O metronidazol é usado no tratamento de infecções causadas por protozoários e naquelas causadas por microrganismos anaeróbicos, no tratamento das formas intestinais e extra-intestinais de amebíase, em tricomoníase e em infecções bacterianas aeróbicas graves. Após administração oral, a absorção é rápida e completa sendo amplamente distribuído, atinge concentração plasmática máxima em 1-2 horas. A meia-vida de eliminação do metronidazol é de 6-12 horas. O objetivo deste trabalho foi avaliar a equivalência terapêutica por meio da bioequivalêmcia entre duas formulações de comprimidos revestidos contendo metronidazol, produzidos por dois fabricantes distintos, permitindo assim a intercambiabilidade entre as formulações. O ensaio de bioequivalência entre o produto teste (FUNED metronidazol) e o produto referência (Flagyl® - Aventis Pharma Ltda.) foi do tipo randomizado, cruzado e aberto. O medicamento foi administrado em dose única de 250 mg de metronidazol aos 24 voluntários sadios. Amostras de sangue foram coletadas até 48 horas após a administração e analisadas por método, desenvolvido e validado, de cromatografia líquida de alta eficiência acoplada à espectrometria de massas (CLAE- MS/MS). As curvas de decaimento plasmático obtidas para o produto teste (FUNED metronidazol) e para o produto referência (Flagyl® - Aventis Pharma Ltda.) foram semelhantes, assim como os parâmetros farmacocinéticos Cmax (teste: 6,66 µg/mL; referência: 6,77 µg/mL), tmax (teste: 1,26 h; referência: 1,90 h), ASC0-t (teste: 73,42 µgxh/mL; referência: 73,56 µgxh/mL), ASC0-∞ (teste: 75,15 µgxh/mL; referência: 75,23 µgxh/mL) e t½el (teste: 8,00 h; referência: 7,76 h). Os intervalos de confiança 90% para a razão de Cmax (92,2 - 106,4 %), ASC0-t (97,2 - 102,5 %) e ASC0-∞ (97,1 - 102,8 %) encontram-se entre 80 e 125 %, intervalo proposto pela ANVISA e FDA para a bioequivalência. A comparação estatística por meio de análise de variância (ANOVA) dos parâmetros Cmax, ASC0-t e ASC0-∞ indica claramente não haver diferença significativa entre os dois produtos contendo 250 mg de metronidazol. Baseado nos resultados farmacocinéticos e estatísticos, conclui-se que os dois produtos são bioequivalentes e, podem ser considerados intercambiáveis na terapêutica. / Metronidazole is used to treat infections caused by protozoa and those caused by anaerobic microorganisms. It is the drug of choice for the treatment of amebiasis in the intestinal and extra-intestinal forms, trichomoniasis and bacterial aerobic diseases. After oral administration, its absorption is rapid, complete and widely distributed, reaching maximum plasma concentration in 1 to 2 hours. The half-life of elimination of metronidazole is 6 to 12 hours. The objective of this study was to evaluate the therapeutic equivalence through bioavalability between two formulations of tablets containing metronidazole, produced by two different manufacturers, thus allowing the interchangeability between the formulations. The bioequivalence assay between the test product (FUNED Metronidazole) and reference product (Flagyl® - Aventis Pharma Ltda.) was a randomized, crossover and open study. The drug was given at a 250 mg metronidazole single dose to 24 healthy volunteers. Blood samples were collected until 48 hours after administration and analyzed by method, developed and validated in high performance liquid chromatography coupled to mass spectrometry (HPLC - MS / MS). The plasma decay curves obtained for the product test (FUNED metronidazole) and the reference product (Flagyl® - Aventis Pharma Ltda.) were statistically similar in the same way as the pharmacokinetic parameters Cmax (test: 6.66 µg/mL, reference: 6.77 µg/mL), tmax (test: 1.26 h; reference: 1.90 h), AUC0-t (test: 73.42 µgxh/mL, reference: 73.56 µgxh/mL) AUC0-∞ (test: 75.15 µgxh/mL, reference: 75.23). The 90% confidence intervals for the ratio of Cmax (92.2 - 106.4%), AUC0-t (97.2 - 102.5%) and AUC0-∞ (97.1 - 102.8%) are between 80 and 125%, range proposed by ANVISA and FDA for bioequivalence assays. This statistical parameters comparison using analysis of variance (ANOVA) Cmax, AUC0-t and AUC0-∞ clearly indicate no significant difference between the two products containing 250 mg of metronidazole. Based on the pharmacokinetic and statistical results, it is possible to conclude that the two products are bioequivalent and could be considered interchangeable in therapy.

