Clozapine: an atypical neuroleptic.

Since it was synthesized in 1960, much has been written about clozapine. Although a number of its properties are those of a neuroleptic, it displays marked differences from classical antipsychotics to the extent that it is currently listed as an atypical neuroleptic. A classical neuroleptic has been defined in man according to its antipsychotic properties, accompanied by extrapyramidal effects, and in animals according to its cataleptic properties, its ability to antagonize apomorphine and amphetamine stereotypies and to suppress the conditioned avoidance response. Moreover, the classical neuroleptic exerted depressive and anhedonic effects in most conditioning schedules. With clozapine, most of these properties are no longer strictly in force to the point that they call in question the validity of the tests carried out to detect the potential of neuroleptics. This article attempts to compare the characteristics of clozapine with those of classical neuroleptics from a toxicological, neuropharmacological, psychopharmacological and clinical point of view.

Nitric oxide: a new messenger in the brain.

The important role played by nitric oxide (NO) in the central nervous system has largely been emphasized in the recent literature. It can originate at least from four different sources: the endothelium of cerebral vessels, the immunostimulated microglia and astrocytes, the nonadrenergic noncholinergic nerve, and the glutamate neuron. NO has been implicated in a large number of pathologies (such as neurotoxicity in Alzheimer's disease and Huntington's disease, cerebral ischemia, stroke, and anxiety) and also in normal physiological functions (such as memory and learning, regulation of the cerebrovascular system, modulation of the wakefulness, mediation of nociception, olfaction, food intake and drinking, regulation of noradrenaline, and dopamine release). The aim of this paper is to review and to integrate the most recent advances in our understanding of the roles of NO in the brain.

Comparative study of the behavioral, neurophysiological, and motor effects of psychotropic drugs in the dog.

The purpose of this study was to present a comparative and detailed picture of behavioral, physiological, and motor effects of some barbiturates, benzodiazepines, and neuroleptics, using an operant procedure of differential reinforcement of response duration (DRRD) in the dog. An increase in response rate with low doses of barbiturates and benzodiazepines is interpreted in terms of behavioral disinhibition, anxiolytic effect, and enhancement of motivation. The depressive effects noted with higher doses of benzodiazepines and barbiturates, as well as with all doses of neuroleptics, can be explained in different ways. For benzodiazepines and barbiturates, ataxic effects more than sedative or motivational effects are implicated. However, for neuroleptics, loss of motivation or "anhedonia" better explains the decrease in operant responding. Differences between the main classes of psychotropic drugs but also within a class between 1,4 benzodiazepines or 1,5 benzodiazepines, hypnotic or nonhypnotic benzodiazepines, and typical or atypical neuroleptics are highlighted and correlated with human observations.

Pyridobenzoxazepine and pyridobenzothiazepine derivatives as potential central nervous system agents: synthesis and neurochemical study.

In order to characterize the pharmacological profile of the different chemical classes of pyridobenzazepine derivatives, a series of N-methylpiperazinopyrido[1,4]- and -[1,5]- benzoxa- and benzothiazepine derivatives were prepared. The affinities for D2, D1, 5-HT2, and cholinergic (M) receptors were measured. In comparison to dibenzazepine reference compounds, a strong decrease of the affinities was observed, less pronounced, however, for the substituted analogues. Oxazepine and thiazepine analogues like clozapine (except 8-chloro-6-(4-methylpiperazin-1-yl)-pyrido[2,3-b][1,4]benzoxazepin e (9) and 8-chloro-6-(4-methylpiperazin-1-yl)pyrido[2,3-b][1,4]- benzothiazepine (11)) were found to be inactive against apomorphine stereotypies. In the open-field test in rats, different molecules showed a high disinhibitory activity as observed with anxiolytic drugs. Moreover, 8-chloro-5-(4-methylpiperazin-1-yl)pyrido[2,3-b][1,5]benzoxazepine (14) presented a clozapine-like profile that was confirmed in the behavioral model in dogs and showed most of the behavioral characteristics described for antipsychotic drugs. Its neurochemical profile, in particular the 5-HT2/D2 ratio, was also compatible with atypical antipsychotic activity.

New pyridobenzodiazepine derivatives as potential antipsychotics: synthesis and neurochemical study.

