Orientadora: Profa. Dra. Daniele Ribeiro de Araújo / Dissertação (mestrado) - Universidade Federal do ABC, Programa de Pós-Graduação em Biossistemas, 2016. / O naproxeno (NPX) é um anti-inflamatório não-esteroidal utilizado no tratamento da artrite reumatóide, osteoartrites e artroses, tanto como terapia única ou em associações farmacoterapêuticas. No entanto, sua administração por via oral está relacionada a efeitos adversos importantes (como toxicidade para o trato gastrintestinal). Nesse contexto, o desenvolvimento de novas formulações para liberação modificada de NPX possibilitaria a redução do número de injeções, aumentando a adesão ao tratamento. A incorporação de nanopartículas constituídas por quitosana-tripolifosfato (QT-TPP) em hidrogéis a base de poloxamers (PL) com diferentes valores de balanço hidrofílicolipofílico, favorece a formação de um sistema duplo de liberação com propriedades termoreversíveis, temperatura de gelificação (transição sol-gel) e razão de liberação do fármaco diferenciadas. Este trabalho visa o desenvolvimento (preparação e caracterização), a avaliação dos perfis de liberação in vitro e da citotoxicidade de novas formulações para liberação controlada de NPX incorporado em nanopartículas QT-TPP dispersas em hidrogéis termorreversíveis, visando a administração por via intra-articular, para o tratamento da artrite. Os diferentes hidrogéis foram preparados a base de poloxamer 407, (18 e 20 %), isolados ou associados ao poloxamer 403 (2 %). Nanopartículas constituídas de QT e TPP foram preparadas, caracterizadas e, posteriormente, dispersas nos hidrogéis termorreversíveis. Para isso, foram realizados estudos da interação fármaco-micelas e fármaconanopartículas (diâmetro hidrodinâmico, índices de polidispersão, eficiência de encapsulação e potencial zeta), para verificar a compatibilidade entre os componentes das formulações, bem como inferir sobre a estabilidade coloidal das mesmas; ii) a determinação da temperatura e da variação de entalpia relacionadas à micelização dos copolímeros isolados e de seus sistemas binários, antes e após a dispersão das nanopartículas, por calorimetria diferencial exploratória; iii) determinação da temperatura de transição sol-gel; iv) avaliação dos perfis de liberação in vitro do fármaco aplicandose diferentes modelos matemáticos. A dispersão das nanopartículas CS/TPP nos hidrogéis alterou as transições de fases e a termoreversibilidade das formulações, especialmente para aquelas com menor concentração de polímero. Por outro lado, a adição de nanopartículas contendo NPX, aos hidrogéis, aumentou as temperaturas relativas à micelização e a variação de entalpia. As análises reológicas mostraram perfis viscoelásticos e os resultados dos ensaios de liberação in vitro de NPX sugerem a contribuição de mecanismos como difusão e relaxamento das cadeias poliméricas. Por fim, com os ensaios de viabilidade celular observou-se a baixa citotoxicidade das formulações, apontando-as como sistemas híbridos promissores para liberação modificada de naproxeno. / Naproxen (NPX) is a non-steroidal anti-inflammatory used in the treatment of rheumatoid arthritis, osteoarthritis and arthrosis, either as sole therapy or pharmacotherapeutic associations. However, the oral administration is related to significant adverse effects (how toxicity to the gastrointestinal tract). Thus, in order to reduce the systemic side effects, intraarticular (IA) injection has been proposed as alternative. In this context, the development of new formulations for modified release NPX brings the possibility to reduce the number of injections, increasing the treatment adherence. The incorporation of nanoparticles of QT-TPP in poloxamers (PL) with different hydrophilic-lipophilic balance values, favors the formation of a dual release system of thermosensitive properties, gelation temperature (Tsol-gel) and due to the different drug release. This work aims the development, evaluation of the release profile in vitro and cytotoxicity of new formulations for controlled release of NPX-QT-TPP nanoparticles dispersed in thermosensitives hydrogels, aiming the injection intraarticular for treatment of arthritis. The different hydrogels were prepared based on poloxamer 407 (18 and 20%), isolated or associated with the poloxamer 403 (2%). Nanoparticles made of QT and TPP were prepared, characterized, and dispersed in the thermosensitives hydrogels. Therefore, studies of drug-micelle interaction and drug-nanoparticles were performed (hydrodynamic diameter, polydispersity, encapsulation efficiency and zeta potential), to verify the compatibility between the components of the formulations, as well as inferences about the colloidal stability of such; ii) determining the temperature and enthalpy change related to micellization of the isolated copolymers and their binary systems before and after dispersion of nanoparticles, by differential scanning calorimetry; iii) determination of the sol-gel transition temperature; iv) evaluation of the release profiles in vitro of the drug by applying different mathematical models. The dispersion of NanoQT/TPP nanoparticles in hydrogels changed phase transitions and thermosensitive formulations, especially those with lower concentrations of polymer. Moreover, the incorporation of NanoQT/TPP containing NPX to the hydrogels increased the micellization temperatures and enthalpy variation. Rheological analyzes showed viscoelastic rprofile and NPX in vitro release assays showed the contribution of mechanisms such as diffusion and relaxation of the polymer chains. Finally, cell viability assays showed low toxicity, indicating the hydrogels as promising hybrid systems for NPX modified release.

