mRNA expression of corticotropin-releasing factor and urocortin 1 after restraint and foot shock together with alprazolam administration

Corticotropin-releasing factor (CRF) is expressed in the paraventricular nucleus of the hypothalamus (PVN), and act centrally to provoke stress-like autonomic and behavioral responses. Urocortins 1-3 are additional ligands to the CRF receptors 1 and 2. Ucn 1 neurons are primarily concentrated in the Edinger-Westphal (EW) nucleus and also have been associated with stress responses. It is also known that UCN 1 respond in different ways depending on the stressor presented. Benzodiazepines can act via the CRF peptidergic system and chronic administration of alprazolam does not interfere with CRF mRNA expression in the PVN, but significantly increase Ucn 1 mRNA expression in the EW. The aim of our study was to investigate the relationship between different stressor stimuli, foot shock (FS) and restraint (R), and the mRNA expression of CRF and Ucn 1 in the PVN and EW using alprazolam (A). We employed fos activation and in situ hybridization. Restraint group presented increased fos-ir and CRF mRNA expression in the PVN compared to FS group. The stress responses of R group were prevented by A. In the EW,fos-ir was higher in the FS group than in the R group, whereas Ucn 1 mRNA expression was higher in the R group than in the FS group. Alprazolam significantly increased fos-ir and Ucn 1 mRNA expression in both groups. Our results show that PVN and EW respond in different ways to the same stressors. Furthermore, EW of stressed animals replies in a complementary way comparing to PVN with the use of Alprazolam. (C) 2010 Elsevier Inc. All rights reserved.

Molecular Complexes of Alprazolam with Carboxylic Acids, Boric Acid, Boronic Acids, and Phenols. Evaluation of Supramolecular Heterosynthons Mediated by a Triazole Ring

A series of molecular complexes, both co-crystals and salts, of a triazole drug-alprazolam-with carboxylic acids, boric acid, boronic acids, and phenols have been analyzed with respect to heterosynthons present in the crystal structures. In all cases, the triazole ring behaves as an efficient hydrogen bond acceptor with the acidic coformers. The hydrogen bond patterns exhibited with aromatic carboxylic acids were found to depend on the nature and position of the substituents. Being a strong acid, 2,6-dihydroxybenzoic acid forms a salt with alprazolam. With aliphatic dicarboxylic acids alprazolam forms hydrates and the water molecules play a central role in synthon formation and crystal packing. The triazole ring makes two distinct heterosynthons in the molecular complex with boric acid. Boronic acids and phenols form consistent hydrogen bond patterns, and these are seemingly independent of the substitutional effects. Boronic acids form noncentrosymmetric cyclic synthons, while phenols form O-H center dot center dot center dot N hydrogen bonds with the triazole ring. (C) 2010 Wiley-Liss, Inc. and the American Pharmacists Association J Pharm Sci 99:3743-3753, 2010.

Drug metabolism in human brain: high levels of cytochrome P4503A43 in brain and metabolism of anti-anxiety drug alprazolam to its active metabolite.

Cytochrome P450 (P450) is a super-family of drug metabolizing enzymes. P450 enzymes have dual function; they can metabolize drugs to pharmacologically inactive metabolites facilitating their excretion or biotransform them to pharmacologically active metabolites which may have longer half-life than the parent drug. The variable pharmacological response to psychoactive drugs typically seen in population groups is often not accountable by considering dissimilarities in hepatic metabolism. Metabolism in brain specific nuclei may play a role in pharmacological modulation of drugs acting on the CNS and help explain some of the diverse response to these drugs seen in patient population. P450 enzymes are also present in brain where drug metabolism can take place and modify therapeutic action of drugs at the site of action. We have earlier demonstrated an intrinsic difference in the biotransformation of alprazolam (ALP) in brain and liver, relatively more alpha-hydroxy alprazolam (alpha-OHALP) is formed in brain as compared to liver. In the present study we show that recombinant CYP3A43 metabolizes ALP to both alpha-OHALP and 4-hydroxy alprazolam (4-OHALP) while CYP3A4 metabolizes ALP predominantly to its inactive metabolite, 4-OHALP. The expression of CYP3A43 mRNA in human brain samples correlates with formation of relatively higher levels of alpha-OH ALP indicating that individuals who express higher levels of CYP3A43 in the brain would generate larger amounts of alpha-OHALP. Further, the expression of CYP3A43 was relatively higher in brain as compared to liver across different ethnic populations. Since CYP3A enzymes play a prominent role in the metabolism of drugs, the higher expression of CYP3A43 would generate metabolite profile of drugs differentially in human brain and thus impact the pharmacodynamics of psychoactive drugs at the site of action.

