Histamine H3 receptor antagonists: from target identification to drug leads.

The successful cloning and functional expression of the histamine H(3) receptor in the late 1990 s has greatly facilitated our efforts to identify small molecule, non-imidazole based compounds to permit the evaluation of H(3) antagonists in models of CNS disorders. High-throughput screening identified several series of lead compounds, including a series of imidazopyridines, which led to JNJ-6379490, a compound with high affinity for the human H(3) receptor. Analysis of structural features common to several series of non-imidazole H(3) receptor ligands resulted in a pharmacophore model. This model led to the design of JNJ-5207852, a diamine-based H(3) antagonist with good in vitro and in vivo efficacy but with an undesirable long half-life. However, further modifications of the template provided an understanding of the effect of structural modifications on pharmacokinetic properties, ultimately affording several additional series of compounds including JNJ-10181457, a compound with an improved pharmacokinetic profile. These compounds allowed in vivo pharmacological evaluation to show that H(3) antagonists promote wakefulness, but unlike modafinil and classical psychostimultants, they do not increase locomotor activity or produce any alteration of the EEG power spectral activity in rats. H(3) antagonists also increase extracellular acetylcholine and norepinephrine but not dopamine in rat frontal cortex and show efficacy in various models of learning-memory deficit. In addition, cFos immunoreactivity studies show H(3) antagonists activate neuronal cells in restricted rat brain regions in contrast to widespread activation after modafinil or amphetamine treatment. Therefore, H(3) antagonists are promising clinical candidates for the treatment of excessive day time sleepiness and/or cognitive disorders.

Acute wake-promoting actions of JNJ-5207852, a novel, diamine-based H3 antagonist.

1 1-[4-(3-piperidin-1-yl-propoxy)-benzyl]-piperidine (JNJ-5207852) is a novel, non-imidazole histamine H3 receptor antagonist, with high affinity at the rat (pKi=8.9) and human (pKi=9.24) H3 receptor. JNJ-5207852 is selective for the H3 receptor, with negligible binding to other receptors, transporters and ion channels at 1 microm. 2 JNJ-5207852 readily penetrates the brain tissue after subcutaneous (s.c.) administration, as determined by ex vivo autoradiography (ED50 of 0.13 mg kg(-1) in mice). In vitro autoradiography with 3H-JNJ-5207852 in mouse brain slices shows a binding pattern identical to that of 3H-R-alpha-methylhistamine, with high specific binding in the cortex, striatum and hypothalamus. No specific binding of 3H-JNJ-5207852 was observed in brains of H3 receptor knockout mice. 3 In mice and rats, JNJ-5207852 (1-10 mg kg(-1) s.c.) increases time spent awake and decreases REM sleep and slow-wave sleep, but fails to have an effect on wakefulness or sleep in H3 receptor knockout mice. No rebound hypersomnolence, as measured by slow-wave delta power, is observed. The wake-promoting effects of this H3 receptor antagonist are not associated with hypermotility. 4 A 4-week daily treatment of mice with JNJ-5207852 (10 mg kg(-1) i.p.) did not lead to a change in body weight, possibly due to the compound being a neutral antagonist at the H3 receptor. 5 JNJ-5207852 is extensively absorbed after oral administration and reaches high brain levels. 6 The data indicate that JNJ-5207852 is a novel, potent and selective H3 antagonist with good in vitro and in vivo efficacy, and confirm the wake-promoting effects of H3 receptor antagonists.

Current understanding of the circadian clock and the clinical implications for neurological disorders.

The changes in behavior that occur on a 24-hour basis to match the 24-hour changes in the physical environment due to the rotation of the earth on its axis are a hallmark of life on the planet Earth. The nervous system of both lower and higher organisms has evolved over millions of years to meet the demands of the dramatic changes in the physical environment that occur in relation to the changes in the light-dark cycle, optimizing the survival and reproductive success of the organism. During the past 50 years, it has been clearly established that the 24-hour nature of life was not simply a response to the 24-hour changes in the physical environment imposed by celestial mechanics, but instead was due to an internal time-keeping system in the brain. Many neurological disorders are associated with abnormal 24-hour rhythms, including the sleep-wake cycle. The recent discovery of the molecular basis of the neural clock in animals offers neurologists new avenues for studying the pathophysiology of neurological disorders.

