Benzodiazepine Receptor Full Agonist Research Articles

Potentiating Effect of Glabridin on GABAA Receptor-Mediated Responses in Dorsal Raphe Neurons

Flavonoid-rich ethanol extracts of licorice root have sedative and anxiolytic effects. Glabridin is a major flavonoid component from licorice which we evaluated by examining GABA responses in acutely isolated dorsal raphe neurons of the rat. Neurons were recorded with patch-clamp methods at a holding potential of - 50 mV. Glabridin potentiated GABA-induced responses by positively modulating GABAA receptor responses with different concentration range. GABA (2 × 10(-6) M)-evoked currents were potentiated in a stepwise pattern increasing glabridin concentration. Between 10(-12) and 10(-8) M glabridin increased GABA responses by about 140 % of the control. At concentrations above 10(-7) M, a much larger, dose-dependent potentiation occurred before reaching a plateau at 3 × 10-6 M glabridin. A hypnotic drug, zolpidem, also induced biphasic concentration-potentiation relationship. The glabridin potentiation ratio was 2.2 times larger than the maximum potentiation to the benzodiazepine receptor full agonist diazepam. Benzodiazepine receptor antagonist, flumazenil (3 × 10(-7) M), failed to inhibit glabridin (3 × 10(-7) M)-induced potentiation. This result implies that glabridin may exhibit sedative and hypnotic effects by potentiating GABAergic inhibition in dorsal raphe neurons by GABAA receptor actions.

Reinforcing effects of compounds lacking intrinsic efficacy at α1 subunit-containing GABAA receptor subtypes in midazolam- but not cocaine-experienced rhesus monkeys.

Benzodiazepines are prescribed widely but their utility is limited by unwanted side effects, including abuse potential. The mechanisms underlying the abuse-related effects of benzodiazepines are not well understood, although α1 subunit-containing GABAA receptors have been proposed to have a critical role. Here, we examine the reinforcing effects of several compounds that vary with respect to intrinsic efficacy at α2, α3, and α5 subunit-containing GABAA receptors but lack efficacy at α1 subunit-containing GABAA receptors ('α1-sparing compounds'): MRK-623 (functional selectivity for α2/α3 subunit-containing receptors), TPA023B (functional selectivity for α2/α3/α5 subunit-containing receptors), and TP003 (functional selectivity for α3 subunit-containing receptors). The reinforcing effects of the α1-sparing compounds were compared with those of the non-selective benzodiazepine receptor partial agonist MRK-696, and non-selective benzodiazepine receptor full agonists, midazolam and lorazepam, in rhesus monkeys trained to self-administer midazolam or cocaine, under a progressive-ratio schedule of intravenous (i.v.) drug injection. The α1-sparing compounds were self-administered significantly above vehicle levels in monkeys maintained under a midazolam baseline, but not under a cocaine baseline over the dose ranges tested. Importantly, TP003 had significant reinforcing effects, albeit at lower levels of self-administration than non-selective benzodiazepine receptor agonists. Together, these results suggest that α1 subunit-containing GABAA receptors may have a role in the reinforcing effects of benzodiazepine-type compounds in monkeys with a history of stimulant self-administration, whereas α3 subunit-containing GABAA receptors may be important mediators of the reinforcing effects of benzodiazepine-type compounds in animals with a history of sedative-anxiolytic/benzodiazepine self-administration.

Differential effects of lorazepam on sleep and activity in C57BL/6J and BALB/cJ strain mice

Compared to C57BL/6 mice, BALB/c mice exhibit greater 'anxiousness' on behavioural tests of anxiety, and can show significantly longer sleep disruptions after exposure to anxiogenic situations. Relative to C57BL/6 mice, BALB/c mice also have reduced benzodiazepine (BZ) receptor densities in the brain and fivefold less BZ receptor density in the amygdala, a region important in anxiety and in the control of arousal. Lorazepam is a BZ receptor full agonist and has been used to treat both anxiety and insomnia. Differences between C57BL/6 and BALB/c mice raise the question of whether BZ agonists would impact sleep and activity differentially in the two strains. We examined the effects of two doses of lorazepam (0.5 and 1.5 mg kg(-1)) or saline alone (0.2 mL) on sleep and activity in C57BL/6 (n = 8) and BALB/c (n = 7) mice. Compared to saline, both doses of lorazepam significantly increased non-rapid eye movement (NREM) and reduced activity in both strains. In C57BL/6 mice, rapid eye movement (REM) was increased at both doses. In BALB/c mice, the 0.5 mg kg(-1) dose had no significant influence on REM, whereas REM was reduced significantly after the 1.5 mg kg(-1) dose. The results demonstrate significant differences between C57BL/6 and BALB/c mice in the effects of lorazepam on REM, whereas the effects on NREM and activity were similar. Strain differences in the number of BZ receptors in the amygdala, but not other brain regions, suggests possible site specificity in the effects of lorazepam on REM. These differences in BZ-binding sites in the amygdala could be a significant factor in differences in the sleep response between C57 and BALB/c mice.

