Transepithelial Transport Properties of Peptidomimetic Thrombin Inhibitors in Monolayers of a Human Intestinal Cell Line (Caco-2) and Their Correlation to in Vivo Data
Tóm tắt
Peptidomimetic thrombin inhibitors (TI), derived from L-Asp-D-Phe were examined in confluent monolayers of a human colon carcinoma cell line (Caco-2) to elucidate their transepithelial transport properties. Effect availabilities, based on activated partial thromboplastin time (aPTT) measurements in rats, after peroral administration of five TI correlated reasonably well with permeability coefficients obtained from in vitro transport studies in Caco-2 monolayers, whereas physicochemical properties, such as molecular mass, solubilities, pKa and octanol-buffer partition coefficients failed to yield meaningful relationships. Substitution of the β-carboxylic group of L-Asp leads to analogues which are mainly transported by passive diffusion, while an unsubstituted carboxylic group favours carrier-mediated active transport. The effects of concentration, temperature, competitive inhibitors and direction dependence on in vitro transport were investigated. The results obtained are compatible with a saturable carrier-mediated transport, operating parallel to a passive paracellular route. The Michaelis-Menten parameters for the active transport component (Km = 1.67 mM, Vmax = 26.5 pmol min−1 mg protein−1) indicate an involvement of the intestinal di/-tripeptide transport system for one of the TI. The Caco-2 transport model may be helpful for the design of perorally active peptidomimetics.
Tài liệu tham khảo
P. L. Smith, D. A. Wall, C. H. Gochoco, and G. Wilson. Routes of delivery: Case studies. (5) Oral absorption of peptides and proteins. Adv. Drug Delivery Rev. 8:253–290 (1992).
J. G. Hardy, S. S. Davis, and C. G. Wilson. Drug delivery to the gastrointestinal tract, Ellis Horwood Limited, New York, 1989.
M. J. Humphrey and P. S. Ringrose. Peptides and related drugs: A review of their absorption, metabolism, and excretion. Drug Met. Rev. 17:283–310 (1986).
L. J. Berliner. Thrombin: structure and function, Plenum Press, New York, 1992.
W. Stueber, R. Koschinsky, D. Dickneite, and C. Kolar. Patent: EP 0513543, (1993).
J. B. F. Bai and G. L. Amidon. Structural specifity of mucosal-cell transport and metabolism of peptide drugs: Implication for oral peptide drug delivery. Pharm. Res. 8:969–978 (1992).
D. I. Friedman and G. L. Amidon. Characterization of the intestinal transport parameters for small peptide drugs. J. Contr. Rel. 13:141–146 (1990).
P. Artursson. Cell culture as models for drug absorption across the intestinal mucosa. Crit. Rev. Ther. Drug Carrier Syst. 8:305–330 (1991).
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).
G. Wilson, I. F. Hassan, C. J. Dix, I. Williamson, and M. Mackay. Transport and permeability properties of human Caco-2 cells. An in vitro model of the intestinal epithelial cell barrier. J. Control. Rel. 11:25–40 (1990).
C. J. Dix, H. Y. Orbay, I. F. Hassan, and G. Wilson. Vitamin B12 transport through polarized monolayers of a colon carcinoma cell line. Biochem. Soc. Trans. 15:443–440 (1987).
A. Blais, P. Bissonnette, and A. Berteloot. Common characteristics for Na-dependent sugar transport in Caco-2 cells and human fetal colon. J. Membr. Biol. 99:113–125 (1987).
A. H. Dantzig and L. Bergin. Uptake of the cephalosporin, cephalexin, by a dipeptide transport carrier in the human intestinal cell line, Caco-2. Biochim. Biophys. Acta 1027:211–217 (1990).
K. I. Inui, M. Yamamoto, and H. Saito. Transepithelial transport of oral cephalosporins by monolayers of intestinal epithelial cell line Caco-2: Specific transport systems in apical and basolateral membranes. J. Pharm. Exp. Ther. 261:195–201 (1992).
W. Stüber, G. Dickneite, R. Koschinsky, M. Reers, D. Hoffmann, and E. P. Paques. Development of highly potent thrombin inhibitors based on amidinophenylalanine derivates. Keystone Symp., Prospects and Progress in Drug Based on Peptides and Proteins. 1993.