L'utilisation de certains principes actifs dans le traitement et prévention de diverses maladies peut être limitée à cause de leur faible biodisponibilité. Cette faible biodisponibilité peut être liée à une solubilité limitée du principe actif et/ou une faible perméabilité à travers les barrières biologiques. La BCS (Biopharmaceutical Classification System), permet de classer les différents principes actifs en fonction de leur solubilité aqueuse et leur perméabilité intestinale. Le but de ce travail est d'augmenter la biodisponibilité de principes actifs peu biodisponibles et appartenant aux classes II et IV. D'une part, des nanosuspensions polymériques chargés en principes actifs peu hydrosolubles ont été préparées et caractérisées. En effet, une faible solubilité aqueuse, empêche le principe actif de se trouver sous forme moléculaire au site d'absorption, limitant ainsi son passage à travers les barrières biologiques. L'augmentation de la solubilité des principes actifs au sein d'un vecteur polymérique permettrait une mise à disposition accrue du principe actif au niveau du site d'absorption et en conséquence une augmentation de la biodisponibilité. L'incorporation de ces principes actifs (celecoxib, diclofenac, econazole, ibuprofène, ivermectine et warfarine) dans la matrice polymérique à permis d'augmenter fortement leur solubilité. Une caractérisation physico-chimique de ces nanosuspensions a été effectuée et des études pharmacocinétiques in vivo ont été réalisées afin de démontrer notre hypothèse. Les résultats obtenus ne permettent pas de confirmer cette hypothèse sauf dans le cas des formulations préparées avec l'Eudragit® RL 30D. D'autre part, un gel d'héparine destiné à l'administration topique a été préparé. Ce gel est obtenu par l'interaction électrostatique entre l'héparine (polyanion) et une suspension polymérique polycationique. Ce gel a démontré une grande capacité à incorporer de l'héparine. Des études de passage cutané ainsi que des études in vivo ont montré que ce gel peut permettre une action locale de l'héparine en évitant ses effets systémiques / Currently, the poor bioavailability of some drugs may limit their use in clinics. The poor bioavailability can be related to a low solubility of the drug and/or a low permeability through biological barriers. BCS (Biopharmacceutical Classifictation System) allows drugs classification as a function of their aqueous solubility and intestinal permeability. The aim of this work is to enhance the bioavailability of poorly available drugs. On one hand, nanosuspensions containing poorly soluble drugs were prepared and characterised. To be absorbed, the drug should be available in its molecular form at the site of absorption; so a sufficient solubility is needed. The hypothesis of our work is to consider that the incorporation of poorly soluble drugs into a polymeric carrier may increase drug solubility and consequently enhance drug bioavailability. The incorporation of different lipophilic drugs (celecoxib, diclofenac, econazole, ibuprofen, ivermectin and warfarin) shows a great enhancement of drug solubility. Physico-chemical characterization as well as in vivo pharmacokinetics studies have been performed. The obtained results, does not allow to confirm our hypothesis except formulations prepared with Eudragit® RL 30D. On the other hand, a heparin gel destined to a topic application has been prepared. The gel is obtained by electrostatical interaction between heparin (polyanion) and a polymeric polycationic nanosuspension. Heparin has been successfully incorporated into the gel and drug may be delivered to obtain a local action of heparin and thus, avoiding its systemic effects

Les produits de glycation avancée (AGEs) sont trouvés en excès dans le plasma et les tissus au cours du diabète, de l'insuffisance rénale et du vieillissement. Ils se forment aussi pendant la cuisson domestique et industrielle des aliments. Actuellement, la plus forte consommation de denrées alimentaires transformées contenant des quantités importantes des AGEs augmente le risque d'exposition de la population aux AGEs et à leurs effets biologiques. C'est ainsi que l'étude de la biodisponibilité des AGEs devient un sujet d'actualité dans la compréhension de la pathogenèse de certaines maladies chroniques, particulièrement le diabète. Deux études cliniques ont été menées dans le but d'étudier l'impact de l'ingestion des AGEs alimentaires sur les fluctuations des concentrations de ces mêmes molécules retrouvées dans les fluides biologiques des individus sains étudiés. Une première étude qui compare la teneur des AGEs ingérés à travers deux régimes alimentaires administré à 62 jeunes adultes. Une deuxième étude qui évalue la teneur en AGEs d'une alimentation basée sur du lait maternel ou du lait de formules infantiles donné à 161 bébés. La quantification de la carboxyméthyllysine (CML) et des AGEs fluorescents comme indicateurs de la réaction de Maillard a été effectuée dans les aliments administrés aux sujets de l'étude ainsi que dans leurs matrices biologiques : plasma, urine et fèces. Les résultats montrent qu'un régime contenant 2,5 fois moins de CML et 1,7 fois moins des AGEs fluorescents, en comparaison avec un régime courant ou standard, peut provoquer des baisses importantes de ces indicateurs dans le plasma, l'urine et la matière fécale des individus sains. Ces diminutions sont plus accentuées au niveau de l'excrétion urinaire. D'autre part, une plus forte présence de la CML dans le lait de formules infantiles correspond à une plus forte quantité de CML dans le plasma et dans l'urine des enfants ayant ingéré cet aliment, à la différence des enfants nourris avec le lait maternel, caractérisé par de faibles quantités de la CML. Chez les adultes soumis au régime bas en AGEs ainsi que les bébés nourris avec le lait maternel, les niveaux de certains paramètres métaboliques tels que le stress oxydatif, les antioxydants importants et des facteurs de l'inflammation cellulaire et systémique ont été normaux mais inférieurs à ceux des adultes et des enfants ayant ingéré de plus fortes quantités d'AGEs. Ces effets indiquent que de plus faibles quantités de produits de Maillard alimentaires pourraient avoir un impact sur la prévention des maladies chroniques. / Advanced Glycation End Products (AGEs) are found in excess in plasma and tissus during diabetes, renal diseases and aging. AGEs are known to form in foods during industrial processes or home cooking. Nowadays, an excessive intake of technologically transformed foods