Role of glutamate ionotropic and benzodiazepine receptors in the ventromedial hypothalamic nucleus on anxiety

The dorsomedial part of the ventromedial hypothalamic nuclei (VMHdm) has been related to the modulation of defensive behavior in mammals. The objective of the present study was to test the hypothesis that administration into the VMHdm of midazolam, a benzodiazepine receptor full agonist, or AP7, a glutamate NMDA receptor antagonist, would produce anxiolytic effects in the elevated plus-maze (EPM) or the Vogel's punished licking tests. Male Wistar rats with unilateral cannulae aimed at the VMHdm received intra-cerebral injections of midazolam (15–60 nmol/0.25 μL), AP7 (0.2–2 nmol/0.3μL) or saline and were submitted to the behavioral tests. Midazolam (30 nmol) increased the percentage of time spent in open arms of the EPM. AP7, on the other hand, decreased open and enclosed arm exploration. In the Vogel test, however, both midazolam (30–60 nmol) and AP7 increased the number of punished licks. Histological control experiments found no significant effects when the drugs were injected into the nearby lateral hypothalamic area. These results suggest that facilitation of gabaergic or antagonism of glutamatergic neurotransmission in the VMHdm can produce anxiolytic-like effects.

The open field as a paradigm to measure the effects of drugs on anxiety-like behaviors: a review

The open field is a very popular animal model of anxiety-like behavior. An overview of the literature on the action elicited by effective or putative anxiolytics in animal subjected to this procedure indicates that classical treatments such as benzodiazepine receptor full agonists or 5-HT 1A receptor full or partial agonists elicit an anxiolytic-like effect in this procedure in most cases (approximately 2/3). However, compounds (triazolobenzodiazepines such as adinazolam and alprazolam, selective serotonin reuptake inhibitors) that have a different spectrum of therapeutic efficacy in anxiety disorders such as panic attacks, generalized anxiety disorder or obsessive-compulsive disorder were poorly effective as anxiolytics in the open field test, suggesting that this paradigm may not model features of anxiety disorders. The procedure is also relevant for the study of compounds endowed with anxiogenic effects, as such effects were detected after treatments with benzodiazepine receptor inverse agonists or with corticotropin releasing factor (CRF) receptor agonists.

Mechanism-based pharmacodynamic modeling of the interaction of midazolam, bretazenil, and zolpidem with ethanol.

The pharmacokinetic and pharmacodynamic interactions of ethanol with the full benzodiazepine agonist midazolam, the partial agonist bretazenil and the benzodiazepine BZ1 receptor subtype selective agonist zolpidem have been determined in the rat in vivo, using an integrated pharmacokinetic-pharmacodynamic approach. Ethanol was administered as a constant rate infusion resulting in constant plasma concentrations of 0.5 g/l. The pharmacokinetics and pharmacodynamics of midazolam, bretazenil, and zolpidem were determined following an intravenous infusion of 5.0, 2.5, and 18 mg/kg respectively. The amplitude in the 11.5-30 Hz frequency band of the EEG was used as measure of the pharmacological effect. For each of the benzodiazepines the concentration-EEG effect relationship could be described by the sigmoid Emax pharmacodynamic model. Significant differences in both EC50 and Emax were observed. The values of the EC50 were 76 +/- 11, 12 +/- 3, and 512 +/- 116 ng/ml for midazolam, bretazenil, and zolpidem respectively. The values of the Emax were 113 +/- 9, 44 +/- 3, and 175 +/- 10 microV/s. In the presence of ethanol the values of the EC50 of midazolam and zolpidem were reduced to approximately 50% of the original value. The values for Emax and Hill-factor were unchanged Due to a large interindividual variability no significant change in EC50 was observed for bretazenil. Analysis of the data on basis of a mechanism-based model showed only a decrease in the apparent affinity constant KPD for all three drugs, indicating that changes in EC50 can be explained entirely by a change in the apparent affinity constant KPD without concomitant changes in the efficacy parameter ePD and the stimulus-effect relationship. The findings of this study show that the pharmacodynamic interactions with a low dose of ethanol in vivo are qualitatively and quantitatively similar for benzodiazepine receptor full agonists, partial agonists, and benzodiazepine BZ1 receptor subtype selective agonists. This interaction can be explained entirely by a change in the affinity of the biological system for each benzodiazepine.

