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Les antipsychotiques atypiques sont des options de traitement de première ligne pour la schizophrénie. Cependant, la prise d’antipsychotiques atypiques, comme l’olanzapine, est associée à des effets secondaires métaboliques : l’augmentation du poids, la dyslipidémie et l’intolérance au glucose. Les mécanismes en lien avec ces effets secondaires sont à ce jour peu connus. Ce mémoire étudie l’évolution de différents paramètres, tant au niveau biométrique (poids, IMC, circonférence abdominale), qu’au niveau sérique (bilan lipidique, glycémie à jeun, insuline, leptine, ghreline) et clinique (mesures des symptômes positifs, négatifs et généraux de la schizophrénie, de même que des comportements alimentaires) chez des sujets schizophrènes, traités pendant 16 semaines avec l’olanzapine. Des examens de résonance magnétique, structurale et fonctionnelle, ont été effectués au début et à la fin du traitement d’olanzapine chez les sujets schizophrènes et chez un groupe de sujets contrôles afin d’identifier les régions cérébrales dont les volumes ou les activations pourraient être associés aux mécanismes d’effets secondaires métaboliques. Nos résultats confirment l’émergence de multiples effets secondaires métaboliques, associés à des modifications des comportements alimentaires, en lien avec la prise d’olanzapine auprès de notre échantillon. Des associations ont été retrouvées entre les changements métaboliques et les volumes de plusieurs régions cérébrales, notamment les hippocampes, les précunei et le gyrus orbitofrontal droit. De plus, des différences en terme d’activations cérébrales entre les sujets contrôles et les patients schizophrènes, qui ont été accentuées par le traitement d’olanzapine, ont aussi été décrites notamment au niveau amygdalien, cérébelleux et des insulas, suggérant l’implication de mécanismes neuronaux dans l’apparition des troubles métaboliques associés aux antipsychotiques atypiques. / Atypical antipsychotics are first line treatment options for schizophrenia. Nevertheless atypical antipsychotics, such as olanzapine, are associated with metabolic adverse effects: weight gain, dyslipidemia and glucose intolerance. Mechanisms underlying these side effects are still poorly understood. This thesis represents a study of the evolution of biometric (body weight, BMI, abdominal circumference), biological (lipid profile, fasting glucose, insulin, leptin, ghrelin) and clinical parameters (positive, negative and general symptoms of schizophrenia as well as eating behavior measures) in schizophrenia patients treated with olanzapine during 16 weeks. Healthy subjects and schizophrenia patients passed structural and functional magnetic resonance examinations (one examination for healthy controls and two examinations for schizophrenia patients, one at the beginning and one after 16 weeks of olanzapine treatment) in order to identify cerebral regions where grey matter volumes or activations could be associated with metabolic side effects mechanisms. Our results confirm that various metabolic adverse effects emerged in our sample of patients during the 16 weeks olanzapine treatment. Eating behavior changes have also been associated with specific metabolic changes. Many associations have been found between cerebral volumes (e.g. bilateral hippocampi and précunei and right orbitofrontal cortex) and metabolic changes. Moreover, cerebral activations differences between healthy controls and schizophrenia patients, that were increased following olanzapine treatment, were also described in the amygdala, cerebellum and insulas, suggesting that neuronal mechanisms were involved in the development of metabolic disorders associated with atypical antipsychotics.

Les antipsychotiques (APs) sont fréquemment prescrits pour les troubles comportementaux associés à la démence. Or, ces produits ont fait l'objet de trois mises en garde (2002, 2004, 2005) en raison d'une augmentation du risque d'événement cérébrovasculaire et de décès. L’objectif de ce mémoire est d’évaluer l'utilisation d’APs dans la population de personnes âgées démentes vivant à domicile, et de déterminer l’effet des mises en garde sur les profils observés. Une cohorte rétrospective de 10,969 personnes âgées démentes ayant débuté un traitement par AP entre le 1er janvier 2000 et le 31 décembre 2009 fut identifiée à partir des banques de données de la Régie de l'assurance maladie du Québec (RAMQ). Des séries chronologiques segmentées ont permis de quantifier l’effet des mises en garde sur l'utilisation d’APs. L'effet de la mise en garde de 2005 sur les caractéristiques des patients traités ainsi que sur les profils d'utilisation (dose et durée) a été évalué, respectivement par des modèles de régression logistique et de régression linéaire multivariés. Le taux délivrance d'APs atypiques a augmenté au cours du temps jusqu'à la mise en garde de 2005 pour ensuite diminuer de 8.96% (IC 95% : -11.91% – -6.02%). L'analyse par produit a révélé la même tendance pour la rispéridone, le seul AP approuvé au Canada pour les personnes âgées démentes. En revanche, le taux de délivrance de quétiapine, qui est hors-indication, a continué d'augmenter. Le taux d'initiation de traitement par APs a cependant diminué au cours du temps pour tous les produits. Les mises en garde ne semblent pas être associées avec un changement dans les caractéristiques des patients traités, ni avec les doses et durées d’utilisation. Le manque d'efficacité des mises en garde est probablement en partie lié à l'absence d'alternatives thérapeutiques pour le traitement des troubles psychologiques et comportementaux chez les patients atteints de démence. / Antipsychotics (APs) are frequently prescribed for the management of behavioural and psychological symptoms of dementia (BPSD). However, three safety warnings have been issued (2002, 2004 and 2005) due to an increased risk of cerebrovascular event and mortality. The aim of this thesis was to evaluate AP usage in the population of community-dwelling elderly with dementia, and to determine the effect of safety warnings on usage patterns. A retrospective cohort of 10,969 elderly with dementia who initiated an AP treatment between January 1st, 2000 and December 31st, 2009 was identified through in the databases of Régie de l’assurance maladie du Québec (RAMQ). Segmented time series analysis was used to quantify the effect of safety warnings on AP dispensing rate. The effect of the 2005 warning on the characteristics of treated patients and on usage patterns (dose and duration) was evaluated, respectively through multivariate logistic and multiple linear regression models. Atypical AP dispensing rates increased until the 2005 safety warning and decreased by 8.96% (95% CI: -11.91% – -6.02%) thereafter. Analysis by individual products yielded similar trends for risperidone, the only AP approved in Canada for elderly with dementia. However, usage of quetiapine, which is off-label, kept increasing. For all products, rate of treatment initiation decreased over time. Safety warnings did not seem to be associated with either changes in prescribed treatment dosage or duration, nor with prescription channeling toward lower risk patients. Apparent lack of efficacy of safety warnings is likely due, in part, to absence of effective treatment alternatives for BPSD.