riches in AGEs, increase the risk of the population to their possible pathophysiological effects. In this contexte, the study of bioavailability of AGEs formed in foods may have a significant impact in understanding the pathogenesis of some chronic diseases as diabetes.Two clinical studies were conducted to study the impact of dietary intake of AGEs on fluctuating levels of these same molecules found in biological fluids of healthy individuals studied. A first study that compared the content of AGEs ingested through two diets administered to 62 young adults. A second study evaluates the AGE content of a diet based on breast milk or infant formula milk given to 161 babies. Quantification of CML and fluorescent AGEs as indicators of the Maillard reaction was performed in the food administered to the subjects and their biological matrices: plasma, urine and feces. The results show that a diet containing less than 2.5 times and 1.7 times of CML and fluorescent AGEs respectively, compared with a current or standard diet, can cause significant declines in these indicators in plasma, urine and fecal matter of healthy individuals. These decreases are more pronounced at the level of urinary excretion. A stronger presence of CML in the milk of infant formula is a higher amount of CML in plasma and urine of children who ingested this food, unlike the children fed with milk breast, characterized by low levels of CML. In adults subjected to diet low in AGEs and babies fed breast milk, the levels of some metabolic parameters such as oxidative stress, antioxidants and important factors of cellular and systemic inflammation, were lower than normal levels of adults and children who ingested higher amounts of AGEs. These effects indicate that lower amounts of dietary Maillard products could have an impact on chronic disease prevention

La zone littorale, de grande importance pour le fonctionnement écologique du lac, est souvent la première zone réceptrice de la contamination provenant du bassin versant. Cette pollution est majoritairement piégée dans les sédiments de surface. Pour autant, les conditions environnementales variables qui règnent en zone littorale sont susceptibles de favoriser la biodisponibilité des contaminants par le jeu des réactions physico-chimiques et biologiques. Cette thèse s’appuie sur l’étude de deux grands lacs péri-alpins, le Lac du Bourget et le Lac d’Annecy, pour mieux comprendre le comportement des polluants sédimentaires (ETM, HAP et PCB) en zone littorale lacustre. Nos travaux visent notamment (1) à caractériser la contamination littorale et l’influence des sources locales dans cette contamination, (2) à évaluer l’écotoxicité des sédiments littoraux et les changements de mobilité et de biodisponibilité des polluants sédimentaires sous l’influence de changements abiotiques de la zone littorale tels que ceux induits par le marnage lacustre. Nos résultats montrent que la contamination des sédiments et organismes autochtones présente une hétérogénéité spatiale significative sur le littoral des deux lacs, tant en intensité qu’en composition (i.e. profil de contamination). L’implication de sources locales (e.g. eaux pluviales drainées sur le bassin-versant) dans cette hétérogénéité spatiale a été mise en évidence. L’écotoxicité des sédiments de la zone littorale, évaluée via la réponse de biomarqueurs enzymatiques (AChE, GST), physiologiques (e.g. taux d’ingestion chez D. magna) et des traits de vie (e.g. taux de croissance, reproduction et survie) présente également une hétérogénéité spatiale significative. La biodisponibilité des ETM sédimentaires a été plus spécifiquement étudiée aux interfaces sédiment-eau-biote. Elle varie selon l’élément, le biote exposé (benthique ou pélagique) et sous l’effet du marnage. L’assèchement et remise en eau du sédiment littoral s’accompagnent d’une réduction de la fraction biodisponible des ETM. L’abaissement de la hauteur d’eau, en revanche, entraîne une accentuation de la remise en suspension de particules sédimentaires susceptibles d’exercer un risque écotoxicologique sur le biote littoral. Nos résultats soulignent la complexité de la contamination littorale et de son transfert dans les grands lacs péri-alpins et encouragent les approches écotoxicologiques dans l’évaluation de l’état écologique. / The littoral zone of lakes is of great ecological importance and yet receives a large array of contaminants from the watershed. This pollution is mainly trapped by the surface sediment, but its bioavailability may be allowed by physico-chemical and biological reactions in this zone of high environmental fluctuations. This work aims at improving knowledge about the behavior of sediment-borne contaminants (trace metals, PAHs and PCBs) in the lake littoral zone by using two large perialpine lakes; Lake Bourget and Lake Annecy. More precisely, our goals were to (1) characterize the littoral contamination and the influence of local sources in this contamination, (2) assess the ecotoxicity of the sediment-borne contamination and the main changes in its mobility and bioavailability under abiotic changes in the littoral zone, such as those related to water-level fluctuations in lakes. The contamination of the sediments and native organisms exhibited a significant spatial heterogeneity along the lake littoral, in both intensity and profile’s composition. The involvement of local sources of contamination (e.g. stormwater runoff) as drivers of this spatial heterogeneity has been highlighted. The ecotoxicity of littoral sediments was assessed through the responses of enzymatic biomarkers (AChE and GST), physiological traits (e.g. ingestion rates in D. magna) and life-history traits (e.g. growth, reproduction and survival rates). Our results also underscored a significant spatial heterogeneity of the littoral sediment ecotoxicity. A greater focus on the bioavailability of trace metals at the sediment-water-biota interfaces showed that bioavailability depends on the metal, the exposed biota (benthic vs pelagic) and fluctuate under water-level fluctuations. Drought and re-immersion of surface sediments were followed with a reduction of the bioavailable fractions of the trace metals. However, decreases in the water-level led to increase suspended sediments in the water column, which may exert an ecotoxicological risk for the littoral biota. Our results underline how complex the littoral contamination and bioavailability are and promote ecotoxicological approaches in the assessment of the ecological status of large lakes.

