Transport of Peptidomimetic Thrombin Inhibitors with a 3-amidino-phenylalanine Structure: Permeability and Efflux Mechanism in Monolayers of a Human Intestinal Cell Line (Caco-2)

Overview

Journal Pharm Res

Specialties Pharmacology
Pharmacy

Date 2001 Oct 6

PMID 11587481

Citations 3

Authors

W KAMM

J Hauptmann

I Behrens

J Sturzebecher

F Dullweber

H Gohlke

M Stubbs

G Klebe

T Kissel

Affiliations

Soon will be listed here.

Abstract

Purpose: Peptidomimetic thrombin inhibitors derived from Nalpha-(2-naphthylsulfonyl)-3-amidino-phenylalanine with different basic and acidic substituents were investigated with respect to their intestinal transport behavior.

Methods: Intestinal permeability coefficients were studied using Caco-2 monolayers and a reversed-phase HPLC method for quantitation.

Results: Apparent permeability coefficients Papp of compounds with a free amidino group were in general low (<10 x 10(-8) cm/s) and independent of the structure of the amide part (C-terminus). Polarized efflux, however, was strongly affected by substituents in the amide moiety yielding the following efflux ratios (ER): methylpiperidide (1) (ER 45) > piperidine carboxylic acid methylester (ER 6-11) > piperidine carboxylic acids (ER 1.9-2.9) > piperazide (ER -0.17). Efflux of (1) was temperature-dependent, but independent of the enantiomeric configuration, accompanied by an increase in transepithelial electrical resistance (TEER), and could be reduced by P-gp inhibitors (PSC 833, Cremophor EL) but not by indomethacin. Replacement of the amidine group of (1) by aminomethyl-, amino-, and oxamidine- moieties drastically increased absorptive permeability (46-68 fold) with ER < 3.4. In contrast, the oxamidine with a C-terminal nipecotic acid residue (8) displayed also a temperature dependent efflux- without altering TEER (ER 22). This efflux was sensitive to PSC833/Cremophor EL and indomethacin.

Conclusions: Basic and acidic residues of amidino-phenylalanine-derived thrombin inhibitors mediate affinity to intestinal efflux pumps. presumably P-gp and MRP. P-gp mediated efflux was related to a net positive charge and accompanied by an increased TEER. Among the methylpiperide (1) promoieties studied the oxamidino group seems to be very promising in overcoming both transport and efflux problems frequently encountered with peptidomimetics containing amidines.

Citing Articles

Biocompatibility Profile and In Vitro Cellular Uptake of Self-assembled Alginate Nanoparticles.

Zhang P, Zhao S, Yu Y, Wang H, Yang Y, Liu C Molecules. 2019; 24(3).

PMID: 30717442 PMC: 6384778. DOI: 10.3390/molecules24030555.

Modeling kinetics of subcellular disposition of chemicals.

Balaz S Chem Rev. 2009; 109(5):1793-899.

PMID: 19265398 PMC: 2682929. DOI: 10.1021/cr030440j.

pH-dependent bidirectional transport of weakly basic drugs across Caco-2 monolayers: implications for drug-drug interactions.

Neuhoff S, Ungell A, Zamora I, Artursson P Pharm Res. 2003; 20(8):1141-8.

PMID: 12948010 DOI: 10.1023/a:1025032511040.

References

Lee K, Thakker D . Saturable transport of H2-antagonists ranitidine and famotidine across Caco-2 cell monolayers. J Pharm Sci. 1999; 88(7):680-7. DOI: 10.1021/js980474k. View

Doppenschmitt S, Spahn-Langguth H, Regardh C, Langguth P . Radioligand-binding assay employing P-glycoprotein-overexpressing cells: testing drug affinities to the secretory intestinal multidrug transporter. Pharm Res. 1998; 15(7):1001-6. DOI: 10.1023/a:1011965707998. View

Prueksaritanont T, Deluna P, Gorham L, Ma B, Cohn D, Pang J . In vitro and in vivo evaluations of intestinal barriers for the zwitterion L-767,679 and its carboxyl ester prodrug L-775,318. Roles of efflux and metabolism. Drug Metab Dispos. 1998; 26(6):520-7. View

Eckhardt U, Stuber W, Dickneite G, Reers M, Petzinger E . First-pass elimination of a peptidomimetic thrombin inhibitor is due to carrier-mediated uptake by the liver. Interaction with bile acid transport systems. Biochem Pharmacol. 1996; 52(1):85-96. DOI: 10.1016/0006-2952(96)00141-4. View