Cette thèse vise à mieux comprendre le rôle du facteur de transcription Nur77 au sein des fonctions physiologiques et pathologiques des voies de neurotransmission dopaminergiques des gaglions de la base. Nous basant sur une implication motrice de Nur77 au sein des dyskinésies induites à la L-DOPA (LIDs), nous avons voulu tester in vivo son implication éventuelle dans les phénomènes moteurs et de sensibilisation associés à l’amphétamine ainsi qu’une implication possible du récepteur nucléaire aux rétinoïdes RXR dont nous avons déjà démontré une implication dans les effets moteurs des antipsychotiques. Un deuxième volet de la recherche à consisté en l’étude de l’implication des extracellular signal-regulated kinase (ERK) et de la protéine kinase C (PKC) dans l’induction de l’ARNm des membres de la famille des Nurs suite à l’activation des cascades de signalisation des récepteurs dopamiergiques D1 et D2 in vivo pour une meilleure compréhension des mécanismes physiopatholoiques liés aux désordres dopaminergiques. Pour ce faire, nous avons, premièrement, soumis des souris sauvages et Nur77-/- à un paradigme de sensibilisation à l’amphétamine ainsi qu’a différents agonistes antagonistes RAR/RXR afin de tester l’implication de Nur77 et d’un éventuel complexe Nur77/RXR dans les effets de l’amphétamine. Deuxièmement, soumis des souris sauvages à une combinaison d’agonistes D1/D2 ou une injection d’antagoniste D2 avec ou sans inhibiteur (ERK1/2 ou PKC) afin de tester l’implication de ces kinases sur l’induction de l’ARNm des membres de la famille des Nurs par hybridation in situ. Nous avons ainsi pu démontrer 1- un rôle moteur de Nur77 dans les effets liés à l’amphétamine notamment avec une absence de stéréotypies et un allongement de la durée de la phase de locomotion chez les souris Nur77-/-; ainsi qu’un rôle éventuel du complexe potentiel Nur77/RXR dans les D1 à mieux définir. 2- un rôle des kinases ERKs et PKCs dans les cascades de signalisation des récepteurs dopaminergiques D1 et D2 menant à l’induction des ARNms de Nur77, Nor-1 et Nurr-1. En perspective, ces résultats nous ouvrent la voie vers une implication éventuelle de Nur77 dans les mécanismes d’apprentissage que sont le Long Terme Potentiation (LTP) et la Long Terme Depotentialisation (LTD) liés aux LIDs et à l’amphétamine. / This thesis aims to better understand the role of the transcription factor Nur77 in the physiological and pathological functions of dopaminergic neurotransmission pathways of the basal gaglia. Based on a motor involvement of Nur77 in L-DOPA induced dyskinesias (LIDs), we wanted to test its possible involvement in the motor mechanisms associated with amphetamine, a drug of abuse that targets the direct neuronal population of the basal ganglia such as L-DOPA. Involvement of extracellular signal-regulated kinase (ERK) and protein kinase C (PKC) has been widely demonstrated in the LIDs and the effects of amphetamine, we wanted to know whether Nur77 mRNA was controlled by these kinases in both the striatonigral and striataux palidal neurons in response to specific activation of the D1 and D2 receptors in vivo. Finally, a possible involvement of RXR has been demonstrated in the motor effects of antipsychotic drugs, so we wanted to test its potential involvement in the effects of amphetamine. To do this, we subjected wild-type mice and Nur77-/- mice to a paradigm of sensitization to amphetamine with or without RXR agonist (DHA) and antagonsite (HX531) to test: 1 - the involvement of Nur77 in the motor effects and the phenomenon of sensitization to amphetamine and 2 - the involvement of any complex Nur77/RXR in these effects. Second, wild-type mice were subjected to a combination of D1/D2 agonists or antagonists D2 with or without ERK inhibitor or PKC inhibitor to test in vivo the induction of the mRNA of the Nurs members family by in situ hybridization. We were able to demonstrate a role of Nur77 in the motor effects associated with amphetamine namely in the absence of stereotypy and a lengthening of the duration of the phase of locomotion in Nur77-/ - mice; and a possible role of the potential complex Nur77/RXR in D1 neurones needed to be better defined. In addition, it is clearly demonstrated that the induction of Nurs members mRNA responds to the activation of signaling cascades of dopaminergic D1 or D2 receptors within the involvement of PKC and ERK kinases is undubtable which opens us the way to a possible involvement of Nur77 in learning mechanisms associated with LIDs and amphetamine that are the Long Terme Potentiation (LTP) and the Long Terme Depotentialisation (LTD).

