MSc (Med) (Child Health Neurodevelopment), Faculty of Health Sciences, University of the Witwatersrand / Background Disruptive behaviour is the most commonly reported mental health problem in individuals with learning disability. Pharmacotherapy is part of a multidisciplinary approach to the treatment of disruptive behaviour. Risperidone, an atypical antipsychotic drug, is the most commonly used treatment for symptom improvement. It is therefore important to establish the efficacy and safety of risperidone therapy in this dependent, vulnerable and young population, given the well documented adverse effects and the potential for long term treatment. Objectives To assess the effects of risperidone for disruptive behaviour in children and adolescents with learning disability. Search strategy The following electronic databases were searched: CENTRAL (Cochrane Central Register of Controlled Trials); MEDLINE; PsycINFO; CINAHL; Clinicaltrials.gov; National Research Register (NRR). In addition, reference lists of relevant publications and narrative reviews were checked; handsearches were done; authors of published trials and pharmaceutical manufacturer of risperidone (Risperdal) were contacted. Selection criteria All randomised or quasi-randomised controlled trials of risperidone versus placebo (or no treatment) for children and adolescents (age less than 18 years) with a diagnosis of learning disability and disruptive behaviour were considered. Data collection and analysis Trial eligibility and data quality were evaluated and analysed by the author and independently verified by an additional reviewer. Unpublished data were considered for inclusion and relevant authors were contacted in the case of incomplete data. Results Four randomised controlled trials involving 279 children and adolescents were identified. The majority of the children were living at home and not institutionalised. Meta-analyses of the primary outcome scales (Nisonger Child Behaviour Rating Form, Aberrant Behaviour Checklist, Behaviour Problem Inventory) measuring several core symptoms of disruptive behaviour, namely conduct problems, self-injury, irritability, aggressive / destructive behaviours and stereotypy suggest statistically significant improvement in disruptive type behaviours in children treated with risperidone compared to placebo. Adverse event data showed that the prevalence of adverse effects viz. weight gain, sedation / somnolence and raised prolactin levels were significantly higher in the children receiving risperidone. Conclusions In the studies included in this review, risperidone treatment for disruptive behaviour in learning disabled children and adolescents appears to have a beneficial effect on certain symptoms of disruptive behaviour. However, the applicability of these findings to wider clinical practice remains unclear, due to poor methodological quality, inadequate study sample size and short duration of treatment of the included studies. Long term safety has not been established and serious adverse effects, affecting growth, are of concern. Further research is required to establish the efficacy and safety of risperidone for disruptive behaviour in learning disabled children and adolescents in clinical practice.

Bioinformatics is a unique field in that it incorporates many different disciplines, including biology, computer science, and statistics, to study biological data. There is a vast array of techniques that utilize bioinformatics, including pangenomics, RNASeq, whole genome metagenomics, and 16S sequencing. To study bacterial interactions, we used a model system of species interactions, Myxococcus xanthus. M. xanthus is a soil bacterium that is a known predator of other bacteria. It has one of the largest repertoires of two component systems (TCS) to respond to external stresses. TCS are a pair of proteins, one that senses environmental stress (histidine kinase, HK) and another that usually acts as a transcriptional regulator (response regulators, RR). We studied a class of RRs, NtrC-like, reliant on an alternative sigma factor, sigma54. The oligomerization of NtrC-like RRs is regulated to modulate activation of the protein, which would change the bacterium’s ability to respond to its environment. We studied HsfA, a NtrC-like RR that regulates specialized metabolites. Specialized metabolites are used in bacterial interactions. In predation interactions they are used to kill prey. Our goal was to find genes that might be involved in specialized metabolite production that would aid in predation. We used prediction tools to find putative binding sites of HsfA to find potentially new metabolites. We used two motifs to attempt to predict if the oligomerization of these response regulators is positively or negatively regulated. We found that the presence of a motif in the receiver domain to be associated with negative regulation of oligomerization, but further studies are needed to experimentally confirm this finding. One environment in which bacterial interactions occur is in the gut. The gut microbiome is the consortium of organisms and their genomic content in the gastrointestinal tract. The gut microbiome is sensitive to aspects of a person’s lifestyle, such as diet and medication. Here we studied the effect of two different diets and two drugs on the gut microbiome. Risperidone, an antipsychotic used to treat schizophrenia and bipolar disorder, has been shown to cause obesity and diabetes. We studied the effect of diet and risperidone usage on weight gain and the microbiome using a C57Bl/6J female mouse model. Our results show that diet has a strong impact on the microbial composition of the gut in response to risperidone. As many mental health patients stop and restart their medication, we examined the effect of stopping and restarting risperidone on the microbiome. When risperidone is stopped the microbiome reverts to a state similar to the control group but diverges into a different microbial composition upon restarting treatment. Interestingly, mice did not gain significantly more weight than their control group upon the second risperidone treatment. Further studies are needed to examine the functional changes occurring with the stop and restart of risperidone to determine the mechanism of mice resisting weight gain during the second round of treatment. Captopril is used to treat hypertension, a very common disease in the United States. Here we studied the effect of captopril on weight gain, metabolic phenotypes, and the gut microbiome. Our results showed that captopril caused an increase in resting metabolic rate (RMR) in mice. This occurred through an increase in energy expenditure. This increase in RMR had the effect of captopril-treated mice being resistant to weight gain. Our group has previously shown that the gut microbiome can directly affect RMR. Therefore, we studied the gut microbiome of captopril-treated mice. We observed a shift in their gut microbiome to organisms Akkermansia muciniphila and Lactobacillus, associated with lean body mass. Captopril therefore has the potential to be a better medication to treat patients with both hypertension and obesity. Further studies are needed to determine the effect of captopril on the microbiome in a hypertension mouse model.