Similar genetic mechanisms may underlie sleep-wake states in neonatal and adult rats.

Genetic differences in the characteristics of sleep-wake states in adult animals offer a potential window for examining how the neonatal and adult behavioural states are related to one another. Our recent finding that adult Wistar-Kyoto (WKY) rats show pronounced genetic differences in sleep-wake patterns relative to the Wistar (WIS) control strain led us to investigate the relationship between these behavioural states in neonates and adults in a longitudinal study in these two strains of rats. Similar pronounced differences in the sleep-wake states were observed between WKY and WIS rats in neonatal and in adult animals. At both ages, WKY rats spent more time in activesleep (AS) and rapid eye movement sleep (REMS) and less time in quiet sleep (QS) and non-REM sleep (NREMS) than WIS rats, and the sleep-wake states were more fragmented in neonatal and adult WKY rats. While it is not known how neonatal AS and QS are physiologically related to adult REMS and NREMS, respectively, the finding of similar differences in the amounts of sleep-wake states in neonatal and adult WKY and WIS rats argues strongly that at some level they are controlled by similar genetic as well as cellular/physiological mechanisms.

The selective neurokinin 1 receptor antagonist R116301 modulates photic responses of the hamster circadian system.

The recent development of selective NK(1) receptor antagonists that are active in vivo provides an important research tool to examine the role of substance P in the regulation of circadian rhythmicity. First, we tested whether R116301 [(2R-trans)-4-[1-[3,5-bis(trifluoromethyl)benzoyl]-2-(phenylmethyl)-4-piperidinyl]-N-(2,6-dimethylphenyl)-1-acetamide (S) hydroxybutanedioate], a new selective NK(1) antagonist, alters the phase-shifting effects of light. Hamsters housed in constant darkness were injected with different doses of R116301, just before being exposed to a light pulse during the subjective night. The results were compared with those obtained with the NK(1) antagonist L-760,735 [2-(R)-(1-(R)-3,5-bis(trifluoromethyl)phenyl)ethoxy)-4-(5-(dimethylaminomethyl)-1,2,3-trioazol-4-yl)methyl-3-(5)-phenyl)morpholine]. Second, the effects of the NK(1) antagonists R116301 or L-760,735 injected immediately after exposure to a light pulse were similarly determined. Third, we investigated whether R116301 or L-760,735 injected during the mid-subjective day or the late subjective night can phase-shift the circadian rhythm of locomotor activity in hamsters housed in constant light. Both compounds reduced, by more than 30%, the phase-advancing effects of a light pulse in hamsters otherwise maintained in constant darkness, only when the drugs were administered before the light pulse. Under constant light conditions, both NK(1) receptor antagonists induced significant phase-advances when injected during the subjective day, but not during the subjective night. The present results indicate that tachykinergic neurotransmission modulates the photic responses of the circadian system upstream of phase resetting mechanisms and suggest that an inhibition of the NK(1) receptor signals "darkness" to the circadian clock.

Role of serotonin in sleep mechanisms.

Since near the time of its discovery over 40 years ago, the serotonergic system has been implicated in the regulation of the sleep-wake cycle. While early studies indicate that serotonin (5-HT) was associated with the initiation and maintenance of sleep, later studies indicate that serotonergic neurons also play a role in inhibiting sleep. As reviewed in this paper, the complex effects of 5-HT in the regulation of sleep is due in part to the fact that 5-HT can act at different areas of the brain that have been associated with the control of sleep and wake. In addition, the recent discovery of multiple 5-HT receptors through the mammalian brain led to the finding that different 5-HT receptors are selectively involved in the regulation of the different sleep states.

Sleep in the Wistar-Kyoto rat, a putative genetic animal model for depression.

