Abstract
In a recent human [11C]-(+)-PHNO positron emission tomography study, olanzapine, clozapine, and risperidone occupied D2 receptors in striatum (STR), but, despite their similar in vitro D2 and D3 affinities, failed to occupy D3 receptors in globus pallidus. This study had two aims: (1) to characterize the regional D2/D3 pharmacology of in vitro and ex vivo [3H]-(+)-PHNO binding sites in rat brain and (2) to compare, using [3H]-(+)-PHNO autoradiography, the ex vivo and in vitro pharmacology of olanzapine, clozapine, risperidone, and haloperidol. Using the D3-selective drug SB277011, we found that ex vivo and in vitro [3H]-(+)-PHNO binding in STR is exclusively due to D2, whereas that in cerebellar lobes 9 and 10 is exclusively due to D3. Surprisingly, the D3 contribution to [3H]-(+)-PHNO binding in the islands of Calleja, ventral pallidum, substantia nigra, and nucleus accumbens was greater ex vivo than in vitro. Ex vivo, systemically administered olanzapine, risperidone, and haloperidol, at doses occupying ∼80% D2, did not occupy D3 receptors. Clozapine, which also occupied ∼80% of D2 receptors ex vivo, occupied a smaller percentage of D3 receptors than predicted by its in vitro pharmacology. Across brain regions, ex vivo occupancy by antipsychotics was inversely related to the D3 contribution to [3H]-(+)-PHNO binding. In contrast, in vitro occupancy was similar across brain regions, independent of the regional D3 contribution. These data indicate that at clinically relevant doses, olanzapine, clozapine, risperidone, and haloperidol are D2-selective ex vivo. This unforeseen finding suggests that their clinical effects cannot be attributed to D3 receptor blockade.
Similar content being viewed by others
Log in or create a free account to read this content
Gain free access to this article, as well as selected content from this journal and more on nature.com
or
References
Bancroft GN, Morgan KA, Flietstra RJ, Levant B (1998). Binding of [3H]PD 128907, a putatively selective ligand for the D3 dopamine receptor, in rat brain: a receptor binding and quantitative autoradiographic study. Neuropsychopharmacology 18: 305–316.
Beckstead RM, Domesick VB, Nauta WJ (1979). Efferent connections of the substantia nigra and ventral tegmental area in the rat. Brain Res 175: 191–217.
Carson RE, Channing MA, Blasberg RG, Dunn BB, Cohen RM, Rice KC et al (1993). Comparison of bolus and infusion methods for receptor quantitation: application to [18F]cyclofoxy and positron emission tomography. J Cereb Blood Flow Metab 13: 24–42.
Farde L, Nordstrom AL, Wiesel FA, Pauli S, Halldin C, Sedvall G (1992). Positron emission tomographic analysis of central D1 and D2 dopamine receptor occupancy in patients treated with classical neuroleptics and clozapine. Relation to extrapyramidal side effects. Arch Gen Psychiatry 49: 538–544.
Freedman SB, Patel S, Marwood R, Emms F, Seabrook GR, Knowles MR et al (1994). Expression and pharmacological characterization of the human D3 dopamine receptor. J Pharmacol Exp Ther 268: 417–426.
Fujita M (2001). In vivo receptor imaging with PET and SPET-pitfalls in quantification. Int Rev Psychiatry 13: 34–39.
Ginovart N, Galineau L, Willeit M, Mizrahi R, Bloomfield PM, Seeman P et al (2006). Binding characteristics and sensitivity to endogenous dopamine of [11C]-(+)-PHNO, a new agonist radiotracer for imaging the high-affinity state of D2 receptors in vivo using positron emission tomography. J Neurochem 97: 1089–1103.
Ginovart N, Willeit M, Rusjan P, Graff A, Bloomfield PM, Houle S et al (2007). Positron emission tomography quantification of [11C]-(+)-PHNO binding in the human brain. J Cereb Blood Flow Metab 27: 857–871.