Orientador: Ivanira Jose Bechara / Tese (doutorado) - Universidade Estadual de Campinas, Instituto de Biologia / Made available in DSpace on 2018-08-11T09:34:23Z (GMT). No. of bitstreams: 1 Bockelmann_PetraKarla_D.pdf: 29179947 bytes, checksum: 8c02cdd38976a29e54d61610a6bfd9fe (MD5) Previous issue date: 2008 / Resumo: As nadadeiras caudais de teleósteos, quando parcialmente amputadas, passam por um rápido processo de regeneração chamado de regeneração epimórfica, caracterizado pela formação de uma massa de células indiferenciadas, diferenciação dessas células, síntese e deposição de matriz extracelular e restauração morfológica. A regeneração da nadadeira é extremamente sensível a fatores físicos e químicos externos, tais como variações na temperatura, intensidade da luz, ação de alguns agentes contaminantes ambientais e ação de algumas drogas que podem interferir na capacidade regenerativa das nadadeiras dos peixes teleósteos. Existem relatos na literatura, que drogas anti-inflamatórias não esteróides, podem interferir de alguma forma na restauração tecidual de diversos organismos, uma vez que inibem a ação da enzima ciclooxigenase e, conseqüentemente, a conversão do ácido araquidônico em prostaglandina, elementos que desempenham funções importantes na proteção celular, crescimento, angiogênese e produção de matriz extracelular. Em vista disso, este trabalho teve como objetivo avaliar os efeitos de drogas antiinflamatórias não esteróides, naproxeno e indometacina, durante o processo regenerativo da nadadeira caudal de peixe teleósteo Cyprinus carpio (carpa). Para isso, foram montados experimentos com cinco grupos: grupo formado com peixes que serviram como controle, grupo formado com peixes que entraram em contato com o naproxeno, na dose de 15,6 mg/L, e três grupos formados por peixes que tiveram contato com a indometacina nas doses 10, 20 e 30 mg/L cada. Os peixes foram anestesiados e suas nadadeiras caudais foram amputadas transversalmente e, após a amputação os peixes foram divididos entre os cinco grupos e permaneceram nos aquários até que a regeneração ocorresse. Os animais foram anestesiados, sacrificados e as nadadeiras em regeneração foram excisadas e fixadas em intervalos de 1, 2, 4, 5, 6, 8, 10 e 12 dias após a amputação. As amostras foram processadas para permitir uma análise histoquímica, ultra-estrutural e morfométrica das possíveis alterações no processo regenerativo da nadadeira caudal de teleósteo em contato com as drogas em questão. Os grupos tratados com o naproxeno e a indometacina utilizada na dose de 10 mg/L apresentaram o processo de regeneração de forma semelhante ao grupo controle, ou seja, não afetaram a formação da capa epidermal, a formação do blastema, a diferenciação das células blastemais, bem como a síntese, deposição, organização e mineralização dos componentes da matriz lepidotriquial e a síntese das actinotriquias durante o processo regenerativo da nadadeira caudal. No entanto, os peixes tratados com a indometacina nas doses de 20 e 30 mg/L apresentaram um atraso no processo de regeneração da lepidotriquia e da actinotriquia quando comparados com os peixes do grupo controle. Estudos mais detalhados sobre os mecanismos de ação das drogas anti-inflamatórias não esteróides e a ação dessas drogas sob a expressão ou a inibição da expressão de alguns genes envolvidos no processo de regeneração da nadadeira caudal de teleósteo talvez possam responder a razão das diferenças de efeitos entre essas duas drogas / Abstract: The fins of teleosts, when partially amputated, they pass for a quick regenerative process called epimorphic regeneration, characterized by the formation of a mass of undifferentiated cells, by the differentiation of these cells, by the synthesis and the deposition of the extracellular matrix and morphological restoration. The regeneration of the fin is extremely sensitive to external physical and chemical factors such as temperature variations, light intensity, the action of some environmental contaminants and the action of some drugs that can interfere in the regenerative capacity of teleost fins. There are some studies that show that nonsteroids anti-inflammatory drugs can interfere somehow in the tissue restoration of many organisms, as they inhibit the action of ciclooxygenase enzyme and, consequently, the conversion of arachidonic acid in prostaglandins, elements that execute important roles in cell protection, growth, angiogenesis and in the production of extracellular matrix. In this sense, this study aimed to evaluate the effects of nonsteroid anti-inflammatory drugs, naproxen and indomethacin, during the regenerative process of the teleost tail fin Cyprinus carpio (carp). Therefore, experiments were undertaken in five groups: the control group fish, group of fish in touch with naproxen in doses of 15.6 mg/L, and three groups of fish in contact with indomethacin in doses of 10, 20 and 30 mg/L each. The fish were anesthetized and their fins transversally amputated and, after amputations the fish were divided among the five groups described above and were left in the aquaria until the occurrence of regeneration. The animals were anesthetized, sacrificed and the regenerating fins excised and fixed in intervals of 1, 2, 4, 5, 6, 8, 10 and 12 days after amputation. The samples were processed in order to permit a histochemical, ultra-structural and morphometric analysis of the possible alterations in the regenerative process of the tail fin of the teleosts in contact with the drugs mentioned above. The group treated with naproxen and indomethacin in a 10 mg/L dose showed a regenerative process similar to the control group, thus, it