La protéine tyrosine phosphatase 1B (PTP 1B) et la glycogène phosphorylase (GP) sont des nouveaux cibles thérapeutiques pour le traitement du diabète (type II) et l'obésité. Dans l'objective de trouver des inhibiteurs actives et sélectives de ces enzymes, des aryl C glycosides tels que les C glycosyl 1,4 naphthaquinones, les 6 O benzoyl C glycosyl benzoquinone /naphthaquinones, les dérivés quinone d'acides glycuroniques et d'amides carboxylés ainsi que les inhibiteurs bidentates C glycosyl quinones ont été conçus et synthétisés. En tout, 178 composés ont été préparés. Le mécanisme et les facteurs d'influence de la réaction de C aryl glycosylation ont également été étudiés. Les essais préliminaires montrent que certaines molécules présentent des activités biologiques très intéressantes. Des études biologiques plus approfondies seront réalisées par la suite.

Herein, the influence of protonation and adsorption upon the redox and electrocatalysis of quinone species - specifically anthraquinone derivatives – is investigated. Through the comparison of the measured rate constants of one-electron reductions of a family of quinones in acetonitrile at both graphite and gold electrodes, it was confirmed that the redox potential indirectly influences the rate of electron transfer in a manner consistent with the potential-dependence of the density of states. In aqueous media, the voltammetric response of both anthraquione-2-sulfonate (AQMS) and anthraquinone-2,6-disulfonate (AQDS) was measured over the full aqueous pH range. A model is provided which is able to describe not just the variation in the formal potential but also the peak height as a function of pH. Importantly, this model predicts that the formal potential for the first (Ef1) and second (Ef2) electron transfers are comparable in magnitude (E^θ _f2−E^_θf1 equals -15mV for AQMS and -36mV for AQDS). This quantitative model is then further extended to consider the situation in which the system is not fully buffered, giving insight into the change of pH at the electrode surface during experimentation. Adsorption to graphitic electrodes can impart a strong influence on the measured voltammetric response. It is demonstrated that through the pre-exposure of a newly prepared graphitic electrode to organic solvents, these adsorption processes can be predominantly blocked. Moreover, it is shown that the electroactivity of the electrode is not significantly altered. This thesis also highlights two cases in which adsorption of the electroactive species may be used to positive effect. First, the surface adsorption of anthraquinone-2-monosulfonate is studied on a graphite electrode, where it is demonstrated that the heterogeneity of the electrode surface may be probed through studying the electrochemical response of the adsorbed species. From this work it is concluded that the rate of electron transfer at the graphitic basal plane is 2-3 orders of magnitude lower than that observed on the edge plane sites. Second, the co-adsorption of DNA and anthraquinone-2-monosulfonate is used as an indirect method to measure the solution phase concentration of DNA (LOD = 8.8μM). The reduced form of anthraquinone is also known to readily reduce oxygen. Through the use of a boron-doped diamond electrode it was possible to directly study the anthraquinone mediated reduction mechanism. Significantly, the voltammetric response indicates the reduction of the oxygen via the semi-quinone intermediate (kf = 4.8 × 10⁹ mol⁻¹ dm³ s⁻¹) is over two orders of magnitude faster than the reaction involving the di-reduced form (kf = 1 × 10⁷ mol⁻¹ dm³ s⁻¹). More importantly, this work provides voltammetric evidence for the existence of the semi-quinone species. This work is subsequently extended through the investigation of the poorly soluble anthraquinone derivative quinizarin. Not only is it possible to detect voltammetrically this biologically relevant species to concentrations as low as 5nM (100ppt), but the methodology also allows the electrochemistry of the quinizarin species to be probed, something which was not previously possible.

