J.E. Leysen

Occupancy of central neurotransmitter receptors by risperidone, clozapine and haloperidol, measured ex vivo by quantitative autoradiography

Risperidone (Risperdal) is a novel antipsychotic drug, with beneficial effects on both positive and negative symptoms of schizophrenia, and with a low incidence of extrapyramidal side effects (EPS). These particular properties have been attributed to the predominant and very potent serotonin 5-HT2 receptor antagonism of the drug combined with less potent dopamine D2 antagonism. In order to provide data on the degree to which various central neurotransmitter receptors are occupied in vivo, we performed ex vivo receptor occupancy studies with risperidone in comparison with clozapine and haloperidol in rats and guinea pigs. Various types of receptors, to which the compounds were known to bind to in vitro, were investigated precisely using receptor autoradiography in sections of the same rat brain except for histamine H1 receptors that were measured in the guinea-pig cerebellum. Risperidone (2 h after s.c. treatment) occupied 5-HT2 receptors at very low doses (ED50 = 0.067 mg/kg). Nearly full occupancy (> 80%) was achieved before H1, D2, alpha 1 and alpha 2 receptors became occupied (ED50 = 0.45, 0.66, 0.75 and 3.7 mg/kg, respectively). Clozapine displayed occupancy of H1 and alpha 1 receptors at low doses (ED50 = 0.15 and 0.58 mg/kg, respectively) and of 5-HT2, 5-HT1C, D2, alpha 2, cholinergic muscarinic and 5-HT1A receptors at higher doses (ED50 = 1.3, 1.8, 9.0, 9.5, 11 and 15 mg/kg, respectively). Haloperidol occupied D2 and alpha 1 receptors at low doses (ED50 = 0.13 and 0.42 mg/kg, respectively) and 5-HT2 receptors at a higher dose (ED50 = 2.6 mg/kg). Occupancy of receptor types occurred with similar ED50-values in various brain areas, e.g. D2 receptors in striatum and mesolimbic areas. The ED50-values for the ex vivo measured occupancy of 5-HT2 and D2 receptors were in good agreement with ED50-values for functional effects putatively mediated by these central receptors. The dose-dependent occupancy of D2 receptors proceeded more gradually with risperidone (slope in the caudate-putamen: 0.85) than with clozapine (slope: 1.44) or haloperidol (slope: 1.51). It has previously been suggested that partial D2 receptor occupancy may suffice to control the positive symptoms of schizophrenia, whereas higher D2 receptor occupancy would induce extrapyramidal symptoms (EPS). The dose ratio for high (75%) vs. low (25%) D2 receptor occupancy in the caudate-putamen, was 37.3 for risperidone, 8.4 for clozapine, and 7.9 for haloperidol.(ABSTRACT TRUNCATED AT 400 WORDS)

Survey on the pharmacodynamics of the new antipsychotic risperidone

This review reports on the pharmacodynamics of the new antipsychotic risperidone. The primary action of risperidone is serotonin 5-HT2 receptor blockade as shown by displacement of radioligand binding (Ki: 0.16 nM), activity on isolated tissues (EC50:0.5 nM), and antagonism of peripherally (ED50: 0.0011 mg/kg) and centrally (ED50:0.014 mg/kg) acting 5-HT2 receptor agonists in rats. Risperidone is at least as potent as the specific 5-HT2 receptor antagonist ritanserin in these tests. Risperidone is also a potent dopamine D2 receptor antagonist as indicated by displacement of radioligand binding (Ki: 1.4 nM), activity in isolated striatal slices (IC50: 0.89 nM), and antagonism of peripherally (ED50: 0.0057 mg/kg in dogs) and centrally acting D2 receptor agonists (ED50: 0.056–0.15 mg/kg in rats). Risperidone shows all effects common to D2 antagonists, including enhancement of prolactin release. However, some central effects such as catalepsy and blockade of motor activity occur at high doses only. Risperidone is 4–10 times less potent than haloperidol as a central D2 antagonist in rats and it differs from haloperidol by the following characteristics: predominant 5-HT2 antagonism; LSD antagonism; effects on sleep; smooth dose-response curves for D2 antagonism; synergism of combined 5-HT2/D2 antagonism; pronounced effects on amphetamine-induced oxygen consumption; increased social interaction; and pronounced effects on dopamine (DA) turnover. Risperidone displays similar activity at pre- and postsynaptic D2 receptors and at D2 receptors from various rat brain regions. The binding affinity for D4 and D3 receptors is 5 and 9 times weaker, respectively, than for D2 receptors; interaction with D1 receptors occurs only at very high concentrations. The pharmacological profile of risperidone includes interaction with histamine H1 and α-adrenergic receptors but the compound is devoid of significant interaction with cholinergic and a variety of other types of receptors. Risperidone has excellent oral activity, a rapid onset, and a 24-h duration of action. Its major metabolite, 9-hydroxyrisperidone, closely mimics risperidone in pharmacodynamics. Risperidone can be characterized as a potent D2 antagonist with predominant 5HT2 antagonistic activity and optimal pharmacokinetic properties.