did not affect the formation of the epidermal layer, the formation of blastema, and the differentiation of blastemal cells, as well as its synthesis, deposition, organization and mineralization of the lepidotrichial matrix and the synthesis of actinotrichia during the process of regeneration of the tail fin. However, the fish treated with indomethacin in doses of 20 and 30 mg/L presented a delay in the regenerative process of the lepidotrichia and actinotrichia when compared to the control group fish. Detailed studies about the mechanisms of nonsteroids anti-inflammatory drugs action and the action of these drugs under the expression or inhibition of expression of some genes involved in the teleost tail fin regenerative process could explain more precisely the reason of the differences of effect between these two drugs / Doutorado / Histologia / Doutor em Biologia Celular e Estrutural

Anti-inflamatórios não esteroidais (AINEs) são frequentemente utilizados para controlar a dor pós-operatória e inflamação em pacientes após a cirurgia oral e maxilofacial, mas eles são muitas vezes associados a problemas gastrointestinais. Recentemente, o AINE naproxeno foi introduzido no mercado em combinação com esomeprazol, um inibidor da bomba de prótons que reduz a formação de ácido do estômago, com o intuito de amenizar esses efeitos colaterais. Usando um design duplo-cego randomizado e cruzado, este estudo teve como objetivo avaliar, pela primeira vez, o controle da dor aguda pós-operatória em 46 pacientes que consumiram um comprimido de NE (naproxeno 500 mg + esomeprazol 20 mg) ou de naproxeno puro (500 mg) a cada 12 horas por 4 dias após a cirurgia dos dois terceiros molares inferiores em posições semelhantes, em dois momentos distintos. As pacientes do sexo feminino relataram significativamente mais dor em 1, 1,5, 2, 3 e 4 horas após a cirurgia e também consumiram sua primeira medicação de socorro (paracetamol) em um tempo significativamente menor quando consumiram NE em comparação com naproxeno puro (3,7 horas e 6,7 horas, respectivamente). Não foram encontradas diferenças significativas nos pacientes do sexo masculino entre cada grupo de pesquisa. Em conclusão, o naproxeno melhorou o manejo da dor aguda pós-operatória quando comparado ao NE em mulheres, embora, ao longo de todo o estudo, a dor foi considerada leve após o uso de ambos os medicamentos em homens e mulheres com escores de dor em média bem abaixo de 40 mm na EAV. / Nonsteroidal anti-inflammatory drugs (NSAIDs) are frequently used to manage postoperative pain and inflammation in patients after oral and maxillofacial surgery, yet they are often associated with gastrointestinal problems. Recently the NSAID naproxen was introduced in combination with esomeprazole, a proton pump inhibitor that reduces stomach acid formation, in an effort to assuage these side effects. Using a double-blinded randomized crossover design, this study aimed to evaluate, for the first time, acute postoperative pain management in 46 volunteers who consumed a tablet of either naproxen (500 mg) with esomeprazole (20 mg) (NE) or only naproxen (500 mg) every 12 hours for 4 days after extractions of the two lower third molars, in two different appointments. Female volunteers reported significantly more pain at 1, 1.5, 2, 3 and 4 hours after surgery and also consumed their first rescue medication (acetaminophen) at a time significantly earlier when consuming NE when compared to naproxen (3.7 hours and 6.7 hours, respectively). No significant differences were found in the males between each research group. In conclusion, naproxen improved acute postoperative pain management when compared to NE in women although, throughout the entire study, pain was mild after the use of both drugs in both men and women with pain scores on average well below 40 mm on the VAS.

Made available in DSpace on 2014-12-17T14:16:25Z (GMT). No. of bitstreams: 1 LilianGSS_Dissert.pdf: 1150151 bytes, checksum: 37f7b5b1cb7bc3900c20765f52024d2e (MD5) Previous issue date: 2010-02-26 / Compounded medicines have been reported by the ANVISA due to decreased of the therapeutic response or toxicity of these formulations. The aim of this work was to investigate the physicochemical quality control among naproxen sodium oral suspensions 25 mg/mL obtained from six compounding pharmacies (A, B, C, D, E and F) and the manufactured suspension (R). In the quality control test, the tests of pH, content, homogeneity, volume and physical and organoleptic characteristics were performed according to the Brazilian Pharmacopoeia. The analytical method for determination of naproxen in suspensions was validate. This method showed excellent precision, accuracy, linearity and specificity. In the content test the suspensions B, C and E showed lower value and the F suspension showed a high value of the content. The products C and E were disapproved in the description of the physical and organoleptic characteristics test. In the pH test, three suspensions were outside specifications (C, E and F). Only the products R, A and D showed satisfactory results in these tests and therefore they were approved for relative bioavailability test. The R, A and D suspensions were orally administered to Wistar rats and the blood samples were taken at time intervals of 10, 20, 40, 60 min, 3, 4, 6, 24 and 48 h. The plasma samples were immediately stored at 80 ?C until analysis of HPLC. The bioanalytical method validation showed specificity, linearity (R2 0.9987), precision, accuracy, good recovery and stability. The chromatographic conditions were: flow rate of 1.2 mL.min-1 with a mobile phase of acetonitrile : sodium phosphate