Thesis (M.A.)--University of North Carolina at Greensboro, 2006. / Title from PDF title page screen. Advisor: Walter Salinger; submitted to the Dept. of Psychology. Includes bibliographical references (p. 76-86).

Thesis (M.A.)--University of North Carolina at Greensboro, 2006. / Title from PDF title page screen. Advisor: Walter Salinger; submitted to the Dept. of Psychology. Includes bibliographical references (p. 76-86).

Στην παρούσα εργασία επιχειρήθηκε η ταυτοποίηση της κρυσταλλικής φάσης της δραστικής ουσίας ρισπεριδόνη στα εμπορικά δισκία αυτής. Να σημειωθεί ότι για τη ρισπεριδόνη είναι γνωστά τρία πολύμορφα. Οι τεχνικές που χρησιμοποιήθηκαν για την υλοποίηση αυτού του σκοπού, ήταν η περίθλαση ακτίνων Χ (XRPD), η φασματοσκοπία υπερύθρου (IR) και η φασματοσκοπία Raman. Η ανάλυση με τις τρεις αυτές τεχνικές κατέδειξε ότι ενώ οι φασματοσκοπίες Raman και IR είχαν χαμηλότερα όρια ανίχνευσης, ως προς τη ρισπεριδόνη, ωστόσο δεν κατάφεραν να αναγνωρίσουν το χρησιμοποιούμενο πολύμορφο. Αυτό κατέστει εφικτό με χρήση της περίθλασης ακτίνων Χ, με τη βοήθεια της οποίας πιστοποιήθηκε ότι η ρισπεριδόνη δεν υπόκεινται σε κανενός είδους κρυσταλλική μετατροπή τόσο κατά τη διαδικασία παραγωγής του δισκίου όσο και κατά την πάροδο ενός χρονικού διαστήματος δύο ετών. / The scope of the current study was the identification of the crystal phase of an active pharmaceutical ingredient, called risperidone, in the commercial tablets. In this point it should be noticed that there are three known polymorphs for risperidone. The techniques used for the materialization of this target were X-ray Powder Diffraction (XRPD), Raman spectroscopy and Infra-red spectroscopy (IR). The analysis with these three techniques showed that although Raman and IR had lower detection limits nevertheless they did not have the ability to identify the existent polymorph. This was attained by using XRPD, by means of which it was certified that risperidone does not undergo any crystalline transformation neither during the manufacturing procedure nor during a period of two years.

Οι κυκλοδεξτρίνες είναι κυκλικοί ολιγοσακχαρίτες αποτελούμενοι από μόρια α-D-γλυκοπυρανόζης συνδεδεμένα με α-1→4 γλυκοζιτικούς δεσμούς. Λόγω στερεοχημικών περιορισμών οι κυκλοδεξτρίνες δεν έχουν απόλυτα κυλινδρικό σχήμα αλλά μοιάζουν περισσότερο με κόλουρο κώνο. Αποτέλεσμα της χαρακτηριστικής των δομής είναι η δημιουργία μιας υδρόφιλης εξωτερικής επιφάνειας και μιας υδρόφοβης εσωτερικής κοιλότητας. Στην εσωτερική κοιλότητα μπορούν να εισέλθουν, μερικά ή ολικά, υδρόφοβα μόρια τα οποία αλληλεπιδρώντας μέσω ασθενών διαμοριακών αλληλεπιδράσεων σχηματίζουν σύμπλοκα έγκλεισης. Ο σχηματισμός συμπλόκων έγκλεισης βιοδραστικών ενώσεων με κυκλοδεξτρίνες αυξάνει την διαλυτότητα δυσδιάλυτων στο νερό βιοδραστικών ενώσεων, βελτιώνει την βιοδιαθεσιμότητά τους, αυξάνει την σταθερότητά τους και μειώνει σημαντικά τυχόν παρενέργειές των. Μια ακόμη τεχνική που χρησιμοποιείται για την αύξηση της διαλυτότητας δυσδιάλυτων στο νερό βιοδραστικών ενώσεων είναι η διαλυτοποίησή τους στο εσωτερικό μικυλλίων επιφανειοδραστικών ενώσεων. Η τεχνική αυτή προσφέρει ανάλογα πλεονεκτήματα όπως και η συμπλοκοποίηση των βιοδραστικών ενώσεων με τις κυκλοδεξτρίνες και χρησιμοποιείται ήδη σε εμπορικά σκευάσματα. Στην παρούσα εργασία μελετήθηκε η επίδραση της φύσης και της συγκέντρωσης κυκλοδεξτρινών (β-CD, Μe-β-CD, HP-β-CD, HP-γ-CD) στην διαλυτότητα της βιοδραστικής ένωσης ρισπεριδόνη (risperidone). Επίσης, προσδιορίσθηκε η μέθοδος παρασκευής ενώσεων έγκλεισης της ρισπεριδόνης με τις παραπάνω κυκλοδεξτρίνες χρησιμοποιώντας κατάλληλες τεχνικές χαρακτηρισμού των σχηματιζόμενων στερεών. Ακόμη, μελετήθηκε η διαλυτοποίηση της ρισπεριδόνης σε μικύλλια τριών επιφανειοδραστικών ενώσεων (SDS, Brij35, Cremophor EL). Η ρισπεριδόνη είναι μια αντιψυχωσική βιοδραστική ένωση που ανήκει σε μια νέα κατηγορία ενώσεων τα παράγωγα της benzisoxazole. Η