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Antitumor Effect of cationic INKKI Peptide from Bovine <i>β</i>-casein on Melanoma B16F10

Journal of cancer Therapy 2012年第4期3卷 237-244页

作者： Ricardo Alexandre Azevedo Adilson Kleber Ferreira Aline Vivian Vatti Auada Kerly Fernanda Mesquita Pasqualoto Rafael Marques-Porto Durvanei Augusto Maria Ivo lebrun Butantan Institute Biochemistry and Biophysical Laboratory Sao Paulo Brazil

cationic peptide with the sequence INKKI 41-45 was isolated from bovine β-casein after tryptic hydrolysis and synthetized. The aim of this work was to evaluate the antiproliferative activity in vitro and antitumor effect in animal model. The in vitro cytotoxicity was evaluated on B16F10 melanoma cells by MTT assay. Detection of apoptosis was measured using the annexin V/PI double staining and cell cycle analysis performed flow cytometry. caspase-3 activity was analyzed with substrate specific fluorogenic DEVD-McA. In vivo, antitumor activity was evaluated in B16F10 melanoma tumor-bearing c57BL/6J mice. The animals were treated with 55 mg/kg INKKI administered into peritumoral region, while control group received saline solution. The following antitumor parameters were examined: tumor volume, number of metastases, tumor delayed time, tumor doubling time. Histological analyses were performed with H & E staining. The results showed that INKKI induced dose-response cytotoxicity selective for B16F10 melanoma cells (Ic50 1.7 μM) and did not present cytotoxic effects for FN1 fibroblast cells. INKKI-induced apoptosis detected trough of annexin V/PI assay and it was accompanied with an increase of sub-G1 apoptotic fractions and significant increase of caspase-3 cleavage. The tumor-bearing mice treated with INKKI showed a significant reduction in tumor volume of 72.62% and decreased of metastasis number loci. In addition, INKKI caused a significant delay in tumor growth and prolonged the tumor doubling time. Histological analysis revealed an increased of necrosis areas and reduction of tumor cells in tumor treated with INKKI, it was a many hallmark of its antitumor effects observed from in vivo experiments. In conclusion, we show that INKKI is a peptide that could be considered a new putative candidate development to anticancer therapy drug.

关键词： β-casein Peptide Apoptosis Metastasis Tumor Growth Melanoma

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Fighting Poor Quality Medicines: Development, Transfer and Validation of Generic HPLc Methods for Analyzing Two WHO Recommended Antimalarial Tablets

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American Journal of Analytical chemistry 2015年第2期6卷 127-144页

作者： Jérémie Kindenge Mbinze Achille Yemoa Pierre lebrun Pierre-Yves Sacré Védaste Habyalimana Nicodème Kalenda André Bigot Eugène Atindehou Philippe Hubert Roland Djang’eing’a Marini University of Liege (ULg) Department of Pharmacy CIRM Laboratory of Analytical Chemistry Li&egrave ge Belgium Laboratoire d’Analyse des M&eacute dicaments D&eacute partement de Gal&eacute nique et d’Analyse des M&eacute dicaments Universit&eacute de Kinshasa Kinshasa XI Democratic Republic of Congo UFR Pharmacie Facult&eacute des Sciences de Sant&eacute Universit&eacute d’Abomey calavi Cotonou B&eacute nin Rwanda Biomedical center Medical Procurement and Production Division Butare Rwanda Universit&eacute F&eacute lix Houphouet Boigny Abidjan C&ocirc te d’Ivoire

As serious but neglected public health problems, poor quality medicines, i.e. for antimalarial medicines, urged to be fought. One of the approaches is to consider the analytical chemistry and separative techniques. In this study, a generic liquid chromatographic method was firstly developed for the purpose of screening 8 antimalarial active ingredients, namely amodiaquine (AQ), piperaquine (PPQ), sulfalene (SL), pyrimethamine (PM), lumefantrine (LF), artesunate (AS), artemether (AM) and dihydroartemisinine (DHA) by applying DoE/DS optimization strategy. Since the method was not totally satisfying in terms of peak separation, further experiments were undergone applying the same development strategy while splitting the 8 ingredients into five groups. Excellent prediction was observed prior to correlation between retention times of predicted and observed separation conditions. Then, a successful geometric transfer was realized to reduce the analysis time focusing on the simultaneous quantification of two WHO’s recommended AcTs in anti-malarial fixed-dose combination (AM-LF and AS-AQ) in tablets. The optimal separation was achieved using an isocratic elution of methanol-ammonium formate buffer (pH 2.8;10 mM) (82.5:17.5, v/v) at 0.6 ml/min through a c18 column (100 mm × 3.5 mm, 3.5 μm) thermostated at 25℃. After a successful validation stage based on the total error approach, the method was applied to determine the content of AM/LF or AS/AQ in seven brands of antimalarial tablets currently marketed in West, central and East Africa. Satisfying results were obtained compared to the claimed contents.

关键词： Antimalarial AcT Simultaneous Determination Poor Quality Substances Design of Experiments Design Space Method Transfer Accuracy Profile

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