Este trabalho apresenta a continuidade da avaliação farmacológica das substâncias LASSBio-579, LASSBio-580 e LASSBio-581, através de ensaios in vitro e in vivo, em busca de um novo protótipo para o desenvolvimento de novos fármacos antipsicóticos de segunda geração. LASSBio-581 se liga a receptores D2-like (Ki=0,95 μM), 5-HT1A (Ki=1,2 μM) e 5-HT2A (Ki=11 μM) com afinidades moderadas. Esta substância é capaz de reduzir a temperatura corporal de roedores, um efeito provavelmente mediado pela ativação de receptores 5-HT1A, e inibir o desenvolvimento de head-twiches e ear-scratches induzidos pela administração de um antagonista de receptores 5-HT2A. Estes efeitos demonstram a capacidade de LASSBio-581 em modular o sistema serotonérgico in vivo e in vitro. Porém, quanto avaliado em modelos animais preditivos de ação antipsicótica, LASSBio-581 foi inativo. LASSBio-580 também é capaz de se ligar a receptores D2-like (Ki=0,73 μM), 5-HT1A (Ki=0,48 μM) e 5-HT2A (Ki=5,7 μM) com afinidades moderadas. Esta substância não foi capaz de inibir o desenvolvimento do comportamento de escalada nem a redução da temperatura corporal de roedores induzidos por apomorfina, não apresentando potencial atividade antipsicótica nos ensaios realizados. Já LASSBio-579 é capaz de modular três diferentes sistemas neurotransmissores envolvidos na patofisiologia da esquizofrenia: a neurotransmissão dopaminérgica, serotonérgica e glutamatérgica. Esta substância se liga a receptores D2-like (Ki=0,11 μM), 5-HT1A (Ki=0,09 μM) e 5-HT2A (Ki=2,2 μM) com afinidades adequadas para uma molécula protótipo que se liga a múltiplos alvos. Apresenta ação antidopaminérgica in vivo, demonstrada em três modelos animais preditivos de atividade antipsicótica (sintomas positivos): inibição da estereotipia anfetamínica (NEVES et al., 2003), bloqueio do comportamento de escalada induzido por apomorfina e hipotermia apomorfínica. A ação agonista 5-HT1A de LASSBio-579 in vivo foi claramente demonstrada através de ensaios de aferição da temperatura corporal, onde o efeito hipotérmico induzido por esta substância é completamente bloqueado pela pré-administração de WAY 100635. Porém, a habilidade de LASSBio-579 em modular a atividade de receptores 5-HT2A in vivo permanece incerta. Ensaios eletrofisiológicos preliminares demonstraram um aumento da liberação de glutamato induzido por LASSBio-579 que parece ser mediado pela ativação de receptores 5-HT2A, porém comportamentos ou efeitos relacionados a ativação deste sub-tipo de receptor serotonérgico não foram identificados em roedores tratados com LASSBio-579. Além disso, a administração de LASSBio-579 não induziu efeitos catatônicos em camundongos em doses até 12 vezes superiores àquela ativa no modelo do bloqueio do comportamento de escalada induzido por apomorfina. Estes resultados demonstram que a estratégia de planejamento de fármacos baseado na estrutura do ligante empregada neste trabalho se mostrou bem sucedida. LASSBio-579 pode ser considerado um novo protótipo de fármaco antipsicótico de segunda geração, ativo em modelos animais de sintomas positivos da esquizofrenia e com baixo potencial de indução de efeitos motores. Porém, algumas limitações em seu perfil farmacológico pode ser identificadas. A afinidade desta substância por receptores dopaminérgicos e serotonérgicos é considerada moderada e inferior a de antipsicóticos atualmente no mercado. Ainda, LASSBio-579 induziu um prejuízo na coordenação motora em roedores e apresentou um perfil farmacocinético pouco adequado a utilização clinica (CONRADO et al., 2008). Estes dados encorajam a busca de substâncias com um perfil farmacológico superior ao de LASSBio-579. Neste sentido, uma triagem farmacológica de 18 derivados N-fenilpiperazínicos análogos a LASSBio-579 foi realizada. Os resultados obtidos nos ensaios de radioligação a receptores D2-like, 5-HT1A e 5-HT2A foram utilizados na proposição de relações qualitativas entre estrutura química das substâncias e a afinidade apresentada pelos diferentes receptores. A partir dos resultados obtidos in vitro, cinco outras substâncias foram selecionadas para avaliação da potencial atividade frente a sintomas positivos da esquizofrenia no modelo de bloqueio do comportamento de escalada induzido por apomorfina. Neste ensaio, apenas LASSBio-664 apresentou atividade, sem induzir catatonia nos animais. Porém, esta substância também induz um importante prejuízo motor nos animais. Ensaios adicionais são necessários a fim de diferenciar o perfil farmacológico de LASSBio-664 e LASSBio-579. Outro objetivo deste trabalho foi iniciar o desenvolvimento de um modelo animal de sintomas da esquizofrenia. Os resultados obtidos até o momento apontam para a possibilidade do desenvolvimento de um modelo relacionado a sintomas negativos/cognitivos da esquizofrenia baseados na esposição à natação forçada repetida. Foi demonstrado que apenas clozapina e não imipramina é capaz de reverter o aumento de imobilidade ao longo dos dias acarretado pela exposição repetida à natação forçada em roedores. Este dado demonstra uma potencial validade preditiva, o primeiro grau de validação necessário para um novo modelo animal. O efeito de LASSBio-579 também foi avaliado neste procolo experimental. / This study strengthened the pharmacological evaluation of the heterocyclic Nphenylpiperazine derivatives LASSBio-579, LASSBio-580 and LASSBio-581 