Abstract
The mechanism of action of cannabidiol, one of the major constituents of cannabis, is not well understood but a noncompetitive interaction with mu opioid receptors has been suggested on the basis of saturation binding experiments. The aim of the present study was to examine whether cannabidiol is an allosteric modulator at this receptor, using kinetic binding studies, which are particularly sensitive for the measurement of allosteric interactions at G protein-coupled receptors. In addition, we studied whether such a mechanism also extends to the delta opioid receptor. For comparison, (-)-Δ9-tetrahydrocannabinol (THC; another major constituent of cannabis) and rimonabant (a cannabinoid CB1 receptor antagonist) were studied. In mu opioid receptor binding studies on rat cerebral cortex membrane homogenates, the agonist 3H-DAMGO bound to a homogeneous class of binding sites with a KD of 0.68±0.02 nM and a Bmax of 203±7 fmol/mg protein. The dissociation of 3H-DAMGO induced by naloxone 10 μM (half life time of 7±1 min) was accelerated by cannabidiol and THC (at 100 μM, each) by a factor of 12 and 2, respectively. The respective pEC50 values for a half-maximum elevation of the dissociation rate constant koff were 4.38 and 4.67; 3H-DAMGO dissociation was not affected by rimonabant 10 μM. In delta opioid receptor binding studies on rat cerebral cortex membrane homogenates, the antagonist 3H-naltrindole bound to a homogeneous class of binding sites with a KD of 0.24±0.02 nM and a Bmax of 352±22 fmol/mg protein. The dissociation of 3H-naltrindole induced by naltrindole 10 μM (half life time of 119±3 min) was accelerated by cannabidiol and THC (at 100 μM, each) by a factor of 2, each. The respective pEC50 values were 4.10 and 5.00; 3H-naltrindole dissociation was not affected by rimonabant 10 μM. The present study shows that cannabidiol is an allosteric modulator at mu and delta opioid receptors. This property is shared by THC but not by rimonabant.
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Abbreviations
- AEBSF:
-
4-(2-aminoethyl)benzenesulfonyl fluoride
- 3H-DAMGO:
-
3H-Tyr-D-Ala-Gly-N-methyl-Phe-Gly-ol
- 3H-NTI:
-
3H-naltrindole
- CBD:
-
(-)-cannabidiol
- THC:
-
(-)-Δ9-tetrahydrocannabinol
References
Akiyama K, Gee KW, Mosberg HI, Hruby VJ, Yamamura HI (1985) Characterization of [3H][2-D-penicillamine, 5-D-penicillamine]-enkephalin binding to δ opiate receptors in the rat brain and neuroblastoma-glioma hybrid cell line (NG 108-15). Proc Natl Acad Sci USA 82:2543–2547
Ameri A (1999) The effects of cannabinoids on the brain. Prog Neurobiol 58:315–348
Bisogno T, Hanuš L, De Petrocellis L, Tchilibon S, Ponde DE, Brandi I, Moriello AS, Davis JB, Mechoulam R, Di Marzo V (2001) Molecular targets for cannabidiol and its synthetic analogues: effect on vanilloid VR1 receptors and on the cellular uptake and enzymatic hydrolysis of anandamide. Br J Pharmacol 134:845–852
Bornheim LM, Correia MA (1989) Effect of cannbidiol on cytochrome P-450 isoenzymes. Biochem Pharmacol 38:2789–2794
Bradford MM (1976) A rapid and sensitive method for the quantitation of microgram quantities of protein utilizing the principle of protein-dye binding. Anal Biochem 72:248–254
Christopoulos A, Kenakin T (2002) G protein-coupled receptor allosterism and complexing. Pharmacol Rev 54:323–374
Consroe P, Martin A, Singh V (1981) Antiepileptic potential of cannabidiol analogs. J Clin Pharmacol 21:S428–S436
Contreras PC, Tam L, Drower E, Rafferty MF (1993) [3H]naltrindole: a potent and selective ligand for labeling delta-opioid receptors. Brain Res 604:160–164
Dhawan BN, Cesselin F, Raghubir R, Reisine T, Bradley PB, Portoghese PS, Hamon M (1996) International Union of Pharmacologists. XII. Classification of opioid receptors. Pharmacol Rev 48:567–592
Guimaraes FS, De Aguiar JC, Mechoulam R, Breuer A (1994) Anxiolytic effect of cannabidiol derivatives in the elevated plus-maze. Gen Pharmacol 25:161–164
