<p>Artemisia Afra, a popular South African traditional herbal medicine is commonly administered as a tea infusion of the leaves. However, clinical trials proving it safety and efficacy are lacking mainly due to the absence of good quality dosage forms and credible placebos for the plant. The objectives of this study were to prepare a standardized preparation of the plant leaves and freeze-dried aqueous extract powder of the leaves, in a tea bag dosage form and to design and prepare credible placebos for these plant materials.</p>

Par l'utilisation de techniques statistiques poussées, l'approche de population rend l'estimation des propriétés pharmacocinétiques (étude des phénomènes d'absorption, distribution, métabolisme et élimination du médicament dans l'organisme) possible chez les patients inclus dans les études de Phase III, en dépit d'un nombre de prélèvements sanguins réduit. Elle permet également d'évaluer, parmi les caractéristiques des patients, les sources de variabilité interindividuelle (covariables) pouvant expliquer les différences potentielles observées dans telle ou telle sous-population.Après un bref rappel bibliographique de la technique, nous présentons les résultats des analyses effectuées sur trois candidats-médicaments en fin de développement clinique. Le premier exemple détaille le processus d'une analyse de population réalisée chez plus de 3000 patients ainsi que les résultats d'une analyse pharmacocinétique/pharmacodynamique. Dans la deuxième partie, nous insistons sur une technique particulière de validation (le bootstrap) et détaillons l'étape de simulation mise en place pour appuyer les conclusions de l'analyse. La dernière analyse présente enfin un modèle pharmacocinétique de population combinant les données d'un produit parent et de son métabolite actif, l'autre originalité de l'analyse étant l'utilisation du randomization test afin de vérifier l'inclusion de covariables dans le modèle.Les conclusions de ces analyses permettent de mieux appréhender la cinétique des trois molécules dans leur population cible par la mise en évidence d'éventuelles sous-populations à risque et, ainsi, de veiller à leur efficacité et à leur absence de toxicité. / Using sophisticated statistical techniques, the population approach allows estimating the pharmacokinetic properties (study of the absorption, distribution, metabolism and elimination phenomena of a drug into the body) in patients included in Phase III studies, despite a low number of blood samples collected. It also allows evaluating, from the patients' characteristics, the sources of interindividual variability (covariates) that may explain the potential differences observed in a given subpopulation.After a brief review of the technical aspects of the population approach, the results of the analyses performed on three drugs in the end of their clinical development will be presented. The first example details the process of a population analysis carried out in more than 3000 patients, together with the results of a pharmacokinetic/pharmacodynamic analysis. In a second part, we underline a validation technique (the bootstrap) and the simulation study performed to emphasize the conclusions of the study. The last example presents a population pharmacokinetic model applied to the simultaneous fit of data from a parent drug and its active metabolite, the other originality of the analysis being the use of the randomization test to verify the covariates inclusion in the model.The conclusions of these analyses allow a better understanding of the kinetics of the three drugs in their target population by emphasizing potential subpopulations at risk and, hence, participate to verify their efficacy and their absence of toxicity.