Drug metabolism in human brain: high levels of cytochrome P4503A43 in brain and metabolism of anti-anxiety drug alprazolam to its active metabolite.

Cytochrome P450 (P450) is a super-family of drug metabolizing enzymes. P450 enzymes have dual function; they can metabolize drugs to pharmacologically inactive metabolites facilitating their excretion or biotransform them to pharmacologically active metabolites which may have longer half-life than the parent drug. The variable pharmacological response to psychoactive drugs typically seen in population groups is often not accountable by considering dissimilarities in hepatic metabolism. Metabolism in brain specific nuclei may play a role in pharmacological modulation of drugs acting on the CNS and help explain some of the diverse response to these drugs seen in patient population. P450 enzymes are also present in brain where drug metabolism can take place and modify therapeutic action of drugs at the site of action. We have earlier demonstrated an intrinsic difference in the biotransformation of alprazolam (ALP) in brain and liver, relatively more alpha-hydroxy alprazolam (alpha-OHALP) is formed in brain as compared to liver. In the present study we show that recombinant CYP3A43 metabolizes ALP to both alpha-OHALP and 4-hydroxy alprazolam (4-OHALP) while CYP3A4 metabolizes ALP predominantly to its inactive metabolite, 4-OHALP. The expression of CYP3A43 mRNA in human brain samples correlates with formation of relatively higher levels of alpha-OH ALP indicating that individuals who express higher levels of CYP3A43 in the brain would generate larger amounts of alpha-OHALP. Further, the expression of CYP3A43 was relatively higher in brain as compared to liver across different ethnic populations. Since CYP3A enzymes play a prominent role in the metabolism of drugs, the higher expression of CYP3A43 would generate metabolite profile of drugs differentially in human brain and thus impact the pharmacodynamics of psychoactive drugs at the site of action.

Phenylboronic acids in crystal engineering: Utility of the energetically unfavorable syn,syn-conformation in co-crystal design