Ginovart N, Wilson AA, Meyer JH, Hussey D, Houle S (2001). Positron emission tomography quantification of [11C]-DASB binding to the human serotonin transporter: modeling strategies. J Cereb Blood Flow Metab 21: 1342–1353.
Graff-Guerrero A, Abi-Saab W, Redden L, Katz D, Houle S, O'Neill A et al (eds) (2008). First demonstration of D3 occupancy in humans: blockade of [11C]-(+)-PHNO PET by ABT-92. Int J Neuropsychopharmacol 11 (Suppl. S1): 223.
Graff-Guerrero A, Mamo D, Shammi CM, Mizrahi R, Marcon H, Barsoum P et al (2009). The effect of antipsychotics on the high-affinity state of D2 and D3 receptors: a positron emission tomography study with [11C]-(+)-PHNO. Arch Gen Psychiatry 66: 606–615.
Ito H, Hietala J, Blomqvist G, Halldin C, Farde L (1998). Comparison of the transient equilibrium and continuous infusion method for quantitative PET analysis of [11C]raclopride binding. J Cereb Blood Flow Metab 18: 941–950.
Joyce JN, Millan MJ (2005). Dopamine D3 receptor antagonists as therapeutic agents. Drug Discov Today 10: 917–925.
Kapur S, VanderSpek SC, Brownlee BA, Nobrega JN (2003). Antipsychotic dosing in preclinical models is often unrepresentative of the clinical condition: a suggested solution based on in vivo occupancy. J Pharmacol Exp Ther 305: 625–631.
Kapur S, Zipursky R, Jones C, Shammi CS, Remington G, Seeman P (2000). A positron emission tomography study of quetiapine in schizophrenia: a preliminary finding of an antipsychotic effect with only transiently high dopamine D2 receptor occupancy. Arch Gen Psychiatry 57: 553–559.
Kapur S, Zipursky RB, Remington G (1999). Clinical and theoretical implications of 5-HT2 and D2 receptor occupancy of clozapine, risperidone, and olanzapine in schizophrenia. Am J Psychiatry 156: 286–293.
Kongsamut S, Roehr JE, Cai J, Hartman HB, Weissensee P, Kerman LL et al (1996). Iloperidone binding to human and rat dopamine and 5-HT receptors. Eur J Pharmacol 317: 417–423.
Lammertsma AA, Bench CJ, Hume SP, Osman S, Gunn K, Brooks DJ et al (1996). Comparison of methods for analysis of clinical [11C]raclopride studies. J Cereb Blood Flow Metab 16: 42–52.
Landwehrmeyer B, Mengod G, Palacios JM (1993). Differential visualization of dopamine D2 and D3 receptor sites in rat brain. A comparative study using in situ hybridization histochemistry and ligand binding autoradiography. Eur J Neurosci 5: 145–153.
Leopoldo M, Berardi F, Colabufo NA, De Giorgio P, Lacivita E, Perrone R et al (2002). Structure-affinity relationship study on N-[4-(4-arylpiperazin-1-yl)butyl]arylcarboxamides as potent and selective dopamine D(3) receptor ligands. J Med Chem 45: 5727–5735.
Levant B (1998). Differential distribution of D3 dopamine receptors in the brains of several mammalian species. Brain Res 800: 269–274.
Levesque D, Diaz J, Pilon C, Martres MP, Giros B, Souil E et al (1992). Identification, characterization, and localization of the dopamine D3 receptor in rat brain using 7-[3H]hydroxy-N,N-di-n-propyl-2-aminotetralin. Proc Natl Acad Sci USA 89: 8155–8159.
McCormick PN, Kapur S, Reckless GE, Wilson AA (2009). Ex vivo [11C]-(+)-PHNO binding is unchanged in animal models displaying increased high-affinity states of the D2 receptor in vitro. Synapse 63: 998–1009.
McCormick PN, Kapur S, Seeman P, Wilson AA (2008). Dopamine D2 receptor radiotracers [11C](+)-PHNO and [3H]raclopride are indistinguishably inhibited by D2 agonists and antagonists ex vivo. Nucl Med Biol 35: 11–17.