Does phenylethylamine act as an endogenous amphetamine in some patients

In brain capillary endothelium and catecholaminergic terminals a single decarboxylation step effected by aromatic amino-acid decarboxylase converts phenylalanine to phenylethylamine, at a rate comparable to that of the central synthesis of dopamine. Phenylethylamine, however, is not stored in intra-neuronal vesicles and is rapidly degraded by monoamine oxidase-B. Despite its short half-life, phenylethylamine attracts attention as an endogenous amphetamine since it can potentiate catecholaminergic neurotransmission and induce striatal hyperreactivity. Subnormal phenylethylamine levels have been linked to disorders such as attention deficit and depression; the use of selegiline (Deprenyl) in Parkinsons disease may conceivably favour recovery from deficient dopaminergic neurotransmission by a monoamine oxidase-B inhibitory action that increases central phenylethylamine. Excess phenylethylamine has been invoked particularly in paranoid schizophrenia, in which it is thought to act as an endogenous amphetamine and, therefore, would be antagonized by neuroleptics. The importance of phenylethylamine in mental disorders is far from fully elucidated but the evolution of phenylethylamine concentrations in relation to symptoms remains a worthwhile investigation for individual psychotic patients.

Endogenous dopamine limits the binding of antipsychotic drugs to D3 receptors in the rat brain: a quantitative autoradiographic study

Summary[3H]7-hydroxy-N,N-di-n-propyl-2-aminotetralin was used as a radioligand for the autoradiographic measurements of dopamine D3 receptors in rat and human brain. Preincubation of the brain sections was necessary to obtain binding of the radioligand in the islands of Calleja and in the nucleus accumbens, but not in cerebellar lobules 9/10 of the rat. D3 receptors were also totally occluded in unwashed sections of the human striatum. The radioligand binding to D3 receptors was maximal after preincubating the sections for at least 10 min. Pretreatment of the animals with reserpine or tetrabenazine, which results in a severe depletion of endogeneous monoamines, strongly reduces the occlusion of D3 receptors in unwashed brain sections. The occlusion of dopamine D3 receptors in brain sections suggests that thein vivo access to D3 receptors may be locally inhibited by endogenous dopamine.Thein vitro binding affinities of 12 antipsychotic drugs for D2 and D3 receptors were evaluated in competition binding experiments, using both rat and cloned human receptors. Most of the compounds showed only a slightly lower affinity for D3 than for D2 receptorsin vitro. Affinities of the antipsychotic drugs for cloned human D2L and D3 receptors were very close to their affinities for the rat receptors.In vivo occupancy of these receptors in the rat brain was measuredex vivo by quantitative autoradiography, 2 hours after subcutaneous drug administration. For most compounds, occupancy of D3 receptors, as compared to D2 receptor occupancy, was lower than expected from the correspondingin vivo affinity ratios. For the new antipsychotic risperidone,in vivo occupancy of D3 receptors was measured both in the islands of Calleja and in the cerebellar lobules 9/10. This compound was three times less potent for the occupancy of D3 receptors in the islands of Calleja than in the cerebellum, an area lacking endogenous dopamine (ED50=28 and 10 mg kg−1, respectively).Based on the observations in the rat brain, it may reasonably be supposed that therapeutic dosages of antipsychotic drugs will induce in patients only a minor occupancy of D3 receptors in brain areas containing high dopamine concentrations. The role of dopamine D3 receptors as a target of antipsychotic drugs may therefore be less important than previously thought.