buffer pH 4.0 (50:50, v/v) at 280 nm, using a C18 column. The confidence interval of 90% for the Cmax and AUCt ratio was within the range of 80 - 125% proposed by the FDA. Only one suspension, obtained from the compounding pharmacy D, was considered bioequivalent to the rate of absorption under the conditions proposed by this study. Thus, the results indicate the need for strict supervision from the relevant authorities to ensure the patient safety and the quality of compounded drugs by pharmacies / Os medicamentos vendidos nas farm?cias de manipula??o t?m sido notificados pela ANVISA quanto ? resposta terap?utica reduzida ou por toxicidade. Disto e visando avaliar a qualidade dos produtos magistrais o presente trabalho objetivou realizar ensaios de controle de qualidade e biodisponibilidade relativa em suspens?es de naproxeno s?dico na concentra??o de 25 mg/mL, obtidas de seis farm?cias de manipula??o na cidade do Natal (A, B, C, D, E e F) e de uma suspens?o de refer?ncia (R). No controle de qualidade f?sico-qu?mico foram realizados ensaios para determina??o do teor, pH, homogeneidade, volume e caracter?sticas organol?pticas. O m?todo utilizado no doseamento mostrou precis?o, exatid?o, linearidade e especificidade. As formula??es obtidas nas farm?cias B, C e E apresentaram um teor abaixo das especifica??es, enquanto que a formula??o F apresentou um teor acima do recomendado pela Farmacop?ia Americana. Em rela??o ao pH as suspens?es C, E e F apresentaram resultados fora das especifica??es farmacop?icas. Desta forma, apenas as suspens?es R, A e D foram selecionadas para o ensaio de biodisponibilidade relativa. As suspens?es R, A e D foram administradas oralmente em ratos Wistar e o sangue foi coletado em intervalos de 10, 20, 40 e 60 min, 3, 4, 6, 24 e 48 h. O plasma obtido foi ent?o armazenado em freezer a 80 ?C para posterior an?lise em CLAE. O m?todo utilizado apresentou especificidade, linearidade (R2 = 0,9987), precis?o, exatid?o, adequada recupera??o e estabilidade. As condi??es cromatogr?ficas utilizadas na metodologia foram: fluxo 1,2 mL/min, a fase m?vel constituiu tamp?o fosfato de s?dio monob?sico 0,01 M pH 4,0 e acetonitrila (50:50, v/v), coluna C18 e comprimento de onda em 280 nm. O ?ndice de confian?a de 90% para as raz?es de Cmax e ASCt se encontrou dentro do intervalo de 80 125% proposto pelo FDA apenas para a suspens?o obtida da farm?cia de manipula??o D, sendo, portanto considerada bioequivalente para a taxa de absor??o nas condi??es propostas por este estudo. Assim, os resultados obtidos indicam a necessidade de uma maior fiscaliza??o pelos ?rg?os competentes de forma a garantir a seguran?a do paciente e qualidade do medicamento produzido pela farm?cia de manipula??o

Anti-inflamatórios não esteroidais (AINEs) são frequentemente utilizados para controlar a dor pós-operatória e inflamação em pacientes após a cirurgia oral e maxilofacial, mas eles são muitas vezes associados a problemas gastrointestinais. Recentemente, o AINE naproxeno foi introduzido no mercado em combinação com esomeprazol, um inibidor da bomba de prótons que reduz a formação de ácido do estômago, com o intuito de amenizar esses efeitos colaterais. Usando um design duplo-cego randomizado e cruzado, este estudo teve como objetivo avaliar, pela primeira vez, o controle da dor aguda pós-operatória em 46 pacientes que consumiram um comprimido de NE (naproxeno 500 mg + esomeprazol 20 mg) ou de naproxeno puro (500 mg) a cada 12 horas por 4 dias após a cirurgia dos dois terceiros molares inferiores em posições semelhantes, em dois momentos distintos. As pacientes do sexo feminino relataram significativamente mais dor em 1, 1,5, 2, 3 e 4 horas após a cirurgia e também consumiram sua primeira medicação de socorro (paracetamol) em um tempo significativamente menor quando consumiram NE em comparação com naproxeno puro (3,7 horas e 6,7 horas, respectivamente). Não foram encontradas diferenças significativas nos pacientes do sexo masculino entre cada grupo de pesquisa. Em conclusão, o naproxeno melhorou o manejo da dor aguda pós-operatória quando comparado ao NE em mulheres, embora, ao longo de todo o estudo, a dor foi considerada leve após o uso de ambos os medicamentos em homens e mulheres com escores de dor em média bem abaixo de 40 mm na EAV. / Nonsteroidal anti-inflammatory drugs (NSAIDs) are frequently used to manage postoperative pain and inflammation in patients after oral and maxillofacial surgery, yet they are often associated with gastrointestinal problems. Recently the NSAID naproxen was introduced in combination with esomeprazole, a proton pump inhibitor that reduces stomach acid formation, in an effort to assuage these side effects. Using a double-blinded randomized crossover design, this study aimed to evaluate, for the first time, acute postoperative pain management in 46 volunteers who consumed a tablet of either naproxen (500 mg) with esomeprazole (20 mg) (NE) or only naproxen (500 mg) every 12 hours for 4 days after extractions of the two lower third molars, in two different appointments. Female volunteers reported significantly more pain at 1, 1.5, 2, 3 and 4 hours after surgery and also consumed their first rescue medication (acetaminophen) at a time significantly earlier when consuming NE when compared to naproxen (3.7 hours and 6.7 hours, respectively). No significant differences were found in the males between each research group. In conclusion, naproxen improved acute postoperative pain management when compared to NE in women although, throughout the entire study, pain was mild after the use of both drugs in both men and women with pain scores on average well below 40 mm on the VAS.