Phenylboronic acids can exist, in principle, in three different conformers (syn,syn; syn,anti and anti,anti) with distinct energy profiles. In their native state, these compounds prefer the energetically favored syn, anti-conformation. In molecular complexes, however, the functionality exhibits conformational diversity. In this paper we report a series of co-crystals, with N-donor compounds, prepared by a design strategy involving the synthons based on the syn, syn-conformation of the boronic acid functionality. For this purpose, we employed compounds with the 1,2-diazo fragment (alprazolam, 1H-tetrazole, acetazolamide and benzotriazole), 1,10-phenanthroline and 2,2'-bipyridine for the co-crystallization experiments. However, our study shows that the mere presence of the 1,2-diazo fragment in the coformer does not guarantee the successful formation of co-crystals with a syn, syn-conformation of the boronic acid. [GRAPHICS] The -B(OH)(2) fragment makes unsymmetrical O-H center dot center dot center dot N heterosynthons with alprazolam (ALP) and 1,10-phenanthroline (PHEN). In the co-crystals of phenylboronic acids with 1H-tetrazole (TETR) and 2,2'-bipyridine (BPY), the symmetrical boronic acid dimer is the major synthon. In the BPY complex, boronic acid forms linear chains and the pyridine compound interacts with the lateral OH of boronic acid dimers that acts as a connector, thus forming a ladder structure. In the TETR complex, each heterocycle interacts with three boronic acids. While two boronic acids interact using the phenolic group, the third molecule generates O-H center dot center dot center dot N hydrogen bonds using the extra OH group, of -B(OH)(2) fragment, left after the dimer formation. Thus, although molecules were selected retrosynthetically with the 1,2-diazo fragment or with nearby hetero-atoms to induce co-crystal formation using the syn,syn-orientation of the -B(OH)(2) functionality, co-crystal formation is in fact selective and is probably driven by energy factors. Acetazolamide (ACET) contains self-complementary functional groups and hence creates stable homosynthons. Phenylboronic acids being weak competitors fail to perturb the homosynthons and hence the components crystallize separately. Therefore, besides the availability of possible hydrogen bond acceptors in the required position and orientation, the ability of the phenyl-boronic acid to perturb the existing interactions is also a prerequisite to form co-crystals. This is illustrated in the table below. In the case of ALP, PHEN and BPY, the native structures are stabilized by weak interactions and may be influenced by the boronic acid fragment. Thus phenylboronic acids can attain co-crystals with those compounds, wherein the cyclic O-H center dot center dot center dot N hydrogen bonds are stronger than the individual homo-interactions. This can lower the lattice energy of the molecular complex as compared with the individual crystals. [GRAPHICS] Phenylboronic acids show some selectivity in the formation of co-crystals with N-heterocycles. The differences in solubility of the components fall short to provide a possible reason for the selective formation of co-crystals only with certain compounds. These compounds, being weak acids, do not follow the Delta pK(a) analysis and hence fail to provide any conclusive observation. Theoretical results show that of the three conformers possible, the syn,anti conformer is the most stable. The relative stabilities of the three conformers syn,anti,syn,syn and anti,anti are 0.0, 2.18 and 3.14 kcal/mol, respectively. The theoretical calculations corroborate the fact that only energetically favorable synthons can induce the formation of heterosynthons, as in ALP and PHEN complexes. From a theoretical and structural analysis it is seen that phenylboronic acids will form interactions with those molecules wherein the heterocyclic and acidic fragments can interrupt the homosynthons. However, the energy profile is shallow and can be perturbed easily by the presence of competing functional groups (such as OH and COOH) in the vicinity. [GRAPHICS] .

Environmental occurrence and fate of psychiatric pharmaceuticals

Os fármacos são importantes contaminantes ambientais. Nas últimas duas décadas, o número de estudos sobre a ocorrência destes poluentes emergentes em matrizes ambientais aumentou significativamente. Esta ocorrência generalizada preocupa a comunidade científica devido a evidências que comprovam a sua capacidade de interferir nos ecossistemas, mesmo em concentrações muito baixas. No caso particular dos fármacos psiquiátricos é expectável que constituam um risco ecológico significativo. Para uma melhor compreensão do impacto real destes poluentes é essencial que se proceda a uma avaliação extensiva da sua persistência e destino em matrizes ambientais. Os estudos apresentados nesta tese pretendem contribuir para melhorar o conhecimento acerca da ocorrência, persistência e destino ambiental de fármacos psiquiátricos. Para este efeito, foram seleccionados, como objecto de estudo, dois grupos de fármacos: anti-epilépticos (carbamazepina) e fármacos com efeitos ansiolíticos e sedativos (as benzodiazepinas diazepam, oxazepam, lorazepam e alprazolam). A fotodegradação é o principal processo que afecta a persistência de poluentes orgânicos em ambientes aquáticos. Consequentemente, a persistência dos cinco fármacos seleccionados foi avaliada através de estudos de fotodegradação directa e indirecta, tendo em consideração a influência de parâmetros relevantes tais como pH, nível de oxigenação e matéria orgânica dissolvida. Os estudos de fotodegradação aqui descritos foram seguidos por cromatografia micelar electrocinética com a aplicação de um capilar com revestimento dinâmico. Adicionalmente, os fotoprodutos resultantes de fotodegradação directa foram identificados por espectrometria de massa. O estudo da carbamazepina no ambiente é particularmente relevante uma vez que esta foi proposta como um potencial marcador de poluição antropogénica. A sua ocorrência em água superficiais, de sub-solo e residuais foi investigada através da implementação de um ensaio imunológico (ELISA), optimizado para a aplicação a triagens ambientais e amostras com matrizes complexas. O destino deste fármaco na interface água/solo foi também investigado usando solos agrícolas submetidos a fertilizações de longo prazo; este estudo permitiu tirar conclusões acerca da contaminação de águas adjacentes por solos contaminados. O trabalho aqui descrito constitui uma abordagem multidisplinar à problemática da ocorrência de fármacos psiquiátricos no ambiente, contribuindo de forma relevante para esta área de estudo.