O chá verde (Camellia sinensis) é reconhecido mundialmente por seu alto teor de polifenóis, em especial as catequinas. As catequinas estão relacionadas à prevenção de várias doenças degenerativas, como o câncer e diabetes, devido ao grande potencial antioxidante. Contudo, vários incovenientes precisam ser superados para aprimorar o uso destes produtos, principalmente em relação à sua biodisponibilidade. O desenvolvimento de carreadores lipídicos na encapsulação de compostos bioativos é uma tecnologia recente capaz de resolver vários problemas de biodisponibilidade apresentadas pelos produtos naturais, produzindo uma estrutura capaz de proteger os compostos ativos. O objetivo deste trabalho foi o desenvolvimento de uma formulação utilizando carreador lipídico, empregando métodos de secagem para aumentar a estabilidade dos compostos bioativos, enfatizando-se processos de preparação, tipo de excipientes e procedimentos para a caracterização físico-química, estabilidade e avaliação da atividade antioxidante in vitro do produto final. Os compostos bioativos foram extraídos a partir das folhas secas e moídas de chá-verde através do processo de maceração dinâmica, sendo filtrado, concentrado e liofilizado. A formulação foi desenvolvida utilizando o sistema de balanço hidrófilo-lipófilo (EHL) com tensoativos nãoiônicos e co-solvente, e caracterizados pelas análises organolépticas, centrifugação, reologia, microscopia óptica, distribuição de tamanho e potencial zeta. Para a etapa de secagem, selecionou-se a formulação mais estável. As formulações foram secas em spray dryer de escala laboratorial a uma vazão de 4,0 g/min, e temperaturas de 100, 120 e 150 °C. O desempenho de secagem foi avaliado pela recuperação do produto. Os pós obtidos por spray drying foram caracterizados quanto ao teor de umidade, atividade da água, densidade, distribuição granulométrica, propriedades de fluxo, cristalinidade, morfologia e redispersibilidade. Após, se avaliou a atividade antioxidante do produto em óleo vegetal (óleo de soja) utilizando o teste acelerado de estabilidade oxidativa em Rancimat®. O EHL, o tipo de tensoativo e a técnica de preparo da formulação influenciaram diretamente na estabilidade do sistema. Para as formulações estudadas, o tensoativo que conferiu uma maior estabilidade foi o Gelucire® 44/14, sendo selecionado para prosseguimento com o processo de secagem das composições. O rendimento do processo atingiu em média 51,3 ±3,5 %, típico para secadores do tipo spray dryer em escala laboratorial. A maioria das partículas apresentou aparência arredondada, sem presença visível de porosidade, independente do adjuvante (lactose ou trealose). O produto apresentou baixa atividade de água (<= 0,22) e teor de umidade (<= 1,79). O aumento da temperatura de secagem provocou um ligeiro aumento no diâmetro médio de partícula quando a lactose foi utilizada como adjuvante de secagem (9,8 ± 5,9 ?m para 13,65 ± 8,4 ?m). Os pós obtidos tiveram baixa densidade, sendo menor em temperaturas mais altas. O índice de Carr e Fator de Hausner (15 % e < 1,25, respectivamente) indicaram boas propriedades de compressão e fluidez. O produto foi prontamente redispersível, recuperando facilmente sua consistência e características originais. A avaliação da estabilidade oxidativa acelerada utilizando Rancimat demonstrou que o processo de encapsulação conferiu uma maior solubilidade e proteção dos compostos bioativos, levando ao aumento da atividade antioxidante. Os resultados aqui relatados confirmam a viabilidade da encapsulação dos compostos bioativos de Camellia sinensis em carreadores lipídios empregando o processo de spray drying. O produto apresenta potencial de aplicação como matéria-prima para a produção de formas orais, inclusão em produtos nutracêuticos e cosméticos / Green tea, a product made from Camellia sinensis leaves, is recognized worldwide by its high polyphenol content, in special the catechins. Tea catechins are linked to the prevention of several degenerative diseases as cancer and diabetes. However, several drawbacks need to be overcome in order to increase the use of this products, being their bioavailability one of the upmost. The development of carrier lipid based containing bioproducts is a recent technology, which can solve several bioavailability problems presented by natural products, producing a structure which confer protection to active compounds. The aim of this work was the development of carrier lipid based compositions containing bioactive compounds of Camellia sinensis (green tea) by spray drying evaluating processes of preparation, type of excipients and characterization of physicochemical properties and evaluation the antioxidant activity in vitro of the product. Bioactive compounds from dried and milled green tea leaves was extracted by dynamic maceration, filtered, concentrated and freeze-dried. The formulation was developed through the utilization of Hydrophilic Lipophilic Balance System (HLB) using the non-ionic surfactants and a co-solvent and characterized by organoleptic analysis, centrifugation, rheology, optical microscopy, size distribution and zeta potential. The most stable compositions were submitted to spray drying. The compositions were dried in a labscale spray dryer at flow rate of 4.0 g/min at temperatures of 100, 120 and 150 °C. The spray drying performance was characterized by determination of the powder production yield. Spray dried powders were characterized by moisture content, water activity, density, size distribution, flow properties, crystallinity, morphology, and redispersibility. After, it was evaluated the antioxidant activity of the product in vegetable oil (soybean oil) using the accelerated oxidative stability test Rancimat®. The HLB, the type of surfactant and the preparation method of the formulation influenced the system stability. For the formulations studied, the surfactant which confers the greater stability was Gelucire® 44/14, which was selected to prepare composition for spray drying. The powder production yield falls around 51.3 ±3.5 %, typical for lab scale spray dryers. Wrinkled and rounded particles, without visible presence porosities were mostly generated, independent of the adjuvant (trehalose or lactose). The product presented low water activity (<= 0.20) and low moisture content (<= 1.79). Increasing in drying temperature caused a slight increase in mean particle diameter, when lactose was used as drying carrier (9.8 ±5.9 ?m to 13.65 ±8.4 ?m). Low density powders were generated, but density tends to be lower at high drying temperatures composition were submitted to spray drying. Carr index and Hausner ratio of the product (< 15% and < 1.25, respectively) were indicative of good compressive and flow properties. The product was promptly redispersible, regaining its original consistency straight forwardly. The accelerated oxidative stability using the Rancimat demonstrated that the encapsulation increased the solubility and protection of bioactive compounds, resulting to increased antioxidant activity of the product. The results here reported confirm the feasibility of entrapment of herbal bioactive compounds of Camellia sinensis in lipid carrier by spray drying. The product has potential to be used as raw material for production of oral dosage forms, inclusion in nutraceutical and cosmetic products.

