Submitted by Luciana Ferreira (lucgeral@gmail.com) on 2014-10-29T14:34:23Z No. of bitstreams: 2 Dissertação - Iury Valentim Jorge Zoghaib - 2013.pdf: 1607539 bytes, checksum: d92b3313edee2773935cb667be5b908d (MD5) license_rdf: 23148 bytes, checksum: 9da0b6dfac957114c6a7714714b86306 (MD5) / Approved for entry into archive by Luciana Ferreira (lucgeral@gmail.com) on 2014-10-30T10:48:45Z (GMT) No. of bitstreams: 2 Dissertação - Iury Valentim Jorge Zoghaib - 2013.pdf: 1607539 bytes, checksum: d92b3313edee2773935cb667be5b908d (MD5) license_rdf: 23148 bytes, checksum: 9da0b6dfac957114c6a7714714b86306 (MD5) / Made available in DSpace on 2014-10-30T10:48:45Z (GMT). No. of bitstreams: 2 Dissertação - Iury Valentim Jorge Zoghaib - 2013.pdf: 1607539 bytes, checksum: d92b3313edee2773935cb667be5b908d (MD5) license_rdf: 23148 bytes, checksum: 9da0b6dfac957114c6a7714714b86306 (MD5) Previous issue date: 2013-10-18 / Conselho Nacional de Pesquisa e Desenvolvimento Científico e Tecnológico - CNPq / The LQFM030 was obtained by molecular simplification of nutlins (inhibitors of p53-MDM2 interaction) and, having demonstrated excellent antineoplastic activity in vitro (cytotoxicity against K-562 cell line with IC50 = 23 M) and in vivo against Ehrlich ascitic tumor with increased survival in treated animals developed the pharmacokinetic study of p.o. in rats. It was used for LC-MS/MS analytical method (Applied Biosystems MDS Sciex API 3200) previously validated together. O LQFM030 was administered to 3 rats (mean weight 186g) in predetermined dose of 100 mg/kg by gavage. After administration, samples were collected from 1.0 mL of blood by cannulation of the left jugular vein with heparinized syringe, the intervals 0-8 h. The blood samples were identified and centrifuged to obtain plasma which was frozen at -20°C until analysis. The kinetic parameters were calculated in the software WinNonlin 5.0 (Pharsight™). Results (mean ± SD) half-life (t1/2) 3.61 ± 0.68 h, total clearance (CLT) 36.49 ± 2.23 mL/min/kg, volume of distribution (Vd) 11,40 ± 1.58 L/kg. The LQFM030 had low value of t1/2, Vd high and high value of CLT. These values allow us to understand that the prototype study demonstrated a good safety profile of tissue distribution and/or has been extensively eliminated. When compared with the kinetic parameters obtained in other studies, it was observed difference in results is justified by the high interspecies variability, mainly in basal metabolic rate and body weight, once the route of administration, orally and intraperitoneally, are kinetically similar. / O LQFM030 foi obtido por simplificação molecular dos nutlins (inibidores da interação MDM2-p53) e, tendo demonstrado excelente atividade antineoplásica in vitro (citotoxicidade contra a linhagem celular K-562 com IC50 = 23 M) e in vivo contra tumor ascítico de Ehrlich, com aumento de sobrevida em animais tratados, desenvolveu-se o estudo do perfil farmacocinético, p.o., em ratos. Empregou-se método analítico em LC-MS/MS (Applied Biosystems MDS Sciex API 3200) previamente validado em colaboração. O LQFM030 foi administrado a 3 ratos (peso médio de 186g), em dose preestabelecida de 100 mg/kg, por gavagem. Após a administração, foram coletadas amostras de 1,0 mL de sangue, por canulação da veia jugular esquerda, com seringa heparinizada, nos intervalos de tempo de 0 a 8 h. As amostras sanguíneas foram identificadas e centrifugadas para obtenção do plasma, que foi congelado a -20ºC até o momento da análise. Os parâmetros cinéticos foram calculados no software Winnonlin 5.0 (Pharsight™). Resultados (média ± DP): meia-vida (t1/2) 3,61 ± 0,68 h; clearance total (CLT) 36,49 ± 2,23 mL/min/kg; volume de distribuição (Vd) 11,40 ± 1,58 L/kg. O LQFM030 apresentou baixo valor de t1/2, elevado Vd e elevado valor de CLT. Estes valores permitem entender que o protótipo estudado demonstrou um bom perfil de distribuição tecidual e/ou foi extensivamente eliminado. Quando comparados com parâmetros cinéticos obtidos em outros estudos, observou-se diferença nos resultados, justificada pela elevada variabilidade interespécies, principalmente na taxa de metabolismo basal e peso corporal, uma vez que as vias de administração, oral e intraperitoneal, são cineticamente semelhantes.