The discovery of a new, safe, atypical antipsychotic remains an important challenge. To achieve this goal, a series of N-methylpiperazinopyrido[2,3-b] [1,4]- and -[1,5]- and -pyrido[4,3-b][1,4]- and -[1,5]- benzodiazepines were synthesized. The dopaminergic (D1, D2), serotonergic (5-HT2), and cholinergic (M) affinities, frequently remarked in the action mechanisms of antipsychotic drugs, were determined using their respective in vitro receptor binding assays. All affinities were reduced for each compound. Optimal substituents were found to be in the 2- or 8-position for the retention of affinities, while substitution at the 5-position by acyl or alkyl groups dramatically diminished binding affinities. Pyridobenzodiazepine derivatives, such as clozapine, were found to be inactive or only weakly effective against apomorphine-mediated stereotypes in rats. In an original and complex behavioral model developed in dogs and successfully used to differentiate distinct classes of psychotropic drugs and to discriminate between typical and atypical neuroleptic drugs, 8-chloro-6-(4-methyl-1-piperazinyl)-11H-pyrido[2,3-b] [1,4]benzodiazepine (9), 8-methyl-6-(4-methyl-1-piperazinyl)-11H-pyrido [2,3-b][1,4]benzodiazepine (12), and 5-(4-methyl-1-piperazinyl)-11H-pyrido[2,3-b][1,5]benzodiazepine (16) showed most of the behavioral characteristics previously described for neuroleptics. Their neurochemical profiles, particularly their 5-HT2/D2 pKi ratios, were compatible with an atypical antipsychotic effect. These compounds were selected for further investigation. The proposed modulations could lead to new possibilities for the pharmacochemistry of diarylazepines.

Hypoxia-induced decrease of brain acetylcholine release detected by microdialysis.

The in-vivo extracellular release of acetylcholine (ACh) from the striatum of rats was measured by intracerebral microdialysis. The ACh levels were first estimated under normoxic conditions, after which the rats were exposed to either 10% or 5% oxygen for 1 h. The average ACh content was 2.9 pmol/20 min. During hypoxia, the ACh levels were decreased to 62% and 44% of the control value with 10% and 5% oxygen, respectively. The data confirmed the high sensitivity of the striatal cholinergic system to hypoxia. The results are discussed in relation to the hypothesis that disruption of Ca2+ homeostasis may underlie the hypoxia-induced deficits in ACh release.

Effects of a long-interval retention on the hypoxia-induced impairment of performance in a fixed-interval schedule.

Retention of the fixed-interval (FI) responding was studied in rats after a chronic hypoxic treatment. The rats were trained over a 25-day period on a FI 60 s schedule. The rats were then given a normoxic or hypoxic (3.5% O2 for 10 min) treatment once a day for 4 days after which they were maintained in the animal quarters for 6 supplementary days. A retention test consisting of 4 daily FI 60 session followed the 10-day interval. The results showed that the performance during the retention test was very similar for both groups of rats. These data contrast with the deficit of retention observed when the retention FI 60 session were undertaken 24 h after the same hypoxic treatment. It is suggested that the results are due to a recovery from hypoxia-induced retrieval deficits rather than to the progressive attenuation of non-specific effects of hypoxia, such as anhedonia, hypoactivity, motivational changes or structural brain damage.

Stimulant effect of the beta-carboline FG 7142 in the open-field test.

The anxiogenic activity of N-methyl-beta-carboline-3-carboxamide (FG 7142) is sometimes difficult to observe in rats. As the open field has recently been applied successfully to test the anxiogenic potential of n-butyl-beta-carboline-3-carboxylate (beta-CCB) in mice, a comparable experiment was performed with FG 7142 (1, 5, 10, 30 mg/kg i.p.) in rats. In contrast to the inhibitory effects measured with beta-CCB, FG 7142 significantly increased the ambulation and rearing scores and induced aggressivity in some animals. A differential sensitivity of mice and rats to beta-carbolines, predominant analeptic properties of FG 7142, and differences in the types of anxiety induced are proposed to account for this discrepancy.

Comparative study of typical neuroleptics, clozapine and newly synthesized clozapine-analogues: correlations between neurochemistry and behaviour.

While neuroleptic therapy with classical compounds has frequently been associated with extrapyramidal side effects, clozapine has revealed an interesting antipsychotic profile without producing any clearcut motor side effects. However, some adverse reactions remained that stimulated the search for improved antipsychotic agents. The aim of this study was to characterize the behavioural and neurochemical profiles of typical neuroleptics (chlorpromazine, haloperidol), clozapine, and four newly synthesized clozapine-analogues. Affinity for dopaminergic (D1,D2), serotonergic (5-HT(2)) and cholinergic (muscarinic) receptors were measured and the ratios of these different binding affinities were determined and correlated with the behavioural effects of the drugs in a complex temporal regulation task in the dog. The four clozapine-analogues showed most of the behavioural characteristics previously described for neuroleptics and their neurochemical profile, particularly their 5-HT(2)/D2 pKi ratio, was compatible with an atypical antipsychotic effect. Among these drugs, JL5 and JL13 showed a high degree of similarity with clozapine. Like clozapine, they did not induce catalepsy and stereotypy/hyperkinesia. Moreover, other motor effects were also reduced (ataxia, akinesia, dystony). and tremor and sialorrhea were completely absent with these two molecules.

