Elsevier

Neuropharmacology

Volume 48, Issue 3, March 2005, Pages 360-371
Neuropharmacology

Peripherally acting NMDA receptor/glycineB site receptor antagonists inhibit morphine tolerance

https://doi.org/10.1016/j.neuropharm.2004.11.005Get rights and content

Abstract

The present study focused on the role of peripheral ionotropic N-methyl-d-aspartate (NMDA) receptors in the development of tolerance to morphine-induced antinociception. An initial experiment revealed that NMDA channel blocker memantine, and NMDA receptor/glycineB site antagonist MRZ 2/576 inhibited maximal electroshock-induced convulsions (MES) in female NMR mice with respective potency of 5.93 and 20.8 mg/kg, while other NMDA receptor/glycineB site antagonists MRZ 2/596 and MDL 105,519 were ineffective, supporting lack of CNS activity of the latter two agents. This observation was also supported by blood–brain barrier experiments in vitro. In male Swiss mice, morphine (10 mg/kg) given for 6 days twice a day (b.i.d.) produced tolerance to its antinociceptive effects in the tail-flick test. The NMDA receptor/glycineB site antagonists, MRZ 2/576 at 0.03, 0.1, 0.3 mg/kg and MRZ 2/596 at 0.1, 0.3, 3 and 10 mg/kg attenuated the development of morphine tolerance. Similarly, in male C57/Bl mice, morphine (10 mg/kg) given for 6 days b.i.d. produced tolerance to its antinociceptive effects in the tail-flick test. Like in Swiss mice, in C57/Bl mice morphine tolerance was attenuated by both MRZ 2/576 and MRZ 2/596. Another NMDA receptor/glycineB site receptor antagonist, MDL 105,519 (that very weakly penetrates to the central nervous system) also inhibited morphine tolerance at the dose of 1 but not 0.1 mg/kg. Moreover, both naloxone hydrochloride (5 and 50 mg/kg) and centrally inactive naloxone methiodide (50 mg/kg) inhibited morphine tolerance suggesting the involvement of peripheral opioid receptors in this phenomenon. The present data suggest that blockade of NMDA receptor/glycineB sites in the periphery may attenuate tolerance to the antinociceptive effects of morphine.

Introduction

Over the last decade research has provided compelling evidence that glutamate receptors are crucially involved in phenomena related to opioid tolerance (see Mao, 1999, Price et al., 2000 for reviews). Antagonists of the ionotropic N-methyl-d-aspartate (NMDA) receptor complex, including memantine, the moderate affinity and highly voltage-dependent clinically used NMDA channel blocker (Parsons et al., 1999) inhibit the development of morphine tolerance (Trujillo and Akil, 1991, Marek et al., 1991, Popik et al., 2000a) (see Bespalov and Trujillo, 2002 for the recent review).

One obstacle to the introduction of NMDA receptor antagonists into clinical practice is undesired “phencyclidine side-effects” profile, that is centrally mediated (Parsons et al., 1999). In turn, NMDA receptor antagonists which weakly penetrate to the brain might have a more favorable profile. However, such compounds may not be applicable to the inhibition of tolerance to morphine-induced antinociception which is believed to be primarily of central origin (McNally and Westbrook, 1998, McNally, 1999, Ueda and Inoue, 1999). On the other hand, recent data reported by Kolesnikov and colleagues demonstrated that local (topical) application of uncompetitive NMDA receptor antagonists, (+)MK-801 or ketamine, inhibited tolerance to topically applied morphine (Kolesnikov et al., 1996, Kolesnikov and Pasternak, 1999b, Kolesnikov and Pasternak, 1999a), suggesting a peripheral component of antinociceptive morphine tolerance. Since systemic rather than local administration of compounds is more favorable from the therapeutic perspective, the aim of the present study was to investigate whether antagonism of NMDA receptors in the peripheral nervous system (PNS) would inhibit tolerance to the antinociceptive effects of systemically administered morphine. As pharmacological tools we used recently developed NMDA receptor antagonists acting at the NMDA receptor/glycineB site. These compounds reach relatively low brain levels after systemic application as compared to plasma values (MRZ 2/576: ∼2% (Hesselink et al., 1999b), MDL 105,519: 0.01–0.08% (Opackajuffry et al., 1998)) or seem to be lacking CNS activity as was the case with MRZ 2/596 (see Sections 3.1.3 BBB permeability studies, 4 Discussion). In contrast, the free brain levels of memantine are over 50% of plasma concentration (Hesselink et al., 1999a).

Section snippets

Receptor binding

Tissue preparation was performed according to Foster and Wong (1987) with some modifications. Male Sprague-Dawley rats (200–250 g, Janvier, Le Genest-Isle, France) were decapitated and their brains were removed rapidly. Tissue was then processed as described previously (Parsons et al., 1997). The amount of protein in the final membrane preparation was determined according to Hartfree (1971) and adjusted to 250–500 μg/ml.

