In vitro permeability of peptidomimetic drugs: The role of polarized efflux pathways as additional barriers to absorption

doi:10.1016/S0169-409X(96)00432-2

Advanced Drug Delivery Reviews

Volume 23, Issues 1–3, 15 January 1997, Pages 143-156

https://doi.org/10.1016/S0169-409X(96)00432-2 Get rights and content

Abstract

Cellular efflux pathways function to remove both endogenous and exogenous substances from the cell. In the case of a polarized cellular barrier, such as the epithelium, these pathways serve an excretory or secretory role in transporting solutes out of tissue. Although well recognized in organs typically associated with drug excretion such as liver and kidney, similar transport pathways have been found in other tissues including the intestinal mucosa and the endothelial cells comprising the blood-brain barrier. Current evidence suggests that these systems may act as barriers to drug absorption into the tissues in which they are found. More recent studies have shown that hydrophobic peptides such as cyclosporin A are substrates for polarized efflux. In this review we examine the evidence for these mechanisms as absorption barriers and the use of in vitro transport models for characterizing this phenomenon. The presence of such pathways may help explain the poor membrane permeability of peptides which, along with metabolism, contributes to their poor in vivo performance.

References (114)

S.G. Schultz
Transport across epithelia: some basic principles
Kidney Int.
(1976)
H.N. Nellans
Paracellular intestinal transport: modulation of absorption
Adv. Drug Deliv. Rev.
(1991)
R.P.J. Oude Elferink et al.
Hepatobiliary secretion of organic compounds; molecular mechanisms of membrane transport
Biochim. Biophys. Acta
(1995)
D.R. Holland et al.
Intestinal secretion of erythromycin base
J. Pharm. Sci.
(1976)
W.T. Beck
The cell biology of multiple drug resistance
Biochem. Pharmacol.
(1987)
U.A. Germann et al.
P-glycoproteins: mediators of multidrug resistance
P.W. Wigler et al.
Inhibition of the multidrug resistance efflux pump
Biochim. Biophys. Acta
(1993)
W.H.M. Peters et al.
Expression of drug-metabolizing enzymes and P-170 glycoprotein in colorectal carcinoma and normal mucosa
Gastroenterology
(1992)
I.A.M. de Lannoy et al.
The mdrl gene product, P-glycoprotein, mediates the transport of the cardiac glycoside, digoxin
Biochem. Biophys. Res. Commun.
(1992)
M. Muller et al.
ATP-dependent transport of amphiphilic cations across the hepatocyte canalicular membrane mediated by mdrl P-glycoprotein
Fed. Eur. Biochem. Soc. Lett.
(1994)

L. Jetté et al.

High levels of P-glycoprotein detected in isolated brain capillaries

Biochim. Biophys. Acta

(1993)

A.H. Schinkel et al.

Disruption of the mouse mdrla P-glycoprotein gene leads to a deficiency in the blood-brain barrier and to increased sensitivity to drugs

Cell

(1994)

N.F.H. Ho et al.

A biophysical model of passive and polarized active transport processes in Caco-2 cells: approaches to uncoupling apical and basolateral membrane events in the intact cell

J. Pharm. Sci.

(1995)

I.J. Hidalgo et al.

Characterization of the human colon carcinoma cell line (Caco-2) as a model system for intestinal epithelial permeability

Gastroenterology

(1989)

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.

(1990)

G. Wilson et al.

Transport and permeability properties of human Caco-2 cells: an in vitro model of the intestinal epithelial cell barrier

J. Control. Rel.

(1990)

P. Wils et al.

Polarized transport of docetaxel and vinblastine mediated by P-glycoprotein in human intestinal epithelial cell monolayers

Biochem. Pharmacol.

(1994)

Y. Takakura et al.

Blood-brain barrier: transport studies in isolated brain capillaries and in cultured brain endothelial cells

Adv. Pharmacol.

(1991)

A. Tsuji et al.

P-glycoprotein as the drug efflux pump in primary cultured bovine brain capillary endothelial cells

Life Sci.