Pharmacometrics is a quantitative science that is rapidly changing the landscape of drug development, and particularly so for the pediatric population. The motivation behind the research underlying this dissertation is to contribute towards the improvement of pediatric drug development by the astute application of pharmacometric methods. Two distinct research areas have been focused upon: 1- improving pediatric pharmacokinetic (PK) trial design and 2- improving pediatric dosing of warfarin by using a genetics-based dosing regimen. The first project examined in detail the feasibility of and simulation-based methodology for implementing a recent regulatory PK quality standard. The focus was on designing pediatric PK trials that employ sparse sampling and population analysis methods, using a simulation-estimation platform. The research provided clarity on the impact of various trial design elements, such as PK sampling, adult data inclusion, PK variability and analysis method on sample size adequacy to honor the standard. The PK quality standard was found to be practically feasible in terms of sample size adequacy. Informative sampling schedule for a given number of PK samples per subject is assumed during trial design. Recommendations are made to: 1- use prior adult or pediatric data for trial design and analysis, wherever possible and 2 - use one-stage population analysis methods and biologically feasible covariate models for designing pediatric PK studies. The second project involved derivation of the first ever pediatric warfarin dosing regimen, including starting dose and titration scheme, based on pharmacogenetics (Cyp2c9 *1/*2/*3 and VKORc1 -1629 G>A polymorphisms). While extensive research and several dosing models for warfarin use in adults exist, there is paucity of data in pediatrics. A validated adult warfarin population PKPD model was bridged using physiological principles and limited pediatric data to arrive at a pediatric PKPD model and dosing regimen. Pediatric data (n=26) from an observational study conducted at the Children’s Hospital Los Angeles (CHLA) was used to qualify the pediatric model. A 2-step pediatric starting dose based on body weight (<20 kg and ≥20 kg) for each of 18 (6 Cyp2c9 x 3 VKORC1) genotype categories is proposed. The titration scheme involves percentage changes relative to previous dose, based on latest patient INR. The dosing regimen targets a major (≥ 60%) proportion of INRs within therapeutic range of 2.0-3.0, by the second week into warfarin therapy. Simulataneously, bleeding and thromboembolic risks are minimized via minimal proportions (≤ 10% and ≤ 20%) of INRs > 3.5 and INRs < 2.0, respectively. In simulations, the proposed dosing regimen performed better on target INR outcomes than the standard-of-care dosing used in the CHLA patients. Given the challeneges in and low likelihood of conducting pediatric warfarin clinical studies, the proposed dosing regimen is believed to be an important advance in pediatric warfarin therapy. Prospective warfarin studies in pediatrics using the proposed dosing regimen are recommended to refine and validate the suggested dosing strategy.

Sulfated low molecular weight lignins (LMWLs) have shown good activity as anticoagulants by allosterically inhibiting thrombin, as well as promising agents for treating emphysema through inhibition of elastolysis, oxidation, and inflammation. Sulfated LMWLs are chemo-enzymatically synthesized from starting monomers caffeic, ferulic, and sinapic acid into sulfated dehydropolymers known as CDS, FDS, and SDS. To further the LMWLs’ development as drugs, their structural composition and physicochemical characteristics were defined in this work. The molecular weight distribution profile of the sulfated LMWLs from size exclusion chromatography performed on a high pressure liquid chromatography system (SEC-HPLC) changed from bimodal when no surfactant is used in the mobile phase of the HPLC to unimodal when surfactant is used in the mobile phase. This indicates that some large molecular weight species, likely an aggregate of smaller molecular weight chains, are disrupted when surfactant is present. The resulting estimates of molecular weight calculated when surfactant is used in the mobile phase resulted in peak average molecular weights of 5700 Da for CDS, 7400 Da for FDS, and 4300 Da for SDS. These molecular weights are 17-45% higher and can be considered more accurate than the previously reported molecular weights (CDS: 3320 Da, FDS: 4120 Da, SDS: 3550 Da) because they were measured directly whereas previous estimates were calculated from GPC-HPLC data of the unsulfated LMWL precursors. Elemental analysis and distribution coefficient measurements were also performed on the LMWL library, revealing information about the level of sulfation and hydrophobic character of the sulfated LMWLs.