ベンゾジアゼピン受容体作動性抗不安薬の青斑核 大脳皮質前頭前野ノルエピネフリン神経系への作用

In this study the effects of benzodiazepine receptor agonistic anxiolytics were examined on the locus coeruleus (LC)-prefrontal cortex (PFC) norepinephrine neurons of freely moving rats, and on stress-induced increase in extracellular norepinephrine in the PFC. While benzodiazepine receptor agonistic anxiolytics were infused via a microdialysis probe in the vicinity of the right LC, the extracellular content of norepinephrine was recorded in the ipsilateral PFC with another microdialysis probe. The differential types of benzodiazepine receptor agonistic anxiolytics midazolam (benzodiazepine receptor full agonist, 10-4M and 5 × 10-4M) and Y-23684 (benzodiazepine receptor partial agonist, 10-4M and 5 × 10-4M) were used. While infusions of Y-23684 (5 × 10-4M) decreased extracelllar norepinephrine in the PFC, midazolam was not effective for the basal values. Next, rats were gently handled for 10 min during infusion of these drugs. The infusion of midazolam (5 × 10-4M) and Y-23684 (5 × 10-4M) inhibited the handling stress-induced increase in extracellular norepinephrine in the PFC. Benzodiazepine receptor antagonist flumazenil (10-4M) attenuated these effects. These indicate that benzodiazepine receptors agonistic anxiolytics play a major role in suppressing stress responses, by acting via benzodiazepine receptors.

Comparative neuroprotective properties of the benzodiazepine receptor full agonist diazepam and the partial agonist PNU-101017 in the gerbil forebrain ischemia model

Recent studies have demonstrated the neuroprotective properties of the novel imidazoquinoline benzodiazepine receptor partial agonist, PNU-101017, in the gerbil forebrain ischemia model. The compound effectively reduces delayed post-ischemic (5 min bilateral carotid occlusion) hippocampal CA 1 neuronal degeneration even when its administration is witheld until 4 h after reperfusion and the effect is unrelated to hypothermia. The purpose of the present study was to determine the comparative abilities of PNU-101017 versus the full agonist diazepam to attenuate post-ischemic CA 1 damage. Male gerbils were treated either 30 min before ischemia induction or immediately after reperfusion with an initial dose of PNU-101017 (30 mg/kg i.p.) or diazepam (10 mg/kg i.p.) with a second dose being given at 2 h after reperfusion. Possible hypothermic effects of either compound were prevented by external heating. In vehicle (0.05 N HCl)-treated gerbils, the loss of hippocampal CA 1 neurons at 5 days was 85%. PNU-101017 pretreatment reduced the loss to 50% ( p<0.05 vs. vehicle) whereas pretreatment with diazepam attenuated damage to only 17% ( p<0.001 vs. vehicle). Delaying treatment with PNU-101017 until just after reperfusion still resulted in a reduction in CA 1 degeneration statistically that was indistinguishable from that seen with pretreatment. In contrast, diazepam post-treatment did not significantly decrease CA 1 neuronal loss. These results suggest that a benzodiazepine receptor partial agonist may have greater neuroprotective practicality than a full agonist for the treatment of global cerebral ischemia. The mechanistic basis for this difference may relate to the partially pro-excitatory neuronal response to endogenous GABA before and after neuronal insult.

Further pharmacological validation of the BALB/c neophobia in the free exploratory paradigm as an animal model of trait anxiety.