Evaluación de la conectividad cerebral bajo fármacos mediante análisis no lineal del EEG

Màster Oficial en Enginyeria Biomèdica

Ethopharmacological evaluation of the rat exposure test: A prey-predator interaction test

The rat exposure test (RET) is a prey (mouse)-predator (rat) situation that activates brain defensive areas and elicits hormonal and defensive behavior in the mouse. Here, we investigated possible correlations between the spatiotemporal [time spent in protected (home chamber and tunnel) and unprotected (surface) compartments and frequency of entries into the three compartments] and ethological [e.g., duration of protected and unprotected stretched-attend postures (SAP), duration of contact with the rat's compartment] measures (Experiment 1). Secondly, we investigated the effects of systemic treatment with pro- or anti-aversive drugs on the behavior that emerged from the factor analysis (Experiment 2). The effects of chronic (21 days) imipramine and fluoxetine on defensive behavior were also investigated (Experiment 3). Exp. 1 revealed that the time in the protected compartment, protected SAP and rat contacts loaded on factor 1 (defensive behavior), while the total entries and unprotected SAP loaded on factor 2 (locomotor activity). Exp. 2 showed that alprazolam (but not diazepam) selectively changed the defensive factor. Caffeine produced a mild proaversive-like effect, whereas yohimbine only decreased locomotor activity (total entries). Fluoxetine (but not imipramine) produced a weak proaversive-like effect. 5-HT1A/5-HT2 receptor ligands did not change any behavioral measure. In Exp. 3, chronic fluoxetine (but not imipramine) attenuated the defensive behavior factor without changing locomotion. Given that the defensive factor was sensitive to drugs known to attenuate (alprazolam and chronic fluoxetine) and induce (caffeine) panic attack, we suggest the RET as a useful test to assess the effects of panicolytic and panicogenic drugs. © 2012 Elsevier B.V.

Ethopharmacological analysis of the open elevated plus-maze in mice

Exposure of rodents to an open elevated plus-maze (oEPM) elicits antinociception and increases plasma corticosterone levels. However, no studies have yet assessed the defensive behaviour repertoire of animals in this modified test. In Experiment 1, factor analysis was employed to characterise the behavioural profile of mice exposed to the oEPM. Experiments 2 and 3 assessed the effects of acute alprazolam (0.5-1.5. mg/kg; diazepam 0.5-1.5. mg/kg), pentylenetetrazole (10.0-30.0. mg/kg), yohimbine (2.0-6.0. mg/kg), mCPP (0.3-3.0. mg/kg), and acute and chronic fluoxetine (10.0-30.0. mg/kg) and imipramine (1.0-15.0. mg/kg) on behaviours identified in Experiment 1. The factor analyses revealed that behaviour in the oEPM can largely (77% total variance) be accounted for in terms of 3 factors: factor 1 ('. depth exploration'; e.g. head-dipping on the arms), factor 2 ('. cautious exploration of arms'; e.g. flatback approach), and factor 3 ('. risk assessment'; stretched attend postures - SAP). Experiments 2 and 3 showed that, over the dose range used, alprazolam selectively attenuated all measures of defensiveness. Similar, though more modest, effects were seen with diazepam. Confirming the intensity of the emotional response to the oEPM (nociceptive, endocrine and behavioural), relatively few significant behavioural changes were seen in response to the anxiogenic compounds tested. Although acute fluoxetine or imipramine treatment failed to modify behaviour in the oEPM, chronic fluoxetine (but not chronic imipramine) attenuated total flat back approach and increased head dipping outside the central square. Together, the results indicate that the oEPM induces behavioural defensive responses that are sensitive to alprazolam and chronic fluoxetine. © 2013 Elsevier B.V.