Chapter1 gives an overview of the basic principles of electrochemistry. A rigorous electrochemical study on the solution phase and solid phase cobalt phthalocyanine (CoPC) is presented in chapter2. The formof CoPC on carbon electrodes was characterized by scanning electron microscope (SEM). The use of CoPC modified edge plane pyrolytic graphite (CoPC-EPPG) for sensing nitrite (NO₂⁻) was also investigated. It was found that the claimed mediator CoPC has no influence on the process. A bare glassy carbon (GC) electrode was successfully applied for the quantitative determination of nitrite as a simple alternative to the modified electrodes reported in the literature (chapter3). Chapter4 compares the voltammetric responses of an edge plane pyrolytic graphite electrode covalently modifed with 2-anthraquinonyl groups (EPPG-AQ2) and solution phase anthraquinone monosulphonate (AQMS) in the presence of a limited concentration of protons. The solution phase and surface bound species show analogous responses resulting in split waves. Digisim™ simulation of the AQMS voltammetry have shown that the pH adjacent to the electrode may be altered by up to 5-6 pH units in low buffered solutions; this is caused by the consumption of protons during the electrochemical reaction. Chapters5 and 6 compare the electrochemical properties of 2-anthraquinonyl groups covalently attached to an edge plane pyrolytic graphite (EPPG) and to a gold electrode. In both cases simulations using newly developedMarcus-Hush-Chidsey theory for a 2e⁻ process assuming a uniform surface did not achieve a good agreement between theory and experiment. Subsequently two models of surface inhomogeneity were investigated: a distribution of formal potentials, E^Ө, and a distribution of electron tunneling distances, r₀. For both EPPG-AQ2 and Au-AQ2 modified electrodes the simulation involving E^Ө distribution turned out to be the most adequate. This is the first time that Marcus-Hush-Chidsey theory has been applied to a 2e⁻ system. Chapter7 briefly summarizes the obtained results.

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Enzimas proteolíticas são comumente empregadas na liberação de peptídeos bioativos com potencial aplicabilidade como aditivos em alimentos. Neste trabalho, estudou-se a capacidade de inibição do escurecimento de maçãs minimamente processadas do soro de leite de búfala hidrolisado por Alcalase em diferentes intervalos, ti (i = 0 h, 0,5 h, 1 h, 2 h, 3 h, 4 h ou 6 h). O processo de hidrólise foi acompanhado por determinação do grau de hidrólise (GH). Para se entender possíveis modos de ação sobre o escurecimento enzimático, o soro, ao longo do tempo de hidrólise, foi submetido às análises de capacidade capacidade antioxidante (capacidade de sequestro do radical ABTS•+, atividade quelante de Fe2+ e poder redutor), reatividade com quinonas e atividade inibitória sobre polifenoloxidases (PFO) extraídas de maçãs Red Delicious. Após os ensaios in vitro, o hidrolisado t4h foi testado quanto à capacidade de inibição do escurecimento de maçãs Red Delicious minimamente processadas, determinada por meio de análises de luminosidade (parâmetro L*) e índice de escurecimento (IE). O soro apresentou aumento significativo (p < 0,05) do GH, da atividade antioxidante, da reatividade com quinonas e da atividade inibitória de PFO em função do tempo de hidrólise. Especificamente para a atividade inibitória de PFO, o hidrolisado t4h alcançou pico de inibição próximo de 50 %. Quanto à capacidade de inibição do escurecimento das maçãs minimamente processadas, o hidrolisado manteve o parâmetro L* das maçãs durante 6 dias de armazenamento, não diferindo significativamente (p > 0,05) do metabissulfito. Além da luminosidade, o soro de leite hidrolisado conseguiu manter melhor o IE das maçãs durante esse tempo de armazenamento, comparativamente ao soro não hidrolisado (p < 0,05). Esses resultados evidenciam informações sobre possíveis aplicações de soro de leite bubalino hidrolisado com Alcalase como substituto natural de aditivos convencionalmente utilizados no controle desse tipo de escurecimento em alimentos. / Proteolytic enzymes are commonly employed in the release of bioactive peptides with potential applicability in food additives. In this work, the ability to inhibit the browning of minimally processed apples from Alcalase hydrolysed buffalo whey at different time, ti (i = 0 h, 0.5 h, 1 h, 2 h, 3 h, 4 h or 6 h). The hydrolysis process was followed by determination of the degree of hydrolysis (DH). In order to understand possible modes of action on the enzymatic browning, the whey was submitted to the analysis of the antioxidant capacity (capacity of sequestration of the ABTS•+ radical, Fe2+ chelating activity and reducing power), reactivity with quinones and inhibitory activity on polyphenoloxidases (PPO) extracted from Red Delicious apples. Following the in vitro assays, the t4h hydrolyzate was tested for the ability to inhibit the browning of minimally processed Red Delicious apples, determined by means of analysis of luminosity (parameter L *) and browning index (BI). Buffalo whey shows significant increase (p < 0.05) in DH, antioxidant activity, reactivity with quinones and PPO inhibitory activity as a function of the hydrolysis time. Specifically for the PPO inhibitory activity, the t4h hydrolyzate reached a near 50% inhibition peak. As for the ability to inhibit the browning of the minimally processed apples, the hydrolyzate kept the L * parameter of the apples during 6 days of storage, not statistically differing (p > 0.05) from the metabisulfite. In addition to the luminosity, the hydrolyzed whey was able to maintain the IE of the apples better during this storage time, compared to the unhydrolyzed whey (p < 0.05). These results evidenced information on possible applications of hydrolysed buffalo whey with Alcalase as a natural substitute for additives conventionally used in the control of this type of browning in foods.