The Wistar-Kyoto (WKY) rat exhibits several behavioral and hormonal abnormalities often associated with depression. One of the hallmarks of depression consists of alterations in the sleep-wake cycle, particularly in rapid eye movement (REM) sleep. If the WKY rat is indeed an animal model for depression, we hypothesized that it should also show sleep abnormalities relative to the control strain, the Wistar (WIS) rat Under baseline conditions, WKY rats showed a 50% increase in total REM sleep time during the 12 h light phase and an increase in sleep fragmentation during both the light and dark phase. The WKY rats also exhibited lower EEG power densities over the entire frequency range (0.2-25.0 Hz) during REM sleep. After a 6 h sleep deprivation, the REM sleep rebound was more pronounced during the dark but not the light phase in the WKY rats. Since the WKY rat represents a genetic model for depression with altered EEG sleep patterns, this strain may be particularly useful for investigating the relationship between depression and sleep abnormalities.

High corticosterone levels in prenatally stressed rats predict persistent paradoxical sleep alterations.

Prenatal stress predisposes rats to long-lasting disturbances that persist throughout adulthood (e.g., high anxiety, dysfunction of the hypothalamo-pituitary-adrenal axis, and abnormal circadian timing). These disturbances parallel to a large extent those found in depressed patients, in which hypercortisolemia and sleep alterations may be related to stress-inducing events. We studied sleep-wake parameters in control and prenatally stressed adult rats (3-4 months old) and examined possible relationships with their corticosterone levels (determined at 2 months of age). Under baseline conditions, prenatally stressed rats showed increased amounts of paradoxical sleep, positively correlated to plasma corticosterone levels. Other changes include increased sleep fragmentation, total light slow-wave sleep time, and a slight decrease in the percentage of deep slow-wave sleep relative to total sleep time. During recovery sleep from acute restraint stress, all sleep changes persisted and were correlated with stress-induced corticosterone secretion. High corticosterone levels under baseline conditions as well as an acute stress challenge may thus predict long-term sleep-wake alterations in rats. Taken together with other behavioral and hormonal abnormalities in prenatally stressed animals, the pronounced changes in sleep-wake parameters that are similar to those found in depressed patients suggest that prenatal stress may be a useful animal model of depression.

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 alpha-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.

Functional activity of 5-HT2 receptors in the modulation of the sleep/wakefulness states.

In the light of recent pharmacological investigations using agonists and antagonists that have potent actions on 5-hydroxytryptamine-2 (5-HT2) receptors, the possible functional role of 5-HT2 receptors in the modulation of the sleep/wakefulness states was examined. Data obtained from animals and from clinical studies suggest that serotonin may exert an inhibitory control on deep slow-wave sleep (SWS) through 5-HT2 receptors. In further investigations, the existence of a diurnal variation in the functional activity of 5-HT2 receptors, that depends on the day/night cycle and/or the melatonin rhythm, was revealed. Questions remain with regard to the physiological significance, of the 5-HT2 receptor-mediated deep SWS regulation, the anatomical site(s) of 5-HT2 receptors involved in this regulation and the mechanism underlying diurnal fluctuations in the functional activity of 5-HT2 receptors.

Effects of ritanserin and chlordiazepoxide on sleep-wakefulness alterations in rats following chronic cocaine treatment.