W. Stueber, R. Koschinsky, C. Kolar, M. Reers, G. Dickneite, D. Hoffmann, J. Czech, K. H. Diehl, and E. P. Paques. Inhibition of thrombin by derivates of the dipeptide aspartic acid-amidinophenylalanine. Peptides: Chemistry and Biology, Proceed. 13th American Peptide Symp., in press.
A Avdeef. Fast simultaneous determination of log P and pKa by potentiometry: para-alkoxyphenol series (methoxy to pentoxy). In C. Silipo, V. Vittoria (eds.), QSAR: rational approaches to the design of bioactive compounds, Elsevier Science Publishers B.V., Amsterdam, 1991, p. 119–122.
E. Walter and T. Kissel. Transepithelial transport and metabolism of thyrotropin-releasing hormone (TRH) in monolayers of a human intestinal cell line (Caco-2): Evidence for an active transport component? Pharm. Res. 11:1576–1581 (1994).
A. B. J. Noach, Y. Kurosaki, M. C. M. Blom-Rosemalen, A. G. de Boer, and D. D. Breimer. Cell-polarity dependent effect of chelation on the paracellular permeability of confluent Caco-2 cell monolayers. Int. J. Pharm. 90:229–237 (1993).
O. H. Lowry, N. J. Rosenbrough, A. L. Farr, and R. J. Randall. Protein measurement with the folin phenol red. J. Biol. Chem. 193:265–275 (1951).
A. R. Hilgers, R. A. Conradi, and P. S. Burton. Caco-2 cell monolayer as a model for drug transport across the intestinal mucosa. Pharm. Res. 9:902–910 (1990).
P. Artursson and J. Karlsson. Correlation between oral drug absorption in humans and apparent drug permeability coefficients in human intestinal epithelial (Caco-2) cells. Biochem. Biophys. Res. Commun. 175:880–885 (1991).
S. Yokohama, K. Yamashita, H. Toguchi, J. Takeuchi, and N. Kitamori. Absorption of thyrotropin-releasing hormone after oral administration of TRH tartrate monohydrate in the rat, dog and human. J. Pharmacobio-Dyn. 7:101–111 (1984).
P. Artursson. Epithelial transport of drugs in cell culture. I: A model for studying the passive diffusion of drugs over intestinal absorptive (Caco-2) cells. J. Pharm. Sci. 79:476–482 (1990).
P. S. Burton, R. A. Conradi, and A. R. Hilgers. (B) Mechanisms of peptide and protein absorption. (2) Transcellular mechanism of peptide and protein absorption: Passive aspects. Advanced Drug Delivery Rev. 7:365–386 (1991).
A. Leo, C. Hansch, and D. Elkins. Partition coefficients and their uses. Chem. Rev. 71:525–616 (1977).
S. H. Yalkowsky, W. Morozowich, and W. I. Higuchi. A physical chemical basis of the design of orally active prodrugs. Drug Design 9:122–185 (1980).
N. F. H. Ho, J. Y. Park, W. Morozowich, and W. I. Higuchi. Physical model approach to the design of drugs with improved intestinal absorption. In E. B. Roche (ed.), Design of biopharmaceutical properties through prodrugs and analogs, American Pharmaceutical Association, Academy of Pharmaceutical Sciences, 1977, p. 136–227.
I. Osiescka, P. A. Porter, R. T. Borchardt, J. A. Fix, and R. C. Gardner. In vitro drug absorption models: brush border membrane vesicles, isolated mucosal cells and everted intestinal rings: characterization and salicylate accumulation. Pharm. Res. 2:284–293 (1983).
P. A. Porter, I. Osiecka, R. T. Borchardt, J. A. Fix, I. Frost, and C. Gardner. In vitro drug absorption models. II. Salicylate, cefoxitin, α-methyldopa and theophylline uptake in cells and rings: correlation with in vivo bioavailability. Pharm. Res. 2:293–297 (1985).
S. Yokohama, T. Yoshioka, K. Yamashita, and N. Kitamori. Intestinal absorption mechanism of thyrotropin-releasing hormone. J. Pharmacobio-Dyn. 7:445–451 (1984).
H. Yuasa, G. L. Amidon, and D. Fleisher. Peptide carrier-mediated transport in intestinal brush border membrane vesicles of rats and rabbits: Cephradine uptake and inhibition. Pharm. Res. 10:400–404 (1993).