Este trabalho apresenta o primeiro estudo dos estados aniônicos de baixa energia da para-Benzoquinona (p-BQ) por meio de cálculos de espalhamento elástico de elétrons pela molécula neutra. Foi utilizado o método multicanal de Schwinger implementado com pseudopotenciais de Bachelet, Hamann e Schlüter, na aproximação de núcleos fixos, e nas aproximações estático-troca e estático-troca mais polarização. As seções de choque integrais foram calculadas em duas geometrias do alvo, otimizadas para o estado fundamental da molécula neutra e do ânion. Os estados aniônicos encontrados foram aracterizados e comparados com resultados experimentais, realizados com a técnica de espectroscopia de transferência eletrônica (ETS). Três estados * foram observados, sendo um ligado e dois metaestáveis, em acordo com os experimentos. Uma ressonância de Feshbach foi encontrada nos cálculos, tal qual é observada experimentalmente e sugerida em diversos estudos teóricos de estado ligado. Acredita-se que a aproximação utilizada para descrever estados excitados do alvo poderá ser reconsiderada, em que é sugerida uma proximação mais sofisticada, a ser utilizada em cálculos futuros. Com este estudo, é possível uma melhor compreensão dos resultados experimentais ETS, assim como é permitida uma interpretação de estudos teóricos sobre os estados aniônicos da p-BQ. Ademais, este trabalho possibilita próximos estudos de espalhamento envolvendo quinonas mais complexas. / This work present the first study of low-energy anionic states in para-benzoquinone (p-BQ) by electron scattering calculations. The elastic scattering calculations were done in the fixed-nuclei approximation, with the Schwinger multichannel method implemented with pseudopotentials of Bachelet, Hamann and Schlüter, in the static-exchange and in the static-exchange plus polarization approximations. The integral cross sections were performed in two target geometries optimized for the neutral and anion ground states. The anionic states found were characterized and compared with experimental results obtained by electron transfer spectroscopy (ETS). Three * states were observed, a bound state and two metastable, in good agreement with experimental results. A Feshbach resonance was found in calculations, also it is observed experimentally and it was suggested in several theoretical studies of bound state. It is believed that the approach used to describe the target excited states can be reconsidered, as it is suggested a more sophisticated approach to be used in future calculations. With this study, a better understanding of experimental results ETS is possible, as it allowed an interpretation of theoretical studies of the anionic states of p-BQ. Moreover, this work enables further scattering studies involving more complex quinones.