(1992)

T. Ohnishi et al.

In vivo and in vitro evidence for ATP-dependency of P-glycoprotein-mediated efflux of doxorubicin at the blood-brain barrier

Biochem. Pharmacol.

(1995)

R.C. Sharma et al.

Peptide transport by the multidrug resistance pump

J. Biol. Chem.

(1992)

D.L. Toppmeyer et al.

Role of P-glycoprotein in dolastatin 10 resistance

Biochem. Pharmacol.

(1994)

P.S. Burton et al.

Evidence for a polarized efflux system for peptides in the apical membrane of Caco-2 cells

Biochem. Biophys. Res. Commun.

(1993)

P.F. Augustijns et al.

Evidence for a polarized efflux system in Caco-2 cells capable of modulating cyclosporin A transport

Biochem. Biophys. Res. Commun.

(1993)

V Phung-Ba et al.

Interaction of pristinamycin I_A with P-glycoprotein in human intestinal epithelial cells

Eur. J. Pharmacol.

(1995)

T. Saeki et al.

Human P-glycoprotein transports cyclosporin A and FK506

J. Biol. Chem.

(1993)

A. Tsuji et al.

Restricted transport of cyclosporin A across the blood-brain barrier by a multidrug transporter, P-glycoprotein

Biochem. Pharmacol.

(1993)

A. Shirai et al.

Transport of cyclosporin A across the brain capillary endothelial cell monolayer by P-glycoprotein

Biochim. Biophys. Acta

(1994)

A. Sakata et al.

In vivo evidence for ATP-dependent and P-glycoprotein-mediated transport of cyclosporin A at the blood-brain barrier

Biochem. Pharmacol.

(1994)

T.W. Loo et al.

Functional consequences of proline mutations in the predicted transmembrane domain of P-glycoprotein

J. Biol. Chem.

(1993)

S.E. Devine et al.

Diversity of multidrug resistance in mammalian cells

J. Biol. Chem.

(1994)

C.E. Herzog et al.

Increased mdr-1/P-glycoprotein expression after treatment of human colon carcinoma cells with P-glycoprotein antagonists

J. Biol. Chem.

(1993)

K.-V Chin et al.

Heat shock and arsenite increase expression of the multidrug resistance (MDR1) gene in human renal carcinoma cells

J. Biol. Chem.

(1990)

G.A. Thomas et al.

Expression of the multidrug resistance-associated protein (MRP) gene in human lung tumours and normal tissue as determined by in situ hybridisation

Eur. J. Cancer

(1994)

C.H.M. Versantvoort et al.

Competitive inhibition by genistein and ATP dependence of daunorubicin transport in intact MRP overexpressing human small cell lung cancer cells

Biochem. Pharmacol.

(1994)

W.D. Stein

Channels, Carriers, and Pumps: An Introduction to Membrane Transport

(1990)

M.J. Jackson

Drug transport across gastrointestinal epithelia

P.S. Burton et al.

Trans-cellular mechanism of peptide and protein absorption: passive aspects

Adv. Drug. Deliv. Rev.

(1991)

W.A. Banks et al.

Carrier-mediated transport of enkaphalins and N-Tyr-MIF-1 across the blood-brain barrier

Am. J. Physiol.

(1986)

Z.H. Israili et al.

Enhancement of xenobiotic elimination: role of intestinal excretion

Drug Metab. Rev.

(1984)

R.K.H. Kinne

Selectivity and direction: plasma membranes in renal transport

Am. J. Physiol.

(1991)

P.G. Dayton et al.

Elimination of drugs by passive diffusion from blood to intestinal lumen: factors influencing nonbiliary excretion by the intestinal tract

Drug. Metab. Rev.

(1983)

J.Y. Mu et al.

Studies of the disposition and metabolism of mefloquine HCl (WR 142490), a quinolinemethanol antimalarial, in the rat

Drug Metab. Dispos.

(1975)

C.C. Lee et al.

Distribution and excretion of radioactivity in rats receiving N-methyl-¹⁴C-erythromycin

J. Pharmacol. Exp. Ther.

(1956)

J.H. Caldwell et al.