Simple gel formulations may be applied to enhance the systemic and local exposure of potential compounds. The aim of this thesis is the development and characterization of controlled release formulations based on thermo-reversible poloxamer gels, which are suitable for novel drug delivery applications.  In particular co-solvents (DMSO, ethanol), mucoadhesive polymers (chitosan, alginate) and salts (sodium tripolyphosphate, CaCl2) have been used to enhance the applications of poloxamer 407 (P407) formulations in preclinical animal studies. The impact of these additives on the micellization and gelation properties of P407 aqueous solutions was studied by calorimetric methods, nuclear magnetic resonance spectroscopy (NMR) and “tube inversion” experiments. The drug release behavior of hydrophobic and hydrophilic drugs was characterized by using a membrane/membrane-free experimental setup. Finally, preliminary pharmacokinetic studies using a mouse model were conducted for screening of selected inhibitors of bacterial type III secretion and for evaluation of different formulations including P407 gel. All additives, used here, reduced the CMTs (critical micelle temperature) of dilute P407 solutions, with the exception of ethanol. The gelation temperature of concentrated P407 solutions was lowered in the presence of CaCl2, DMSO, TPP and alginate. 1H MAS (Magic Angle Spinning) NMR studies revealed that DMSO influences the hydrophobicity of the PPO segment of P407 polymers. Low concentrations of DMSO did not show any major effect on the drug release from P407 gels and may be used to improve the exposure of lead compounds in poloxamer gels. A newly developed in situ ionotropic gelation of chitosan in combination with TPP in P407 gels showed an enhanced resistance to water and reduced the release rates of model drugs. From preliminary pharmacokinetic studies in mice it was revealed that poloxamer formulations resulted in an increased plasma half-life of the lead compound.

Submitted by Luciana Ferreira (lucgeral@gmail.com) on 2016-08-09T14:22:46Z No. of bitstreams: 2 Dissertação - Alarisse Arçari Fachetti Zoghaib - 2015.pdf: 1188040 bytes, checksum: 9ab52dc612351ab19ab98e90e7ce6ec9 (MD5) license_rdf: 0 bytes, checksum: d41d8cd98f00b204e9800998ecf8427e (MD5) / Approved for entry into archive by Luciana Ferreira (lucgeral@gmail.com) on 2016-08-09T14:24:14Z (GMT) No. of bitstreams: 2 Dissertação - Alarisse Arçari Fachetti Zoghaib - 2015.pdf: 1188040 bytes, checksum: 9ab52dc612351ab19ab98e90e7ce6ec9 (MD5) license_rdf: 0 bytes, checksum: d41d8cd98f00b204e9800998ecf8427e (MD5) / Made available in DSpace on 2016-08-09T14:24:14Z (GMT). No. of bitstreams: 2 Dissertação - Alarisse Arçari Fachetti Zoghaib - 2015.pdf: 1188040 bytes, checksum: 9ab52dc612351ab19ab98e90e7ce6ec9 (MD5) license_rdf: 0 bytes, checksum: d41d8cd98f00b204e9800998ecf8427e (MD5) Previous issue date: 2015-09-28 / The ruthenium (II)/amino acid complex (rutrpII) demonstrated high anticancer activity against murine breast cancer (tumor of Ehrlich) in vitro and in vivo, and increased the median survival of animals. There was then necessary to investigate the pharmacokinetic parameters of the drug prototype as a requirement to pre-clinical knowledge of it. This study aimed to obtain the pharmacokinetic profile of ruthenium (II) / tryptophan administered to rats intraperitoneally in a single dose by quantifying this substance in plasma using validated analytical methodology in UV-VIS spectrophotometer. The methodology consisted of the administration of the prototype rutrpII to 3 rats at a dose of 6 mg/kg, intraperitoneally. Samples of blood 1.0 ml were collected by cannulation of the left