by means of in vitro and in vivo pharmacological assays searching a new second generations antipsychotic lead compound. LASSBio-581 presented moderate affinitties for D2-like (Ki=0.95 μM), 5-HT1A (Ki=1.2 μM) e 5-HT2A (Ki=11 μM). This compound induced an hypothermic effect in rodents probably mediated by 5-HT1A receptor activation. LASSBio-581 administration inhibited the induction of head-twiches and ear-scratches by a 5-HT2A agonist. These results shown that LASSBio-581 modulates serotonergic neurotransmission in vivo and in vitro. However, it was inactive on animal models predictive of antipsychotic activity. LASSBio-580 presented moderate affinitties for D2-like (Ki=0.73 μM), 5-HT1A (Ki=0.48 μM) e 5-HT2A (Ki=5.7 μM). This compound did not inhibited apomophine-induced climbing nor apomorphine-induced hypothermia. Thus, among the three compounds initially evaluated, LASSBio-579 was the only one that exhibited promising results. This derivative was able to modulate three neurotransmitter systems involved in schizophrenia’s pathophysiology: dopaminergic, serotonergic and glutamateric ones. As a multi-target lead compound, LASSBio-579 presented adequated affinities for D2-like, 5-HT1A and 5-HT2A receptors (Ki D2-like = 0.11 μM, 5-HT1A = 0.093 μM and 5-HT2A = 2.2 μM). Its antidopaminergic in vivo effect was demonstrated in three animal models of positive symptons of schizophrenia: amphetamineinduced stereotypy (NEVES et al., 2003), apomorphine-induced climbing behavior and apomorphine-induced hypothermia. Regarding the serotonergic system, LASSBio-579 was considered a 5-HT1A receptor agonist, since the hypothermia induced by this compound was blocked by WAY 100,635 pre-administration. The ability of LASSBio-579 to modulate 5-HT2A receptors was not fully characterized. Electrophysiological assays demonstrated an increase on synaptic glutamate release induced by LASSBio-579 that may be related to 5-HT2A receptor activation, however this compound did not induce any behavior related to 5-HT2A activation in rodents. In addition, LASSBio-579 did not induce catalepsy in mice at doses 12 folds higher than those active at apomorphine-induced climbing test. Thus, LASSBio-579 represents a new antipsychotic lead compound active in animal models of positive symptoms of schizophrenia and with a mild propensity to induce motor side effects. However, some limitations on LASSBio-579’s pharmacological profile could be identified. The affinity of LASSBio-579 for dopamine and serotonin receptors is moderate and lower than those presented by second generation antipsychotics. Furthermore, this compound induced a mild decrease on locomotion and exploratory behavior, a meaningful motor coordination impairment and presents a limited oral bioavailability and a low brain penetration (CONRADO et al., 2008). Considering this, a pharmacologycal screening of 18 N-phenylpiperazine derivatives were done in order to achieve an optimized analogue of LASSBio-579. Structural features of this molecular scaffold were discussed regarding binding affinity and selectivity for D2-like, 5-HT1A and 5-HT2A receptors. Among the compounds prepared, LASSBio-664 exhibited an adequate binding profile and a potential for schizophrenia positive symptoms treatment without cataleptogenic effects. However, the motor coordination impairment remained. Additional pharmacological characterization of LASSBio-664 is still needed to find differences from LASSBio-579’s profile. Another aim of this study was to start the development of an animal model of schizophrenia symptons. It was shown that clozapine but not imipramine presented an anti-immobility effect in animals repeated exposed to forced swimming. This result points to the usefullness of repeated forced swimming protocol for developing new animal models predictive of antipsychotic action. The effect of LASSBio-579 in this protocol was also evaluated.

Coordenação de Aperfeiçoamento de Pessoal de Nível Superior / Schizophrenia is a psychiatric disorder that affects 1% of world population and its pharmacologic treatment consists in the use of antipsychotics. It is known that chronic treatment with classical antipsychotics, such as haloperidol, can cause motors disturbers, among which stands out the tardive dyskinesia (TD). The TD pathophysiology has been associated with oxidative stress increase in areas of the brain related to the movements control. So, studies have proposed the use of natural compounds with antioxidants properties to decrease the production of reactive species on TD or on animals models of orofacial dyskinesia (OD). Bauhinia forficata (B. forficata), a plant used on folk medicine such as hypoglycemic, and presents antioxidant properties, could be used to reduce the oxidative stress present on OD. Thus, the first aim of this study was evaluate the effect of B. forficata on in vitro lipid peroxidation induced by pro-oxidants and also, the possible protector effect on OD, through of the quantifications of vacuous chewing movements (VCM), and on locomotor and exploratory activities decrease induced by haloperidol in rats (manuscript 1). B. forficata prevented the lipid peroxidation by pro-oxidant agents nitroprusside sodium, Fe2+/EDTA and Fe2+. Furthermore, adult male rats received haloperidol (38 mg/kg) each 28 days for 16 weeks and B. forficata decoction (2.5 g/L) on the place of drinking