φαρμακολογική δράση της εστιάζεται στους υποδοχείς της D2 (dopamine) και 5-ΗΤ2 (serotonin) στον εγκέφαλο. Πρόκειται για μια δυσδιάλυτη στο νερό βιοδραστική ένωση με αποτέλεσμα να δημιουργούνται προβλήματα στις φαρμακοτεχνικές μορφές της. Είναι ασθενής βάση με δύο στάδια πρωτονίωσης με συνέπεια η διαλυτότητά της να μειώνεται με την αύξηση του pH. Παρασκευάσθηκαν στερεά μίγματα της ρισπεριδόνης με τις κυκλοδεξτρίνες (β-CD, Μe-β-CD, HP-β-CD, HP-γ-CD) χρησιμοποιώντας διαφορετικές τεχνικές(απλή ανάμιξη, λειοτρίβηση με τον σχηματισμό πάστας, συγκαταβύθιση και λυοφιλοποίηση) προκειμένου να προσδιορισθεί η τεχνική ή οι τεχνικές που οδηγούν στο σχηματισμό συμπλόκου έγκλεισης. Για την μελέτη των στερεών μιγμάτων και την πιστοποίηση του σχηματισμού συμπλόκων έγκλεισης μεταξύ της ρισπεριδόνης και των υπό μελέτη κυκλοδεξτρινών χρησιμοποιήθηκαν οι τεχνικές α) Περίθλαση Ακτίνων-Χ, β) Διαφορική Θερμιδομετρία Σάρωσης και γ) Ηλεκτρονική Μικροσκοπία Σάρωσης. Επίσης μελετήθηκε η στερεοχημεία των σχηματιζόμενων συμπλόκων της ρισπεριδόνης με τις υπό μελέτη κυκλοδεξτρίνες με την τεχνική του Πυρηνικού Μαγνητικού Συντονισμού (NMR). Για την παρασκευή των διαλυμάτων χρησιμοποιήθηκε ο δευτεριωμένος διαλύτης d6-DMSO. Πραγματοποιήθηκαν μελέτες διαλυτότητας της ρισπεριδόνης σε υδατικά διαλύματα παρουσία διαφορετικών συγκεντρώσεων των β-CD, Μe-β-CD, HP-β-CD, HP-γ-CD σε θερμοκρασίες 25, 30 και 35°C σε διαλύματα PBS με pH=10. Η μελέτη της επίδρασης των επιφανειοδραστικών ενώσεων έγινε στους 25°C και σε pH=10. Οι τεχνικές της διαφορικής θερμιδομετρίας σάρωσης και περίθλασης ακτίνων-Χ είναι οι καταλληλότερες για την μελέτη και τον χαρακτηρισμό των στερεών συμπλόκων της ρισπεριδόνης με τις κυκλοδεξτρίνες. Συγκεκριμένα, η απουσία της ισχυρής ενδόθερμης κορυφής που αντιστοιχεί στο σημείο τήξεως της ρισπεριδόνης στα θερμογραφήματα των στερεών μιγμάτων που παρασκευάσθηκαν με την μέθοδο της συγκαταβύθισης αποτελεί σαφή ένδειξη για τον σχηματισμό συμπλόκων έγκλεισης της ρισπεριδόνης με τις κυκλοδεξτρίνες. Ανάλογα, στα φάσματα περίθλασης ακτίνων-X η απουσία των χαρακτηριστικών κορυφών της ρισπεριδόνης στα μίγματα που παρασκευάσθηκαν με την μέθοδο της συγκαταβύθισης αποτελεί μια επιπλέον ένδειξη για τον σχηματισμό συμπλόκων ενώσεων μεταξύ της ρισπεριδόνης και των υπό μελέτη κυκλοδεξτρινών. Αντίθετα, η τεχνική της ηλεκτρονικής μικροσκοπίας σάρωσης δεν αποτελεί από μόνη της ασφαλή τεχνική για την ταυτοποίηση των συμπλόκων έγκλεισης μεταξύ τη ρισπεριδόνης και των κυκλοδεξτρινών. Από την μελέτη των φασμάτων NMR των διαλυμάτων που περιείχαν σε διάφορες αναλογίες ρισπεριδόνη και τις υπό μελέτη κυκλοδεξτρίνες προσδιορίσθηκε η στερεοχημεία των σχηματιζόμενων συμπλόκων. Από την μελέτη των ισοθέρμων διαλυτότητας διαπιστώθηκε ότι: α) η διαλυτότητα της ρισπεριδόνης αυξάνει με την συγκέντρωση της κυκλοδεξτρίνης σε όλες τις θερμοκρασίες για όλες τις κυκλοδεξτρίνες που μελετήθηκαν και β) οι ισόθερμοι διαλυτότητας είναι της μορφής AL στην οποία όλα τα σχηματιζόμενα σύμπλοκα είναι ευδιάλυτα και έχουν στοιχειομετρία 1:1. Στους 25°C, την μεγαλύτερη συμπλεκτική ικανότητα παρουσίασε η β- CD. Οι Me-β-CD και HP-β-CD παρουσίασαν μικρότερη συμπλεκτική ικανότητα ενώ η συμπλεκτική ικανότητα για την HP-γ-CD ήταν σημαντικά μειωμένη. Με βάση την συμπλεκτική ικανότητά τους, οι υπό μελέτη κυκλοδεξτρίνες μπορούν να ταξινομηθούν με την ακόλουθη σειρά: β-CD > Me-β-CD > HP-β-CD >> HP-γ-CD Η αύξηση της θερμοκρασίας δεν επηρέασε την μορφή των ισοθέρμων διαλυτότητας, επηρέασε την διαφορά της συμπλεκτικής ικανότητας των β-CD και Me-β-CD. Στους 30°C, η συμπλεκτική ικανότητα της Me-β-CD ήταν ελάχιστα μικρότερη από αυτή της β-CD ενώ στους 35°C έγινε λίγο μεγαλύτερη σε σύγκριση με την β-CD. Για τις άλλες δύο κυκλοδεξτρίνες HP-β-CD και HP-γ-CD διατηρήθηκε η ίδια κατάσταση και στις δύο θερμοκρασίες. Από την κλίση των ισοθέρμων διαλυτότητας προσδιορίσθηκαν οι σταθερές σχηματισμού των συμπλόκων έγκλεισης της