Orientador: Maria Julia Marques / Dissertação (mestrado) - Universidade Estadual de Campinas, Instituto de Biologia / Made available in DSpace on 2018-08-11T07:04:11Z (GMT). No. of bitstreams: 1 Albuquerque_TerezaCristinaPessoade_M.pdf: 1316775 bytes, checksum: 93039fe90ce98e8b3c47f1e5074f823b (MD5) Previous issue date: 2008 / Resumo: Em fibras musculares distróficas de camundongos mdx e na distrofia muscular de Duchenne (DMD), a deficiência da distrofina provoca lesão do sarcolema e degeneração muscular. Esta deficiência está associada a alterações na estabilidade do sarcolema e aumento dos níveis intracelulares de cálcio, que podem provocar mionecrose. O mecanismo exato que provoca a lesão da membrana e a mionecrose são desconhecidos. Uma das hipóteses é que a resposta inflamatória endógena pode aumentar a lesão do sarcolema devido à ausência da distrofina. O processo inflamatório está associado com a regeneração muscular e com o reparo tecidual. Antiinflamatórios não-esteróides (AINE) podem reduzir a inflamação que ocorre como conseqüência de lesão muscular através da inibição das ciclo-oxigenases, provocando redução da síntese de prostaglandinas. Estudos anteriores mostraram que houve atraso da regeneração muscular após a utilização de AINE. Neste estudo, foi verificado se os AINE naproxeno e nimesulida reduzem a mionecrose no camundongo mdx, um dos modelos experimentais da DMD. Camundongos mdx (n= 48) com 21 dias de idade receberam injeções intraperitoneais de naproxeno (10 mg/kg de peso corporal) ou de nimesulida (25 mg/kg de peso corporal) durante 15 ou 30 dias consecutivos. Camundongos mdx não tratados receberam injeção intraperitoneal de solução salina. Para a observação e contagem de fibras em degeneração, três animais de cada grupo receberam injeção intraperitoneal de azul de Evans (AE), marcador que indica alterações de permeabilidade do sarcolema. Secções transversais dos músculos esternomastóide (STN) e tibial anterior (TA) foram coradas com hematoxilina-eosina. A secção total transversa foi dividida em área de infiltrado inflamatório, com fibras em estágios iniciais de regeneração (áreas INFL/REG), e áreas com fibras em estágio avançado de regeneração (áreas REG). Estas áreas foram expressas como porcentagens da área seccional transversa total. O número de fibras regeneradas (fibras com núcleo central), fibras com núcleo periférico e fibras em degeneração (positivas ao corante AE) foram quantificadas. O naproxeno mostrou-se mais efetivo que a nimesulida na diminuição da mionecrose, influenciando de maneira diferenciada os ciclos de degeneração/regeneração de músculos distróficos, principalmente quando utilizado nos estágios iniciais destes ciclos. Concluímos que a utilização de inibidores da produção de prostaglandinas não prejudica a regeneração muscular de camundongos mdx, podendo ser uma alternativa para o tratamento de miopatias, especialmente se associada a outros fármacos de ação mais direta na proteção à mionecrose / Abstract: In dystrophin-deficient fibers of mdx mice and in Duchenne muscular dystrophy (DMD), the lack of dystrophin leads to sarcolemma breakdown and muscle degeneration. The lack of dystrophin is associated with changes in membrane stability and increased levels of calcium in the muscle fiber, which leads to myonecrosis. The exact mechanism determining the sarcolemmal lesion and myonecrosis is unknown. One hypothesis is that the endogenous inflammatory response exacerbates the muscle fiber membrane damage due to the lack of dystrophin. Inflammation has also been associated to muscle regeneration and repair. Non-steroidal anti-inflammatory drugs (NSAID) can reduce inflammation following injury by inhibiting cyclooxygenase and leading to a reduction in prostaglandin synthesis. Previous experimental studies have indicated delayed muscle regeneration after NSAID. In this work, we verified the effects of the cyclooxygenase inhibitors, naproxen and nimesulide, on the extent of myofiber necrosis in the mdx mice model of DMD. Mdx mice (n=48) at 21 days after birth received daily intraperitoneal injections of naproxen (10 mg/kg body weight) or nimesulide (25 mg/kg body weight) for 15 or 30 days. Untreated mdx mice were injected with saline. To measure muscle fiber damage, some mice were injected with Evans blue dye (EBD; n=3), a marker of sarcolemmal lesion. Cryostat cross-sections of the sternomastoid (STN) and tibialis anterior (TA) muscles were stained with hematoxylin and eosin. The whole cross-sectional area of the muscles was divided into areas of inflammatory infiltrate with early regenerating fibers (INFL/REG areas) and areas with late regenerated fibers (REG areas). These areas were expressed as a percentage of the total cross-section area. The number of regenerated fibers (central nucleated fibers), fibers with peripheral cell nuclei and degenerated fibers (positive to EBD) was counted. Naproxen was more effective than nimesulide in decreasing myonecrosis. We concluded that the use of prostaglandin inhibitors does not impair muscle regeneration in the mdx mice. They could be useful therapeutic alternatives in the treatment of DMD, but should be accompanied by other strategies directed against muscle degeneration / Mestrado / Anatomia / Mestre em Biologia Celular e Estrutural