Orientador : Hérida Regina Nunes Salgado / Coorientador: Cristina Helena dos Reis Serra / Banca: Marlus Chorilli / Banca: Marcello Garcia Trevisan / Banca: Fernanda Nervo Raffin / Banca: Ana Dóris de Castro / Resumo: O darunavir, antirretroviral inibidor de protease, compõe a terapia considerada altamente eficaz no tratamento da síndrome da imunodeficiência humana adquirida, e é um dos fármacos disponibilizados gratuitamente pelo Sistema Único de Saúde (SUS). Os produtos genéricos têm contribuído para a manutenção e abrangência desta política de distribuição gratuita de medicamentos, ao reduzir os custos desses produtos. Neste propósito, este estudo busca fornecer informações sobre o comportamento físicoquímico e estabilidade do fármaco e da formulação, diante de diferentes condições climáticas, bem como, sobre sua solubilidade e liberação a partir da formulação. Além disso, pretende apresentar ferramentas in vitro e in silico que contribuam, no futuro próximo, para a produção de novos produtos contendo darunavir que apresentem alta qualidade, especialmente, genéricos do medicamento referência. Para tanto, a solubilidade do fármaco foi investigada em diferentes meios. A baixa solubilidade aquosa do fármaco foi verificada, e os meios adequados aos testes de dissolução foram escolhidos. O perfil de dissolução in vitro do darunavir e de absorção in vivo a partir papel dos comprimidos de Prezista® contendo 300 mg de fármaco foram investigados. Obtevese um perfil de dissolução discriminativo para o produto e um método de quantificação por espectrofotometria derivada foi desenvolvido e validade. Uma correlação entre os dados de fração dissolvida in vitro e fração absorvida in vivo do darunavir foi alcançada (r2= 0,95). Este é um método preditivo para avaliar o desempenho in vivo do darunavir a partir de comprimidos de liberação imediata, com base em dados in vitro, e é importante para minimizar o uso de animais e seres humanos em experimentos durante o desenvolvimento novos produtos farmacêuticos contendo darunavir. Além disso, uma ferramenta in silico foi desenvolvida e validada para a previsão da ... / Abstract: Darunavir, a protease inhibitor, constitute the highly active antiretroviral therapy against the acquired immunodeficiency syndrome. It is one of agents given free by the Brazilian Health System (SUS). Generic products have contributed to the maintenance and the policy of free distribution of medicines by the decreasing of costs of these products. In this way, this study seeks to provide information on the physical and chemical behavior and stability of the drug and formulation, facing different weather conditions, as well as provide information on their solubility and release from the formulation. It also aims to provide in vitro and in silico tools to contribute in the near future, for the production of new products containing darunavir, especially generic medicines. Therefore, the solubility of the drug was investigated in different media. The low aqueous solubility of the drug was observed, and suitable media for dissolution testing were chosen. The in vitro dissolution profile of darunavir and the in vivo absorption from Prezista® (tablets containing 300 mg of drug) were investigated. It was obtained a discriminating dissolution profile of the product and a method of quantification by spectrophotometry derivative was developed and validated. A correlation between the data of in vitro dissolved fraction and in vivo absorbed fraction of darunavir was achieved (r2 = 0.95). This is a predictive method to evaluate the in vivo performance of darunavir from immediate-release tablets, based on in vitro data, and it is important to minimize the use of animals and humans in experiments during the development of new medicines containing darunavir. In addition, an in silico tool was developed and validated for predicting the oral bioavailability of darunavir formulations, from their dissolution profile. For this, physiologically based pharmacokinetic modeling was applied through Simcyp® software. These, in vitro and in silico tools, assist in ... / Doutor