The present experiments were aimed at investigating the ability of established or putative anxiolytics to reduce the neophobia exhibited by BALB/c mice in the free exploratory paradigm. Results confirm the anxiolytic effects of the benzodiazepine receptor full agonist chlordiazepoxide (2.5-7.5 mg/kg), of meprobamate (15-60 mg/kg) and of ethanol (0.5-1.5 g/kg) and extend the pharmacological action of these compounds to a test situation devoid of anxiogenic components, that is to trait anxiety. The non-competitive NMDA antagonist MK 801 (0.04-0.16 mg/kg) elicited very similar behavioural effects. However, the alpha 2-adrenoceptor antagonists yohimbine (0.5-2 mg/kg) and idazoxan (0.3-2.7 mg/kg), the barbiturate pentobarbital (3.75-30 mg/kg), the mixed 5HT2 receptor antagonist ritanserin (0.25-4 mg/kg) and the D2 dopaminergic antagonist sulpiride (8-32 mg/kg) failed to decrease neophobia in BALB/c mice. The discussion focuses on the adequacy of this animal model of human pathology. The BALB/c neophobia may not model panic attacks because of the absence of worsening by the panic-provoking agent yohimbine and the lack of attenuation by CCK-B receptor antagonists. Because of its chronicity, this paradigm may model generalized anxiety, a pathology that has been suggested to overlap trait anxiety.

Enhancement by flumazenil of dopamine release in the nucleus accumbens of rats repeatedly exposed to diazepam or imidazenil.

The effect of long-term treatment (three times daily for 3 weeks) with a behaviorally relevant dose of the benzodiazepine receptor partial agonist imidazenil (0.5 mg/kg, IP) on basal dopamine release in the nucleus accumbens of freely moving rats was compared with that of diazepam (3 mg/kg, IP), a benzodiazepine receptor full agonist. Challenge doses of imidazenil and diazepam decreased the extracellular dopamine concentration in the nucleus accumbens by approximately the same extent in animals repeatedly exposed to vehicle or to the respective drug. Moreover, the abrupt discontinuation of long-term treatment with diazepam or imidazenil failed to affect basal dopamine release in this brain area during the first 5 days of withdrawal. In contrast, administration of the benzodiazepine receptor antagonist flumazenil (4 mg/kg, IP) elicited a marked increase (95 or 60%) in dopamine release in the nucleus accumbens 6 h after withdrawal of diazepam or imidazenil, respectively. Flumazenil induced a similar but smaller effect (50% increase) 5 days after diazepam withdrawal but had no effect 5 days after discontinuation of imidazenil treatment. The results support an involvement of the mesoaccumbens dopaminergic neurons in the withdrawal syndrome precipitated by flumazenil and allow further differentiation of benzodiazepine receptor partial and full agonists with respect to dependence liability of dopaminergic neurons in the nucleus accumbens.

Blockade of Behavioral Effects of Bretazenil by Flumazenil and ZK 93,426 in Pigeons

Benzodiazepine receptor partial agonists manifest full efficacy in preclinical tests of anxiolytic drug action but do not fully reproduce the discriminative stimulus effects of benzodiazepine receptor full agonists in pigeons. The partial agonist, bretazenil, binds to both diazepam-sensitive and diazepam-insensitive GABA A receptors. Previous studies have suggested a role for each of these receptor populations in some behavioral effects of bretazenil in pigeons. A possible role for these receptor subtypes in the behavioral effects of bretazenil was further investigated through drug interaction studies with the benzodiazepine receptor antagonists, flumazenil and ZK 93,426. Whereas flumazenil binds with high affinity to both receptor isoforms, ZK 93,426 binds preferentially to diazepam-sensitive binding sites. Bretazenil markedly increased punished responding of pigeons without significantly affecting nonpunished responding. In pigeons discriminating the full benzodiazepine receptor agonist, midazolam, from saline, bretazenil produced only 60–75% maximal effect. Flumazenil and ZK 93,426 neither increased punished responding nor substituted for midazolam, but dose-dependently blocked the effects of bretazenil on punished responding. Flumazenil also dose-dependently blocked the effects of bretazenil in midazolam-discriminating pigeons, whereas ZK 93,426 only attenuated this effect. These results indicate that bretazenil's actions as a partial agonist at diazepam-sensitive benzodiazepine receptors mediate increases in punished responding and substitution for the discriminiative stimulus effects of midazolam in pigeons. The differences in the effects of flumazenil and ZK 93,426 on the discriminative stimulus effects of bretazenil suggest a potential contribution of diazepam-insensitive sites to this behavioral effect.