A dissolução de fármacos administrados por via oral é pré-requisito para sua absorção e eficácia clínica. Esta constatação fornece a base para as tentativas de estabelecer correlações entre a dissolução in vitro e a biodisponibilidade in vivo de medicamentos, denominadas correlações in vitro - in vivo (CIVIV). As correlações in vitro-in vivo referem-se ao estabelecimento de uma relação racional entre propriedades biológicas, ou parâmetros derivados destas, produzidas por uma forma farmacêutica, e suas propriedades ou características físico-químicas. O estabelecimento desse tipo de correlação de dados pode possibilitar a substituição dos estudos in vivo, necessários para avaliação da biodisponibilidade de medicamentos, por estudos in vitro, com grandes vantagens éticas e econômicas. A probabilidade de obtenção de correlação in vitro-in vivo é maior quando o processo limitante da absorção do fármaco é a dissolução, o que ocorre para medicamentos contendo fármacos de classe biofarmacêutica II (baixa solubilidade e alta permeabilidade) e para sistemas de liberação controlada de fármacos. O diclofenaco de sódio (DFS) é um fármaco que possui ampla absorção através do intestino (próximo a 100%) e baixa solubilidade em pH abaixo de 6,0, o que o inclui na classe II do SCB. Caracteriza-se como antiinflamatório não esteroidal, com atividades analgésica e antipirética. Considerando as características físico-químicas e farmacológicas do DFS, é objetivo deste trabalho desenvolver e realizar uma validação interna e externa de correlação CIVIV para formulações contendo este fármaco. Para o desenvolvimento e validação interna de CIVIV, foram empregados comprimidos matriciais de DFS contendo o excipiente hidroxipropilmetilcelulose (HPMC) como agente controlador da liberação do fármaco. Três formulações de DFS contendo HPMC em diferentes concentrações (com diferentes taxas de liberação do fármaco) foram avaliadas em estudo de dissolução. Os dados de fração absorvida, obtidas pelo método de deconvolução (MOURÃO, 2009), foram plotados diretamente com os dados de fração dissolvida, obtendo um CIVIV com determinação linear de 0,9738. Para a validação externa, o modelo de CIVIV obtido (intercepto e inclinação) foi utilizado para prever os dados de fração absorvida a partir dos dados de dissolução, empregando três formulações comerciais de DFS. Os resultados de absorção foram extrapolados pelo método de convolução para a obtenção da curva plasmática prevista. Os dados da curva plasmática prevista foram comparados aos dados da curva real de DFS, obtidas no estudo de biodisponibilidade, e os erros de predição (EP%) entre as curvas, foram estabelecidos para a validação externa. Uma CIVIV de nível C foi estabelecida entre diversos parâmetros, empregando as formulações matriciais de DFS e as formulações comerciais. De acordo com os resultados de nível C obtidos, a metodologia de dissolução proposta pode ser empregada para o desenvolvimento de novas formulações contendo o fármaco DFS. / The dissolution of drugs administered orally is a pre-requisite for its absorption and clinical effectiveness. This finding provides the basis for attempts to establish correlations between in vitro dissolution and in vivo bioavailability of drugs, called correlations in vitro-in vivo (IVIVC). The correlation between in vitro-in vivo refers to the establishment of a rational relationship between biological properties, or parameters derived from those produced by a pharmaceutical form and its properties or physicochemical characteristics. The establishment of this type of correlation data may allow the replacement of in vivo studies necessary to evaluate the bioavailability of drugs, for in vitro studies with high ethical and economic advantages. The probability of obtaining correlation in vitro-in vivo is greater when the process of limiting drug absorption is dissolution, which occurs for drug products containing drugs biopharmaceutical class II (low solubility and high permeability) and controlled release systems drugs. Diclofenac sodium (DFS) is a nonsteroidal anti-inflammatory drug with analgesic and antipyretic properties. The objective of this work is to develop and implement an internal and external validation correlation IVIVC for formulations of DFS. For the development and internal validation of IVIVC were employees of DFS matrix tablets containing the excipient hydroxypropyl methylcellulose (HPMC) as a controlling agent drug release. Three formulations containing DFS in HPMC at different concentrations (with different rates of drug release) were evaluated in a study of dissolution. The absorbed fraction data obtained by the deconvolution method (MOURÃO, 2009), were plotted directly with the dissolved fraction data, obtaining a linear IVIVC with determination of 0.9738. For external validation, the obtained IVIVC model (intercept and slope) was used to predict the fraction absorbed data from dissolution data using three commercial formulations of DFS. The absorption results were extrapolated by the method of convolution to obtain the expected plasma curve. The plasma curve data were compared to data provided in real DFS curve, obtained in the bioavailability study, and the prediction error (PE%) between the curves were established for external validation. A level C IVIVC has been established between various parameters, using the matrix formulations of DFS and commercial formulations. According to the results obtained from C-level, the methodology proposed dissolution can be used to develop new drug formulations containing the DFS drug.