Membranes were suspended and incubated in 50 mM Tris–HCl, pH 8.0 for 45 min at 4

Receptor binding

In Scatchard analysis [3H]MDL 105,519 had a Kd of 6.03 ± 0.14 nM and a Bmax of 4.03 pmol/mg protein. MRZ 2/576 and MRZ 2/596 displaced [3H]MDL-105,519 binding to rat cortical membranes with Kis of 126 and 160 nM, respectively (Table 1, IC50 corrected according to the Cheng-Prussoff relationship for 2 nM [3H]MDL 105,519). MDL-105,519 displaced its own binding with a Ki of 11.6 nM. Non-specific binding determined with glycine 100 μM was only 15–20% and standard compounds displaced binding to non-specific

Discussion

NMDA receptor binding and patch clamp studies confirmed that MRZ 2/576 and MRZ 2/596 bind with moderate (nanomolar) affinity to NMDA receptor/glycineB site whereas MDL 105,519 was some 5–10-fold more potent. Because of the use of glycine (1 μM) in the patch clamp studies, IC50 values obtained in these experiments were 4–9-fold higher than the Ki values obtained in binding experiments. However, correction of these IC50 values to Kbs taking into account a Kd for glycine in these patch clamp

Acknowledgements

The study was supported in part by Institute of Pharmacology Statutory Activity.

References (47)

  • G.P. McNally

    Pain facilitatory circuits in the mammalian central nervous system: their behavioral significance and role in morphine analgesic tolerance

    Neuroscience and Biobehavioral Reviews

    (1999)
  • T. Olivar et al.

    Differential effects of N-methyl-d-aspartate receptor blockade on nociceptive somatic and visceral reflexes

    Pain

    (1999)
  • J. Opackajuffry et al.

    Preliminary evaluation of the glycine site antagonists [11C]L 703,717 and [3H]MDL 105,519 as putative PET ligands for central NMDA receptors: in vivo studies in rats

    Quantitative Functional Brain Imaging with Positron Emission Tomography

    (1998)
  • C.G. Parsons et al.

    Memantine is a clinically well tolerated N-methyl-d-aspartate (NMDA) receptor antagonist – a review of preclinical data

    Neuropharmacology

    (1999)
  • M. Quartaroli et al.

    GV196771A, an NMDA receptor/glycine site antagonist, attenuates mechanical allodynia in neuropathic rats and reduces tolerance induced by morphine in mice

    European Journal of Pharmacology

    (2001)
  • V. Raghavendra et al.

    Reversal of morphine tolerance and dependence by melatonin: possible role of central and peripheral benzodiazepine receptors

    Brain Research

    (1999)
  • H. Ueda et al.

    Peripheral morphine analgesia resistant to tolerance in chronic morphine-treated mice

    Neuroscience Letters

    (1999)
  • R.J. Valentino et al.

    Receptor binding, antagonist, and withdrawal precipitating properties of opiate antagonists

    Life Science

    (1983)
  • G.L. Wenk et al.

    Neuroprotection by novel antagonists at the NMDA receptor channel and glycine(b) sites

    European Journal of Pharmacology

    (1998)
  • B.M. Baron et al.

    [3H]MDL 105,519, a high-affinity radioligand for the N-methyl-d-aspartate receptor-associated glycine recognition site

    Journal of Pharmacology and Experimental Therapeutics

    (1996)
  • I.V. Belozertseva et al.

    Short-acting NMDA receptor antagonist MRZ 2/576 produces prolonged suppression of morphine withdrawal in mice

    Naunyn-Schmiedeberg's Archives of Pharmacology

    (2000)
  • A. Bespalov et al.

    Drug tolerance

  • J. Booher et al.

    Growth and cultivation of dissociated neurons and glial cells from embryonic chick, rat and human brain in flask cultures

    Neurobiology

    (1972)
  • Cited by (22)

    • Tolerance develops to the antiallodynic effects of the peripherally acting opioid loperamide hydrochloride in nerve-injured rats

      2013, Pain
      Citation Excerpt :

      Because MDL 105,519 did not prevent systemic loperamide tolerance, and local MK-801 did not prevent tolerance to intraplantar loperamide-induced antiallodynia, NMDA receptors in the PNS seem not to be essential for loperamide tolerance in SNL rats. However, this notion contradicts previous observations that local MK-801 treatment blocks topical morphine antinociceptive tolerance [21] and that the peripherally acting NMDA receptor/glycine-B site antagonist MRZ 2/596 attenuates systemic morphine antinociceptive tolerance in mice [7]. The inconsistencies between studies could be attributable to differences in animal species, drug treatment (e.g., dose, route, and timing of administration), tolerance-inducing paradigm, behavioral tests, and importantly, animal conditions (e.g., uninjured versus nerve-injured animals).

    • Glutamatergic substrates of drug addiction and alcoholism

      2008, Biochemical Pharmacology
      Citation Excerpt :

      Since this study, numerous others have shown that morphine tolerance and/or withdrawal symptoms (including naloxone-precipitated CPA) are reduced by NMDA receptors antagonists [270,305,319–329]; AMPA/KA receptor antagonists [322,330];

    View all citing articles on Scopus

    The referees of the present paper suggested to assess brain penetration of MRZ 2/596 using brain microdialysis. It was not possible at the time of manuscript preparation but such opportunity appeared after manuscript acceptance. The study revealed, that in rats after application of MRZ 2/596 at the dose of 30 mg/kg ip maximal concentrations in brain reach c.a. 100 nM (N=3) concentrations corrected for in vitro recovery. Thus, it is unlikely that dose used in the present study (0.05–1 mg/kg) resulted in brain levels of MRZ 2/596 that would affect NMDA receptors assuming lack of differences in blood brain barrier between rat and mice.

    View full text