water or water to drink everyday for 16 weeks. The haloperidol treatment increased the VCM and reduced the locomotor and exploratory activities relative to control group on open field test. B. forficata co-treatment was not able to prevent the motor alterations induced by haloperidol, as well as only partially prevented VCM in rats. On the other hand, B. forficata caused an increase on locomotor activity. These results showed that B. forficata has antioxidant potential, but this effect is associated partially to protection against VCM induced by haloperidol in rats (manuscript 1). Some studies have showed that chronic treatment with classic antipsychotics can cause metabolic side effects. B. forficata which is known on folk medicine by glycemia decrease, could prevent the metabolic side effects caused by use of antipsychotics. So, the other propound of this study was to investigate the weight gain, glycemic levels and other metabolic parameters in rats chronically treated with haloperidol and the possible effect of B. forficata on these metabolic side effects (manuscript 2). After 16 weeks of treatment, the animals treated with haloperidol showed high glycemic prevalence and high glucose levels. B. forficata co-treatment elevated glucose and triglycerides levels. No difference was observed on cholesterol, urea, creatinine, alanine aminotransferase (AST), aspartate aminotransferase (ALT) and lactate dehydrogenase (LDH). Haloperidol treatment caused weight gain and promoted a significant decrease on brain/body weight rate. In conclusion, B. forficata co-treatment and haloperidol can increase the number of hyperglycemic animals. Thus, is necessary to emphasize the importance of more toxicology studies on chronic treatment with B. forficata to avoid health implications of population. / A esquizofrenia é uma desordem psiquiátrica que atinge cerca de 1% da população mundial e seu tratamento farmacológico consiste na utilização de antipsicóticos. Sabe-se que o tratamento crônico com antipsicóticos clássicos, como o haloperidol, pode causar distúrbios motores, dentre os quais se destaca a discinesia tardia (DT). A fisiopatologia da DT tem sido associada ao aumento do estresse oxidativo em áreas do cérebro relacionadas ao controle dos movimentos. Desta forma, estudos têm proposto o uso de compostos naturais com propriedades antioxidantes para diminuir a produção de espécies reativas na DT ou em modelos animais de discinesia orofacial (DO). A Bauhinia forficata (B. forficata), uma planta utilizada na medicina popular como hipoglicemiante, e que apresenta propriedades antioxidantes, poderia ser utilizada para reduzir o estresse oxidativo presente na DO. Logo, o primeiro objetivo deste estudo foi avaliar o efeito da B. forficata na peroxidação lipídica in vitro induzida por diferentes pró-oxidantes, e também o possível efeito protetor da B. forficata na DO, através da quantificação dos movimentos de mascar no vazio (MMV), e na diminuição da atividade locomotora e exploratória induzidos por haloperidol em ratos (manuscrito 1). A B. forficata preveniu a formação de peroxidação lipídica induzida pelos agentes pró-oxidantes nitroprussiato de sódio, Fe2+/EDTA e Fe2+. Além disso, ratos adultos machos receberam haloperidol (38 mg/kg) a cada 28 dias por 16 semanas, e o decocto de B. forficata (2,5 g/L) no lugar da água de beber ou água para beber todos os dias por 16 semanas. O tratamento com o haloperidol aumentou o número de MMV, e reduziu as atividades locomotora e exploratória dos animais em relação ao grupo controle no teste do campo aberto. O co-tratamento com B. forficata não foi capaz de prevenir as alterações motoras induzidas por haloperidol, bem como preveniu apenas parcialmente os MMV em ratos. Por outro lado, a B. forficata sozinha causou um aumento na atividade locomotora. Os resultados desse estudo mostraram que a B. forficata tem potencial antioxidante, mas esse efeito está associado apenas parcialmente à proteção contra os MMV induzidos pelo haloperidol em ratos (manuscrito 1). Alguns estudos têm mostrado que o tratamento crônico com antipsicóticos clássicos pode causar efeitos colaterais metabólicos. A B. forficata, conhecida na medicina popular por diminuir a glicemia, poderia prevenir os efeitos colaterais metabólicos causados pelo uso de antipsicóticos. Assim, outra proposta deste estudo foi investigar o ganho de peso, os níveis glicêmicos e outros parâmetros metabólicos em ratos tratados cronicamente com haloperidol e o possível efeito da B. forficata nesses efeitos colaterais metabólicos (manuscrito 2). Após 16 semanas de tratamento, os animais tratados com haloperidol apresentaram alta prevalência glicêmica e altos níveis séricos de glicose. O co-tratamento com B. forficata elevou os níveis de glicose e de triglicerídeos. Nenhuma diferença foi observada nos níveis de colesterol, uréia, creatinina, alanina aminotransferase, aspartato aminotransferase (AST) e lactato desidrogenase (LDH). O tratamento com haloperidol causou aumento no ganho de peso corporal e promoveu significante diminuição na taxa peso do cérebro/corpo. Em conclusão, o co-tratamento com B. forficata e haloperidol pode aumentar o número de animais hiperglicêmicos. Assim, é necessário enfatizar a importância de mais estudos toxicológicos sobre o tratamento crônico com B. forficata para evitar implicações na saúde da população.