Ilex paraguariensis (erva mate) é uma planta amplamente usada na América Latina sob a forma de infusão aquosa. Dentre as propriedades atribuídas a esta planta encontra-se a atividade antioxidante que sugere um papel importante desta droga vegetal na prevenção e tratamento de doenças associadas ao estresse oxidativo como a aterosclerose, fotocarcinogênese e fotoenvelhecimento, entre outras. No entanto alguns compostos presentes nesta planta se encontram sob a forma esterificada, o que poderia dificultar a sua adequada absorção. Uma maneira de aumentar a biodisponibilidade de antioxidantes em extratos vegetais é promover a sua hidrólise visando à liberação dos compostos ativos. O objetivo deste trabalho foi estudar comparativamente o extrato hidroetanólico de Ilex paraguariensis antes e após hidrólise enzimática quanto à composição fitoquímica, atividade antioxidante in vitro e in vivo, biodisponibilidade de compostos antioxidantes e distribuição tecidual destes compostos em animais de experimentação. Para tanto o extrato foi obtido por percolação etanol:água (50% v/v) e sua hidrólise realizada por reação enzimática. A caracterização fitoquímica foi realizada por cromatografia líquida de alta eficiência (CLAE) e espectrofotometria e a atividade antioxidante dos extratos pelos métodos de DPPH e ORAC. Para os ensaios in vivo os extratos (hidrolisado e não hidrolisado) foram administrados oralmente (por gavage) a ratos Wistar machos em sistema de dose única ou doses repetidas (30 dias). Coletou-se o sangue, pele, fígado e cérebro, analisando-se a concentração dos compostos de interesse e a atividade antioxidante destes tecidos pelo método de ORAC. O extrato apresentou boa atividade antioxidante e conteúdo fenólico, sendo que estes valores não foram significativamente alterados pela hidrólise. No entanto, a hidrólise possibilitou uma maior absorção dos compostos de interesse, aumentando a atividade antioxidante plasmática. / Ilex paraguariensis (yerba mate) is a plant broadly used in Latin America as an aqueous infusion. Among its biological properties is the antioxidant activity, which suggests a important role in prevention and treatment of oxidative stress associated diseases, such as atheroclerosis, photocarcinogenesis and photoaging among other. However some of the compounds responsible for that activity are, in crude plant extract, under esterified form, which could make absorption more difficult. An approach to increase the bioavailability of antioxidants from plant extracts is to submit the extract to hydrolysis in order to release the active compounds. The goal of this work was comparatively evaluate the hydroethanolic extract of Ilex paraguariensis before and after enzimatic hydrolysis concerning phytochemical composition, in vivo and in vitro antioxidant activity, bioavailability and tissue distribution of antioxidant compounds in rats. The extract was obtained by percolation with ethanol:water (50% v/v) and the hydrolysis performed by enzymatic reaction. The phytochemical characterization was performed by high performance liquid chromatography (HPLC) and spectrophotometry and the antioxidant activity by DPPH and ORAC methods. Hydrolyzed and non hydrolyzed extracts were given orally (by gavage) to male Wistar rats in single and multiple dose (30 days) regimen. Blood, skin, liver and brain were removed, and the concentration of antioxidant compounds and antioxidant activity by ORAC method were evaluated. The crude hydroethanolic extract showed antioxidant activity and phenolic content, but these values were not significantly changed by hydrolysis. However the hydrolysis increased the absorption of the compounds and the plasma antioxidant activity.