Phase-transfer catalysts derived from the cinchona alkaloids cinchonine and cinchonidine are widely used in the asymmetric alkylation of substrates bearing moieties that resonance stabilize their enolates. The investigation of α-oxygenated esters revealed decreased α-proton acidity, indicating the oxygen's overall destabilizing effect on enolates by electron-pair repulsion. Alkylation of α-oxygenated aryl ketones with various alkyl halides proved successful with a cinchonidine catalyst, giving products with high yield and enantioselectivity. The resulting compounds were converted to esters through modified Baeyer-Villiger oxidation. Alkylation with indolyl electrophiles gave products that underwent decomposition under Baeyer-Villiger conditions. Alternative N-methylimidazolyl ketones were explored. Alkylated imidazolyl ketones, obtained in high yield and enantioselectivity, could be converted to esters through treatment with methyl triflate and basic methanol. This technique has the advantage of not requiring stoichiometric addition of chiral reagents, which is requisite when employing traditional chiral auxiliaries. This method's utility is demonstrated in the total asymmetric syntheses of (+)-kurasoin B and analogs, and 12-(S)-HETE. Kurasoin B is a fungal-derived natural compound possessing moderate farnesyl transfer (FTase) inhibitive activity (IC50 = 58.7 μM). FTase catalyzes post-translation modifications of membrane-bound Ras proteins, which function in signal cell transduction that stimulates cell growth and division. The oncogenic nature of mutated Ras proteins is demonstrated by their commonality in human tumors. Thus, FTase inhibitors like (+)-kurasoin B possess potential as cancer chemotherapy leads. Derivatization may enable structure-activity-relationship studies and greater FTase inhibition activity to be found. 12-(S)-HETE, a metabolite from a 12-lipoxygenase pathway from arachidonic acid, has been found to participate in a large number of physiological processes. Its transient presence in natural tissues makes total synthesis an attractive avenue for obtaining sufficient quantities for further study. Five asymmetric syntheses of 12-(S)-HETE have been reported. Three require chiral resolutions of racemates, with the undesired enantiomers being discarded or used for other applications. Asymmetric PTC alkylation is also described for aryl acetates, whose products were enantioenriched through recrystallization. This technique is applied to a total synthesis of the anti-inflammatory drug (S)-Naproxen.

Le sel sodique du carboxyméthylamidon à haute teneur en amylose, HASCA (Amylose 60%-amylopectine 40%), est un polymère hydrophile ionique utilisé pour obtenir des comprimés matriciels à libération prolongée. Il est caractérisé par la formation d'un hydrogel lors de la mise en contact avec le milieu de dissolution. La libération du principe actif (PA) à travers ce polymère est principalement contrôlée par la diffusion fickienne et la relaxation du réseau polymérique. De plus, la solubilité du PA est un facteur qui permet de moduler la libération, cependant, la solubilité d’un médicament dépend de la forme utilisée et du pH. Les bases ou les acides libres présentent une solubilité moins élevée que leurs sels. Nous proposons d’étudier l’effet d’une combinaison entre le sel et la forme acide ou le sel et la forme alcaline dans le même comprimé matriciel d’HASCA. Comme objectif de ce travail, nous étudions l’influence de la nature du polymère sur le profil de libération de PA dans un milieu aqueux en gradient de pH à cause de la nature des matrices à base d’HASCA caractérisées par la présence de groupement carboxyliques, ionisés ou non selon l’acidité du milieu de dissolution. Nous étudions également l’influence de la nature du PA (base libre ou son sel vs acide libre ou son sel) sur le profil de libération, ceci en sélectionnant des PAs modèles représentatifs de chaque catégorie. L’influence de changement de proportions entre la forme libre (acide ou base) et son sel sur le profil de libération est aussi investiguée. Pour ce, des comprimés à base de HASCA avec des proportions différentes de, soit le naproxène acide et le naproxène de sodium (PA acide et son sel), soit la tétracycline et le chlorhydrate de tétracycline (PA alcalin et son sel) ont été utilisés. Ceux-ci sont évalués lors des tests de dissolution dans un milieu simulant le milieu gastro-intestinal, selon les normes de l’USP (spectrophotométrie UV). Nous avons également menés des études de vitesse de dissolution intrinsèque sur les PAs purs, afin de déterminer leur solubilité et vitesse de libération dans les même pH de dissolution étudiés. Nous avons réussit d’obtenir des comprimés matriciels à base de HASCA convenables à être administrés une fois par jour en utilisant une combinaison du naproxène acide et son sel comme PA. Tandis que les résultats ont montré qu’en utilisant la tétracycline et son sel, les temps de libération étaient trop élevés pour permettre la réalisation d’une formulation convenable pour une administration quotidienne unique / HASCA, High amylose sodium carboxymethyl starch (amylose-amylopectin 60% 40%), a hydrophilic ionic polymer used in sustained release matrix tablets, is characterized by the formation of a hydrogel when in contact with the dissolution medium. Fickian diffusion and relaxation of the polymer chains are the main mechanisms controlling the release of the active ingredients (AI). On the other hand, a common method for sustaining the release of a drug is to decrease its solubility. It’s known that the solubility of a drug depends on the form used, either the free base versus its salt or the free acid versus its salt. A combination of the salt and acid forms or the salt and base forms in the same HASCA matrix tablet was proposed. The objective of this work is to study the effect of the polymer’s nature on the release profile of AI in an aqueous medium with a pH gradient because HASCA is characterized by the presence of a carboxylic group, ionized or not according to the acidity of the dissolution medium. The influence of the nature of the AI, the salt or free base versus the free acid or its salt, on the release profile is also studied by selecting models of AI in each category. The influence of the proportion between the free form (acid or base) and the combination with its salt on the release profile is also investigated. Therefore tablets containing HASCA with different proportion of either naproxen acid and naproxen sodium (acid AI and its salt) or tetracycline and tetracycline hydrochloride (basic AI and it salt) are used. This will be evaluated during dissolution tests in a medium similar to the gastrointestinal environment according to the standards of USP (UV spectrophotometry). Intrinsic dissolution tests will also be conducted on pure AI for different studied pH. This study shows that using HASCA, it’s possible to obtain sustained release tablets for administration once a day using a combination of acid and salt as AI.