Intestinal secretion of digoxin in the rat

Naunyn-Schmiedeberg's Arch. Pharmacol.

(1980)

C.F. George et al.

Elimination of drugs by active intestinal transport

J. Pharm. Pharmacol.

(1979)

F. Lauterbach

Absorption of cardiac glycosides

Acta. Pharmacol. Toxicol.

(1971)

F. Lauterbach

Resorption und Sekretion von arzneistoffen durch die mukosaepithelien des gastrointestinaltraktes

Arzneim. Forsch.

(1975)

F. Lauterbach

Intestinal secretion on ions and drugs

S.M. Simon et al.

Cell biological mechanisms of multidrug resistance in tumors

Cited by (27)

Oral colon delivery of insulin with the aid of functional adjuvants
2012, Advanced Drug Delivery Reviews
Citation Excerpt :
However, this is mostly evoked by specific receptor/ligand interactions. In addition to the difficulties encountered in crossing the mucosal barrier, polarized efflux systems located in the apical membrane could participate in limiting the absorption of these drugs [97,98]. Mainly hydrophobic peptides, e.g. the immunosuppressant cyclosporin A and cyclopeptidic macrolactone antibiotic pristinamycin IA, might indeed be removed from the cell by P-glycoprotein and multi-drug resistance-associated protein, which would be responsible for non-biliary intestinal excretion of a variety of structurally unrelated compounds.
Oral colon delivery is currently considered of importance not only for the treatment of local pathologies, such as primarily inflammatory bowel disease (IBD), but also as a means of accomplishing systemic therapeutic goals. Although the large bowel fails to be ideally suited for absorption processes, it may indeed offer a number of advantages over the small intestine, including a long transit time, lower levels of peptidases and higher responsiveness to permeation enhancers. Accordingly, it has been under extensive investigation as a possible strategy to improve the oral bioavailability of peptide and protein drugs. Because of a strong underlying rationale, most of these studies have focused on insulin. In the present review, the impact of key anatomical and physiological characteristics of the colon on its viability as a protein release site is discussed. Moreover, the main formulation approaches to oral colon targeting are outlined along with the design features and performance of insulin-based devices.
Synthesis of new class dipeptide analogues with improved permeability and antithrombotic activity
2006, Bioorganic and Medicinal Chemistry
3-(S)-1,2,3,4-Tetrahydro-β-carboline-3-carboxylic acid isolated from A. Chinese G. Don was found to possess moderate anti-aggregation activity, but with poor bioavailability. To improve its pharmacological property, we designed and synthesized a series of novel dipeptide analogues by incorporating tetrahydro-β-carboline-3-carboxylic acid skeleton as an amino acid surrogate (∗Trp). It turned out these dipeptide analogues exhibited good membrane permeability based on in vitro Caco-2 cell monolayers permeability assay. As a result, the overall biological properties of these molecules were significantly improved depending on the nature of the amino acid residues introduced onto the 3-position of the tetrahydro-β-carboline moiety. It was very interesting to notice that these dipeptide analogues (5b,c,h,i,n,o,p,q) displayed a remarkable dual antiaggregatory activity in both of ADP- and PAF-induced platelet aggregation assay, and their aggregation response was significantly higher than that of aspirin (p < 0.01). In addition, these dipeptide analogues were observed for the dose-dependent antithrombotic effect using in vivo rat arterial thrombosis model. The potency of antithrombotic activity of 5h,i,n,p was significantly higher than that of aspirin (n = 12, p < 0.01) at equal dose (5 μmol/kg).
Influence of efflux transporters on the intestinal passage of drugs
2002, Revue Francaise des Laboratoires
La P-glycoprotéine (P-gp) est une glycoprotéine membranaire impliquée dans le phénotype de multidrug resistance et entraîne une diminution des concentrations intracellulaires des médicaments cytotoxiques. Après un récapitulatif de ses caractéristiques, cette revue présente les effets de la P-gp au niveau des tissus sains. Son implication dans la biodisponibilité de médicaments substrats est évoquée à travers divers exemples d'études menées in vitro et in vivo chez l'homme. La synergie entre la P-gp et le métabolisme intestinal pré-systémique assuré par le cytochrome P450 3A4 renforce le rôle de la P-gp dans de nombreuses interactions pharmacocinétiques entre médicaments.
P-glycoprotein (P-gp) is a membrane glycoprotein which causes multidrug resistance phenotypes and leads to a decrease of the intracellular concentration of cytotoxic drugs. After a summary of its main properties, this review will focus on the effects of P-gp in normal tissues. Several examples of in vitro and in vivo studies will show the involvement of P-gp in the oral bioavailability of drugs. The cooperation existing between P-gp and the pre-systemic intestinal metabolism mediated by cytochrome P4503A4 shows that P-gp has a role in numerous drug-drug pharmacokinetic interactions.