Análise etofarmacológica do teste de exposição ao rato (interação presa-predador) para o estudo da ansiedade em camundongos

Com o objetivo de propor mecanismos que auxiliem na compreensão da neurobiologia do medo e da ansiedade, diversos modelos animais vem sendo utilizados. Dentre eles destaca-se o Teste de Exposição ao Rato (RET), que é um modelo etológico de interação presa (camundongo) – predador (rato), conhecido por eliciar a expressão de diversos tipos de comportamentos defensivos no camundongo, tais como avaliação de risco, esquiva, congelamento, entre outros. Com o intuito de aumentar a aversividade do modelo proposto por Yang e colaboradores (2004), foi proposto um novo modelo de RET, semelhante ao modelo padrão, porém com alterações de algumas variáveis como aumento da luminosidade, ausência de maravalha, alteração do local de inserção do camundongo no aparato, substituição da tela de arame (usada na separação da superfície) por uma tela de acrílico e arame e uma divisão virtual da superfície em duas partes, uma distante da tela por 10 cm (superfície proximal) e outra com 13 cm até chegar ao túnel (superfície distal). Um estudo piloto realizado comparando o novo teste de exposição ao rato com o modelo padrão demonstrou que no novo modelo de exposição há uma maior preferência pela presa às áreas protegidas (toca e túnel) e uma maior expressão de avaliação de risco, indicando talvez uma maior sensibilidade à expressão de comportamentos defensivos de camundongos e possivelmente a fármacos que atuem no sistema de defesa. No presente estudo foram investigados os efeitos dos (i) benzodiazepínicos diazepam e alprazolam, (ii) dos fármacos ansiogênicos cafeína e pentilenotetrazol e (iii) do antidepressivo fluoxetina (aguda e cronicamente). O tratamento com alprazolam resultou num efeito antiaversivo mais evidente quando comparado ao diazepam. O tratamento agudo 10 com cafeína, por sua vez, apresentou um efeito ansiogênico bastante sutil, diferente... (Resumo completo, clicar acesso eletrônico abaixo)

Efeitos de benzodiazepinas sobre o fenômeno da aversão condicionada por lugar induzida por estímulo nociceptivo em camundongos

Medo e ansiedade são emoções que se originam nas reações de defesa de caráter adaptativo que animais exibem perante ameaças que podem comprometer sua integridade física ou sua própria sobrevivência. Tais situações resultam em um repertório de comportamentos defensivos espécie-específicos, alterações autonômicas e inibição da dor. A dor é descrita por uma variedade de sensações e experiências emocionais associadas e constituída de dois principais componentes: o perceptivo-discriminativo, que permite ao indivíduo reconhecer o estímulo como doloroso e localizá-lo numa região do corpo, e o aversivo-cognitivo-motivacional, que compreende uma série de respostas defensivas, objetivando a autopreservação imediata. O modelo de condicionamento por lugar, para medir efeitos não discriminativos da estimulação nociceptiva, vem sendo utilizado em abordagens recentes para o estudo do componente emocional relacionado à dor. Existem paradigmas baseados na aprendizagem associativa entre o estado afetivo e de dicas ambientais que precedem a aplicação de um estímulo, o qual pode ter propriedade aversiva ou prazerosa, dependendo da resposta que se quer estudar. O presente estudo investigou a ação do fármaco triazolobenzodiazepínico, Alprazolam, sobre o fenômeno da ACL em camundongos, que foram separados em grupos experimentais. No primeiro dia, parte dos animais sofreu estimulação nociceptiva, através da injeção de formalina a 2,5% na pata traseira direita, e foi confinado nos ambientes diferenciados de uma Plataforma Elevada fechada (um lado com grade no assoalho e outro sem grade). No segundo dia, parte dos animais 7 (de acordo com o grupo de estudo) recebeu tratamento farmacológico (Alprazolam ou salina) 30 minutos antes de ser reexposto à plataforma, registrando-se o tempo gasto nos dois ambientes da plataforma elevada fechada... (Resumo completo, clicar acesso eletrônico abaixo)