L'exposition aux mélanges de contaminants (environnementaux, alimentaires ou thérapeutiques) soulève de nombreuses interrogations et inquiétudes vis-à-vis des probabilités d'interactions toxicocinétiques et toxicodynamiques. Une telle coexposition peut influencer le mode d’action des composants du cocktail et donc de leur toxicité, suite à un accroissement de leurs concentrations internes. Le bisphénol A (4 dihydroxy-2,2-diphenylpropane) est un contaminant chimique répandu de manière ubiquitaire dans notre environnement, largement utilisé dans la fabrication des plastiques avec l’un des plus grands volumes de production à l’échelle mondiale. Il est un perturbateur endocrinien par excellence de type œstrogèno-mimétique. Cette molécule est biotransformée en métabolites non toxiques par un processus de glucuronidation. L'exposition concomitante à plusieurs xénobiotiques peut induire à la baisse le taux de glucuronidation du polluant chimique d'intérêt, entre autres la co-exposition avec des médicaments. Puisque la consommation de produits thérapeutiques est un phénomène grandissant dans la population, la possibilité d’une exposition simultanée est d’autant plus grande et forte. Sachant que l'inhibition métabolique est le mécanisme d'interaction le plus plausible pouvant aboutir à une hausse des niveaux internes ainsi qu’à une modulation de la toxicité prévue, la présente étude visait d'abord à confirmer et caractériser ce type d'interactions métaboliques entre le bisphénol A et le naproxène, qui est un anti-inflammatoire non stéroïdiennes (AINS), sur l'ensemble d'un organe intact en utilisant le système de foie de rat isolé et perfusé (IPRL). Elle visait ensuite à déterminer la cinétique enzymatique de chacune de ces deux substances, seule puis en mélange binaire. Dans un second temps, nous avons évalué aussi l’influence de la présence d'albumine sur la cinétique métabolique et le comportement de ces deux substances étudiées en suivant le même modèle de perfusion in vivo au niveau du foie de rat. Les constantes métaboliques ont été déterminées par régression non linéaire. Les métabolismes du BPA et du NAP seuls ont montré une cinétique saturable avec une vélocité maximale (Vmax) de 8.9 nmol/min/ mg prot de foie et une constante d'affinité de l'enzyme pour le substrat (Km) de 51.6 μM pour le BPA et de 3 nmol/min/mg prot de foie et 149.2 μM pour le NAP. L'analyse des expositions combinées suggère une inhibition compétitive partielle du métabolisme du BPA par le NAP avec une valeur de Ki estimée à 0.3542 μM. Les résultats obtenus montrent que l’analyse de risque pour les polluants environnementaux doit donc prendre en considération la consommation des produits pharmaceutiques comme facteur pouvant accroitre le niveau interne lors d’une exposition donnée. Ces données in vivo sur les interactions métaboliques pourraient être intégrées dans un modèle pharmacocinétique à base physiologique (PBPK) pour prédire les conséquences toxicococinétique (TK) de l'exposition d'un individu à ces mélanges chimiques. / Exposure to mixtures of contaminants (environmental, alimentary or therapeutic) poses many questions and concerns about the probability of toxicokinetic and toxicodynamic interactions. Such co-exposure may influence the mode of action of the cocktail of components and therefore their toxicity, as a result of an increase of their internal concentrations. Bisphenol A (2,2-dihydroxy-4 diphenylpropane) is a ubiquitous chemical contaminant in our environment, widely used in the manufacture of plastics, with one of the largest production volumes globally. It is an endocrine disruptor of oestrogen-mimetic type. This molecule is metabolized into non-toxic metabolites by glucuronidation process. Several factors including the co-exposure to other xenobiotics may reduce the glucuronidation rate of the chemical pollutant of interest, such as a co-exposure with drugs. Given that the consumption of therapeutic products is a growing phenomenon in the population, simultaneous exposures of pollutants with medicinal drugs are becoming a concern for health risk assessment. Knowing that metabolic inhibition is the most plausible mechanism of interaction that could result in an increase of the internal levels and a modulation of the expected toxicity, the present study aimed at confirming and characterizing this type of metabolic interactions between bisphenol A and naproxen, which is a non-steroidal anti-inflammatory drug (NSAID), on the set of an intact organ using isolated and perfused rat liver system (IPRL). The study also served to determine the enzyme kinetics metabolism of each substance alone and in binary mixture. In a second step, we also evaluated the influence of the presence of albumin on the metabolic kinetics and behavior of the two substances studied using the same in vivo perfusion model of rat liver. Metabolic constants were determined by nonlinear regression. The metabolism of BPA and NAP alone demonstrated saturable kinetics with maximal velocity (Vmax) and affinity constant (Km) equal to 8.9 nmol/min/mg prot liver and 51.6 μM for BPA, and 3 nmol/min/mg prot liver and 149.2 μM for NAP. The analysis of combined exposures suggests a partial competitive inhibition of BPA metabolism by NAP with a Ki value estimated at 0.3542 µM. From these results, it appears that the risk assessment of environmental pollutants must therefore consider the consumption of pharmaceutical products as an important agent that can increase the internal levels for a given exposure. Accordingly, these in vivo data on metabolic interactions can be incorporated into a physiologically based pharmacokinetic models (PBPK) to predict the toxicokinetic consequences (TK) of the exposure by an individual to these chemical mixtures.