INTRODUCTION: Pancreatic Ductal Adenocarcinoma (PDAC) is an aggressive form of cancer with a high mortality rate, primarily due to lack of effective treatment options. Current therapeutic approaches are limited to surgical resection of the pancreas during early stages of the disease and to the use of non-specific chemotherapeutic drugs such as gemcitabine, neither of which has successfully improved the 5-year survival rate of PDAC. Both the lack of effective treatments and the high mortality of the disease call for the urgent need to develop new therapeutic options. OBJECTIVES: This thesis project focuses on an endogenous inhibitor of the neuropilin 2 receptor (NRP2) called semaphorin 3F (SEMA3F) and its use as a potential new drug in the fight against pancreatic cancer. By binding the transmembrane receptor neuropilin 2 (NRP2), SEMA3F can inhibit angiogenesis and cellular proliferation. Interestingly, given its role as a guidance molecule, it is also a potent mediator of cellular repulsion. All three of these effects will be analyzed in the context of this study. METHODS: Syngeneic pancreatic cancer cells were injected orthotopically in two separate groups of mice. One group involved the use of transgenic Nrp2-/- mice, and served as a way to analyze the absence of the receptor on the vasculature and how that affects the growth of the primary tumor and the formation of metastases in the liver. The other group received intravenous injections of SEMA3F-expressing and control adenovirus, and served to explore the effect of SEMA3F as a potential therapy against the growth of the primary tumor in the pancreas and distant metastases in the liver. RESULTS: We observed a decrease in pancreatic tumor and metastatic growth in the absence of Nrp2 in our transgenic mouse model compared to the WT control. Mice injected with SEMA3F-expressing adenovirus also showed a decrease in primary tumor growth as well as a reduction in the formation of metastases in the liver compared to the control. CONCLUSION: Nrp2 mediates angiogenesis in pancreatic cancer, which facilitates the growth of the primary tumor as well as the formation of metastases. Our results indicate that the anti-angiogenic, anti-proliferative and repulsive actions of SEMA3F could be used to develop an effective treatment option for PDACpancreatic ductal adenocarcinoma. / 2020-07-24T00:00:00Z

Pode-se afirmar que no século XX originou-se uma indústria farmacêutica multinacional com extraordinária capacidade de pesquisa e desenvolvimento para produzir novos fármacos. Porém, contradizendo este potencial inovador, o número de fármacos introduzidos no mercado vem declinando desde 1960 e as oportunidades abertas com os avanços da biologia molecular, da genômica, da bioinformática e da química ainda não trouxeram os resultados esperados. No Brasil, a pesquisa científica tem obtido resultados de grande relevância, mas encontra muitas dificuldades para levar novos produtos ao mercado. O objetivo principal deste trabalho é discutir estratégias de fomento para a pesquisa e desenvolvimento de fármacos no país, procurando conciliar os requisitos técnicos e econômicos deste processo com as competências preexistentes. O referencial metodológico adotado enfatiza o papel determinante das relações econômicas na pesquisa e desenvolvimento de fármacos e procura encontrar caminhos compatíveis com a realidade nacional. Para tanto, discute as características técnicas e econômicas desta atividade, bem como a estratégia das empresas inovadoras e algumas experiências brasileiras nesta área do conhecimento. Fundamentado na análise de 766 novos fármacos introduzidos no mercado mundial entre 1984-2003, nos pilares econômicos do processo de inovação e no contexto político-institucional da pesquisa e desenvolvimento de fármacos, propõe alternativas para aprimorar o desenvolvimento científico e tecnológico do setor farmacêutico brasileiro. Muitas atividades relacionadas com a pesquisa e desenvolvimento de fármacos são realizadas no país, mas encontram-se dispersas nas principais universidades, centros e institutos de pesquisa. O mapeamento destas competências representa o ponto de partida para a criação de uma rede de inovação no setor farmacêutico. Um dos gargalos identificados neste trabalho é a fragilidade do suporte institucional para negociações de alta tecnologia, do qual fazem parte as patentes, os acordos de cooperação e a transferência de tecnologia. Para viabilizar, no Brasil, a incorporação de ferramentas de alta tecnologia empregadas no desenvolvimento de fármacos foi proposta a criação do Laboratório Nacional de Pesquisa e Desenvolvimento de Fármacos. Além de garantir sofisticação técnica, este Laboratório Nacional atuaria como instituição aberta, multidisciplinar, assumindo o papel de um centro de articulação das iniciativas voltadas para o desenvolvimento de fármacos, podendo gerar recursos para financiar a pesquisa e contribuir para