Walter E, Kissel T, Reers M, Dickneite G, Hoffmann D, Stuber W . Transepithelial transport properties of peptidomimetic thrombin inhibitors in monolayers of a human intestinal cell line (Caco-2) and their correlation to in vivo data. Pharm Res. 1995; 12(3):360-5. DOI: 10.1023/a:1016244316584. View

Arimori K, Nakano M . Drug exsorption from blood into the gastrointestinal tract. Pharm Res. 1998; 15(3):371-6. DOI: 10.1023/a:1011959828103. View

Draper M, Martell R, Levy S . Indomethacin-mediated reversal of multidrug resistance and drug efflux in human and murine cell lines overexpressing MRP, but not P-glycoprotein. Br J Cancer. 1997; 75(6):810-5. PMC: 2063393. DOI: 10.1038/bjc.1997.145. View

Saitoh H, Gerard C, Aungst B . The secretory intestinal transport of some beta-lactam antibiotics and anionic compounds: a mechanism contributing to poor oral absorption. J Pharmacol Exp Ther. 1996; 278(1):205-11. View

Lang V, Langguth P, Ottiger C, Wunderli-Allenspach H, Rognan D, Rothen-Rutishauser B . Structure-permeation relations of met-enkephalin peptide analogues on absorption and secretion mechanisms in Caco-2 monolayers. J Pharm Sci. 1997; 86(7):846-53. DOI: 10.1021/js960387x. View

10.

Hauptmann J, Sturzebecher J . Influence of indocyanine green on plasma disappearance and biliary excretion of a synthetic thrombin inhibitor of the 3-amidinophenyl-alanine piperazide-type in rats. Pharm Res. 1998; 15(5):751-4. DOI: 10.1023/a:1011927204847. View

11.

Weller T, Alig L, Beresini M, Blackburn B, Bunting S, Hadvary P . Orally active fibrinogen receptor antagonists. 2. Amidoximes as prodrugs of amidines. J Med Chem. 1996; 39(16):3139-47. DOI: 10.1021/jm9509298. View

12.

Sturzebecher J, Prasa D, Hauptmann J, Vieweg H, Wikstrom P . Synthesis and structure-activity relationships of potent thrombin inhibitors: piperazides of 3-amidinophenylalanine. J Med Chem. 1997; 40(19):3091-9. DOI: 10.1021/jm960668h. View

13.

Hunter J, Jepson M, Tsuruo T, Simmons N, Hirst B . Functional expression of P-glycoprotein in apical membranes of human intestinal Caco-2 cells. Kinetics of vinblastine secretion and interaction with modulators. J Biol Chem. 1993; 268(20):14991-7. View

14.

Aszalos A, Thompson K, Yin J, Ross D . Combinations of P-glycoprotein blockers, verapamil, PSC833, and cremophor act differently on the multidrug resistance associated protein (MRP) and on P-glycoprotein (Pgp). Anticancer Res. 1999; 19(2A):1053-64. View

15.

Hauptmann J, Paintz M, Kaiser B, Richter M . Reduction of a benzamidoxime derivative to the corresponding benzamidine in vivo and in vitro. Pharmazie. 1988; 43(8):559-60. View

16.

Rewinkel J, Lucas H, Smit M, Noach A, Van Dinther T, Rood A . Design, synthesis and testing of amino-bicycloaryl based orally bioavailable thrombin inhibitors. Bioorg Med Chem Lett. 1999; 9(19):2837-42. DOI: 10.1016/s0960-894x(99)00483-7. View

17.

Gan L, Yanni S, Thakker D . Modulation of the tight junctions of the Caco-2 cell monolayers by H2-antagonists. Pharm Res. 1998; 15(1):53-7. DOI: 10.1023/a:1011944602662. View

18.

Annaert P, van Gelder J, Naesens L, De Clercq E, van den Mooter G, Kinget R . Carrier mechanisms involved in the transepithelial transport of bis(POM)-PMEA and its metabolites across Caco-2 monolayers. Pharm Res. 1998; 15(8):1168-73. DOI: 10.1023/a:1011923420719. View

19.

Bear C . Drugs transported by P-glycoprotein inhibit a 40 pS outwardly rectifying chloride channel. Biochem Biophys Res Commun. 1994; 200(1):513-21. DOI: 10.1006/bbrc.1994.1478. View

20.

Aungst B, Saitoh H . Intestinal absorption barriers and transport mechanisms, including secretory transport, for a cyclic peptide, fibrinogen antagonist. Pharm Res. 1996; 13(1):114-9. DOI: 10.1023/a:1016093704095. View