Programa de Pós-Graduação em Ciências Farmacêuticas
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Doctoral Thesis Obtenção, caracterização e avaliação da atividade antifúngica de nanopartículas de quitosana contendo o peptídio TistH identificado na peçonha do escorpião Tityus stigmurus(2019-05-23) Rêgo, Manoela Torres do; Pedrosa, Matheus de Freitas Fernandes; Silva Júnior, Arnobio Antonio da; ; ; ; Machado, Paula Renata Lima; ; Andrade, Vânia Sousa; ; Martins, Alice Maria Costa; ; Azevedo, Eduardo Pereira de;The Tityus stigmurus scorpion venom has been studied in recent years and revealed many toxins (peptides) with high potential for biotechnological applications. The transcriptomics approach from the venom glands of T. stigmurus, realized by our scientific group, identified several bioactive peptides with pharmacological potential. Among the molecules identified, the activity of peptide like-hypotensins class (TistH, Tityus stigmurus hypotensin), reported in the literature, evidenced the TistH with bradykinin-potentiating peptide, in addition exert antifungal effect. The maximum efficacy of this class of compounds can be achieved by immobilizing it in specific and suitable biomaterials or suitable carriers. This study objective to obtained and characterized physiochemically TistH-loading cross-linked chitosan nanoparticles for incorporation and adsorption, evaluate the biocompatibility as well as improve of antifungal effect. The nanossystems containing the TistH by incorporation (CN-TistH-Inc) and absorption (CN-TistH-Ads) methods at concentration of 0.5 and 1.0% were obtained by ionic gelation technique. The nanosystems at 0.5 and 1.0% were analyzed by dynamic light scattering showed positive zeta potential, with size particles less than 160 nm and polydispersity index less than 0.3. The encapsulation efficiency were greater than 96.5% and the success was confirmed by polyacrylamide gel electrophoresis and Fourier transform infrared spectroscopy. The CNTistH-Inc and CN-TistH-Ads at 0.5 and 1.0% were analyzed by atomic force microscopy and scanning electronic microscopy evidenced nanometric particles, spherical shape with smooth surface and homogeneous aspects. The stability was evaluated by particle size and polydispersity index can possible observed stable colloidal dispersions for 4 months. According in vitro release profile was observed for all formulations prolonged and sustained release for 24 hours, with kinetic parabolic diffusion mechanism. The biocompatibility was evaluated in erythrocytes, normal cells originated from murine macrophage (RAW 264.7) and kidney of African green monkeys (Vero E6) suggested biocompatibility that both nanosystems, in concentration used for antifungal assay, not demonstrated be hemolytic and cytotoxic, respectively. The antifungal activity was evaluated through of determination of the Minimum inhibitory concentration (MIC), minimum fungicidal concentration (MFC) and antibiofilm activity used yeast of genus Candida. The CN-TistH-Inc and CN-TistH-Ads showed a minimal inhibitory concentration of 89.2 µg/mL against Candida albicans, 11.1 µg/mL for C. parapsilosis and C. tropicalis, confirmed by minimum fungicidal concentrations assay. Moreover, the TistH-loaded cross-linked chitosan nanoparticles significantly reduced the biofilm formation of clinical yeast sepsis of C. tropicalis and C. krusei, as well as clinical yeasts of vulvovaginal candidiasis of C. albicans. Thus, this study demonstrated that chitosan nanoparticles are promising for carrying and delivering the peptides TistH from Tityus stigmurus, with improve antifungal activity against genus Candida spp. may eventually be applied in the future as a possible antifungal agent.Master Thesis Avaliação da atividade anti-inflamatória do extrato aquoso, frações e compostos identificados nos frutos de Hancornia Speciosa Gomes (Apocynaceae)(Universidade Federal do Rio Grande do Norte, 2015-06-19) Rêgo, Manoela Torres do; Pedrosa, Matheus de Freitas Fernandes; Bitencourt, Mariana Angélica Oliveira; ; http://lattes.cnpq.br/0893922249078027; ; http://lattes.cnpq.br/2929963416385218; ; http://lattes.cnpq.br/0735782214033188; Guerra, Gerlane Coelho Bernardo; ; http://lattes.cnpq.br/5677318431530876; Soares, Luiz Alberto Lira; ; http://lattes.cnpq.br/4290808161139329Hancornia speciosa Gomes (Apocynaceae), popularly known as ‘mangabeira’, has been used in folk medicine to treat inflammatory disorders, hypertension, dermatitis, diabetes, liver diseases and stomach disorders. Regarding the Hancornia speciosa fruits, the ethnobotany indicates its use especially for treating inflammation and tuberculosis. However, no study has been done so far to prove such biological activities. The objective was evaluation anti-inflammatory activity from the fruits of Hancornia speciosa Gomes (mangabeira). Aqueous extract was prepared by decoction, subsequently submitted the liquid-liquid fractionation. The secondary metabolites were identified by high performance liquid chromatography coupled with detector diode array (HPLC-DAD) and liquid chromatography diode array detector coupled with mass spectrometry (LC-DAD-MS). The anti-inflammatory properties of the aqueous extract, dichloromethane (CH2Cl2), ethyl acetate (EtOAc) and n-butanol (n-BuOH) fractions of the fruits from H. speciosa, as well as rutin and chlorogenic acid were investigated using in vitro and in vivo models. In vivo tests comprised the xylene-induced ear edema that was measured the formation of edema, carrageenan-induced peritonitis was evaluated the total leukocytes at 4h and zymosan-induced air pouch was measured the total leukocytes and differential cell count at 6, 24 and 48 hours, whereas in vitro tests were evaluated levels of cytokines IL-1β, IL-6, IL-12 and TNF-α using ELISA obtained of carrageenan-induced peritonitis model. The results showed the presence of rutin and chlorogenic acid were detected in the aqueous extract from H. speciosa fruits by HPLC-DAD and LC-DAD-ME. Furthermore, the aqueous extracts and fractions, as well as rutin and chlorogenic acid significantly inhibited the xilol-induced ear edema and reduced cell migration in the animal models such as carrageenan-induced peritonitis and zymosan-induced air pouch. In addition, reduced levels of cytokines IL-1β, IL-6, IL-12 and TNF-α were observed. This is the first study that demonstrated the anti-inflammatory effect of aqueous extract from Hancornia speciosa fruits against different inflammatory agents in animal models, suggesting that their bioactive molecules, especially rutin and chlorogenic acid contributing, at least in part, to the anti-inflammatory effect of aqueous extract. These findings support the widespread use of Hancornia speciosa in popular medicine and demonstrate that this aqueous extract has therapeutic potential for the development of a herbal drugs with anti-inflammatory properties.