o desenvolvimento científico e tecnológico. Acordos de cooperação com empresas inovadoras e organismos internacionais fazem parte das estratégias para captação de recursos. Linhas de pesquisa alinhadas com as necessidades do Sistema Único de Saúde e com outras políticas do setor público também devem nortear a pesquisa e desenvolvimento de fármacos no país. A exploração de novos alvos moleculares, articulada com projetos genômicos, inovações incrementais, doenças negligenciadas e produtos naturais são apontados como áreas estratégicas. A afirmação de que o Brasil reúne as condições necessárias para participar do processo de desenvolvimento de fármacos - hipótese primária deste trabalho - encontra sustentação nos argumentos apresentados e revela que as condições para a inovação tecnológica nunca foram tão favoráveis quanto agora. Estimular o debate acerca de estratégias que possam fomentar o desenvolvimento científico e tecnológico da pesquisa e desenvolvimento de fármacos no país representa a contribuição almejada por este trabalho. / It can be asserted that in the twentieth century a multinational pharmaceutical industry with extraordinary research and development capacity to produce new drugs arose. However, contradicting this innovative potential, the number of new chemical entities introduced in the market is declining since 1960 and the opportunities open with the progresses of molecular biology, genomics, bioinformatics and chemistry haven\'t brought the expected results yet. In Brazil, the scientific research has been obtaining results of great relevance, but a lot of difficulty is found to introduce new products into the market. The main purpose of this work is to discuss fomentation strategies for the drug research and development in the country, trying to reconcile technical and economic requirements of this process with the pre-existent competences. The adopted methodological referential emphasizes the decisive role of economic relationships in drug research and development and tries to find out compatible ways with the national reality. For that, it discusses the technical and economic characteristics of this activity, as well as the strategy of innovative companies and some Brazilian experiences in this knowledge area. Based on the analysis of 766 new chemical entities introduced in world market among 1984 to 2003, in economic pillars of the innovation process and in political-institutional context of drug research and development, alternatives are proposed to straighten out the scientific and technological development of Brazilian pharmaceutical sector. Many activities related to the drug research and development are accomplished in the country, but they are scattered in the main universities, research centers and institutes. The charting of these competences represents the start up to create an innovation net in the pharmaceutical section. One of the bottlenecks identified in this work is the fragility of institutional support for high technology negotiations, of which patents, cooperation agreements and technology transference make part. To make it possible, in Brazil, incorporation of high technology tools used in drug development, creation of National Laboratory of Drug Research and Development was proposed. Besides guaranteeing technical sophistication, this National Laboratory would act as an open institution, multidisciplinary, shouldering the role of an articulation center of initiatives aiming drug development, being able to generate resources to finance research and to contribute to scientific and technological development. Cooperation agreements with innovative companies and international organisms are part of strategies to raise funds research fields aligned with the needs of the Brazilian Unique Health System (SUS) and with other policies of Public Sector also must direct the drug research and development in the country. New molecular targets evaluation articulated in genomic process, incremental innovations, neglected diseases and natural products are pointed out as strategic areas. The statement that Brazil has conditions of participating in the process of drug development - primary hypothesis of this work - finds back-up in reported arguments and reveals that conditions for technological innovation have never been as favorable as now. To stimulate the debate concerning strategies that can foment scientific and technological development of drug research and development in the country represents the contribution aimed for this work.

