Advertisement

Pharmaceutical Research

, Volume 22, Issue 5, pp 721–727 | Cite as

Experimental and Computational Studies of Epithelial Transport of Mefenamic Acid Ester Prodrugs

  • Kamonthip Wiwattanawongsa
  • Vimon TantishaiyakulEmail author
  • Luelak Lomlim
  • Yon Rojanasakul
  • Sirirat Pinsuwan
  • Sanae Keawnopparat
Research Paper

Abstract

Purpose.

A series of ester derivatives of mefenamic acid were synthesized with the aim of suppressing local gastrointestinal toxicity of mefenamic acid. A computational method was used to assist the design of the prodrug and to gain insights into the structure relationship of these compounds as P-glycoprotein (P-gp) substrates. The prodrugs were studied for their enzymatic stability, bidirectional permeability across Caco-2 monolayer, and their potential as transporter modulators

Methods.

Bidirectional transport studies were performed using Caco-2 cells. Compounds exhibiting an efflux ratio of ≥2 were further examined for their potential interaction with P-gp and multidrug resistance–associated protein (MRP) using verapamil and indomethacin. Calcein efflux inhibition studies were conducted to investigate the efflux mechanism of these compounds. Geometry optimization of the esters was performed, and the spatial separation of two electron donor groups of each prodrug was measured.

Results.

Morpholinoethyl ester (3) and pyrrolidinoethyl ester (4) of mefenamic acid showed evidence of efflux mechanism. Inhibition by verapamil had a pronounced effect on the transport of 3 and 4. Indomethacin, however, completely inhibited the apical efflux of 3 but enhanced the efflux ratio of 4. Both compounds increased the ratio of cellular calcein accumulation by 3- to 5-fold over control. Consistent with the experimental data, the computational results suggest the involvement of P-gp or its interaction in 3 and 4 transport.

Conclusions.

Apical efflux of 3 is associated with P-gp and MRP, but the efflux of 4 involves P-gp and/or MRP. The computational approach used in this study provided the basis for P-gp substrates of compounds 3 and 4 from their electron donor subunits spatial separation.

Key Words:

Caco-2 MRP P-gp prodrugs transport 

Preview

Unable to display preview. Download preview PDF.

Unable to display preview. Download preview PDF.