Permeability of the peptidic GH secretagogues hexarelin and EP 51389, across rat jejunum
2001, Peptides
The intestinal permeability of hexarelin and EP 51389, two growth hormone releasing hexa- and tri- peptide analogues, was assessed in vitro with side-by-side diffusion chambers in the apical-to-basolateral (AP-to-BL) and in the basolateral-to-apical (BL-to-AP) direction using excised rat jejunal segments. The effect of EP 51389 on P-glycoprotein (P-gp) was evaluated by rhodamine 123 accumulation on monolayers of CH^RC5 cells with increasing concentrations of EP 51389. Hexarelin and EP 51389 permeability were found to be < 1%. Permeability coefficients (P_app) were 18.87 ± 2.86 (×10⁻⁷ cm/s) and 5.87 ± 0.45 (×10⁻⁷ cm/s) for hexarelin and EP 51389, respectively. Bidirectional studies revealed that hexarelin transport was similar in both directions. EDTA did not influence hexarelin permeability. Permeability was predominantly secretory for EP 51389 as P_app in the BL-to-AP direction [32.56 ± 6.11 (×10⁻⁷ cm/s)] was greater than AP-to-BL. Confirming involvement of a secretory transport system, chlorpromazine inhibited EP 51389 transport across the jejunum. EP 51389 inhibited P-gp in a dose dependent manner resulting in the intracellular accumulation of rhodamine in CH^RC5 cells. These results suggest that: 1) the intestinal permeability of hexarelin and EP 51389 is poor; 2) the passage of hexarelin is mainly via a transcellular passive pathway since the contribution of paracellular permeability to the overall permeability is rather low; 3) P-gp may act as a potential barrier for the intestinal absorption of EP 51389.
Improvement of oral peptide bioavailability: Peptidomimetics and prodrug strategies
1997, Advanced Drug Delivery Reviews
Clinical development of orally active peptide drugs has been restricted by their unfavorable physicochemical properties, which limit their intestinal mucosal permeation and their lack of stability against enzymatic degradation. Successful oral delivery of peptides will depend, therefore, on strategies designed to alter the physicochemical characteristics of these potential drugs, without changing their biological activity, in order to overcome the physical and biochemical barrier properties of the intestinal cells. This manuscript will focus on the physiological limitations for oral peptide delivery and on various strategies using chemical modifications to improve oral bioavailability of peptide-based drugs.
Mechanistic roles of neutral surfactants on concurrent polarized and passive membrane transport of a model peptide in Caco-2 cells
1997, Journal of Pharmaceutical Sciences
The transport of the model peptide Acf(NMef)₂NH₂ across Caco-2 cell monolayers was studied in the apical (AP) to basolateral (BL) and the BL to AP direction in the presence of Polysorbate 80 or Cremophore EL in the AP compartment. Increasing surfactant concentrations resulted in increasing AP → BL peptide permeability and decreasing BL → AP permeability. In either direction, limiting permeabilities were achieved at concentrations less than the critical micellar concentrations (cmc’s) of the surfactants, and remained constant at much higher concentrations. These plateau permeabilities were not equivalent in the two directions. This residual assymetry was abolished by increasing the peptide concentration. Altogether, the observations support the presence of at least two pumps in Caco-2 cells for this peptide, polarized in the BL → AP direction. These experimental results were analyzed within the context of a quantitative biophysical model incorporating concurrent passive diffusion across the Ap and BL membranes accompanied by surfactant-inhibitable active polarized efflux across the AP membrane. The model was also used to locate the additional transport activity at the BL membrane as an uptake pump. Under conditions of complete inhibition, the intrinsic passive diffusional permeability of Acf(NMef)₂NH₂ was found to be 13 × 10^- 6 cm/s, essentially identical with results reported earlier with this peptide utilizing verapamil as an inhibitor. With respect to the mechanism of surfactant inhibition of the apical efflux tranport, the monomeric species was found to be responsible with no contribution from micelles. Modeling the mode of inhibition as a noncompetitive MichaelisMenten process gave identical K_i’s of 0.5 μΜ for the two surfactants. Finally, increase of either surfactant beyond 750 μΜ resulted in a decrease of peptide permeability in the AP → BL direction. This was attributed to weak association of the peptide with micelles in the AP compartment, which effectively decreased the thermodynamic activity of the peptide at surfactant concentrations greater than 20 times their cmc. Both the experimental approach and accompanying theoretical model demonstrated in this work will allow for further characterization of the inhibitory potencies of surfactants for the nonpassive efflux pathway in vitro and in vivo.