The rising use of natural products creates an imperative need for an enhanced awareness of the safety of current and new products making their way into the marketplace. An important example is natural products containing lignans as the principal active component. Despite their structural similarity the lignan of creosote bush can cause hepato- and renal toxicity while the lignans of flaxseed have no reported serious toxicity. This dissertation aimed to investigate the oxidative metabolism of such lignans to determine whether reversible, competitive interactions and/or bioactivation may explain the differences in their apparent toxicity.<p> The first objective was to study the metabolism and bioactivation of nordihydroguaiaretic acid (creosote bush) and secoisolariciresinol (flaxseed). Nordihydroguaiaretic acid metabolism in rat liver microsomes led to the production of three glutathione adducts formed via ortho¬-quinone reactive intermediates. This metabolism was independent of NADPH and thus attributed to autoxidation. Secoisolariciresinol metabolism yielded lariciresinol and no glutathione adducts suggesting an absence of bioactivation to reactive quinone intermediates.<p> The second objective was to study the autoxidation of nordihydroguaiaretic acid. The major autoxidation product was a unique, stable schisandrin-like cyclolignan which was the result of nordihydroguaiaretic acid cyclization. The half-life of nordihydroguaiaretic acid in aqueous solution, pH 7.4, 37ºC is 3.14 hours suggesting the cyclolignan may be responsible for some of the biological effects of nordihydroguaiaretic acid.<p> The third objective was to study the inhibition of cytochrome P450 isoforms 1A2, 2B, 2C11 and 3A by lignans derived from creosote bush and flaxseed. None of the lignans caused irreversible inhibition. Both creosote bush and flaxseed lignans caused reversible inhibition of P450 enzyme activity that involved competitive or mixed-type inhibition, however the inhibition was present at nonphysiologically relevant concentrations. Activation of cytochrome P450 isoforms was also observed at low lignan concentrations. The results suggest that P450-mediated bioactivation or reversible inhibition cannot explain the differences in toxicity noted between the lignans of creosote bush and flaxseed.<p> This work suggests a minimal risk for drug-lignan interactions at P450 enzymes. Further studies are warranted to determine the presence and biological and toxicological role of the nordihydroguaiaretic acid cyclolignan in herbal preparations.

The rising use of natural products creates an imperative need for an enhanced awareness of the safety of current and new products making their way into the marketplace. An important example is natural products containing lignans as the principal active component. Despite their structural similarity the lignan of creosote bush can cause hepato- and renal toxicity while the lignans of flaxseed have no reported serious toxicity. This dissertation aimed to investigate the oxidative metabolism of such lignans to determine whether reversible, competitive interactions and/or bioactivation may explain the differences in their apparent toxicity.<p> The first objective was to study the metabolism and bioactivation of nordihydroguaiaretic acid (creosote bush) and secoisolariciresinol (flaxseed). Nordihydroguaiaretic acid metabolism in rat liver microsomes led to the production of three glutathione adducts formed via ortho¬-quinone reactive intermediates. This metabolism was independent of NADPH and thus attributed to autoxidation. Secoisolariciresinol metabolism yielded lariciresinol and no glutathione adducts suggesting an absence of bioactivation to reactive quinone intermediates.<p> The second objective was to study the autoxidation of nordihydroguaiaretic acid. The major autoxidation product was a unique, stable schisandrin-like cyclolignan which was the result of nordihydroguaiaretic acid cyclization. The half-life of nordihydroguaiaretic acid in aqueous solution, pH 7.4, 37ºC is 3.14 hours suggesting the cyclolignan may be responsible for some of the biological effects of nordihydroguaiaretic acid.<p> The third objective was to study the inhibition of cytochrome P450 isoforms 1A2, 2B, 2C11 and 3A by lignans derived from creosote bush and flaxseed. None of the lignans caused irreversible inhibition. Both creosote bush and flaxseed lignans caused reversible inhibition of P450 enzyme activity that involved competitive or mixed-type inhibition, however the inhibition was present at nonphysiologically relevant concentrations. Activation of cytochrome P450 isoforms was also observed at low lignan concentrations. The results suggest that P450-mediated bioactivation or reversible inhibition cannot explain the differences in toxicity noted between the lignans of creosote bush and flaxseed.<p> This work suggests a minimal risk for drug-lignan interactions at P450 enzymes. Further studies are warranted to determine the presence and biological and toxicological role of the nordihydroguaiaretic acid cyclolignan in herbal preparations.

No description available.

No description available.