Hampson AJ, Grimaldi M, Axelrod J, Wink D (1998) Cannabidiol and (-)Δ9-tetrahydrocannabinol are neuroprotective. Proc Natl Acad Sci USA 95:8268–8273
Howlett AC, Barth F, Bonner TI, Cabral G, Casellas P, Devane WA, Felder CC, Herkenham M, Mackie K, Martin BR, Mechoulam R, Pertwee RG (2002) International Union of Pharmacology. XXVII. Classification of cannabinoid receptors. Pharmacol Rev 54:161–202
Kenakin T (1993) Pharmacological analysis of drug-receptor interaction. Raven Press, New York
Leppik RA, Lazareno S, Mynett A, Birdsall NJ (1998) Characterization of the allosteric interactions between antagonists and amiloride analogues at the human alpha2A-adrenergic receptor. Mol Pharmacol 53:916–925
Leppik RA, Mynett A, Lazareno S, Birdsall NJ (2000) Allosteric interactions between the antagonist prazosin and amiloride analogs at the human alpha1A-adrenergic receptor. Mol Pharmacol 57:436–445
Malfait AM, Gallily R, Sumariwalla PF, Malik AS, Andreakos E, Mechoulam R, Feldmann M (2000) The nonpsychoactive cannabis constituent cannabidiol is an oral anti-arthritic therapeutic in murine collagen-induced arthritis. Proc Natl Acad Sci USA 97:9561–9566
Martin AR, Consroe P, Kane VV, Shah V, Singh V, Lander N, Mechoulam R, Srebnik M (1987) Structure-anticonvulsant activity relationships of cannabidiol analogs. NIDA Res Monogr 79:48–58
Pertwee RG (1999) Pharmacology of cannabinoid receptor ligands. Curr Med Chem 6:635–664
Pertwee RG (2005) Inverse agonism and neutral antagonism at cannabinoid CB1 receptors. Life Sci 76:1307–1324
Pertwee RG, Ross RA, Craib SA, Thomas A (2002) (-)-Cannabidiol antagonizes cannabinoid receptor agonists and noradrenaline in the mouse vas deferens. Eur J Pharmacol 456:99–106
Potter LT, Ferrendelli CA, Hanchett HE, Holliefield MA, Lorenzi MV (1989) Tetrahydroaminoacridine and other allosteric antagonists of hippocampal M1 muscarine receptors. Mol Pharmacol 35:652–660
Rinaldi-Carmona M, Barth F, Heaulme M, Shire D, Calandra B, Congy C, Martinez S, Maruani J, Neliat G, Caput D, Ferrara P, Soubrie P, Breliere JC, Le Fur G (1994) SR 141716A, a potent and selective antagonist of the brain cannabinoid receptor. FEBS Lett 350:240–244
Rog DJ, Nurmikko TJ, Friede T, Young CA (2005) Randomized, controlled trial of cannabis-based medicine in central pain in multiple sclerosis. Neurology 65:812–819
Schlicker E, Kathmann M (2001) Modulation of transmitter release via presynaptic cannabinoid receptors. Trends Pharmacol Sci 22:565–572
Srivastava MD, Srivastava BI, Brouhard B (1998) Delta9 tetrahydrocannabinol and cannabidiol alter cytokine production by human immune cells. Immunopharmacology 40:179–185
Tränkle C, Weyand O, Voigtländer U, Mynett A, Lazareno S, Birdsall NJ, Mohr K (2003) Interactions of orthosteric and allosteric ligands with [3H]dimethyl-W84 at the common allosteric site of muscarinic M2 receptors. Mol Pharmacol 64:180–190
Ulibarri I, Garcia-Sevilla JA, Ugedo L (1987) Modulation of brain α2-adrenoceptor and μ-opioid receptor densities during morphine dependence and spontaneous withdrawal in rats. Naunyn-Schmiedeberg’s Arch Pharmacol 336:530–537
Vaysse PJJ, Gardener EL, Zukin RS (1987) Modulation of rat brain opioid receptors by cannabinoids. J Pharm Exp Ther 241:534–539
Zhao GM, Qian X, Schiller PW, Szeto HH (2003) Comparison of [Dmt1]DALDA and DAMGO in binding and G protein activation at μ, δ and κ opioid receptors. J Pharm Exp Ther 307:947–954
Acknowledgements
This study was supported by grants from the Deutsche Forschungsgemeinschaft (Schl 266/5-5 and Graduiertenkolleg 246 TP 01). We are also indebted to Mrs. P. Zeidler for her skilled technical assistance and to GW Pharmaceuticals and Sanofi-Aventis for gifts of drugs.
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Kathmann, M., Flau, K., Redmer, A. et al. Cannabidiol is an allosteric modulator at mu- and delta-opioid receptors. Naunyn Schmied Arch Pharmacol 372, 354–361 (2006). https://doi.org/10.1007/s00210-006-0033-x
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DOI: https://doi.org/10.1007/s00210-006-0033-x