Identification of 5-HT2 receptors, α1-adrenoceptors and amine release sites in rat brain by autoradiography with [125I]7-amino-8-iodo-ketanserin

Using autoradiographical techniques, we found that [125I]7-amino-8-iodo-ketanserin ([125I]AMIK) labelled with high affinity 5-HT2 receptors, alpha 1-adrenoceptors and sites involved in the release of biogenic amines and metabolites. The three binding sites for [125I]AMIK were identified by using selective inhibitors, i.e. BW 501 for 5-HT2 receptors, prazosin for alpha 1-adrenoceptors and tetrabenazine for the sites involved in the release of biogenic amines and metabolites. Using quantitative image analysis, we calculated the inhibition curves for the drugs in areas containing one of the receptor binding sites, i.e. the claustrum for 5-HT2 receptors, the thalamic nuclei for the alpha 1-adrenoceptors and the rostral part of the caudate putamen for the release sites. By using appropriate combinations of [125I]AMIK and drugs to occlude other binding sites, we could selectively label 5-HT2 receptors (in the presence of 0.1 microM prazosin and 1 microM tetrabenazine), alpha 1-adrenoceptors (in the presence of 1 microM BW 501 and 1 microM tetrabenazine) and release sites (in the presence 0.1 microM prazosin and 1 microM BW 501). Most binding to 5-HT2 receptors was observed in the cortical areas, the caudal caudate-putamen and the substantia nigra, while moderate binding was noticed in the nucleus accumbens, the olfactory bulb and the olfactory tubercle. There were many alpha 1-adrenoceptors in the numerous thalamic nuclei and few in the cortical areas, the periaqueductal grey matter and the substantia nigra.(ABSTRACT TRUNCATED AT 250 WORDS)

Differential Regulation of Dopamine-D2 and Serotonin-S2 Receptors by Chronic Treatment with the Serotonin-S2 Antagonists, Ritanserin, and Setoperone

Serotonin-S2 receptor sites were first described in studies in which 3H-spiperone was used in frontal cortical membrane preparations (Leysen et al. 1978; Peroutka and Snyder 1979). The receptor binding sites were further characterized and could be better distinguished by using the more selective serotonin antagonist 3H-ke-tanserin (Leysen et al. 1982). Serotonin-S2 receptors were shown to have a role in serotonin agonist-induced behavioral excitation and discriminative stimulus effects in rodents, and also to mediate serotonin-induced vasoconstriction and platelet function. Recently it has been shown that inositol phospholipid turnover forms part of the signal transducing system coupled to serotonin-S2 receptor sites (for review see Leysen et al. 1984).

The tachykinin NK1 receptor antagonist, RP67580, inhibits the bradykinin-induced rise in intracellular Ca2+ concentration in bovine pulmonary artery endothelial cells

The bradykinin-induced rise in intracellular Ca2+ concentration ([Ca2+]i) and the bradykinin receptor involved in this response were characterized in bovine pulmonary artery endothelial cells. It was found that bradykinin induces an intracellular biphasic Ca2+ response, consisting of a transient peak followed by an elevated plateau phase. Both bradykinin and the bradykinin B1 receptor agonist, des-Arg9-bradykinin, induced a concentration-dependent increase in [Ca2+]i, but the bradykinin-induced rise was much greater. Moreover, the bradykinin-induced [Ca2+]i rise could be inhibited by the bradykinin B2 receptor antagonists, D-Arg0[Hyp3, Thi(5,8), D-Phe7]bradykinin and Hoe 140 (D-Arg[Hyp3, Thi5, D-Tic7, Oic8]bradykinin), but not by the bradykinin B1 receptor antagonist, des-Arg9-[Leu8]bradykinin. From these results it can be concluded that a bradykinin B2 receptor is involved in this response. Furthermore, we found that the tachykinin NK1 receptor antagonist, RP67580 ([imino 1 (methoxy-2-phenyl)-2 ethyl]-2 diphenyl 7,7 perhydroisoindolone-4 (3aR, 7aR)), and its negative enantiomer, RP68651 (2-[1-imino 2-(2 methoxy phenyl) ethyl] 7,7 diphenyl 4-perhydroisoindolone (3aS-7aS)), could inhibit the bradykinin-induced [Ca2+]i response, although no functional tachykinin NK1 receptors were found. Binding studies evidenced no binding of RP67580 or RP68651 to the bradykinin receptor. We conclude that RP67580 inhibits the bradykinin-induced rise in [Ca2+]i via a bradykinin B2 receptor-independent mechanism.