Orientador: Hérida Regina Nunes Salgado / Banca: Leticia Scherer Koester / Banca: Ian Castro Gamboa / Banca: Luis Vitor Silva do Sacramento / Banca: Marlus Chorilli / Resumo: Objetivou-se, neste trabalho, realizar um estudo químico-farmacológico, bem como o desenvolvimento de uma formulação galênica das folhas de Spondias dulcis Forst., conhecida popularmente como cajarana. Está planta apresenta uso popular como antiinflamatório e antisséptico. Inicialmente, foi feita uma avaliação farmacognóstica e microbiológica da droga. Em seguida, foram produzidos extratos hidroalcoólicos utilizando delineamento experimental e desenvolvido e validado um método para quantificação de polifenóis totais. O extrato hidroalcoólico foi avaliado em ensaios in vitro (atividade antimicrobiana e antioxidante) e in vivo (atividade laxante e toxicidade aguda). O componente majoritário foi identificado e isolado, utilizando técnicas de cromatografia em contracorrente e de espectroscopia. Por fim, foi desenvolvida e caracterizada uma dispersão sólida obtida com extrato e o polímero PVP K30 e avaliada sua estabilidade, utilizando cinética não isótermica. A droga vegetal foi caraterizada pela densidade aparente, tamanho de partícula, perda por dessecação, pH e teor de extrativos e controle microbiológico. A espectroscopia na região do infravermelho permitiu a identificação de grupamentos específicos provenientes de diversos metabólitos e a análise térmica identificou seis etapas de degradação. O extrato escolhido foi obtid por turboextração, utilizando a solução de 50% de etanol com 20% de droga vegetal. O método desenvolvido e validado para polifenóis totais apresentoi-se de... (Resumo completo, clicar acesso eletrônico abaixo) / Abstract: The objective of this work was to conduct a chemical-pharmacological study, as well as the development of a galenic formulation of the leaves of Spondias dulcis Forst., known as cajarana. This plant presents popular use as antiinflammatory and antiseptic. Initially, a pharmacological and microbiological evaluation of the herbal drug. Hydroalcoholic extracts were produced using experimental design and developed and validated a method for quantification of total polyphenols. The hydroalcoholic extract was evaluated in vitro tests (antimicrobial and antioxidant activity) and in vivo (laxative activity and acute toxicity). The main component, identified and isolated, countercurrent chromatography and spectroscopy techniques. Finally, a solid dispersion obtained with extract and the polymer PVP K30 was developed and characterized and its stability evaluated using non-isothermal kinetics. The herbal drug was characterized by apparent density, part size, desiccation loss, pH and extractive content and microbiological control. The infrared spectroscopy identified of specific data of several metabolites and a thermal analysis identified six stages of degradation. The extract were produced by turboextraction, use a solution of 50% ethanol with 20% plant drug. The method developed and validated for total polyphenols presented within the conformities in the figures of merit (selectivity, linearity, precision, accuracy and robustness). The pharmacological activity verified in the case of ... (Complete abstract click electronic access below) / Doutor

New drugs are needed for better cancer management. Clinical trials are currently underway to assess the use of flavonoids (natural polyphenols) as anticancer agents. Among them, myricetin has been shown to induce cell cycle arrest and apoptosis in pre-clinical cancer models. We hypothesised that myricetin-derived novel flavonoids designed to enhance this natural potential and improve on the drug-likeness limitations of myricetin might have increased potential for their application in the management of breast and ovarian cancer. The effect of a library of novel flavonoids was screened on 3 panels of breast and ovarian cancer cell lines, representing different molecular subtypes and phenotypes, to assess their potency. The second-generation bi-methoxylated analogue AO-1530-OMe (Oncamex) was identified as the most effective candidate in the library, with sub-micromolar concentrations exerting a strong antiproliferative effect across almost all models studied. Results suggested that changes in the hydroxylation profile, the addition of methoxylations and a decyl alkyl chain were some of the structure-activity relationships contributing to this improved efficacy. Plate assays showed 8 h treatment with Oncamex reduced cell viability and induced cytotoxicity and apoptosis, concomitant with caspase activation and PARP cleavage. Pre-incubation with an antioxidant partially blocked these effects, suggesting the possible involvement of ROS modulation in the mechanism of action of Oncamex. Fluorescence microscopy reported the quick and stable delivery of