Narendran R, Slifstein M, Guillin O, Hwang Y, Hwang DR, Scher E et al (2006). Dopamine (D2/3) receptor agonist positron emission tomography radiotracer [11C]-(+)-PHNO is a D3 receptor preferring agonist in vivo. Synapse 60: 485–495.
Newman AH, Grundt P, Nader MA (2005). Dopamine D3 receptor partial agonists and antagonists as potential drug abuse therapeutic agents. J Med Chem 48: 3663–3679.
Nobrega JN, Seeman P (1994). Dopamine D2 receptors mapped in rat brain with [3H](+)PHNO. Synapse 17: 167–172.
Nordstrom AL, Farde L, Nyberg S, Karlsson P, Halldin C, Sedvall G (1995). D1, D2, and 5-HT2 receptor occupancy in relation to clozapine serum concentration: a PET study of schizophrenic patients. Am J Psychiatry 152: 1444–1449.
Nordstrom AL, Farde L, Wiesel FA, Forslund K, Pauli S, Halldin C et al (1993). Central D2-dopamine receptor occupancy in relation to antipsychotic drug effects: a double-blind PET study of schizophrenic patients. Biol Psychiatry 33: 227.
Paxinos G, Watson C (1986). The Rat Brain in Stereotaxic Coordinates. Academic Press: New York.
Rabiner E, Slifstein M, Nobrega J, Plisson C, Huiban M, Raymond R et al (2009). In vivo quantification of regional dopamine-D3 receptor binding potential of (+)-PHNO: studies in non-human primates and transgenic mince. Synapse 63: 782–793.
Schotte A, Janssen PF, Bonaventure P, Leysen JE (1996a). Endogenous dopamine limits the binding of antipsychotic drugs to D3 receptors in the rat brain: a quantitative autoradiographic study. Histochem J 28: 791–799.
Schotte A, Janssen PF, Gommeren W, Luyten WH, Van Gompel P, Lesage AS et al (1996b). Risperidone compared with new and reference antipsychotic drugs: in vitro and in vivo receptor binding. Psychopharmacology (Berl) 124: 57–73.
Schotte A, Janssen PF, Gommeren W, Luyten WH, Leysen JE (1992). Autoradiographic evidence for the occlusion of rat brain dopamine D3 receptors in vivo. Eur J Pharmacol 218: 373–375.
Seabrook GR, Patel S, Marwood R, Emms F, Knowles MR, Freedman SB et al (1992). Stable expression of human D3 dopamine receptors in GH4C1 pituitary cells. FEBS Lett 312: 123–126.
Sokoloff P, Giros B, Martres MP, Bouthenet ML, Schwartz JC (1990). Molecular cloning and characterization of a novel dopamine receptor (D3) as a target for neuroleptics. Nature 347: 146–151.
Swanson LW (1982). The projections of the ventral tegmental area and adjacent regions: a combined fluorescent retrograde tracer and immunofluorescence study in the rat. Brain Res Bull 9: 321–353.
Tang L, Todd RD, Heller A, O'Malley KL (1994). Pharmacological and functional characterization of D2, D3 and D4 dopamine receptors in fibroblast and dopaminergic cell lines. J Pharmacol Exp Ther 268: 495–502.
Tsukada H, Harada N, Nishiyama S, Ohba H, Sato K, Fukumoto D et al (2000). Ketamine decreased striatal [11C]raclopride binding with no alterations in static dopamine concentrations in the striatal extracellular fluid in the monkey brain: multiparametric PET studies combined with microdialysis analysis. Synapse 37: 95–103.