The effects of ritanserin, a 5-hydroxytryptamine-2 (5-HT2) receptor antagonist, and chlordiazepoxide, a benzodiazepine agonist, on sleep-wakefulness disturbances in rats after acute administration of cocaine and after discontinuation of chronic cocaine treatment were examined. Intraperitoneal (IP) injection of chlordiazepoxide (10 mg/kg) but not ritanserin (0.63 mg/kg) prevented the increase of wakefulness (W) and the reduction of light slow wave sleep (SWS1) and deep slow wave sleep (SWS2) induced by an acute injection of cocaine (20 mg/kg IP). Daily injection of cocaine (20 mg/kg for 5 days, then 30 mg/kg for 5 days IP) at the onset of the light phase elicited an increase of W and a concomitant decrease of SWS1, SWS2 and paradoxical sleep (PS) in the light phase, followed by a rebound in SWS2 and PS in the subsequent dark phase. Following cocaine discontinuation, the circadian distribution of sleep-wakefulness states remained disturbed in saline-treated rats for at least 5 days. Both ritanserin (0.63 mg/kg IP/day) and chlordiazepoxide (10 mg/kg IP/day) reduced the alteration in the distribution of W and SWS2 throughout the light-dark cycle from the first day of administration on, but failed to prevent PS alterations. The mechanisms by which both compounds exert their effect are probably quite different. For chlordiazepoxide sedative and sleep-inducing properties probably play a major role. In contrast, for ritanserin SWS2-increasing properties and its ability to reverse preference for drugs of abuse without inducing aversion might be key factors.

Sleep-wakefulness patterns in the helpless rat.

Sleep-wakefulness patterns were analyzed during a 15-day period in the rat, in relation to induction of helplessness. After a session of inescapable electric footshocks, rats did exhibit escape deficits in avoidance conditioning as classically described, and their spontaneous sleep-wakefulness patterns were not different from those of controls. However, reduced PS latency and increased PS amounts were observed in the helpless group after shuttle-box sessions, especially during the initial period after the induction of helplessness. Such modifications of PS latency and PS amounts are evocative of the sleep impairments classically observed in endogenous depression.

Differential effects of the new antipsychotic risperidone on sleep and wakefulness in the rat.

The effects of risperidone, a new antipsychotic with potent 5-hydroxytryptamine2 (5-HT2) and dopamine-D2 (DA-D2) antagonistic properties, were studied on sleep-wakefulness patterns in rats. Administration of low doses (0.01-0.16 mg/kg i.p.) resulted in a significant increase of deep slow wave sleep (SWS2) and a decrease of wakefulness (W) and light slow wave sleep (SWS1). High doses (0.63-2.5 mg/kg) produced opposite effects. Paradoxical sleep (PS) was significantly reduced over the dose range tested. The increase of SWS2 after low doses of risperidone could be related to a predominant and potent 5-HT2 receptor blocking activity.

The light-dark cycle modulates the effects of ritanserin on sleep-wakefulness patterns in the rat.

The 5-HT2 receptor antagonist ritanserin (0.63 mg/kg IP) produced differential effects on sleep-wakefulness patterns in rats when administered during the light or dark period: an increase in the duration of deep slow wave sleep at the expense of light slow wave sleep, paradoxical sleep and wakefulness when injected during the light period, and no major sleep alteration when given at dark onset. Since circadian variations in serotonin receptor density might modulate the sleep response, we examined the effects of ritanserin on sleep in rats exposed to continuous light for 10 days, and whether 5-HT2 receptors were affected in separate groups of rats exposed to similar conditions. No significant changes in the KD- and Bmax -values of various receptors were found. However, ritanserin produced greater effects in continuous light conditions than when given during the light period in the 12-hr light-dark condition. This suggests a possible role of 5-HT2 receptors in the organization of sleep when the environmental photoperiod is disturbed.

Melatonin modulates the sensitivity of 5-hydroxytryptamine-2 receptor-mediated sleep-wakefulness regulation in the rat.

The interaction between melatonin and two 5-hydroxytryptamine (5-HT2) compounds was studied on sleep patterns in rats. Administration of the 5-HT2 receptor antagonist ritanserin (0.63 mg/kg, i.p.) resulted in a significant increase of deep slow wave sleep (SWS2) and a decrease of paradoxical sleep (PS). The 5-HT2 receptor agonist DOM (1-(2,5-dimethoxy-4-methylphenyl)-2-aminopropane) (0.63 mg/kg, i.p.) produced a significant reduction of both SWS2 and PS. Melatonin (1 mg/kg, i.p.) alone did not alter sleep but counteracted the sleep effects induced by ritanserin as well as DOM. It is proposed that melatonin modulates the sensitivity of 5-HT2 receptor-mediated sleep response probably by an indirect route.