A nonbenzodiazepine partial agonist. s-(+)-DN-2327, hab minimal physical dependence-producing liability, but shows cross-dependence on barbital in rats

1. 1. Physical dependence and cross-physical dependence on barbital of the benzodiazepine receptor partial agonist S-(+)-DN-2327 and the benzodiazepine receptor full agonist diazepam were compared in male Fischer 344 rats. 2. 2. In the physical dependence study, rats were treated with S-(+)-DN-2327 (30, 100, 300 and 1000 mg/kg/day) or diazepam (30, 100 and 300 mg/kg/day) for 4 weeks by the drug admixed with food method. After stopping the treatment, the body weight and food consumption in the diazepam 100 and 300 mg/kg groups tended to decrease or decreased to values lower than those in the control group, whereas these parameters in the S-(+)-DN-2327 30,100 and 300 mg/kg groups were comparable to the control group values. 3. 3. In the cross-dependence study, rats were treated with increasing doses of barbital by admixing the drug with food for 4 weeks, after which the diet admixed with barbital was replaced by basal diet alone or admixed with S-(+)-DN-2327 or diazepam (target doses: 100 and 300 mg/kg/day for each compound). During the substitution period, the decreases in body weight and food consumption in both S-(+)-DN-2327 and both diazepam groups were suppressed compared with those in the basal diet group. 4. 4. These results suggest that S-(+)-DN-2327 possesses minimal physical dependenceproducing liability, but shows cross-dependence on barbital, as do benzodiazepine receptor full agonists.

Evaluation of the dependence liability of quinolizinones acting as partial agonists at the benzodiazepine receptor

AbstractThe quinolizinones Ro 19‐5663, Ro 19‐5686 and Ro 41‐3696 bind with high affinity to the benzodiazepine receptor (BZR) in vitro in the rat brain. These compounds exhibit anxiolytic‐like effects in an operant punishment task in mice and rats and anticonvulsant effects by fully protecting against pentylenetetrazol‐induced tonic seizures in mice and rats and fully protecting against audiogenic seizures in genetically susceptible mice. However, they produced at most only minimal motor impairment in the rotarod task in rodents. These compounds attenuated flunitrazepaminduced sleep in squirrel monkeys, thus antagonizing the effects of a BZR full agonist. In a precipitated withdrawal paradigm, chronic oral treatment in seizure‐prone young DBA/2J mice or adult squirrel monkeys was followed by iv challenge with the BZR antagonist sarmazenil. Following the completion of a chronic treatment regiment with high doses of these quinolizinones, no signs of precipitated withdrawal were observed in mice and only weak signs of sarmazenil‐induced withdrawal were observed in monkeys. In contrast, sarmazenil challenge after chronic treatment with the BZR full agonists triazolam and alprazolam elicited marked withdrawal reactions in mice and monkeys, respectively. In sum, these quinolizinones are BZR partial agonists which offer potential anxiolytic and anticonvulsant efficacy concomitant to reduced adverse effects, including a clear reduction of dependence liability, in comparison to BZR full agonists. © 1995 Wiley‐Liss, Inc.

Antagonism of isoniazid-induced convulsions by abecarnil in mice tolerant to diazepam

The ability of the benzodiazepine receptor full agonist diazepam, the selective agonist abecarnil, and the partial agonist imidazenil to antagonize convulsions induced by isoniazid (200 mg/kg, SC) was studied in mice chronically treated with diazepam (3 mg/ kg, IP, three times daily) or abecarnil (0.1 or 1 mg/kg, IP, three times daily or 6 mg/kg, SC, daily). Diazepam induced tolerance to its own anticonvulsant effect. In contrast, chronic treatment with abecarnil failed to induce tolerance to its own anticonvulsant activity. Animals treated with abecarnil at 0.1 mg/kg developed cross-tolerance to imidazenil, whereas those treated with 1 mg/kg became less sensitive to diazepam. Mice chronically treated with abecarnil at 6 mg/kg showed almost complete tolerance to diazepam. Abecarnil was able to antagonize the convulsions elicited by isoniazid in diazepam-tolerant mice. These data indicate that chronic administration of abecarnil, unlike that of classical benzodiazepines, does not induce tolerance to its anticonvulsant effect, and that abecarnil overcomes tolerance induced by long-term treatment with the full agonist diazepam.

Enhancement of acetylcholine release by flumazenil in the hippocampus of rats chronically treated with diazepam but not with imidazenil or abecarnil.

