Differential Involvement of P-Glycoprotein (ABCB1) in Permeability, Tissue Distribution, and Antinociceptive Activity of Methadone, Buprenorphine, and Diprenorphine: In Vitro and In Vivo Evaluation

doi:10.1002/jps.21770

Journal of Pharmaceutical Sciences

Volume 98, Issue 12, December 2009, Pages 4928-4940

https://doi.org/10.1002/jps.21770 Get rights and content

Abstract

Conclusions based on either in vitro or in vivo approach to evaluate the P-gp affinity status of opioids may be misleading. For example, in vitro studies indicated that fentanyl is a P-gp inhibitor while in vivo studies indicated that it is a P-gp substrate. Quite the opposite was evident for meperidine. The objective of this study was to evaluate the P-gp affinity status of methadone, buprenorphine and diprenorphine to predict P-gp-mediated drug-drug interactions and to determine a better candidate for management of opioid dependence. Two in vitro (P-gp ATPase and monolayer efflux) assays and two in vivo (tissue distribution and antinociceptive evaluation in mdr1a/b (−/−) mice) assays were used. Methadone stimulated the P-gp ATPase activity only at higher concentrations, while verapamil and GF120918 inhibited its efflux (p < 0.05). The brain distribution and antinociceptive activity of methadone were enhanced (p < 0.05) in P-gp knockout mice. Conversely, buprenorphine and diprenorphine were negative in all assays. P-gp can affect the PK/PD of methadone, but not buprenorphine or diprenorphine. Our report is in favor of buprenorphine over methadone for management of opioid dependence. Buprenorphine most likely is not a P-gp substrate and concerns regarding P-gp-mediated drug-drug interaction are not expected. © 2009 Wiley-Liss, Inc. and the American Pharmacists Association J Pharm Sci 98:4928–4940, 2009

Section snippets

INTRODUCTION

The blood–brain barrier (BBB) acts as a regulator of the flux of xenobiotics between the systemic circulation and the central nervous system (CNS). The permeability of drugs across the BBB has been traditionally considered as a factor of molecular weight, protein binding, H-bonding, degree of ionization and lipid solubility.¹ However, a convincing body of evidence suggests that the efflux transporter, P-glycoprotein (P-gp, Abcb1), may also modulate drug disposition into the brain.2, 3, 4, 5, 6,

Drug-Stimulated P-gp ATPase Activity

Drug stimulated P-gp ATPase activity was estimated by Pgp-GIO assay system (Promega, Madison, WI). This method relies on the ATP dependence of the light-generating reaction of firefly luciferase. ATP consumption is detected as a decrease in luminescence. In a 96 well plate, recombinant human P-gp (25 µg) was incubated with P-gp-GIO assay buffer™ (20 µL) (control, n = 4), verapamil (200 µM) (n = 4), sodium orthovanadate (100 µM) (n = 4), methadone (5–100 µM) (n = 3/conc.), buprenorphine (5–100 µM) (n =

Effect of Modulators on P-gp ATPase Activity

Different concentrations of methadone, buprenorphine, and diprenorphine (Fig. 2) were examined to determine their effects on the P-gp ATPase activity. Each opioid together with a known excess of ATP was incubated with recombinant human P-gp. ATP consumption due to P-gp stimulation by each opioid was detected as a decrease in luminescence, that is, the higher the potency of a compound to stimulate the P-gp ATPase activity, the lower the luminescence signal. The rate of ATP consumption due to

DISCUSSION

The objective of this study was to evaluate the P-gp affinity status of methadone, buprenorphine and diprenorphine to predict P-gp-mediated drug–drug interactions and to determine a better candidate for management of opioid dependence. Classification of opioids or any other compound as a P-gp substrate, nonsubstrate, inducer or inhibitor based on only one assay may be misleading.¹^,²¹^,²³^,³² As a result, we used two in vitro (P-gp ATPase and monolayer efflux assays) and two in vivo (opioids'

Acknowledgements

This study was supported in part by University of Maryland Intramural Research grant (A.C.), a Predoctoral Fellowship from the Egyptian Ministry of Higher Education (H.E.H.) and Ralph Shangraw Predoctoral Fellowship from University of Maryland (H.E.H.).

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Research ArticlesDifferential Involvement of P-Glycoprotein (ABCB1) in Permeability, Tissue Distribution, and Antinociceptive Activity of Methadone, Buprenorphine, and Diprenorphine: In Vitro and In Vivo Evaluation

Abstract

Section snippets

INTRODUCTION

Drug-Stimulated P-gp ATPase Activity

Effect of Modulators on P-gp ATPase Activity

DISCUSSION

Acknowledgements

J Pharm Sci

J Pharm Sci

J Pharm Sci

J Pharm Sci

Biochem Pharmacol

J Pharm Sci

Adv Drug Deliv Rev

Biochem Pharmacol

Bioorg Med Chem Lett

J Pharm Sci

J Biol Chem

J Pharm Sci

J Chromatogr B Analyt Technol Biomed Life Sci

Pain

Biochim Biophys Acta

J Pharmacol Toxicol Methods

J Biol Chem

Adv Drug Deliv Rev

J Pharm Sci

Eur J Pharmacol

Passive permeability and P-glycoprotein-mediated efflux differentiate central nervous system (CNS) and non-CNS marketed drugs

J Pharmacol Exp Ther

Normal viability and altered pharmacokinetics in mice lacking mdr1-type (drug-transporting) P-glycoproteins

Proc Natl Acad Sci USA

Absence of the mdr1a P-Glycoprotein in mice affects tissue distribution and pharmacokinetics of dexamethasone, digoxin, and cyclosporin A

J Clin Invest

Role of P-glycoprotein in pharmacokinetics: Clinical implications

Clin Pharmacokinet

Dose-ranging study of oxycodone for chronic pain in advanced cancer

J Clin Oncol

Research Articles
Differential Involvement of P-Glycoprotein (ABCB1) in Permeability, Tissue Distribution, and Antinociceptive Activity of Methadone, Buprenorphine, and Diprenorphine: In Vitro and In Vivo Evaluation