jugular vein with heparinized syringe, the intervals from 0 to 9 hr. After centrifugation the plasma was frozen at -20 until the time of analysis. The bioanalytical method to quantify rutrpII was developed and validated in UV-VIS at a wavelength of 417 nm. From the construction of the concentration versus time curve, the kinetic parameters were calculated (Software WinNonlin 5.0 (Pharsight ™) Results were: Tmax = 8 h, Cmax = 169.86 mg/mL, t1/2 = 1.04 ± 0.02 h, ClT/F = 1.32 ± 0.05 mL/min/kg, Vd/F = 1,98 ± 0,05 L/kg. The developed and validated bioanalytical method was suitable for the detection and quantification of RutrpII in rat plasma. The values of pharmacokinetic profiles found, and extrapolated on allometric scaling allow us to understand that the compound studied showed a slow tissue distribution profile and / or was eliminated slowly (Vd low and low value CLT), due to a possible affinity between RutrpII and plasma proteins. This affinity may be explained by the similarity of their chemical structure with iron, enabling it to be transported by biomolecules such as transferrin, albumin or any other, and having as a target the DNA of cancer cells. In general, the compounds of ruthenium (II) / amino acids may be promising drugs for the amino acids present in the complex, may facilitate the recognition of the complex as a whole by DNA, and thus less toxic. / O complexo de rutênio(II)/aminoácido (rutrpII) demonstrou alta atividade antineoplásica contra carcinoma de mama murino (tumor de Ehrlich) in vitro e in vivo, sendo que neste último, aumentou a média de sobrevida dos animais. Fez-se necessária então a investigação dos parâmetros farmacocinéticos deste protótipo de fármaco como requisito ao conhecimento pré-clínico do mesmo. O objetivo deste trabalho foi obter o perfil farmacocinético do rutênio(II)/triptofano administrado a ratos, via intraperitoneal, em dose única por meio da quantificação desta substância em plasma utilizando metodologia analítica validada em espectrofotômetro UV-VIS. A metodologia consistiu na administração do protótipo rutrpII a 3 ratos em dose de 6 mg/kg, por via intraperitonal. Foram coletadas amostras de 1,0 mL de sangue, por canulação da veia jugular esquerda, com seringa heparinizada, nos intervalos de tempo de 0 a 9 h. Após centrifugação, o plasma foi congelado a -20ºC até o momento da análise. O método bioanalítico de quantificação do rutrpII foi desenvolvido e validado em espectrofotômetro UV-VIS no comprimento de onda de 417 nm. A partir da construção da curva de concentração versus tempo, foram calculados os parâmetros cinéticos (Software Winnonlin 5.0 (Pharsight™)). Os resultados encontrados foram: Tmax= 8h, Cmax= 169,86 µg/mL, t(1/2) =1,04 ± 0,02 h,ClT/F = 1,32 ± 0,05 mL/min/kg, Vd/F = 1,98 ± 0,05 L/kg. O método bioanalítico desenvolvido e validado foi adequado para a detecção e quantificação do RutrpII em plasma de rato. Os valores dos perfis farmacocinéticos encontrados, e extrapolados em escala alométrica, permitem entender que o composto estudado apresentou um perfil lento de distribuição tecidual e/ou foi eliminado lentamente (baixo Vd e baixo valor de CLT), devido a uma possível afinidade entre o RutrpII e as proteínas plasmáticas. Esta afinidade pode ser explicada pela semelhança de sua estrutura química com o ferro, possibilitando que ele seja transportado por biomoléculas como a transferrina, albumina ou alguma outra, e tendo como alvo, o DNA da células cancerosas. De maneira geral os compostos de rutênio(II)/ aminoácidos podem ser fármacos promissores pois os aminoácidos presentes nos complexos, podem facilitar o reconhecimento do complexo como um todo pelo DNA, sendo assim menos tóxicos.