Tsukada H, Ohba H, Harada N, Kakiuchi T (2008). Effects of anesthesia on kinetics of [11C]MNPA and its response to methamphetamine in the monkey brain. Neuroimage 41: T40.
van der Zwaal EM, Luijendijk MC, Adan RA, la Fleur SE (2008). Olanzapine-induced weight gain: chronic infusion using osmotic minipumps does not result in stable plasma levels due to degradation of olanzapine in solution. Eur J Pharmacol 585: 130–136.
van Vliet LA, Rodenhuis N, Dijkstra D, Wikstrom H, Pugsley TA, Serpa KA et al (2000). Synthesis and pharmacological evaluation of thiopyran analogues of the dopamine D3 receptor-selective agonist (4aR,10bR)-(+)-trans-3,4,4a,10b-tetrahydro-4-n-propyl-2H,5H [1]b enzopyrano[4,3-b]-1,4-oxazin-9-ol (PD 128907). J Med Chem 43: 2871–2882.
Willeit M, Ginovart N, Kapur S, Houle S, Hussey D, Seeman P et al (2006). High-affinity states of human brain dopamine D2/3 receptors imaged by the agonist [11C]-(+)-PHNO. Biol Psychiatry 59: 389–394.
Wilson AA, McCormick P, Kapur S, Willeit M, Garcia A, Hussey D et al (2005). Radiosynthesis and evaluation of [11C]-(+)-4-propyl-3,4,4a,5,6,10b-hexahydro-2H-naphtho[1,2-b][1,4]oxazin-9 -ol as a potential radiotracer for in vivo imaging of the dopamine D2 high-affinity state with positron emission tomography. J Med Chem 48: 4153–4160.
Yildiz A, Eryilmaz M, Gungor F, Erkilic M, Karayalcin B (2000). Regional cerebral blood flow in schizophrenia before and after neuroleptic medication. Nucl Med Commun 21: 1113–1118.
Acknowledgements
We thank Drs Gary Remington, Jeffrey Meyer, and Neil Vasdev for valuable discussion, and Dr Bernard Le Foll for his generous donation of the D3-selective drug SB277011. This work was financially supported by the Canadian Institutes of Health Research (grant numbers MOP 74702 and MOP 44051).
Author information
Authors and Affiliations
Corresponding author
Ethics declarations
Competing interests
Patrick N McCormick, Alan A Wilson, José Nobrega, and Roger Raymond have no conflicts of interest to disclose. Ariel Graff-Guerrero has received professional services compensation from Abbott Laboratories, grant support from Janssen, and speaker compensation from Eli Lilly. Shitij Kapur has received grant support from AstraZeneca, Bristol-Myers Squibb, and Glaxo Smith Kline, and has acted as consultant/scientific advisor or received speaker compensation from AstraZeneca, Bioline, Bristol-Meyers Squibb, Eli Lilly, Janssen, Lundbeck, Otsuka, Organon, Pfizer, Servier, and Solvay Wyeth.
Additional information
Supplementary Information accompanies the paper on the Neuropsychopharmacology website
Supplementary information
Rights and permissions
About this article
Cite this article
McCormick, P., Kapur, S., Graff-Guerrero, A. et al. The Antipsychotics Olanzapine, Risperidone, Clozapine, and Haloperidol Are D2-Selective Ex Vivo but Not In Vitro. Neuropsychopharmacol 35, 1826–1835 (2010). https://doi.org/10.1038/npp.2010.50
Received:
Revised:
Accepted:
Published:
Issue date:
DOI: https://doi.org/10.1038/npp.2010.50
Keywords
This article is cited by
-
Cariprazine alleviates core behavioral deficits in the prenatal valproic acid exposure model of autism spectrum disorder
Psychopharmacology (2021)
-
Evaluation of dopamine D3 receptor occupancy by blonanserin using [11C]-(+)-PHNO in schizophrenia patients
Psychopharmacology (2021)
-
Effects of the T-type calcium channel antagonist Z944 on paired associates learning and locomotor activity in rats treated with the NMDA receptor antagonist MK-801
Psychopharmacology (2018)
-
The effects of buspirone on occupancy of dopamine receptors and the rat gambling task
Psychopharmacology (2017)
-
Occupancy of Dopamine D3 and D2 Receptors by Buspirone: A [11C]-(+)-PHNO PET Study in Humans
Neuropsychopharmacology (2016)