View all citing articles on Scopus

View full text

In vitro permeability of peptidomimetic drugs: The role of polarized efflux pathways as additional barriers to absorption

Abstract

Kidney Int.

Adv. Drug Deliv. Rev.

Biochim. Biophys. Acta

J. Pharm. Sci.

Biochem. Pharmacol.

Biochim. Biophys. Acta

Gastroenterology

Biochem. Biophys. Res. Commun.

Fed. Eur. Biochem. Soc. Lett.

Biochim. Biophys. Acta

Cell

J. Pharm. Sci.

Gastroenterology

J. Pharm. Sci.

J. Control. Rel.

Biochem. Pharmacol.

Adv. Pharmacol.

Life Sci.

Biochem. Pharmacol.

J. Biol. Chem.

Biochem. Pharmacol.

Biochem. Biophys. Res. Commun.

Biochem. Biophys. Res. Commun.

Eur. J. Pharmacol.

J. Biol. Chem.

Biochem. Pharmacol.

Biochim. Biophys. Acta

Biochem. Pharmacol.

J. Biol. Chem.

J. Biol. Chem.

J. Biol. Chem.

J. Biol. Chem.

Eur. J. Cancer

Biochem. Pharmacol.

Channels, Carriers, and Pumps: An Introduction to Membrane Transport

Drug transport across gastrointestinal epithelia

Trans-cellular mechanism of peptide and protein absorption: passive aspects

Adv. Drug. Deliv. Rev.

Carrier-mediated transport of enkaphalins and N-Tyr-MIF-1 across the blood-brain barrier

Am. J. Physiol.

Enhancement of xenobiotic elimination: role of intestinal excretion

Drug Metab. Rev.

Selectivity and direction: plasma membranes in renal transport

Am. J. Physiol.

Elimination of drugs by passive diffusion from blood to intestinal lumen: factors influencing nonbiliary excretion by the intestinal tract

Drug. Metab. Rev.

Studies of the disposition and metabolism of mefloquine HCl (WR 142490), a quinolinemethanol antimalarial, in the rat

Drug Metab. Dispos.

Distribution and excretion of radioactivity in rats receiving N-methyl-14C-erythromycin

J. Pharmacol. Exp. Ther.

Intestinal secretion of digoxin in the rat

Naunyn-Schmiedeberg's Arch. Pharmacol.

Elimination of drugs by active intestinal transport

J. Pharm. Pharmacol.

Absorption of cardiac glycosides

Acta. Pharmacol. Toxicol.

Resorption und Sekretion von arzneistoffen durch die mukosaepithelien des gastrointestinaltraktes

Arzneim. Forsch.

Intestinal secretion on ions and drugs

Cell biological mechanisms of multidrug resistance in tumors

Distribution and excretion of radioactivity in rats receiving N-methyl-¹⁴C-erythromycin