The effects of long-term treatment (three times a day for 3 weeks) with pharmacologically active doses of the novel anxiolytics and anticovulsants abecarnil (0.5 mg/kg, IP) and imidazenil (0.5 mg/kg, IP) on basal hippocampal acetylcholine release in freely moving rats were compared with those of diazepam (3 mg/kg, IP). Challenge doses of diazepam, abecarnil, and imidazenil decreased the extracellular acetyl-choline concentration in the hippocampus by the same extent in animals chronically treated with the respective drug or vehicle. Moreover, the abrupt discontinuation of long-term treatment with diazepam, abecarnil, or imidazenil failed to affect hippocampal acetylcholine release during the first 5 days of withdrawal. In contrast, the acute administration of the benzodiazepine receptor antagonist flumazenil (1 mg/kg, IP) 2 days after diazepam withdrawal elicited a marked increase (65%) in acetylcholine release in the hippocampus. Flumazenil failed to induce the same effect 5 days after diazepam withdrawal or 2 or 5 days after discontinuation of long-term treatment with abecarnil or imidazenil. These results indicate that (i) the inhibitory effects of full (diazepam), partial (imidazenil), and selective (abecarnil) benzodiazepine receptor agonists on acetylcholine output in rat hippocampus are not affected by repeated drug administration; (ii) discontinuation of long-term treatment with each type of agonist does not affect hippocampal cholinergic mechanisms; and (iii) flumazenil increases acetylcholine release only in the hippocampus of rats chronically treated with diazepam. Together, these data further differentiate the pharmacology of benzodiazepine receptor full agonists from that of partial and selective agonists.

Bidirectional modulation of cortical acetylcholine efflux by infusion of benzodiazepine receptor ligands into the basal forebrain

In a previous in vivo microdialysis study in rats, it was found that cortical acetylcholine (ACh) efflux was reliably increased by a multimodal appetitive stimulus (onset of darkness with presentation of palatable food). Furthermore, this stimulated ACh efflux was significantly enhanced by systemic administration of a benzodiazepine receptor (BZR) weak inverse agonist and significantly reduced by a BZR full agonist. These effects contrasted with the minimal effects of BZR ligands on basal cortical ACh efflux in resting animals. The aim of the present study was to determine whether this modulation of stimulated cortical ACh efflux by BZR ligands was mediated within the basal forebrain. ACh efflux, measured with in vivo microdialysis, was stimulated by onset of darkness, an event which predicted delivery of palatable food. The BZR full inverse agonist, β-CCM (3.0 μg/hemisphere) or the full agonist chlordiazepoxide (40.0 μg/hemisphere) was infused into the basal forebrain just prior to the darkness/food stimulus. Similar to previous results with systemic administration, the BZR full inverse agonist enhanced, while the full agonist reduced, stimulated cortical ACh efflux. These results demonstrate that the action of BZR ligands in the basal forebrain is sufficient for their modulation of cortical ACh release.

Characterization of novel ligands for wild-type and natural mutant diazepam-insensitive benzodiazepine receptors

A series of benzodiazepine receptor ligands with different chemical structures were evaluated for their affinities at diazepam-sensitive and diazepam-insensitive binding sites for [ 3H]Ro 15-4513 (ethyl-8-azido-5,60dihydro-5-methyl-6-oxo-4H-imidazo-[1,5a][1,4]benzodiazepine-3-carboxylate) in cerebellar GABA A receptors. Rats of Wistar strain and of alcohol-sensitive (ANT) and alcohol-insensitive (AT) lines were used. The ANT rats possess a single point mutation in their GABA A receptor α6 subunit, which makes their diazepam-insensitive sites sensitive to benzodiazepine agonists, unlike those of AT and Wistar rats. All compounds evaluated displayed high-affinity binding to diazepam-sensitive sites ( K i < 50 nM). In contrast, a wider range of affinities were observed at diazepam-insensitive sites which depended upon the basic structure and substitutions. The 7- and 8-halogen substituted imidazobenzodiazepines and 12-halogen substituted diimidazoquinazolines displayed the highest affinities ( K i < 15 nM), while intermediate to low affinities (100 < K i < 4000 nM) were displayed by imidazoquinazolines, thienopyrimidines, one oxoimidazoquinoxaline, and some cyclopyrrolones. The imidazoquinoxalines evaluated displayed the lowest affinity ( K i > 10000 nM). The oxoimidazoquinoxaline, 6-chloro-3-(5-cyclopropyl-1,2,4-oxadiazol-3-yl)-4,5-dihydro-5-isopropyl-4-oxo-imidazol[1,5-a]quinoxaline (NNC 14-0578) and suriclone represent the first benzodiazepine receptor full agonists to bind with relatively high affinity ( K i ∼ 100 nM) to diazepam-insensitive sites. The 5 position substituted methoxybenzyl, dimethylallyl, and 4-fluorobenzyl oxoimidazoquinoxaline analogs demonstrated a 58–336-fold higher affinity for ANT than AT diazepam-insensitive sites. Classical benzodiazepines having a 5-phenyl substituent have demonstrated a similar preference for ANT sites, suggesting that all these structures bind to diazepam-insensitive sites in the same orientation. The other compounds evaluated demonstrated only a more modest selectivity (1–12-fold), indicating different structural requirements for binding to mutant ANT and wild-type AT and Wistar receptors. These results expand the range of ligands which display high affinity for diazepam-insensitive sites. Such compounds shoudl be helpful in determining intrinsic actions of high-affinity ligands at these sites and in assessing the contribution of these sites in enhanced sedative sensitivity of cerebellar function in the ANT rats.