Effect of chronic hypoxic treatment on the retention of fixed-interval responding.

The present experiments aimed to study the effects of hypoxic treatment (10 min on 3.5% oxygen) on the stabilized performance of rats in a fixed-interval schedule (FI 60 s). The hypoxic treatment was given once a day, for 3 days, immediately after the FI session. The results showed that the operant performance was disturbed from the very first posthypoxia FI session. There was a sharp drop in response rate, and changes in the temporal distribution of responses occurred. The data obtained greatly limit the importance of nonspecific factors in explaining these changes in performance. The results are discussed with reference to the retrieval hypothesis. In particular, it is suggested that the decreases of performance may be due to an effect of hypoxia on the retrieval mechanisms associated with long-term memory.

Enhanced resistance effect of piracetam upon hypoxia-induced impaired retention of fixed-interval responding in rats.

Rats were trained on a fixed-interval schedule of 60 s (FI 60). After stabilization of performance, rats were chronically submitted to hypoxic treatment (3.5% O2, 10 min) once a day, immediately after the daily FI 60 session. Hypoxia disturbed the retention of FI responding. It was mainly characterized by a decrease in response rate and in pause duration, and by changes in the temporal distribution of responses. Animals receiving piracetam (100 mg/kg, IP) 30 min before each FI session followed by hypoxia were significantly less affected than saline-treated animals. Results are discussed with reference to the effects of hypoxia and piracetam on nonspecific factors and on memory function. It is suggested that the effects of piracetam are due to alleviation of hypoxia-induced memory retrieval deficit rather than to a protection against hypoxic brain cell injury.

Effects of low doses of neuroleptics on temporal regulation in a differential reinforcement of response duration (DRRD) schedule in the dog.

It has been shown that low doses of neuroleptics could disinhibit behaviour in animals as well as in man. This study aims to measure the effects of low doses of haloperidol (0.01, 0.05, 0.1 mg/kg) and sulpiride (5, 10, 15 mg/kg) in the dog using a differential reinforcement of response duration (DRRD) schedule with positive and negative external stimuli. Together with a decrease in response rate, a leftward shift in the temporal distribution of response duration is measured. These results are discussed in terms of a deregulation of the internal clock, a lessening in the ability to wait for the reward, a reduction in the frustration of not obtaining reinforcements when errors are made and an increase in the sensibility to reinforcement through appetite stimulation or decrease in the satiety level.

Effects of specific dopaminergic agonists and antagonists in the open-field test.

It has been found that dopaminergic transmission could be involved in some aspects of anxiety. The present study aims to explore this hypothesis further, using specific DA1 (SKF 38393) and DA2 (bromocriptine) agonists or DA1 (SCH 23390), and DA2 (zetidoline) antagonists in the open-field test. The results confirm previous studies indicating that DA1 and DA2 agonists predominantly increase locomotor activity, while DA1 and DA2 antagonists predominantly decrease it. However, at low doses, the four drugs increase the peripheral ambulation score significantly and, with the exception of zetidoline, also increase the central ambulation score. The observations made with zetidoline confirm the hypothesis that a specific presynaptic DA2 antagonism could be determinant for the disinhibitory effects of low doses of neuroleptics. A collateral action on 5HT transmission is also suggested to explain an hypothetic anxiolytic action of DA agonists and SCH 23390 at lower doses.

Anxiolytic potential of sulpiride, clozapine and derivatives in the open-field test.

Recently acquired data question the sharp dichotomy between anxiolytics and neuroleptics, since disinhibitory effects have been measured in the rat with very low doses of haloperidol and higher doses of atypical neuroleptics in FI and DRL schedules, but also in the open-field test. That the DA transmission in certain brain regions is involved in some aspects of anxiety has recently been suggested. The present study confirms this hypothesis particularly with high doses of sulpiride (80 mg/kg) and clozapine (24 mg/kg) when tested in the open-field test. Moreover, the results show how a slight chemical modification of clozapine can give a direction to pharmacological activity with one derivative still resembling clozapine and the second one resembling haloperidol. As neuroleptics do not seem to influence the synthesis and utilization of GABA, the higher entry score observed with them would seem to depend above all on DA antagonism in the mesolimbic system.

Differentiation of haloperidol and clozapine using a complex operant schedule in the dog.