ρισπεριδόνης με τις υπό μελέτη κυκλοδεξτρίνες. Η διαφοροποίηση της συμπλεκτικής ικανότητας των β-CD, Me-β-CD, HP-β-CD και HP-γ-CD μπορεί να αποδοθεί α) στο διαφορετικό μέγεθος της υδρόφοβης κοιλότητας και β) στην διαφορετική υδροφοβικότητα των υποκατεστημένων κυκλοδεξτρινών. Από την μεταβολή των σταθερών σχηματισμού των συμπλόκων με την αύξηση της θερμοκρασίας υπολογίσθηκε η μεταβολή της ενθαλπίας (ΔΗ) και η μεταβολή της εντροπίας (ΔS) για τον σχηματισμό των συμπλόκων. Σε όλες τις κυκλοδεξτρίνες υπολογίσθηκε αρνητική μεταβολή της ενθαλπίας και θετική αλλά μικρή μεταβολή της εντροπίας. Οι μεταβολές αυτές συνδέονται άμεσα με την απελευθέρωση των μορίων νερού ‘υψηλής ενθαλπίας’ που βρίσκονται στην υδρόφοβη κοιλότητα της κυκλοδεξτρίνης και με υδρόφοβες αλληλεπιδράσεις ανάμεσα στην ρισπεριδόνη και στην κυκλοδεξτρίνη. Οι δύο αυτές αλληλεπιδράσεις ευνοούν τον σχηματισμό και την σταθερότητα των σχηματιζόμενων συμπλόκων. Τέλος, από τα πειράματα διαλυτότητας της ρισπεριδόνης παρουσία των επιφανειοδραστικών (SDS, Brij35 και Cremophor EL) διαπιστώθηκε ότι α) την μεγαλύτερη αύξηση διαλυτότητας προκάλεσε το SDS και β) η διαλυτοποιητική ικανότητα του Brij35 ήταν λίγο μικρότερη από αυτή του Cremophor EL. / Cyclodextrins are cyclic oligosaccharides composed of a-D-glucopyranose units connected with a-1→4 glucosidic bonds. Because of stereochemical restrictions, cyclodextrins do not have absolutely a cylindrical shape but they resemble a truncated cone. As a result of their characteristic structure is the formation of a hydrophilic external surface and a hydrophobic internal cavity. In the internal cavity can enter partly or wholly hydrophobic molecules which by interacting through weak intermolecular interactions form inclusion complexes. The inclusion complex formation of bioactive compounds, improves their bioavailability, enhances their stability and reduces considerably the side effects after administration. Another technique which is used for increasing the solubility of water sparingly soluble bioactive compounds is their solubilization in the micelles of surfactants. This technique offers analogous advantages such as the complexation of bioactive compounds with cyclodextrins and is used already in commercial drugs. In this study, the influence of the nature and concentration of cyclodextrins (β-CD, Μe-β-CD, HP-β-CD, HP-γ-CD) was studied on the solubility of the bioactive compound risperidone. It was also determined the inclusion compounds preparation method of risperidone with the above cyclodextrins by using appropriate characterization techniques of the formed solids. In addition, the solubilization of risperidone in micelles of three surfactant compounds (SDS, Brij35, Cremophor EL) was studied. Risperidone is an antipsychotic bioactive compound which belongs to a new chemical class, the benzisoxazole derivatives. Its pharmacological action focuses on the D2 (dopamine) and 5-HT2 (serotonin) receptors in the brain. It is about a water sparingly soluble bioactive compound resulting in problems on its pharmaceutical formulations. It is a weak base, with two protonization stages, and as such its solubility decreases with pH increase. Risperidone solid mixtures with the cyclodextrins (β-CD, Μe-β-CD, HP-β-CD, HP-γ-CD) were prepared by using different techniques (physical mixture, kneading, coprecipitation, lyophilisation) in order to determine the technique or the techniques that lead to the formation of inclusion complex. For the study of solid mixtures and the assessment of inclusion complex formation between the risperidone and the cyclodextrins under investigation the following techniques were used: a) X-Ray Diffraction (XRD) b) Differential Scanning Calorimetry (DSC) and c) Scanning Electron Microscopy (SEM). Furthermore, the stereochemistry of the formed inclusion complexes of risperidone with the specific cyclodextrins was also