Proactive aggression: A two part study

Aggressive behavior can be divided into the subtypes: reactive and proactive. Reactive aggressive acts occur in response to a stimulus or provocation. Proactive aggressive acts occur without provocation and are goal-directed. A number of findings have suggested that individuals displaying proactive aggression may be discerned from individuals not displaying proactive aggression on measures of personality, psychopathology and psychopathy, as well as on aggressive histories and type and severity of aggressive behaviors committed. The current study was conducted in two phases; phase 1 and 2. This was because phase 1 compared proactive aggressive, reactive aggressive and non-aggressive subjects on questionnaire measures, while phase 2 observed the acute effects of the benzodiazepine alprazolam on only proactive aggressive subjects. The phase 1 hypotheses were that proactive aggressive subjects would show greater numbers of personality disorders and have greater psychopathy relative to reactive and non-aggressive subjects. To verify these hypotheses subjects were recruited from the community and classified as proactive (n = 20), reactive (n = 20) or non-aggressive (n = 10) via laboratory behavioral testing. Classified subjects were administered a battery of questionnaires pertaining to personality disorders (SCID-II, OMNI-IV), psychopathy (PCL-R) and aggression history. The results of these questionnaire measures were subjected to statistical analyses, which confirmed the hypotheses. In the second phase, the acute effects of three doses of the benzodiazepine alprazolam were evaluated in proactive aggressive subjects on proactive aggressive responding in the computer-based Point Subtraction Aggression Paradigm (PSAP). In phase 2 it was hypothesized that alprazolam would produce dose dependent decreases in aggressive responding. Subjects were never provoked in this phase, and aggressive responding was classified as proactive. Studies of drugs acting on the GABA system have frequently found decreases in aggression in animals and humans, although there have also been findings of increased (paradoxical) aggression. The hypothesis was tested by statistical analysis of proactive aggressive responding under placebo vs. under alprazolam. The hypothesis was supported by six of seven subjects. Aggressive responding was significantly, decreased under alprazolam relative to placebo in six subjects. One subject showed increases in aggressive responding.^

Predictors of benzodiazepine self -administration behavior in anxious patients

The purpose of this study was to examine factors that may be associated with benzodiazepine (BZ) self-administration and risks of dependence in anxious patients. Preliminary work included examination of psychosocial characteristics and subjective drug response as potential predictors of medication use. Fifty-five M, F patients with generalized anxiety or panic disorder participated in a 3-week outpatient Choice Procedure in which they self-medicated “as needed” with alprazolam (Alz) and placebo. Findings showed that a large amount of variance in alprazolam preference, frequency, and quantity of use could be predicted by measures of anxiety, drug liking, and certain personality characteristics. The primary study extended this work by examining whether individual differences in Alz sensitivity also predict patterns of use. Twenty anxious patients participated in the study, which required 11 weekly clinic visits. Ten of these also participated in a baseline assessment of HPA-axis function that involved 24-hour monitoring of cortisol and ACTH levels and a CRH Stimulation Test. This assessment was conducted on the basis of prior evidence that steroid metabolites exert neuromodulatory effects on the GABA A receptor and that HPA-axis function may be related to BZ sensitivity and long-term disability in anxious patients. Patients were classified as either HIGH or LOW users based on their p.r.n. patterns of Alz use during the first 3 weeks of the study. They then participated in a 4-week dose response trial in which they received prescribed doses of medication (placebo, 0.25, 0.5, and 1.0mg Alz), each taken TID for 1 week. The dose response trial was followed by a second 3-week Choice Procedure. Findings were not indicative of biological differences in Alz sensitivity between the HIGH and LOW users. However, the HIGH users had higher baseline anxiety and greater anxiolytic response to Alz than the LOW users. Anxiolytic benefits of p.r.n. and prescribed dosing were shown to be comparable, and patients' conservative patterns of p.r.n. medication use were not affected by the period of prescribed dosing. Although there was not strong evidence to suggest relationships between HPA-axis function and Alz use or sensitivity, interesting findings emerged about the relationship between HPA-axis function and anxiety. ^