Oncamex to the mitochondria. Fluorescent probes showed that Oncamex can induce mitochondrial superoxide production at concentrations associated with its antiproliferative effects. Study of the electrochemical properties of Oncamex by cyclic voltammetry supported this. Differential gene expression analysis following a microarray experiment showed Oncamex induces changes in the expression of genes controlling cell cycle and apoptosis. Together with previous results, the findings from this analysis led to the postulation of a model for the mechanism of action of Oncamex: due to its enhanced reactivity and mitochondrial targeting, Oncamex can generate mitochondrial superoxide, leading to mitochondrial dysfunction, membrane permeabilisation and the activation of the JNK pathway and the transcription factor FOXO3, which together contribute to the induction of intrinsic apoptosis and the inhibition of proliferation. Further proliferation assays on cell culture models also reported enhanced effect of Oncamex when administered in combination with paclitaxel and TRAIL. These improved responses were observed in breast and ovarian cancer models, including cells lines characterised by their treatment-resistant phenotype. Cotreatment with Oncamex also improved the effect of tamoxifen on anti-oestrogen resistant LCC9 breast cancer cells. Results from preliminary in vivo studies in mice implanted with the MDA-MB-231 breast cancer xenograft were consistent with an antiproliferative effect of Oncamex (25mg/kg/day) in vivo, as treatment inhibited tumour growth and reduced the expression of the marker of proliferation Ki-67 without signs of systemic toxicity. Tissues from this experiment also allowed for preliminary in vivo validation of the proposed mechanism of action of Oncamex by immunohistochemistry. The in vivo cytostatic effect of Oncamex was confirmed in a second in vivo experiment, which also investigated the effect of Oncamex at higher doses or in combination with paclitaxel. In conclusion, the novel flavonoid Oncamex has shown a promising antiproliferative effect in pre-clinical models of breast and ovarian cancer, including models of treatment-resistant cancers. Preliminary in vivo studies have demonstrated a partial recapitulation of the effect of Oncamex. A mechanistic model has been proposed by which Oncamex induces intrinsic apoptosis through its redox reactivity and mitochondrial targeting. These results support the potential of this prototypic candidate, although possible work in the structure and formulation of this candidate and further study and validation of its mechanism of action is needed for its continued development as an anticancer agent.

Diseases which affect the central nervous system present a huge burden to sufferers and caregivers. In tandem with longevity, prevalences of age-related neurodegenerative diseases are increasing. However, despite the evident necessity for pharmaceutical interventions, there has been a distinct lack of drug development to combat these disorders. This is largely attributed to high financial costs of using rodent models. Thus the validation of a more cost-effective in vivo system would facilitate pharmaceutical screening. The work presented in this thesis addresses this issue by assessing the utility of zebra fish in two costly areas of translational neurobiology { lead identi cation and safety pharmacology. An aversive classical conditioning assay was developed and automated as a behavioural screening method. This robust assay allows fast assessment of cognition and cognitive decline. The effect of neurotoxin treatment on aversive learning was then assessed using this assay, demonstrating its efficacy as a screening tool for neurodegeneration research. Subsequently, a transgenic zebra fish line - expressing a mutated form of the Alzheimer's-associated human amyloid precursor protein - was assessed, demonstrating an age-related cognitive impairment. Additionally new genetic zebra fish lines were generated, which over-express genes (both endogenous and transgenic) related to Alzheimer's-like pathologies. Whilst these were not assessed within this thesis, they present promising tools for possible future investigations. Regarding safety pharmacology, regulatory bodies require all CNS-penetrant drugs be assessed for abuse potential. Zebra fish display reward responses to several common drugs of abuse (e.g. amphetamine, cocaine, morphine). Thus, the latter sections of this thesis evaluated the utility of zebra fish for assessing human abuse potential. A CPP paradigm was utilised to test a range of drugs, with the sensitivity and specificity of zebra fish compared to previous reports using rodent. Additionally, the development of a zebra fish drug discrimination assay was attempted. However the paradigms utilised failed to develop an efficacious assay.