assessed with the technique of Nuclear Magnetic Resonance (NMR). As a solvent for the preparation of the solutions it was used the deuterated solvent d6-DMSO. Solubility studies in aqueous solutions of risperidone with the cyclodextrins were performed in 25, 30 and 35°C in PBS (phosphate buffered saline) at pH=10. The study of the influence of the surfactant compounds (SDS, Brij35, Cremophor EL) was realised at 25°C and pH=10. The techniques of DSC and XRD are the most appropriate for the investigation and characterization of solid complexes of risperidone with cyclodextrins. Particularly, the absence of the strong endothermic peak which corresponds to the melting point of risperidone in the DSC thermograms of the solid mixtures which were prepared by the coprecipitation method constitutes a clear indication of the inclusion complex formation of risperidone with the cyclodextrins. Accordingly, in the XRD spectra the absence of the characteristic peaks of risperidone in the mixtures prepared by the coprecipitation method comprises an additional indication for the formation of inclusion complexes between risperidone and the cyclodextrins under investigation. On the contrary, the SEM does not constitute per se a safe method for the identification of the inclusion complexes between risperidone and the cyclodextrins. The stereochemistry of the formed inclusion compounds was assessed through the spectra received by the application of NMR on solutions which contained risperidone and the specific cyclodextrins in different proportions. In particular, it was determined which part of risperidone molecule enters the hydrophobic cavity of cyclodetxrin. The study of solubility isotherms showed that: a) solubility of risperidone increases with the concentration of cyclodextrins at all temperatures studied, for all the cyclodextrins studied and b) solubility isotherms have a graphical representation of the AL type, which shows that all formed complexes are water soluble with 1:1 stoichiometry. At 25°C, β-CD presented the greatest complexive ability, Me-β-CD and HP-γ-CD presented a smaller complexive ability whereas that of HP-γ-CD was significantly decreased. Based on the complexive ability, the cyclodextrins under discussion can be classified in the following order: β-CD>Me-β-CD>HP-β-CD>> HP-γ-CD The temperature increase did not affect the solubility isotherms but affected the difference of complexation ability of β-CD and Me-β-CD. At 30°C, the complexation ability of Me-β-CD was slightly smaller than β-CD’s, whereas at 35°C became slightly greater compared to that of β-CD. For the other two cyclodextrins; namely HP-β-CD and HP-γ-CD the same behavior was maintained at both temperatures. From the gradient of the solubility isotherms the inclusion complex formation constants were determined. The differentiation of the complexation ability of cyclodextrins can be attributed a) to the different size of the hydrophobic cavity and b) to the different hydrophobicity of the substituted cyclodextrins. The enthalpy change (ΔH) and the entropy change (ΔS) during complex formation were calculated from the change of the complexation constants with the increase of the temperature. In all of the cyclodextrins it was found negative enthalpy change and positive but small entropy change. These alterations are directly connected to the release of ‘high enthalpy’ water molecules which are located in the cyclodextrin’s hydrophobic cavity and to hydrophobic interactions among risperidone and cyclodextrin. These two interactions favor the complex formation and stability. Finally, from the solubility experiments of risperidone in the presence of surfactant compounds SDS, Brij35 and Cremophor EL, it was established that a) the greatest solubility increase was caused by the SDS and b) the solubilization ability of Brij35 was a little smaller than Cremophor’s one.