This study aimed to differentiate chronically administered typical (haloperidol) and atypical (clozapine) neuroleptics in the dog using a complex temporal regulation schedule combining operant, voluntary, and involuntary motor parameters. Although clozapine and haloperidol showed some characteristics of neuroleptics, justifying their adherence to the same class of compounds, differences have also been highlighted and compared to the clinical observations. Haloperidol induced catalepsy, tremor, dystony, hyperkinesia, and stereotypy. Subjects produced anticipated responses before any stimulus. Incomplete and delayed responses were also produced. An interpretation in terms of akathisia and anhedonia has been suggested. Clozapine induced tremor, exploration, dystony, and hypersalivation. Subjects produced disinhibitory responses to the negative stimulus and incomplete responses but these latter were submitted to tolerance. The simultaneous presence of tranquilizing and disinhibitory effects has been reported on the clinical potential of clozapine both in cases of positive and negative schizophrenic symptomatologies.

Amineptine improves the performance of dogs in a complex temporal regulation schedule.

Amineptine is a tricyclic antidepressant with activating properties, that stimulates spontaneous locomotor activity in rodents and elevates mood in humans. It mainly inhibits dopamine uptake and weakly increased dopamine release. Formulating the hypothesis that this drug would decrease waiting capacity, we decided to test amineptine in a Differential Reinforcement of Response Duration schedule (DRRD 9 s Limited Hold 1.5 s) in the dog. The drug was administered orally at 2.5, 5.0, 7.5, 10 and 20 mg/kg, 1 h before the experimental session. Between 2.5 and 10 mg/kg, amineptine improved the performance by increasing the response (nonsignificantly) and reinforcement (significantly) rates and by increasing the peak of correct responses (significantly). The inverse effect was measured for the reinforcement rate (nonsignificantly) and for the peak of correct responses (significantly) at the dose of 20 mg/kg. These results were compared to those obtained with other classes of drugs, like neuroleptics, barbiturates or anxiolytics, that disturbed the performance, and particularly with low doses of neuroleptics, which also increase the dopamine release. The positive effects of amineptine on performance (2.5-10 mg/kg) were related to its inhibitory effect on dopamine uptake and discussed in terms of improved vigilance and attention, increase of waiting capacity, improved anticipation, and cognitive enhancement.

Transformation of a waiting schedule into a temporal regulation schedule (DRRD) by addition of external stimuli in the dog.

Waiting schedules do not impose temporal regulation but condition the animal to give the operant response during a given time. At the end of the required delay, a positive discriminative stimulus is presented. The suspension of the response while the discriminative stimulus is being given suspension of the response while the discriminative stimulus is being given is accompanied by reinforcement. The transformation of a waiting schedule into a temporal regulation schedule is generally achieved by suppressing the external facilitating factors or by physically modifying them. Our study reveals that a similar transformation can be achieved in the dog by the addition of a further stimulus. This stimulus, which is physically exactly the same as the excitatory stimulus and which punctuates the waiting period, is randomly introduced into the temporal delay. The absence of reinforcement in response to the added stimulus should force the animal to regulate its behavior in time and the additional negative discriminative stimulus favours the expression of the active nature of the inhibation. The results show that subjects can differentiate their response durations according to stimuli that only differ according to temporal location. Thus this pattern resembles a DRRD schedule. The peak of responses at the time of the inhibition stimulus reveals considerable behavioral conflict : either the response must be maintained or the inhibition suppressed. The positive or negative resolution of this conflict reveals noteworthy aspects of the behavioural inhibition process.

Comparative study of pirlindole, a selective RIMA, and its two enantiomers using biochemical and behavioural techniques.

The interaction with monoamine oxidase A (MAO-A) and B has been shown to be sensitive to the absolute configuration of molecules. Therefore, the aim of this study was to compare the effects of the racemic pirlindole (a selective and reversible MAO-A inhibitor) and its two enantiomers using biochemical techniques (in vitro and ex vivo determination of rat brain MAO-A and MAO-B activity) and behavioural models (forced swimming test and reserpine-induced hypothermia and palpebral ptosis test). In vitro, the MAO-A IC50 of (+/-)-pirlindole, R-(-)-pirlindole and S-(+)-pirlindole were 0.24, 0.43 and 0.18 microM, respectively. Ex vivo, their ID50 were 24.4, 37.8 and 18.7 mg/kg i.p. The differences between the three compounds were not significant, with a ratio between the two enantiomers [R-(-)/S-(+)] of 2.2 in vitro and 2.0 ex vivo. MAO-B was only slightly inhibited. In the forced swimming test and the reserpine-induced hypothermia and ptosis model, the three compounds had an antidepressant profile. In the forced swimming test, the minimal effective dose ratio between the R-(-) and the S-(+) was again around 2.0. The behavioural observations were thus clearly in accordance with the biochemical data.