Este trabalho apresenta a continuidade da avaliação farmacológica das substâncias LASSBio-579, LASSBio-580 e LASSBio-581, através de ensaios in vitro e in vivo, em busca de um novo protótipo para o desenvolvimento de novos fármacos antipsicóticos de segunda geração. LASSBio-581 se liga a receptores D2-like (Ki=0,95 μM), 5-HT1A (Ki=1,2 μM) e 5-HT2A (Ki=11 μM) com afinidades moderadas. Esta substância é capaz de reduzir a temperatura corporal de roedores, um efeito provavelmente mediado pela ativação de receptores 5-HT1A, e inibir o desenvolvimento de head-twiches e ear-scratches induzidos pela administração de um antagonista de receptores 5-HT2A. Estes efeitos demonstram a capacidade de LASSBio-581 em modular o sistema serotonérgico in vivo e in vitro. Porém, quanto avaliado em modelos animais preditivos de ação antipsicótica, LASSBio-581 foi inativo. LASSBio-580 também é capaz de se ligar a receptores D2-like (Ki=0,73 μM), 5-HT1A (Ki=0,48 μM) e 5-HT2A (Ki=5,7 μM) com afinidades moderadas. Esta substância não foi capaz de inibir o desenvolvimento do comportamento de escalada nem a redução da temperatura corporal de roedores induzidos por apomorfina, não apresentando potencial atividade antipsicótica nos ensaios realizados. Já LASSBio-579 é capaz de modular três diferentes sistemas neurotransmissores envolvidos na patofisiologia da esquizofrenia: a neurotransmissão dopaminérgica, serotonérgica e glutamatérgica. Esta substância se liga a receptores D2-like (Ki=0,11 μM), 5-HT1A (Ki=0,09 μM) e 5-HT2A (Ki=2,2 μM) com afinidades adequadas para uma molécula protótipo que se liga a múltiplos alvos. Apresenta ação antidopaminérgica in vivo, demonstrada em três modelos animais preditivos de atividade antipsicótica (sintomas positivos): inibição da estereotipia anfetamínica (NEVES et al., 2003), bloqueio do comportamento de escalada induzido por apomorfina e hipotermia apomorfínica. A ação agonista 5-HT1A de LASSBio-579 in vivo foi claramente demonstrada através de ensaios de aferição da temperatura corporal, onde o efeito hipotérmico induzido por esta substância é completamente bloqueado pela pré-administração de WAY 100635. Porém, a habilidade de LASSBio-579 em modular a atividade de receptores 5-HT2A in vivo permanece incerta. Ensaios eletrofisiológicos preliminares demonstraram um aumento da liberação de glutamato induzido por LASSBio-579 que parece ser mediado pela ativação de receptores 5-HT2A, porém comportamentos ou efeitos relacionados a ativação deste sub-tipo de receptor serotonérgico não foram identificados em roedores tratados com LASSBio-579. Além disso, a administração de LASSBio-579 não induziu efeitos catatônicos em camundongos em doses até 12 vezes superiores àquela ativa no modelo do bloqueio do comportamento de escalada induzido por apomorfina. Estes resultados demonstram que a estratégia de planejamento de fármacos baseado na estrutura do ligante empregada neste trabalho se mostrou bem sucedida. LASSBio-579 pode ser considerado um novo protótipo de fármaco antipsicótico de segunda geração, ativo em modelos animais de sintomas positivos da esquizofrenia e com baixo potencial de indução de efeitos motores. Porém, algumas limitações em seu perfil farmacológico pode ser identificadas. A afinidade desta substância por receptores dopaminérgicos e serotonérgicos é considerada moderada e inferior a de antipsicóticos atualmente no mercado. Ainda, LASSBio-579 induziu um prejuízo na coordenação motora em roedores e apresentou um perfil farmacocinético pouco adequado a utilização clinica (CONRADO et al., 2008). Estes dados encorajam a busca de substâncias com um perfil farmacológico superior ao de LASSBio-579. Neste sentido, uma triagem farmacológica de 18 derivados N-fenilpiperazínicos análogos a LASSBio-579 foi realizada. Os resultados obtidos nos ensaios de radioligação a receptores D2-like, 5-HT1A e 5-HT2A foram utilizados na proposição de relações qualitativas entre estrutura química das substâncias e a afinidade apresentada pelos diferentes receptores. A partir dos resultados obtidos in vitro, cinco outras substâncias foram selecionadas para avaliação da potencial atividade frente a sintomas positivos da esquizofrenia no modelo de bloqueio do comportamento de escalada induzido por apomorfina. Neste ensaio, apenas LASSBio-664 apresentou atividade, sem induzir catatonia nos animais. Porém, esta substância também induz um importante prejuízo motor nos animais. Ensaios adicionais são necessários a fim de diferenciar o perfil farmacológico de LASSBio-664 e LASSBio-579. Outro objetivo deste trabalho foi iniciar o desenvolvimento de um modelo animal de sintomas da esquizofrenia. Os resultados obtidos até o momento apontam para a possibilidade do desenvolvimento de um modelo relacionado a sintomas negativos/cognitivos da esquizofrenia baseados na esposição à natação forçada repetida. Foi demonstrado que apenas clozapina e não imipramina é capaz de reverter o aumento de imobilidade ao longo dos dias acarretado pela exposição repetida à natação forçada em roedores. Este dado demonstra uma potencial validade preditiva, o primeiro grau de validação necessário para um novo modelo animal. O efeito de LASSBio-579 também foi avaliado neste procolo experimental. / This study strengthened the pharmacological evaluation of the heterocyclic Nphenylpiperazine derivatives LASSBio-579, LASSBio-580 and LASSBio-581 by means of in vitro and in vivo pharmacological assays searching a new second generations antipsychotic lead compound. LASSBio-581 presented moderate affinitties for D2-like (Ki=0.95 μM), 5-HT1A (Ki=1.2 μM) e 5-HT2A (Ki=11 μM). This compound induced an hypothermic effect in rodents probably mediated by 5-HT1A receptor activation. LASSBio-581 administration inhibited the induction of head-twiches and ear-scratches by a 5-HT2A agonist. These results shown that LASSBio-581 modulates serotonergic neurotransmission in vivo and in vitro. However, it was inactive on animal models predictive of antipsychotic activity. LASSBio-580 presented moderate affinitties for D2-like (Ki=0.73 μM), 5-HT1A (Ki=0.48 μM) e 5-HT2A (Ki=5.7 μM). This compound did not inhibited apomophine-induced climbing nor apomorphine-induced hypothermia. Thus, among the three compounds initially evaluated, LASSBio-579 was the only one that exhibited promising results. This derivative was able to modulate three neurotransmitter systems involved in schizophrenia’s pathophysiology: dopaminergic, serotonergic and glutamateric ones. As a multi-target lead compound, LASSBio-579 presented adequated affinities for D2-like, 5-HT1A and 5-HT2A receptors (Ki D2-like = 0.11 μM, 5-HT1A = 0.093 μM and 5-HT2A = 2.2 μM). Its antidopaminergic