O objetivo desta tese de doutorado foi testar psicofármacos tradicionais em aplicações diferentes das originais, tendo por base o conhecimento de seus mecanismos de ação e uma visão clínica focada no temperamento. A primeira parte da tese é constituída pelo ensaio clínico de flunarizina comparado com haloperidol no tratamento de pacientes com esquizofrenia. A segunda parte é composta de três relatos de caso oriundos da prática clínica e que, apesar de freqüentes, não costumam ser objetos de pesquisa. A flunarizina, um bloqueador de canal de cálcio empregado para tratar enxaqueca e vertigem foi escolhida pelo fato dela produzir sinais e sintomas extrapiramidais em pacientes idosos, o que posteriormente foi relacionado a sua propriedade como antagonista dos receptores dopaminérgicos do tipo D2, além da existência de dados em modelos animais que indicavam o seu potencial efeito antipsicótico. A flunarizina foi eficaz como antipsicótico e apresentou um perfil de efeitos adversos favorável, além de ter uma meiavida longa e baixo custo. A segunda parte da tese apresenta relatos de casos clínicos freqüentes, mas pouco estudados no âmbito científico. Nesse sentido propomos uma avaliação do paciente com base no temperamento afetivo e emocional, de acordo com o modelo de temperamento proposto por Lara e Akiskal (2006). O primeiro aborda o uso de baixas doses de risperidona no tratamento de ciúme patológico. O segundo se refere à comorbidade do transtorno obsessivocompulsivo e transtorno bipolar, tratados com divalproato de sódio e lamotrigina. O último artigo aponta para o uso de baixas doses de quetiapina em pacientes com sintomas sublimiares e temperamento hipertímico ou ciclotímico que apresentam desregulação emocional. Em conjunto, nossos resultados reforçam a idéia de que as medicações existentes podem sem mais exploradas para o tratamento de quadros clínicos diferentes das suas indicações originais. / The aim of this thesis was to test traditional drugs in applications different from the original, on the basis the knowledge of the mechanisms of action and a vision clinic focused on temperament. The first part of thesis consists in clinical trial of flunarizine compared with haloperidol in the treatment of patients with schizophrenia. The second part is composed by three case reports from clinical practice and that, despite frequent, not usually are objects of research. Flunarizine, a calcium channel blocker used to treat migraine and vertigo, was chosen because it induces extrapyramidal signs in elder patients which was later related to antagonist properties at dopamine D2 receptors, beyond the existence of data in animal models, which indicated its potential antipsychotic effect. Flunarizine was effective as antipsychotic and it presents a favorable side effects profile, beyond the long half-life and low cost. The second part of the thesis presents reports of frequent cases, but little studied under scientific scope. Therefore, we propose an evaluation of the patient with basis on the affective and emotional temperament, according to the temperament model proposed by Lara and Akiskal (2006). The first deals with the use of low doses of risperidone in the treatment of pathological jealousy. The second refers to the comorbidity of obsessive-compulsive disorder and bipolar disorder, treated with divalproex sodium and lamotrigine. The last article points to the use of low doses of quetiapine in patients with subthreshold symptoms and hyperthymic or cyclothymic temperaments presenting emotional deregulation. Together, our results strengthen the idea that the existing medications can be further explored for the treatment of clinical indications different from their originals.

O objetivo desta tese de doutorado foi testar psicofármacos tradicionais em aplicações diferentes das originais, tendo por base o conhecimento de seus mecanismos de ação e uma visão clínica focada no temperamento. A primeira parte da tese é constituída pelo ensaio clínico de flunarizina comparado com haloperidol no tratamento de pacientes com esquizofrenia. A segunda parte é composta de três relatos de caso oriundos da prática clínica e que, apesar de freqüentes, não costumam ser objetos de pesquisa. A flunarizina, um bloqueador de canal de cálcio empregado para tratar enxaqueca e vertigem foi escolhida pelo fato dela produzir sinais e sintomas extrapiramidais em pacientes idosos, o que posteriormente foi relacionado a sua propriedade como antagonista dos receptores dopaminérgicos do tipo D2, além da existência de dados em modelos animais que indicavam o seu potencial efeito antipsicótico. A flunarizina foi eficaz como antipsicótico e apresentou um perfil de efeitos adversos favorável, além de ter uma meiavida longa e baixo custo. A segunda parte da tese apresenta relatos de casos clínicos freqüentes, mas pouco estudados no âmbito científico. Nesse sentido propomos uma avaliação do paciente com base no temperamento afetivo e emocional, de acordo com o modelo de temperamento proposto por Lara e Akiskal (2006). O primeiro aborda o uso de baixas doses de risperidona no tratamento de ciúme patológico. O segundo se refere à comorbidade do transtorno obsessivocompulsivo e transtorno bipolar, tratados com divalproato de sódio e lamotrigina. O último artigo aponta para o uso de baixas doses de quetiapina em pacientes com sintomas sublimiares e temperamento hipertímico ou ciclotímico que apresentam desregulação emocional. Em conjunto, nossos resultados reforçam a idéia de que as medicações existentes podem sem mais exploradas para o tratamento