TMS- EEG Signatures of GABAergic Neurotransmission in the Human Cortex

Combining transcranial magnetic stimulation (TMS) and electroencephalography (EEG) constitutes a powerful tool to directly assess human cortical excitability and connectivity. TMS of the primary motor cortex elicits a sequence of TMS-evoked EEG potentials (TEPs). It is thought that inhibitory neurotransmission through GABA-A receptors (GABAAR) modulates early TEPs (<50 ms after TMS), whereas GABA-B receptors (GABABR) play a role for later TEPs (at ∼100 ms after TMS). However, the physiological underpinnings of TEPs have not been clearly elucidated yet. Here, we studied the role of GABAA/B-ergic neurotransmission for TEPs in healthy subjects using a pharmaco-TMS-EEG approach. In Experiment 1, we tested the effects of a single oral dose of alprazolam (a classical benzodiazepine acting as allosteric-positive modulator at α1, α2, α3, and α5 subunit-containing GABAARs) and zolpidem (a positive modulator mainly at the α1 GABAAR) in a double-blind, placebo-controlled, crossover study. In Experiment 2, we tested the influence of baclofen (a GABABR agonist) and diazepam (a classical benzodiazepine) versus placebo on TEPs. Alprazolam and diazepam increased the amplitude of the negative potential at 45 ms after stimulation (N45) and decreased the negative component at 100 ms (N100), whereas zolpidem increased the N45 only. In contrast, baclofen specifically increased the N100 amplitude. These results provide strong evidence that the N45 represents activity of α1-subunit-containing GABAARs, whereas the N100 represents activity of GABABRs. Findings open a novel window of opportunity to study alteration of GABAA-/GABAB-related inhibition in disorders, such as epilepsy or schizophrenia.

7-Chloro-3-methyl-3,4-dihydro-2H-1,2,4-benzothiadiazine S,S-dioxide (IDRA 21), a congener of aniracetam, potently abates pharmacologically induced cognitive impairments in patas monkeys.

We report here on the ability of IDRA 21 and aniracetam, two negative allosteric modulators of glutamate-induced DL-alpha-amino-3-hydroxy-5-methyl-4-isoxazolepropionic acid (AMPA) receptor desensitization, to attenuate alprazolam-induced learning deficit in patas monkeys working in a complex behavioral task. In one component of a multiple schedule (repeated acquisition or "learning"), patas monkeys acquired a different four-response chain each session by responding sequentially on three keys in the presence of four discriminative stimuli (geometric forms or numerals). In the other component (performance) the four-response chain was the same each session. The response chain in each component was maintained by food presentation under a fixed-ratio schedule. When alprazolam (0.1 or 0.32 mg/kg p.o.) was administered alone, this full allosteric modulator of gamma-aminobutyric acid type A (GABAA) receptors produced large decreases in the response rate and accuracy in the learning component of the task. IDRA 21 (3 or 5.6 mg/kg p.o.) and aniracetam (30 mg/kg p.o.) administered 60 min before alprazolam, having no effect when given alone, antagonized the large disruptive effects of alprazolam on learning. From dose-response studies, it can be estimated that IDRA 21 is approximately 10-fold more potent than aniracetam in antagonizing alprazolam-induced learning deficit. We conclude that IDRA 21, a chemically unrelated pharmacological congener of aniracetam, improves learning deficit induced in patas monkeys by the increase of GABAergic tone elicited by alprazolam. Very likely IDRA 21 exerts its behavioral effects by antagonizing AMPA receptor desensitization.