in vivo effect was demonstrated in three animal models of positive symptons of schizophrenia: amphetamineinduced stereotypy (NEVES et al., 2003), apomorphine-induced climbing behavior and apomorphine-induced hypothermia. Regarding the serotonergic system, LASSBio-579 was considered a 5-HT1A receptor agonist, since the hypothermia induced by this compound was blocked by WAY 100,635 pre-administration. The ability of LASSBio-579 to modulate 5-HT2A receptors was not fully characterized. Electrophysiological assays demonstrated an increase on synaptic glutamate release induced by LASSBio-579 that may be related to 5-HT2A receptor activation, however this compound did not induce any behavior related to 5-HT2A activation in rodents. In addition, LASSBio-579 did not induce catalepsy in mice at doses 12 folds higher than those active at apomorphine-induced climbing test. Thus, LASSBio-579 represents a new antipsychotic lead compound active in animal models of positive symptoms of schizophrenia and with a mild propensity to induce motor side effects. However, some limitations on LASSBio-579’s pharmacological profile could be identified. The affinity of LASSBio-579 for dopamine and serotonin receptors is moderate and lower than those presented by second generation antipsychotics. Furthermore, this compound induced a mild decrease on locomotion and exploratory behavior, a meaningful motor coordination impairment and presents a limited oral bioavailability and a low brain penetration (CONRADO et al., 2008). Considering this, a pharmacologycal screening of 18 N-phenylpiperazine derivatives were done in order to achieve an optimized analogue of LASSBio-579. Structural features of this molecular scaffold were discussed regarding binding affinity and selectivity for D2-like, 5-HT1A and 5-HT2A receptors. Among the compounds prepared, LASSBio-664 exhibited an adequate binding profile and a potential for schizophrenia positive symptoms treatment without cataleptogenic effects. However, the motor coordination impairment remained. Additional pharmacological characterization of LASSBio-664 is still needed to find differences from LASSBio-579’s profile. Another aim of this study was to start the development of an animal model of schizophrenia symptons. It was shown that clozapine but not imipramine presented an anti-immobility effect in animals repeated exposed to forced swimming. This result points to the usefullness of repeated forced swimming protocol for developing new animal models predictive of antipsychotic action. The effect of LASSBio-579 in this protocol was also evaluated.

A não adesão ao tratamento farmacológico é um fator determinante ao agravamento das doenças crônicas e está intimamente ligada à recaída de quadros psicóticos. Mesmo sendo uma opção de incentivo à adesão e prevenção de recaída, o Decanoato de Haloperidol por si só não garante o sucesso do tratamento. O objetivo deste trabalho foi descrever as características sócio-demográficas e clínicas de pacientes com prescrição de Decanoato de Haloperidol assistidos em um serviço ambulatorial de saúde mental. Utilizamos um questionário estruturado para a coleta dos dados via prontuário dos pacientes e listagens mensais de retirada da medicação na farmácia do serviço. Foram analisados 167 prontuários de pacientes. A maioria dos pacientes apresentou um maior risco para ocorrência de recaídas frente à retiradas irregulares da medicação no serviço de saúde. De modo geral, o uso irregular da medicação esteve associado a maiores intervalos de dias prescritos para administração de Decanoato de Haloperidol, maior tempo de tratamento/prescrição e diagnóstico de esquizofrenia. Os profissionais de saúde mental precisam estar mais atentos à complexidade do fenômeno da adesão. Ao conhecer melhor sua clientela, estes profissionais podem refletir acerca da assistência psicossocial que um serviço de saúde mental comunitário deve a sua clientela e de ações que minimizem o risco de recaídas nessa população. / No adherence to pharmacological treatment is a determinant factor in the severity of chronic illnesses and is closely related to relapses. Indicated as option of better control the adhesion and fallen again prevention, Haloperidol Decanoate by itself does not guarantee the success of the treatment. The objective was to describe demographic and clinical characteristics of patients with lapsing of Haloperidol Decanoate attended in outside service of mental health. We used structure questionary to tax data in handbooks and withdrawals medication lists of pharmacy service. We analyse 167 handbooks of patients. The analysis of the data showed that the majority of patients presented a bigger risk to occurrence of fallen again front the irregular withdrawals of the medication in health service. In general way, the irregular use of the medication was associates the biggest intervals of days prescribed for administration of Haloperidol Decanoate, greater treatment/indicate time and diagnosis of schizophrenia. Professionals of mental health need to be more intent the complexity adhesion phenomenon. When knowing better its clientele, these professionals can reflect concerning the psicossocial assistance that a communitarian service of mental health must its clientele and of actions that minimize the risk of fallen again into this population.

OBJECTIVES: The aim of our study was to assess clinical presentation of early-onset schizophrenia spectrum disoders (EO-SSD), the time to first improvement and efficacy associated with selected atypical (AAPs) and typical (TAPs) antipsychotics, as well as two main side effects - weight gain and treatment-emergent extrapyramidal symptoms (EPSs) during the treatment in patients with EO-SSD. METHODS: This was a systematic chart review of all patients receiving routine clinical care in our department, with selected AAPs (risperidone, olanzapine, ziprasidone, quetiapine and clozapine) and TAPs (haloperidol, perphenazine and sulpiride), for schizophrenic psychoses, between 1997 and 2007. During this period, our review identified 173 patients (85 males, 88 females; mean age 15.8±1.6 years); their treatment included 297 treatment trials. Data on premorbid adjustment, prodromal symptoms and psychopathology at admission, as well as comorbidity were evaluated based on the patients' medical records. The time to first improvement could be estimated in 258 treatment trials; of these, 195 (76%) comprised AAPs and 63 (24%) TAPs. The time to first improvement was assessed in agreement with the methodology established for retrospective studies as the number of treatment days prior to the first record of improvement...