Functional role of 5-HT2 receptors in the regulation of sleep and wakefulness in the rat.

Recently developed agents specifically acting on different 5-hydroxytryptamine (5-HT) receptor populations were used to analyze the functional role of 5-HT2 receptor subtypes in the sleep-wakefulness cycle of the rat. The 5-HT2 receptor antagonist ritanserin injected intraperitoneally (IP) (0.04-2.5 mg/kg) induced an increase in deep slow wave sleep (SWS2) duration at the expense of wakefulness (W), light slow wave sleep (SWS1) and paradoxical sleep (PS). The stimulation of 5-HT2 receptors by 1-(2,5-dimethoxy-4-methylphenyl)-2-aminopropane (DOM) produced a dose-related increase in W and a dose-dependent decrease in both SWS2 and PS. Pretreatment with ritanserin (0.16-2.5 mg/kg) or with cinanserin (2.5-5 mg/kg), another 5-HT2 receptor antagonist, dose-dependently reversed the W enhancement and the SWS2 deficit produced by DOM, but not the PS deficit. Sleep-wakefulness alterations (increase in W and SWS1 combined with a suppression of SWS2 and PS) observed after IP injection of two putative 5-HT1 receptor agonists, 8-hydroxy-2-(di-n-propylamino) tetralin (8-OH-DPAT) (2.5 mg/kg) and 5-methoxy-3-(1,2,3,6-tetrahydro-4-pyridinyl)-1H-indole (RU 24969) (0.63 mg/kg), were not modified by ritanserin pretreatment (0.16-2.5 mg/kg). These results further support the hypothesis that the serotonergic system plays an active role in the regulation of the sleep-wakefulness cycle in the rat and that 5-HT2 receptors are involved in this action. In addition, it is suggested that 5-HT1 receptor subtypes are unlikely to interact with 5-HT2 receptors in the sleep-wakefulness modulation mediated through 5-HT2 receptors.

Time course of night sleep EEG in the first year of life: a description based on automatic analysis.

The aim of this study is to describe the time course of night sleep in the first year of life. Forty-eight infants aged between 1 and 54 weeks were polygraphically recorded for 1 night. The central occipital EEG derivation was processed with a lab computer in order to obtain every 30 sec an EEG parameter value. The parameter is based on the joint frequency-amplitude distribution of the EEG and displays fluctuations between 2 extreme levels, high voltage low frequency (HVLF) and low voltage high frequency (LVHF). The range of the fluctuations between HVLF and LVHF increases from the period of 1-6 weeks to the period of 7-14 weeks. A further increase of the parameter range occurs after 24 weeks, which remains restricted to the first half of the night. The recurrence time of LVHF and HVLF episodes (possibly corresponding to quiet sleep paradoxical sleep cycles) is about 56 min and does not change with age in the first year of life.

Effects of lithium on dopamine behavioural supersensitivity induced by rapid eye movement sleep deprivation.

The effects of lithium on the potentiation of d-amphetamine-induced hyperlocomotion were evaluated in rapid eye movement (REM) sleep deprived rats. Under control conditions, pretreatment with lithium during 7 days did not modify the hyperlocomotion produced by d-amphetamine. REM sleep deprivation induced a pronounced potentiation of the locomotor response to d-amphetamine. In a stress control group this potentiation also occurred, but to a lesser degree than in the REM sleep deprived group. Lithium pretreatment prevented the increased response to d-amphetamine in both REM sleep deprived and stress control animals. The effects of lithium in REM sleep deprived rats are in accordance with reports that lithium is able to prevent the development of dopamine receptor supersensitivity. However, it cannot be excluded that in both REM sleep deprived and stress control groups the increased response to d-amphetamine is related to noradrenergic changes and/or noradrenergic-dopaminergic interactions. REM sleep deprivation seems to be an interesting model to study the underlying mechanisms of manic-depressive illness.