovich, J. Biber, W. Clauss, H. Murer, and H. Daniel. Expression cloning and functional characterization of the kidney cortex high-affinity proton-coupled peptide transporter. Proc. Natl. Acad. Sci. 93:284-289 (1996).

    PubMed  Google Scholar 

  6. M. Boll, D. Markovich, W.-M. Weber, H. Korte, H. Daniel, and H. Murer. Expression cloning of a cDNA from rabbit small intestine related to proton-coupled transport of peptides, β-lactam antibiotics and ACE-inhibitors. Pflügers Arch—Eur. J. Physiol. 429:146-149 (1994).

    Google Scholar 

  7. W. Akarawut, C.-J. Lin, and D. E. Smith. Noncompetitive inhibition of glycylsarcosine transport by quinapril in rabbit renal brush border membrane vesicles: Effect on high-affinity peptide transporter. J. Pharmacol. Exp. Ther. 287:684-690 (1998).

    PubMed  Google Scholar 

  8. P.L. Jørgensen and J. C. Skou. Purification and characterization of (Na+-K+)-ATPase in preparations from the outer medulla of rabbit kidney. Biochim. Biophys. Acta 233:366-380 (1971).

    PubMed  Google Scholar 

  9. C.H. Fiske and Y. Subbarow. The colorimetric determination of phosphorous. J. Biol. Chem. 66:375-400 (1925).

    Google Scholar 

  10. M.M. Bradford. A rapid and sensitive method for the quantitation of microgram quantities of protein utilizing the principle of protein-dye binding. Anal. Biochem. 72:248-254 (1976).

    Article  PubMed  Google Scholar 

  11. U. Hopfer, K. Nelson, J. Perrotto, and K. J. Isselbacher. Glucose transport in isolated brush-border membrane from rat small intestine. J. Biol. Chem. 248:25-32 (1973).

    PubMed  Google Scholar 

  12. V. Ganapathy, G. Burckhardt, and F. H. Leibach. Characteristics of glycylsarcosine transport in rabbit intestinal brush-border membrane vesicles. J. Biol. Chem. 259:8954-8959 (1984).

    PubMed  Google Scholar 

  13. S. Silbernagl, V. Ganapathy, and F. H. Leibach. H+ gradient-driven dipeptide reabsorption in proximal tubule of rat kidney: Studies in vivo and in vitro. Am. J. Physiol. 253:F448-F457 (1987).

    PubMed  Google Scholar 

  14. J. C. Dearden and G. M. Bresnen. The measurement of partition coefficients. Quant. Struct.-Act. Relat. 7:133-144 (1988).

    Google Scholar 

  15. S. A. Ranadive, A. X. Chen, and A. T. M. Serajuddin. Relative lipophilicities and structural-pharmacological considerations of various angiotensin-converting enzyme (ACE) inhibitors. Pharm. Res. 9:1480-1486 (1992).

    PubMed  Google Scholar 

  16. A. R. Kugler, S. C. Olson, and D. E. Smith. Determination of quinapril and quinaprilat by high-performance liquid chromatography with radiochemical detection, coupled to liquid scintillation counting spectrometry. J. Chromatogr. B 666:360-367 (1995).

    Google Scholar 

  17. Y.-C. Cheng and W. H. Prusoff. Relationship between the inhibition constant (Ki) and the concentration which causes 50 percent inhibition (150) of an enzymatic reaction. Biochem. Pharmacol. 22:3099-3108 (1973).

    Article  PubMed  Google Scholar 

  18. D. E. Smith, A. Pavlova, U. V. Berger, M. A. Hediger, T. Yang, Y. G. Huang, and J. B. Schnermann. Tubular localization and tissue distribution of peptide transporters in rat kidney. Pharm. Res. 15:1244-1249 (1998).

    PubMed  Google Scholar 

  19. C.-J. Lin and D. E. Smith. Competitive inhibition of GlySar by enalapril in rabbit renal BBMV: Effect on high-affinity peptide transporter. Pharm. Res. 14(Suppl):S332-S333 (1997).

    Google Scholar 

  20. C. Evers, W. Haase, H. Murer, and R. Kinne. Properties of brush border vesicles isolated from rat kidney cortex by calcium precipitation. Membrane Biochem. 1:203-219 (1978).

    Google Scholar 

  21. T. D. McKinney and M. E. Kunnemann. Procainamide transport in rabbit renal cortical brush border membrane vesicles. Am. J. Physiol. 249:F532-F541 (1985).

    PubMed  Google Scholar 

  22. D. A. Griffiths, S. D. Hall, and P. P. Sokol. Effect of 3′-azido-3′-deoxythymidine (AZT) on organic ion transport in rat renal brush border membrane vesicles. J. Pharmacol. Exp. Ther. 260:128-133 (1992).

    PubMed  Google Scholar 

  23. Y. Miyamoto, J. L. Coone, V. Ganapathy, and F. H. Leibach. Distribution and properties of the glycylsarcosine-transport system in rabbit renal proximal tubule: Studies with isolated brush-border-membrane vesicles. Biochem. J. 249:247-253 (1988).

    PubMed  Google Scholar 

  24. T. Zhu, A. Steel, X.-Z. Chen, M. A. Hediger, and D. E. Smith. Differential recognition of ACE inhibitors in Xenopus oocytes expressing the intestinal and renal peptide transporters PepT1 and PepT2. PharmSci 1(Suppl):S393-S394 (1998).

    Google Scholar 

  25. H. Daniel, E. L. Morse, and S. A. Adibi. Determinants of substrate affinity for the oligopeptide/H+ symporter in the renal brush border membrane. J. Biol. Chem. 267:9565-9573 (1992).

    PubMed  Google Scholar 

  26. H. Daniel and S. A. Adibi. Transport of β-lactam antibiotics in kidney brush border membrane: Determinants of their affinity for the oligopeptide/H+ symporter. J. Clin. Invest. 92:2215-2223 (1993).

    PubMed  Google Scholar 

  27. S. A. Adibi. Renal assimilation of oligopeptides: Physiological mechanisms and metabolic importance. Am. J. Physiol. 272:E723-E736 (1997).

    PubMed  Google Scholar 

  28. P. W. Swaan, M. C. Stehouwer, and J. J. Tukker. Molecular mechanism for the relative binding affinity to the intestinal peptide carrier. Comparison of three ACE-inhibitors: Enalapril, enalaprilat, and lisinopril. Biochim. Biophys. Acta 1236:31-38 (1995).

    PubMed  Google Scholar 

  29. P. W. Swaan and J. J. Tukker. Molecular determinants of recognition for the intestinal peptide carrier. J. Pharm. Sci. 86:596-602 (1997).

    PubMed  Google Scholar 

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Correspondence to David E. Smith.

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Lin, CJ., Akarawut, W. & Smith, D.E. Competitive Inhibition of Glycylsarcosine Transport by Enalapril in Rabbit Renal Brush Border Membrane Vesicles: Interaction of ACE Inhibitors with High-Affinity H+/Peptide Symporter. Pharm Res 16, 609–615 (1999). https://doi.org/10.1023/A:1018847818766

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  • DOI: https://doi.org/10.1023/A:1018847818766

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