Former mining areas are well-known globally to be a significant anthropogenic source of contaminants being dispersed into the surrounding environment. Various human activities, including ore mineral mining, industrial activities, domestic waste production, and the agricultural application of fertilisers and pesticides, are likely to contribute to the release of huge amounts of potentially toxic metals into the ecosystem, which have harmful effects on the flora and fauna and on human health. Therefore, the main aim of this study is to evaluate the contamination that arises from some selected heavy metals (Cu, Pb, Zn, Mn, Cr, Ni, and V) in topsoil and floodplain samples from the Ecton mining area. Ecton Hill is located in the southern part of the Peak District, Staffordshire, England, and bounded by the River Manifold from the west. This area has been mined for sulphide minerals, which were extracted extensively from the 16th century until the mid-19th century; the area is currently being used for cattle rearing and agricultural purposes. Therefore, it would be worth finding out the extent to which the area has been polluted by the aforementioned metals. To this end, topsoil and floodplain samples were collected and analysed for their total concentrations using XRF technique and different granulometric classes (i.e. clay, silt, and sand) using a (Malvern Mastersizer Long Bed) laser granulometer with a presentation unit of MS-17. In addition, soil specific factors, including organic matter content, organic carbon, pH, Eh, and cation exchange capacity were also measured. Spatial distribution maps were constructed using a GIS approach for the metals studied over the study area. Contamination and ecological risk assessments were carried out via the geoaccumulation index (Igeo) and enrichment factors (EF) respectively. Moreover, collected soils for different land uses have been assessed using the UK government soil guidelines (i.e. ICRCL and CLEA's SGVs soil values). Furthermore, the bioavailability, leachability, and fractionation (using five-steps sequential extraction) of the metals in various soil phases were characterised using correlation matrix and principal component analysis (PCA) approaches. The GIS- based spatial analysis maps reveal that elevated concentrations of the metals are located around the sites of the mining waste in the area. The contamination assessment results indicate that Cu, Pb, Zn have a contamination degree ranging between strongly contaminated (class 4) and extremely contaminated (class 6). The results of the ecological assessment by enrichment factor (EF) show that Pb has the highest enrichment factor. The bioavailability results of the heavy metals under study, via EDTA, show that Cu, Pb, Zn have the highest bioavailable fractions. The regression analysis demonstrates that Mn gives the best fit regression equation with the highest R2 value of 0.825. The leachability results reveal that, of the seven heavy metals, Zn has the highest leachable value, whereas the lowest leachable was recorded for Cr. Speciation was measured using the five-steps procedure, and the results show that Cu, Pb and Zn are mainly associated with the organic matter fraction, whilst, Cr, Ni and V are associated with the residual fraction. The principal component analysis (PCA) revealed that oxides of Fe/Al, organic matter, and the clay and silt fractions are the main soil parameters responsible for binding heavy metals to the soil surfaces of the study area. Changing the redox potential conditions and acidification was investigated and the results indicate that such changes have significant effects on the release of heavy metals from the soil particles at Ecton Hill.

A correlação in vitro - in vivo (CIVIV) refere-se ao estabelecimento de uma relação racional entre uma propriedade in vitro de uma forma farmacêutica (FF) e uma característica biológica, ou parâmetros derivados destas, produzidas a partir da absorção do fármaco, liberado por uma FF. Para o desenvolvimento de uma CIVIV, são necessárias três ou mais formulações, as quais são avaliadas em relação ao comportamento de dissolução e à biodisponibilidade (BD), e por meio do cálculo de deconvolução, estimam-se as frações absorvidas. A furosemida, fármaco modelo, é um diurético usado no tratamento de hipertensão. Este fármaco é classificado como classe IV do sistema de classificação biofarmacêutico (SCB) (Amidon et al., 1995). O objetivo do presente trabalho foi estabelecer uma CIVIV para formas farmacêuticas (FFs) de liberação modificada contendo complexo de furosemida e hidroxipropil-&#946-ciclodextrina (HP-&#946-CD), a partir de ensaios de dissolução e estudos de BD. O complexo de furosemida e HP-&#946-CD foi obtido por liofilização e caracterizado por análise térmica, solubilidade e permeabilidade. A partir do complexo, foram produzidas cinco formulações de comprimidos de liberação modificada, com diferentes concentrações de hidroxipropilmetilcelulose (HPMC) (10-30%). Estas foram submetidas aos estudos de dissolução com o aparato II. Destas, foram selecionadas três formulações com perfis distintos e submetidas ao estudo com o aparato IV e posteriormente ao estudo de BD. A partir destes resultados foi estabelecida uma CIVIV e esta foi avaliada por meio da validação interna. Foi realizado o estudo in silico de previsão das curvas de decaimento plasmático com emprego dos programas, STELLA® e Simcyp®, a partir dos dados: solubilidade da furosemida; dissolução a partir das formulações e dados farmacocinéticos obtidos a partir da injeção intravenosa do medicamento referência. Quanto à caracterização do complexo, os ensaios termoanalíticos sugerem que a furosemida forme complexo de inclusão com a HP-&#946;-CD pela técnica da liofilização. Observou-se o aumento da solubilidade em relação ao fármaco puro. Entretanto, quanto à permeabilidade, avaliada por meio do PAMPA (permeabilidade em membrana artificial paralela), os resultados foram semelhantes entre o fármaco puro e