Characteristics of an atypical benzodiazepine, Ro 5-4864.

Ro 5-4864 is a 1,4 benzodiazepine which, atypically, does not bind to the classical CNS benzodiazepine receptors, but has high affinity for the peripheral type of binding site found both in the periphery and in the brain. Biochemical evidence for alternative sites of action for this compound is discussed. We review the behavioral profile of Ro 5-4864 (sedative, convulsant and anxiogenic in rodents) and also describe the behavioral effects of combining Ro 5-4864 treatment with benzodiazepines (e.g., diazepam, chlordiazepoxide) and with other drugs that modify the activity of benzodiazepines (Ro 15-1788, CGS 8216, picrotoxin, PK 11195, phenytoin). In the light of these interactions and electrophysiological evidence we conclude that the actions of Ro 5-4864 are most likely to be mediated at the GABA-benzodiazepine receptor complex in the CNS.

Beta-carbolines can enhance or antagonize the effects of punishment in mice.

Six beta-carboline ligands at central benzodiazepine (BZ) receptors were tested for their anxiolytic or anxiogenic properties in mice in the four-plate test. ZK 93 423 and ZK 91 296 increased activity which had been suppressed by punishment (1 mA, 60 ms footshock) at doses which exerted no effect on unpunished locomotion. ZK 93 426, ZK 90 886, FG 7142, and DMCM exerted no antipunishment activity themselves, and antagonized the ability of diazepam to increase both punished and unpunished locomotor activity. DMCM, FG 7142, and ZK 90 886, but not ZK 93 426, also enhanced the ability of a reduced level of footshock (0.3 mA) to suppress activity. This propunishment activity of DMCM and ZK 90 886 took place at doses which had no effect on unpunished locomotion. The nature of the effect of the individual beta-carbolines on punishment was related to the nature of their interaction with the BZ/GABA receptor/chloride channel complex (GBC complex). Thus the antipunishment properties of ZK 93 423 and ZK 91 296 were associated with their ability to increase binding of 35S-t-butylbicyclo-phosphorothionate (TBPS) to its binding site associated with the chloride channel, whereas DMCM, FG 7142 and ZK 90 886, which exerted propunishment effects, reduced TBPS binding. ZK 93 426, which was neutral with respect to punished activity, had the weakest effect on TBPS binding. These results are discussed in the context of a possible role of GBC complex in anxiety.

Safety signal withdrawal: a behavioural paradigm sensitive to both "anxiolytic" and "anxiogenic" drugs under identical experimental conditions.

A new method involving the blockade of operant behaviour induced by the withdrawal of a conditioned signal for safety without presentation of a punishment signal has been developed for studying drugs with anxiolytic or anxiogenic properties. For this purpose, rats were trained under two alternating components of a multiple schedule of reinforcement FR8 (food)/FR1 (food) + RR 50% (shocks randomly delivered with 50 +/- 15% of the presses). The nonpunished and punished periods were signalled by one cue light above the right lever (safety signal) or the left lever (punishment signal), respectively. On the test session (safety signal withdrawal), the safety signal was turned off at the end of the first nonpunished period, but the punishment signal was not presented (every press was food rewarded and no shocks were delivered). During this period (4 min), rats exhibited a strong blockade of responding that lessened over time. This suppression seemed not to be caused by intervening events such as novelty, temporal conditioning, schedule of food delivery or ambiguity of the signal presented. The behavioural blockade induced by withdrawal of the safety signal was reduced by benzodiazepines: diazepam (0.5-4 mg/kg), chlordiazepoxide (4-8 mg/kg), nitrazepam (0.25-2 mg/kg), alprazolam (0.25-1 mg/kg), and partial agonists at benzodiazepine receptors: bretazenil (0.125-8 mg/kg) and ZK 91296 (32-64 mg/kg). Various 5-HT-related drugs also lessened the behavioural blockade:pCPA (3 x 150 mg/kg) and the 5-HT1A receptor agonists, buspirone (0.25-2 mg/kg), gepirone (0.25-1 mg/kg) but not 8-OH-DPAT. Compounds that may cause anxiety in humans further enhanced the blockade of lever pressing induced by the safety signal withdrawal at doses that did not modify baseline responding: d-amphetamine (0.125-0.5 mg/kg), caffeine (16 mg/kg) and picrotoxin (1 mg/kg). FG 7142 (8 mg/kg) and CGS 8216 (2-8 mg/kg) decreased responding during both components of the session. Therefore, the present paradigm seems sensitive to both "anxiolytic" and "anxiogenic" effects of drugs under identical procedural conditions.

Intrinsic actions of the benzodiazepine receptor antagonist Ro 15-1788.

The imidazodiazepine Ro 15-1788 is a benzodiazepine receptor antagonist that was initially reported to be lacking in intrinsic activity in a variety of test situations in which benzodiazepine-like effects can be identified. However, many recent studies have shown that this compound does indeed have intrinsic activity in a variety of behavioural, neurological, electrophysiological and biochemical preparations in both animals and man. The purpose of the present review is firstly to describe these intrinsic actions, and secondly to consider to what extent these intrinsic actions of Ro 15-1788 have implications for current concepts of the functioning of the benzodiazepine receptor.