de quadros clínicos diferentes das suas indicações originais. / The aim of this thesis was to test traditional drugs in applications different from the original, on the basis the knowledge of the mechanisms of action and a vision clinic focused on temperament. The first part of thesis consists in clinical trial of flunarizine compared with haloperidol in the treatment of patients with schizophrenia. The second part is composed by three case reports from clinical practice and that, despite frequent, not usually are objects of research. Flunarizine, a calcium channel blocker used to treat migraine and vertigo, was chosen because it induces extrapyramidal signs in elder patients which was later related to antagonist properties at dopamine D2 receptors, beyond the existence of data in animal models, which indicated its potential antipsychotic effect. Flunarizine was effective as antipsychotic and it presents a favorable side effects profile, beyond the long half-life and low cost. The second part of the thesis presents reports of frequent cases, but little studied under scientific scope. Therefore, we propose an evaluation of the patient with basis on the affective and emotional temperament, according to the temperament model proposed by Lara and Akiskal (2006). The first deals with the use of low doses of risperidone in the treatment of pathological jealousy. The second refers to the comorbidity of obsessive-compulsive disorder and bipolar disorder, treated with divalproex sodium and lamotrigine. The last article points to the use of low doses of quetiapine in patients with subthreshold symptoms and hyperthymic or cyclothymic temperaments presenting emotional deregulation. Together, our results strengthen the idea that the existing medications can be further explored for the treatment of clinical indications different from their originals.

O objetivo desta tese de doutorado foi testar psicofármacos tradicionais em aplicações diferentes das originais, tendo por base o conhecimento de seus mecanismos de ação e uma visão clínica focada no temperamento. A primeira parte da tese é constituída pelo ensaio clínico de flunarizina comparado com haloperidol no tratamento de pacientes com esquizofrenia. A segunda parte é composta de três relatos de caso oriundos da prática clínica e que, apesar de freqüentes, não costumam ser objetos de pesquisa. A flunarizina, um bloqueador de canal de cálcio empregado para tratar enxaqueca e vertigem foi escolhida pelo fato dela produzir sinais e sintomas extrapiramidais em pacientes idosos, o que posteriormente foi relacionado a sua propriedade como antagonista dos receptores dopaminérgicos do tipo D2, além da existência de dados em modelos animais que indicavam o seu potencial efeito antipsicótico. A flunarizina foi eficaz como antipsicótico e apresentou um perfil de efeitos adversos favorável, além de ter uma meiavida longa e baixo custo. A segunda parte da tese apresenta relatos de casos clínicos freqüentes, mas pouco estudados no âmbito científico. Nesse sentido propomos uma avaliação do paciente com base no temperamento afetivo e emocional, de acordo com o modelo de temperamento proposto por Lara e Akiskal (2006). O primeiro aborda o uso de baixas doses de risperidona no tratamento de ciúme patológico. O segundo se refere à comorbidade do transtorno obsessivocompulsivo e transtorno bipolar, tratados com divalproato de sódio e lamotrigina. O último artigo aponta para o uso de baixas doses de quetiapina em pacientes com sintomas sublimiares e temperamento hipertímico ou ciclotímico que apresentam desregulação emocional. Em conjunto, nossos resultados reforçam a idéia de que as medicações existentes podem sem mais exploradas para o tratamento de quadros clínicos diferentes das suas indicações originais. / The aim of this thesis was to test traditional drugs in applications different from the original, on the basis the knowledge of the mechanisms of action and a vision clinic focused on temperament. The first part of thesis consists in clinical trial of flunarizine compared with haloperidol in the treatment of patients with schizophrenia. The second part is composed by three case reports from clinical practice and that, despite frequent, not usually are objects of research. Flunarizine, a calcium channel blocker used to treat migraine and vertigo, was chosen because it induces extrapyramidal signs in elder patients which was later related to antagonist properties at dopamine D2 receptors, beyond the existence of data in animal models, which indicated its potential antipsychotic effect. Flunarizine was effective as antipsychotic and it presents a favorable side effects profile, beyond the long half-life and low cost. The second part of the thesis presents reports of frequent cases, but little studied under scientific scope. Therefore, we propose an evaluation of the patient with basis on the affective and emotional temperament, according to the temperament model proposed by Lara and Akiskal (2006). The first deals with the use of low doses of risperidone in the treatment of pathological jealousy. The second refers to the comorbidity of obsessive-compulsive disorder and bipolar disorder, treated with divalproex sodium and lamotrigine. The last article points to the use of low doses of quetiapine in patients with subthreshold symptoms and hyperthymic or cyclothymic temperaments presenting emotional deregulation. Together, our results strengthen the idea that the existing medications can be further explored for the treatment of clinical indications different from their originals.