o complexo. Quanto ao comportamento de dissolução, avaliado com emprego dos aparatos II e IV, observou-se que as formulações apresentaram perfis de dissolução distintos. Os resultados do estudo de BD indicaram que a concentração do HPMC tem impacto relevante na absorção da furosemida. Foram obtidas correlações lineares a partir dos dados de fração absorvida e de dissolução, com coeficiente de determinação de 0,7662 para o aparato II e de 0,96017 para o IV. A validação interna da CIVIV empregando o aparato IV indicou que a correlação foi satisfatória. O estudo in silico de previsão das curvas de decaimento plasmático demonstrou que, nas condições empregadas, o modelo desenvolvido com o STELLA® foi mais preditivo do que o obtido pelo Simcyp®. / The in vitro - in vivo correlation (IVIVC) refers to the establishment of a rational relationship between a in vitro property of a pharmaceutical form (PF) and a biological characteristic or parameters derived from those, produced from the absorption of a drug released from a PF. For the development of an IVIVC, it is necessary three or more formulations, which are evaluated in relation to the dissolution behavior and for bioavailability (BA), calculating by deconvolution, an estimated absorbed fractions. Furosemide, a model drug, is a diuretic used in the treatment of hypertension. This drug is classified as class IV from biopharmaceutical classification system (BCS) (Amidon et al., 1995). The objective of this study was to establish an IVIVC for pharmaceutical forms (PFs) with modified release containing furosemide complexed with hydroxypropyl-&#946;-cyclodextrin (HP-&#946;-CD), from dissolution tests and BA studies. The complex of furosemide and HP-&#946;-CD was obtained by freeze-drying and characterized by thermal analysis, the solubility and the permeability. From the complex were produced five modified release tablet formulations, with different concentrations of hydroxypropylmethylcellulose (HPMC) (10-30%). These formulations were subjected to dissolution studies with the apparatus II. From these, three formulations with distinct profiles were selected and subjected to dissolution study with apparatus IV and subsequently to the BA study. From these results, an IVIVC was established and this was evaluated by internal validation. The in silico study was conducted to predict plasma decay curves with employment programs, STELLA® and Simcyp®, from the following data: furosemide solubility, dissolution from the formulations evaluated and pharmacokinetic data obtained from intravenous drug reference. From characterization of the complex, the thermoanalytical tests suggest that furosemide form inclusion complex with HP-&#946;-CD by freeze-drying technique. It was observed an increased solubility compared to the pure drug. However, permeability results, as assessed by the PAMPA (Parallel artificial membrane permeability), were similar for both furosemide and the complex. As for the dissolution behavior, evaluated with apparatus II and IV, so it was observed that the formulations showed an distintict profile. it was observed that the formulations produced showed different dissolution profiles. The results form BA assays indicated that the HPMC concentration has an important impact on the furosemide absorption. It was obtained a linear correlation from absorption fraction and dissolution data, with the determination coefficient of 0.7662 to apparatus II and 0.96017 from apparatus IV. Internal validation, with the IVIVC obtainted from apparatus IV, indicated that the correlation obtained was satisfactory. The in silico study predicted plasma decay curves, showed that under the conditions used, the model developed with STELLA® was more predictive than the model obtained by Simcyp®.

Foram utilizados 504 pintos de corte, machos, da linhagem Cobb, de 1 dia de idade, distribuídos aleatoriamente em 18 tratamentos com 4 repetições pelo período de 21 dias, para determinar a biodisponibilidade relativa do fósforo (BRP) em milho, cevada e farelo de soja sem e com a adição de fitase microbiana à dieta. Foram adicionados dois níveis de fitase (0 ou 750 FTU/kg de ração) e dois níveis de fósforo suplementar (0,05 ou 0,10%) à dieta basal semi-purificada com baixo nível de fósforo total (0,40%) sendo a fonte padrão de fósforo o fosfato bicálcico. Ao término do experimento, foram removidas as tíbias esquerdas e os dedos médios direitos para posterior análise de cinzas ósseas. A BRP para cada fonte orgânica foi calculada pelo método do slope ratio através da regressão dos valores de peso de cinzas na tíbia em função dos níveis de fósforo suplementar ingerido (mg/dia). Os valores de BRP para milho, cevada e farelo de soja sem a adição de fitase foram 55,4; 122,4 e 70,4%, respectivamente. Com a adição de fitase estes valores foram para 170,1; 153,6 e 156,40%, respectivamente / Five hundred and four male Cobb chicks were used in a 21 days trial to determine the relative bioavailability of phosphorus (RBP) in corn, barley and soybean meal, with and without addition of microbial phytase (0 or 750 FYT/kg of diet) and two levels of suplemmentary phosphorus (0.05 ou 0.10%). Suplemmentary phosphorus and microbial phytase were added to the basal diet, which had been semi-purified and had had low levels of total phosphorus (0.40%), being dicalcium phosphate the pattern source of phosphorus. Animals were randomly assigned into 18 treatments with four repetitions each one. By the end of the experiment, animals left tibiae and right medium fingers were removed for later ashes analysis. A RBP for each organic phosphorus source was calculated by the slope ratio method, through regression analysis of tibiae ashes weight data in function of ingested levels of suplemmentary phosphorus (mg/day) data. The values of RBP for corn, barley and soybean meal without phytase were 55.4, 122.4 and 70.4%, respectively. However, the RBP values for corn, barley and soybean meal with phytase were 170.1, 153.6 and 156.40%, respectively