References

  1. 1.
    1. E. L. Semble and W. C. Wu. Anti-inflammatory drugs and gastric mucosal damage. Semin. Arthritis Rheum. 16:271–286 (1987).CrossRefPubMedGoogle Scholar
  2. 2.
    2. V. K. Tammara, M. M. Narurkar, A. M. Crider, and M. A. Khan. Morpholinoalkyl ester prodrugs of diclofenac: synthesis, in vitro and in vivo evaluation. J. Pharm. Sci. 83:644–648 (1994).PubMedGoogle Scholar
  3. 3.
    3. A. Seelig. A general pattern for substrate recognition by P-glycoprotein. Eur. J. Biochem. 251:252–261 (1988).CrossRefGoogle Scholar
  4. 4.
    4. J. E. Penzotti, M. L. Lamb, E. Evensen, and P. D. J. Grootenhuis. A computational ensemble pharmacophore model for identifying substrates of P-glycoprotein. J. Med. Chem. 45:1737–1740 (2002).CrossRefPubMedGoogle Scholar
  5. 5.
    5. P. Augustijns, P. Annaert, P. Heylen, G. Van den Mooter, and R. Kinget. Drug absorption studies of prodrug esters using the Caco-2 model: evaluation of ester hydrolysis and transepithelial transport. Int. J. Pharm. 166:45–53 (1998).CrossRefGoogle Scholar
  6. 6.
    6. D. P. Olson, B. J. Taylor, and S. P. Ivy. Detection of MRP functional activity: calcein AM but not BCECF AM as a multidrug resistance-related protein (MRP1) substrate. Cytometry 46:105–113 (2001).CrossRefPubMedGoogle Scholar
  7. 7.
    7. S. Benzaria, H. Pelicano, R. Johnson, G. Maury, J. L. Imbach, A. M. Aubertin, G. Obert, and G. Gosselin. Synthesis, in vitro antiviral evaluation, and stability studies of bis(S-acyl-2-thioethyl) ester derivatives of 9-[2-(phosphonomethoxy)ethyl]adenine (PMEA) as potential PMEA prodrugs with improved oral bioavailability. J. Med. Chem. 39:4958–4965 (1996).CrossRefPubMedGoogle Scholar
  8. 8.
    8. T. Hirohashi, H. Suzuki, X. Y. Chu, I. Tamai, A. Tsuji, and Y. Sugiyama. Function and expression of multidrug resistance-associated protein family in human colon adenocarcinoma cells (Caco-2). J. Pharmacol. Exp. Ther. 292:265–270 (2000).PubMedGoogle Scholar
  9. 9.
    9. L. M. S. Chan, S. Lowes, and B. H. Hirst. The ABCs of drug transport in intestine and liver: efflux proteins limiting drug absorption and bioavailability. Eur. J. Pharm. Sci. 21:25–51 (2004).CrossRefPubMedGoogle Scholar
  10. 10.
    10. N. Petri, C. Tannergren, D. Rungstad, and H. Lennernas. Transport characteristics of fexofenadine in the Caco-2 cell model. Pharm. Res. 21:1398–1404 (2004).CrossRefPubMedGoogle Scholar
  11. 11.
    11. H. M. Prime-Chapman, R. A. Fearn, A. E. Cooper, V. Moore, and B. H. Hirst. Differential multidrug resistance-associated protein 1 through 6 isoform expression and function in human intestinal epithelial Caco-2 cells. J. Pharmacol. Exp. Ther. 311:476–484 (2004).CrossRefPubMedGoogle Scholar
  12. 12.
    12. L. Homolya, Z. Hollo, U. A. Germann, I. Pastan, M. M. Gottesman, and B. Sarkadi. Fluorescent cellular indicators are extruded by the multidrug resistance protein. J. Biol. Chem. 268:21493–21496 (1993).PubMedGoogle Scholar
  13. 13.
    13. O. Legrand, G. Simonin, J. Y. Perrot, R. Zittoun, and J. P. Marie. Pgp and MRP activities using calcein-AM are prognostic factors in adult acute myeloid leukemia patients. Blood 91:4480–4488 (1998).PubMedGoogle Scholar
  14. 14.
    14. C. H. M. Versantvoort, T. Bagrij, K. A. Wright, and P. R. Twentyman. On the relationship between the probenecid-sensitive transport of daunorubicin or calcein and the glutathione status of cells overexpressing the multidrug resistance-associated protein (MRP). Int. J. Cancer 63:855–862 (1995).PubMedGoogle Scholar
  15. 15.
    15. M. Essodaigui, H. J. Broxterman, and A. Garnier-Suillerot. Kinetic analysis of calcein and calcein-acetoxymethylester efflux mediated by the multidrug resistance protein and P-glycoprotein. Biochemistry 37:2243–2250 (1998).CrossRefPubMedGoogle Scholar
  16. 16.
    16. N. Feller, H. J. Broxterman, D. C. R. Wahrer, and H. M. Pinedo. ATP-dependent efflux of calcein by the multidrug resistance protein (MRP): no inhibition by intracellular glutathione depletion. FEBS Lett. 368:385–388 (1995).PubMedGoogle Scholar
  17. 17.
    17. S. Kajiji, J. A. Dreslin, K. Grizzuti, and P. Gros. Structurally distinct MDR modulators show specific patterns of reversal against P-glycoproteins bearing unique mutations at serine939/941. Biochemistry 33:5041–5048 (1994).CrossRefPubMedGoogle Scholar
  18. 18.
    18. I. Tamai and A. R. Safa. Azidopine noncompetitively interacts with vinblastine and cyclosporin A binding to P-glycoprotein in multidrug resistant cells. J. Biol. Chem. 266:16796–16800 (1991).PubMedGoogle Scholar
  19. 19.
    19. I. Tamai and A. R. Safa. Competitive interaction of cyclosporins with the vinca alkaloid-binding site of P-glycoprotein in multidrug resistant cells. J. Biol. Chem. 265:16509–16513 (1990).PubMedGoogle Scholar
  20. 20.
    20. T. Saeki, K. Ueda, Y. Tanigawara, R. Hori, and T. Komano. P-glycoprotein mediated transcellular transport of MDR-reversing agents. FEBS Lett. 324:99–102 (1993).CrossRefPubMedGoogle Scholar
  21. 21.
    21. S. P. Cole, K. E. Sparks, K. Fraser, D. W. Loe, C. E. Grant, G. M. Wilson, and R. G. Deeley. Pharmacological characterization of multidrug resistant MRP-transfected human tumor cells. Cancer Res. 54:5902–5910 (1994).PubMedGoogle Scholar
  22. 22.
    22. T. R. Stouch and O. Gudmundsson. Progress in understanding the structure-activity relationships of P-glycoprotein. Adv. Drug Deliv. Rev. 54:315–328 (2002).PubMedGoogle Scholar
  23. 23.
    23. V. Tantishaiyakul, K. Wiwattanawongsa, S. Pinsuwan, S. Kasiwong, N. Phadoongsombut, S. Kaewnopparat, N. Kaewnopparat, and Y. Rojanasakul. Characterization of mefenamic acid-guaiacol ester: stability and transport across Caco-2 cell monolayers. Pharm. Res. 19:1013–1018 (2002).PubMedGoogle Scholar

Copyright information

© Springer Science+Business Media, Inc. 2005

Authors and Affiliations

  • Kamonthip Wiwattanawongsa
    • 1
  • Vimon Tantishaiyakul
    • 1
    Email author
  • Luelak Lomlim
    • 1
  • Yon Rojanasakul
    • 2
  • Sirirat Pinsuwan
    • 3
  • Sanae Keawnopparat
    • 3
  1. 1.Department of Pharmaceutical Chemistry, Faculty of Pharmaceutical SciencesPrince of Songkla UniversityHat-Yai, SongkhlaThailand
  2. 2.Department of Basic Pharmaceutical Sciences, School of PharmacyWest Virginia UniversityMorgantownUSA
  3. 3.Department of Pharmaceutical Technology, Faculty of Pharmaceutical SciencesPrince of Songkla UniversityHat-Yai, SongkhlaThailand

Personalised recommendations