Le sel sodique du carboxyméthylamidon à haute teneur en amylose, HASCA (Amylose 60%-amylopectine 40%), est un polymère hydrophile ionique utilisé pour obtenir des comprimés matriciels à libération prolongée. Il est caractérisé par la formation d'un hydrogel lors de la mise en contact avec le milieu de dissolution. La libération du principe actif (PA) à travers ce polymère est principalement contrôlée par la diffusion fickienne et la relaxation du réseau polymérique. De plus, la solubilité du PA est un facteur qui permet de moduler la libération, cependant, la solubilité d’un médicament dépend de la forme utilisée et du pH. Les bases ou les acides libres présentent une solubilité moins élevée que leurs sels. Nous proposons d’étudier l’effet d’une combinaison entre le sel et la forme acide ou le sel et la forme alcaline dans le même comprimé matriciel d’HASCA. Comme objectif de ce travail, nous étudions l’influence de la nature du polymère sur le profil de libération de PA dans un milieu aqueux en gradient de pH à cause de la nature des matrices à base d’HASCA caractérisées par la présence de groupement carboxyliques, ionisés ou non selon l’acidité du milieu de dissolution. Nous étudions également l’influence de la nature du PA (base libre ou son sel vs acide libre ou son sel) sur le profil de libération, ceci en sélectionnant des PAs modèles représentatifs de chaque catégorie. L’influence de changement de proportions entre la forme libre (acide ou base) et son sel sur le profil de libération est aussi investiguée. Pour ce, des comprimés à base de HASCA avec des proportions différentes de, soit le naproxène acide et le naproxène de sodium (PA acide et son sel), soit la tétracycline et le chlorhydrate de tétracycline (PA alcalin et son sel) ont été utilisés. Ceux-ci sont évalués lors des tests de dissolution dans un milieu simulant le milieu gastro-intestinal, selon les normes de l’USP (spectrophotométrie UV). Nous avons également menés des études de vitesse de dissolution intrinsèque sur les PAs purs, afin de déterminer leur solubilité et vitesse de libération dans les même pH de dissolution étudiés. Nous avons réussit d’obtenir des comprimés matriciels à base de HASCA convenables à être administrés une fois par jour en utilisant une combinaison du naproxène acide et son sel comme PA. Tandis que les résultats ont montré qu’en utilisant la tétracycline et son sel, les temps de libération étaient trop élevés pour permettre la réalisation d’une formulation convenable pour une administration quotidienne unique / HASCA, High amylose sodium carboxymethyl starch (amylose-amylopectin 60% 40%), a hydrophilic ionic polymer used in sustained release matrix tablets, is characterized by the formation of a hydrogel when in contact with the dissolution medium. Fickian diffusion and relaxation of the polymer chains are the main mechanisms controlling the release of the active ingredients (AI). On the other hand, a common method for sustaining the release of a drug is to decrease its solubility. It’s known that the solubility of a drug depends on the form used, either the free base versus its salt or the free acid versus its salt. A combination of the salt and acid forms or the salt and base forms in the same HASCA matrix tablet was proposed. The objective of this work is to study the effect of the polymer’s nature on the release profile of AI in an aqueous medium with a pH gradient because HASCA is characterized by the presence of a carboxylic group, ionized or not according to the acidity of the dissolution medium. The influence of the nature of the AI, the salt or free base versus the free acid or its salt, on the release profile is also studied by selecting models of AI in each category. The influence of the proportion between the free form (acid or base) and the combination with its salt on the release profile is also investigated. Therefore tablets containing HASCA with different proportion of either naproxen acid and naproxen sodium (acid AI and its salt) or tetracycline and tetracycline hydrochloride (basic AI and it salt) are used. This will be evaluated during dissolution tests in a medium similar to the gastrointestinal environment according to the standards of USP (UV spectrophotometry). Intrinsic dissolution tests will also be conducted on pure AI for different studied pH. This study shows that using HASCA, it’s possible to obtain sustained release tablets for administration once a day using a combination of acid and salt as AI.