Benzodiazepine receptor ligands have no specific action on working memory in a delayed conditional discrimination task in rats

We investigated effects of benzodiazepine (BDZ) receptor ligands on working memory in a delayed conditional discrimination (DCD) task. The BDZ receptor full agonist midazolam (0.1-1.0mg/kg) dose-dependently impaired performance independent of delay, indicating no specific effect on working memory. The non-sedative BDZ receptor partial agonist bretazenil (0.06-0.6mg/kg), the inverse agonist beta-CCM (0.05-0.45mg/kg), the partial inverse agonist FG 7142 (0.5-5.0mg/kg), the antagonist flumazenil (1-10mg/kg), and the antagonist ZK 93 426 (1-10mg/kg) did not significantly affect performance. It is concluded that BDZ ligands do not affect working memory in a positively-motivated DCD task. Midazolam also impaired performance in the no-delay condition, suggesting loss of stimulus control, possibly through an attentional impairment.

The benzodiazepine receptor antagonist flumazenil increases acetylcholine release in rat hippocampus

The benzodiazeoine receptor antagonist flumazenil (2.5–20 mg/kg i.p.) increased acetylcholine (ACh) release by up to 85% in the hippocampus of freely moving rats. In contrast, the benzodiazepine receptor full agonist diazepam (2.5–10 mg/kg i.p.) decreased ACh release up to a maximum of 45% in the same brain area. Injection of flumazenil (10 pmol) or diazepam (10 pmol) into the medial septum increased (95%) or reduced (50%), respectively, ACh release in the hippocampus. The maximum effect produced by those drugs was of the same magnitude as that observed after systemic injection. The changes in hippocampal cho cholinergic function elicitec by activation and blockade of benzodiazepine receptors in the medial septum may thus play a crucial role in the alterations of the cognitive processes elicited by benzodiazepine receptor ligands.

Effects of benzodiazepine receptor ligands on simultaneous visual discriminations of variable difficulty

Benzodiazepine receptor (BZR) ligands have been demonstrated to affect the performance in tasks measuring attentional abilities. In such tasks, subjects typically are required to discriminate visual and/or auditory stimuli. The possibility that the effects of BZR ligands on the performance in tasks measuring attention are primarily due to effects on discriminative processes has not been tested systematically. Rats were trained to discriminate between simultaneously presented pairs of visual stimuli flashing either at 5 Hz versus 4.17, 3.75, 2.5, 1.67 or 1.25 Hz (group 1; FAST), or at 1.25, 1.46, 1.67, 2.5 or 3.33 Hz versus 5 Hz (group 2; SLOW) for 4.8 s (20 trials per discrimination type; sequence of pairs was randomized). In both groups, response accuracy depended significantly on the discriminability of the stimuli, with near perfect accuracy in response to most different pairs of stimuli and near chance-level accuracy in response to least different pairs of stimuli. Administration of the BZR full agonist chlordiazepoxide (1.56, 6.25, 9.38 mg/kg; i.p.) potently increased the number of errors of omission which, following the higher doses, confounded the effects on absolute numbers of correct and incorrect responses. However, the available data do not suggest that the agonist affected the animals' abilities to discriminate between the stimuli. Similarly, administration of the BZR ligands ZK 93 426 and MDL 26,479 (which negatively modulate GABAergic transmission) produced no systematic effects. These data suggest that the effects of BZR ligands on the performance in tasks measuring attentional abilities are not primarily due to effects on the animals' ability to discriminate sensory stimuli.