A risperidona é um medicamento antipsicótico que quando metabolizada da origem a dois metabólitos hidroxilados quirais, a 7-hidroxirisperidona (7-RispOH) e a 9-hidroxirisperidona (9-RispOH). A 9-RispOH apresenta as mesmas propriedades farmacológicas que a risperidona e já é comercializada como fármaco, com o nome genérico de paliperidona. Estudos em humanos mostram que existem diferenças na disposição cinética dos enantiômeros da 9-RispOH, com uma maior prevalência do enantiômero (+)-9-RispOH em plasma. Dessa forma, este trabalho teve como finalidade avaliar a capacidade de algumas espécies de fungos e também de catalisadores de Jacobsen em (bio)transformar enantiosseletivamente a risperidona em seu metabólito ativo 9-RispOH. Para tanto, foi desenvolvido um método de separação para a risperidona e seus metabólitos utilizando cromatografia líquida de alta eficiência quiral. Este método foi então aplicado nos estudos de biotransformação enantiosseletivo e nos estudos de catálise assimétrica. A separação cromatográfica foi realizada empregando a coluna Chiralcel OJ-H e metanol:etanol (50:50, v/v) + 0,2% de trietilamina como fase móvel. A vazão e temperatura utilizadas foram 0,8 mL min-1 e 25oC, respectivamente. Para extração dos analitos do meio de cultura, foi empregada a microextração em fase sólida (SPME) como técnica de preparação de amostra. O processo de SPME foi realizado utilizando uma fibra C18 mergulhando diretamente a fibra na amostra por 30 minutos e dessorvendo a fibra diretamente na fase móvel por 5 minutos. A validação e estudos de biotransformação foram realizados empregando a cromatografia líquida acoplada à espectrometria de massas (LC-MS/MS). O método foi validado e todos os parâmetros encontram-se de acordo com as recomendações da literatura. O estudo de biotransformação foi realizado com diferentes espécies de fungos e somente os fungos do gênero Cunninghamella foram capazes de biotransformar a risperidona em seu metabólito ativo. O fungo Cunninghamella echinulata foi capaz de biotransformar estereosseletivamente a risperidona no seu metabólito ativo (+)-9-RispOH com excesso enantiomérico de 100% e o fungo Cunninghamella elegans foi também capaz de biotransformar estereosseletivamente a risperidona nos dois enantiômeros da 9-RispOH em diferentes proporções. Os estudos preliminares de catálise assimétrica foram realizados empregando a cromatografia líquida e detecção por UV-Vis, injetando diretamente alíquotas no sistema cromatográfico. Esses estudos mostraram que na condição de reação 1:50:50 (em número de mols, catalisador:oxidante:substrato) houve uma catálise assimétrica da risperidona que demonstrou ser enantiosseletiva para o metabólito 7-RispOH (E1). / Risperidone is an atypical antipsychotic drug. Its metabolism yields in two hydroxylated chiral metabolites, 7-hydroxyrisperidone (7-RispOH) and 9-hydroxyrisperidone (9-RispOH). The 9-RispOH metabolite presents the same pharmacologic activity of the parent drug risperidone. This led this drug to be marketed as drug under the generic name paliperidone. Studies have shown differences in the kinetic disposition of the 9-RispOH enantiomers with higher prevalence of the (+)-9-RispOH enantiomer in plasma. Thus, this work aimed to evaluate the ability of some species of fungi and Jacobsen catalysts in the enantioselective (bio)transformation of risperidone into its active chiral metabolite 9-RispOH. To accomplish that, it was developed a separation method to analyze risperidone and its metabolites by chiral high-performance liquid chromatography. This method was employed in enantioselective biotransformation studies and in asymmetric catalysis studies. The chromatographic separation was performed on a Chiralcel OJ-H column using methanol:ethanol (50:50, v/v) plus 0.2% triethylamine as the mobile phase at a flow rate of 0.8 mL min-1. The SPME process was performed by immersing directly a C18 probe fiber in the culture medium during 30 min. The analytes were desorbed from the fiber directly in the mobile phase during 5 min. The method validation and the biotransformation studies were performed by high-performance liquid chromatography coupled with tandem mass spectrometry (LC-MS/MS). The method was completely validated and all parameters were in agreement with the literature recommendations. The biotransformation studies were performed employing different species of fungi and only the Cunninghamella genus was able to biotransform risperidone into its active metabolite. The Cunninghamella echinulata fungus was able to biotransform risperidone into the active metabolite, (+)-9-RispOH, resulting in 100% of enantiomeric excess. The Cunninghamella elegans fungus was also able to biotransform stereoselectively risperidone into (+)- and ()-9-RispOH enantiomers at different rates. Preliminary studies of asymmetric catalysis were performed using high-performance liquid chromatography with UV-Vis detector (HPLC-UV). The aliquots were directly injected in the chromatography system. These studies showed that the reaction with 1:50:50 (catalyst:oxidant:substrate, in number of mols, in this sequence) presented an asymmetric catalysis of risperidone and that showed to be enantioselective to 7-RispOH (E1) metabolite.