Notre travail porte sur l’évaluation de l’utilisation et des risques associés aux neuroleptiques en population générale. Nous avons organisé notre travail selon deux approches. D’abord, de façon classique en pharmaco-épidémiologie, une « approche « pragmatique » a permis d’étudier en « vie réelle », (i) l’utilisation des neuroleptiques dans la population générale en France à partir des données de l’Echantillon Généraliste des Bénéficiaires (EGB) et de la base Midi-Pyrénées de Pharmacovigilance (BMPPV), ainsi que dans une population de femmes enceintes à partir de la base de données EFEMERIS (Evaluation chez la Femme Enceinte des MEdicaments et de leurs RISques); (ii) les risques associés à cette utilisation à partir de la BMPPV, de la base mondiale de pharmacovigilance, VigiBase® et de la base de données EFEMERIS. Deuxièmement, nous avons testé une approche originale « explicative » à partir de VigiBase®, la méthode PE-PD (PharmacoEpidémiologie-PharmacoDynamie) pouvant potentiellement mettre en évidence le mécanisme d’action de certains effets indésirables. Nous avons étudié pour cela l’exemple largement connu des mouvements anormaux induits par les neuroleptiques. Pour l’approche « pragmatique », l’analyse montre une modification de l’utilisation des neuroleptiques en population générale et chez la femme enceinte, avec un report des prescriptions des neuroleptiques de première génération (NL1G) vers ceux de deuxième génération (NL2G). Si l’utilisation reste globalement stable, on note un accroissement de l’utilisation des NL2G dans deux populations vulnérables vis-à-vis des effets indésirables : les enfants/adolescents et les femmes enceintes. Malgré le report NL1G vers les NL2G, la prescription des médicaments à fort poids atropinique est élevée et stable au cours des dernières années. Concernant les risques associés, nous n’avons pas pu confirmer dans la base EFEMERIS, le signal retrouvé dans VigiBase de malformations congénitales gastro-intestinales avec les neuroleptiques. Pour les mouvements anormaux, le risque apparait moins important avec la clozapine qu’avec les autres neuroleptiques. Des différences existent entre les classes d’âge, avec, par exemple, moins d’akathisie, mais plus de dystonie aiguë chez les moins de 18 ans. Pour l’approche « explicative », la méthode PE-PD a permis de mettre en évidence l’un des mécanismes potentiels responsables de la réduction des mouvements anormaux sous neuroleptique, à savoir l’action antagoniste sur les récepteurs sérotoninergique 5-HT2A et muscariniques M1. Notre travail de thèse nous a permis à travers l’étude des neuroleptiques de discuter, (1) d’abord au niveau méthodologique, de la complémentarité entre les bases médicamenteuses (type EGB et EFEMERIS) et les bases de pharmacovigilance pour les études d’utilisation des médicaments et des risques associés, avec, en particulier, l’opportunité d’étudier le risque malformatif des médicaments avec les bases de pharmacovigilance. Ensuite, (2), au niveau pharmacologique, de mettre en évidence la problématique du poids atropinique des prescriptions associées aux neuroleptiques sur les bénéfices et/ou les dommages. Enfin, (3), dans une approche de pharmacologie sociale, en partant des résultats de la méthode PE-PD, de discuter de la place du mécanisme d’action des médicaments dans leur évaluation, de la pharmacologie fondamentale, en passant par la pharmacologie clinique, et de son éventuelle intégration en pharmaco-épidémiologie. / Our works deals with the use of antipsychotic drugs and risks associated with their use in general population. First, a “pragmatic” approach of pharmacoepidemiology was performed to evaluate in real life, (i) the use of antipsychotics in the French general population by using data from EGB (Echantillon Généraliste des Bénéficiaires) and from the Midi-Pyrénées Pharmacovigilance database (BMPPV), as well as in pregnant women with data from EFEMERIS (Evaluation chez la Femme Enceinte des MEdicaments et de leurs RISques); (ii) the risks associated with antipsychotics by using the BMPPV, VigiBase® the World Health Organization (WHO) Global Individual Case Safety Report (ICSR) and data from EFEMERIS. Secondly, we examined an original “explanatory” approach with VigiBase®, the PE-PD method (Pharmacoepidemiologic-Pharmacodynamic method) to investigate potential mechanisms of adverse drug reactions (ADRs). We deliberately selected an ADR with a well-established mechanism – i.e. movement disorders – in order to test the validity of the PE-PD method. For the “pragmatic approach”, our analysis found marked changes in antipsychotics use both in the general population and in pregnant women, with a switch from first generation antipsychotics (FGAPs) to second generation antipsychotics (SGAPs). The use of antipsychotics in general population remained relatively constant but a raise was observed for SGAPs in two vulnerable populations concerning ADRs: children/adolescents and pregnant women. Despite the switch (from FGAPs to SGAPs), associated use of drugs with high atropinic properties was highly prevalent and constant during the recent years. Concerning risks, no association was found in EFEMERIS data, not allowing to confirm the signal of gastrointestinal congenital malformations associated with antipsychotic use described in VigiBase®. For movement disorders, the risk found was lower with clozapine than with other antipsychotics. Risks of movement disorders differed according to age classes, with less akathisia, but more acute dystonia in children and adolescents (< 18 years). For the “explanatory approach”, the PE-PD method found an inverse correlation between serotonergic 5-HT2A or muscarinic M1 receptor occupancies and reports of movement disorders involving APs. Our thesis work allowed, through the study of neuroleptics (1) first, discussing, from a methodological point of view, the complementarity between French drugs databases (EGB and EFEMERIS) and pharmacovigilance databases for studying drug utilization and risks associated, in particular the opportunity to study the risk of malformations associated with drug use; then, (2) at the pharmacological level, highlighting the issues on the benefits and / or harms of the atropinic burden of prescriptions associated with antipsychotics; finally, (3) in a social pharmacology approach, starting from the results of the PE-PD method, discussing the place of the mechanism of drug action in drug evaluation, from basic to clinical pharmacology, and its possible integration into pharmacoepidemiology.