Clonazepam-induced hyperphagia in nondeprived rats: tests of pharmacological specificity with Ro5-4864, Ro5-3663, Ro15-1788 and CGS 9896.

Nondeprived male rats were familiarised with daily 60 min access to a highly palatable diet, consisting of powdered rat diet, sweetened condensed milk and water. Clonazepam (0.625-5.0 mg/kg, IP) produced a substantial increase in food consumption within the first 30 min of access. The increase was similar across all dose conditions, suggesting that a maximal effect may have been achieved with a dose as small as 0.625 mg/kg. The hyperphagia induced by clonazepam was reversed by the benzodiazepine receptor antagonist, Ro15-1788 (5.0-20.0 mg/kg), indicating that the effect was benzodiazepine receptor-mediated. Treatments with the peripheral-type benzodiazepine agonist, Ro5-4864, did not induce a hyperphagic response. Instead, food consumption was significantly depressed following the administration of Ro5-4864 at 20 and 40 mg/kg, IP. A comparison of the clonazepam and Ro5-4864 data suggests that benzodiazepine-induced hyperphagia is mediated by central-type benzodiazepine binding sites. The pyrazoloquinoline, CGS 9896, binds with high affinity to benzodiazepine sites and has recently been described as a nonsedating anxiolytic. CGS 9896 (2.5-20.0 mg/kg, administered either IP or PO) did not affect consumption of the highly palatable diet. In consequence, anxiolytic and hyperphagic effects of drug actions at benzodiazepine receptors may be dissociated in the case of this compound. The atypical 1,4-benzodiazepine, Ro5-3663, a GABA antagonist which may act at the picrotoxinin site, produced a dose-related reduction in food consumption. Comparison with the results for Ro5-4864 rules out an interpretation for the anorexia in terms of anxiogenic effects.(ABSTRACT TRUNCATED AT 250 WORDS)

Gamma-aminobutyric acid, its related enzymes and receptor-binding sites in the human ovary and fallopian tube.

The concentration of gamma-aminobutyric acid (GABA), the activities of related enzymes, i.e. glutamate decarboxylase and GABA transaminase, as well as the level of specific GABA binding sites were determined in ovaries and fallopian tubes obtained surgically from 31 women. None of the biochemical parameters examined showed a correlation with the age and hormonal background (serum estradiol and progesterone levels) of the patients. The respective ovarian and tubal values did not differ significantly in groups operated on because of uterine myoma and carcinoma. In organs from pregnant women, however, most GABAergic markers altered significantly. These findings indicate some gestation-related role for the ovarian and tubal GABA systems in humans.

The radiosynthesis of [18F]PK 14105 as an alternative radioligand for peripheral type benzodiazepine binding sites.

A method has been developed for labelling PK 14105 [N-methyl-N-(1-methyl-propyl)-1(2-fluoro-5-nitrophenyl)isoquinoline-3- carboxamide], a ligand that has high affinity and selectivity for peripheral type benzodiazepine binding sites (PBBS), with NCA fluorine-18 (t1/2 = 109.8 min, beta + = 96.9%). The method involves treating the 2-chloro-analogue with cyclotron-produced NCA [18F]fluoride in dimethyl sulphoxide, with rubidium carbonate as base, at 140 degrees C for 20 min. Purification is achieved by separation on a reverse phase Sep-Pak followed by PHLC on a silica gel column, to give chemically and radiochemically pure product with a specific activity of ca 7.4 GBq/mumol (200 mCi/mumol), decay-corrected to the end of radionuclide production (EOB). The radiosynthesis requires 210 min. giving a radiochemical yield of 10-20%, decay-corrected to EOB. [18F]PK 14105 was found to bind avidly to sites associated with kainic acid-induced unilateral lesions of rat striata. Such binding was blocked by pre-dosing the rat with PK 11195, so providing evidence for specific binding to PBBS. These results suggest that [18F]PK 14105 has potential for studying phenomena associated with PBBS in man by PET.

Interrelaçäo dos benzodiazepínicos com receptores do Gaba-ionoforo CL- / Interrelation of the benzodiazepines and gaba-receptor-ionophore complex

O mecanismo de açäo dos benzodiazepínicos no complexo macromolecular constituído por receptor do GABA, receptor dos benzodiazepínicos, e ionoforo C1- é revisto. É enfatizado que a interaçäo dos benzodiazepínicos com seus receptores facilita o efeito do GABA como neurotransmissor ao nível do sistema nervoso central. O processo molecular desencadeado por essa interaçäo ainda näo está completamente esclarecido, embora o resultado final da ativaçäo dos receptores GABAérgicos seja a abertura dos canais de cloro. A açäo de outros agonistas, de drogas antagonistas, e de possíveis ligantes endógenos é também discutida