Characterization of methadone as a β-arrestin-biased μ-opioid receptor agonist

Mol Pain. 2016 Jun 17;12:1744806916654146. doi: 10.1177/1744806916654146. Print 2016.


Background: Methadone is a unique µ-opioid receptor agonist. Although several researchers have insisted that the pharmacological effects of methadone are mediated through the blockade of NMDA receptor, the underlying mechanism by which methadone exerts its distinct pharmacological effects compared to those of other µ-opioid receptor agonists is still controversial. In the present study, we further investigated the pharmacological profile of methadone compared to those of fentanyl and morphine as measured mainly by the discriminative stimulus effect and in vitro assays for NMDA receptor binding, µ-opioid receptor-internalization, and µ-opioid receptor-mediated β-arrestin recruitment.

Results: We found that fentanyl substituted for the discriminative stimulus effects of methadone, whereas a relatively high dose of morphine was required to substitute for the discriminative stimulus effects of methadone in rats. Under these conditions, the non-competitive NMDA receptor antagonist MK-801 did not substitute for the discriminative stimulus effects of methadone. In association with its discriminative stimulus effect, methadone failed to displace the receptor binding of MK801 using mouse brain membrane. Methadone and fentanyl, but not morphine, induced potent µ-opioid receptor internalization accompanied by the strong recruitment of β-arrestin-2 in µ-opioid receptor-overexpressing cells.

Conclusions: These results suggest that methadone may, at least partly, produce its pharmacological effect as a β-arrestin-biased µ-opioid receptor agonist, similar to fentanyl, and NMDA receptor blockade is not the main contributor to the pharmacological profile of methadone.

Keywords: Methadone; NMDA receptor; internalization; µ-opioid receptor.

Publication types

  • Research Support, Non-U.S. Gov't

MeSH terms

  • Animals
  • CHO Cells
  • Cricetinae
  • Cricetulus
  • Dizocilpine Maleate / pharmacology
  • Endocytosis / drug effects
  • Fentanyl / pharmacology
  • Gastrointestinal Transit / drug effects
  • Male
  • Methadone / pharmacology*
  • Mice, Inbred ICR
  • Nociception / drug effects
  • Rats, Inbred F344
  • Receptors, N-Methyl-D-Aspartate / metabolism
  • Receptors, Opioid, mu / agonists*
  • beta-Arrestins / metabolism*


  • Receptors, N-Methyl-D-Aspartate
  • Receptors, Opioid, mu
  • beta-Arrestins
  • Dizocilpine Maleate
  • Methadone
  • Fentanyl