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Study of the binding interaction of tyrosine kinase inhibitors with serum albumin
Rodová, Marie ; Indra, Radek (advisor) ; Heidingsfeld, Olga (referee)
Sunitinib and vandetanib are anti-cancer medications prescribed for medullary thyroid cancer (in the case of vandetanib) and for renal cell carcinoma, gastrointestinal stromal tumor, and pancreatic cancer (in the case of sunitinib). They belong to the group of tyrosine kinase inhibitors and act by exhibiting anti-angiogenic effects and by inhibiting tumor cell proliferation and survival through VEGFR. Additionally, vandetanib also inhibits tumor cell survival via EGFR and RET. In the presented thesis, we investigated the binding interaction between serum albumin and the TKIs vandetanib and sunitinib using BSA, HSA, and blood plasma. We examined the differences in interaction between the TKIs and various serum albumins, including pure BSA, pure HSA, and blood plasma, as well as the nature and location of the binding interaction. Additionally, we studied the influence of other ligands on this interaction and the photosensitivity of sunitinib itself. Utilizing spectroscopic techniques, including UV-VIS absorption and fluorescence quenching, we have determined the Stern-Volmer and binding constants, as well as the thermodynamic parameters, for the binding interactions of sunitinib and vandetanib with BSA and HSA. Our results indicate that complex formation occurs between BSA and sunitinib, BSA and...
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Study of effects of tyrosine kinase inhibitors and their metabolites on tumour cell lines
Kolárik, Matúš ; Indra, Radek (advisor) ; Vinklářová, Lucie (referee)
Vandetanib, lenvatinib and cabozantinib are inhibitors of receptor tyrosine kinases approved to treat locally advanced or metastatic thyroid gland, kidney and liver cancers. These multi- kinase inhibitors, inhibit phosphorylation of tyrosine moieties of protein, thus modulate cell signalization in cancer cells. Metabolites of vandetanib, lenvatinib and cabozantinib were detected in vitro as well as in vivo in blood and urine. Cytochromes P450 and flavin monooxygenases were identified as primary enzymes participating in metabolism of these drugs. Literature lacks information regarding pharmacological efficacy of vandetanib, lenvatinib and cabozantinib metabolites. The aim of this diploma thesis was the investigation of pharmacological efficacy of N-oxides of vandetanib, lenvatinib and cabozantinib. The viability measurement under normoxic and hypoxic conditions was employed to determined their efficacy. The expression of enzymes of the first phase of xenobiotics metabolism (CYP 450 1A1, 1B1, 3A4 a CYP 450 oxidoreductase) and receptor tyrosine kinases RET and VEGFR2, as well as mechanism of changes in their expression were investigated using western blotting and flow cytometry. High performance liquid chromatography was utilised to investigate possible metabolism of tyrosine kinase inhibitors and...
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The effect of NADPH:cytochrome P450 reductase and cytochrome b5 on metabolism of vandetanib by cytochrome P450 3A5
Škriabová, Simona ; Indra, Radek (advisor) ; Mrízová, Iveta (referee)
There are several ways for cancer treatment. One of them is chemotherapy, when cancer patients are given a cytostatic or a combination of multiple types of drugs. The aim of this bachelor thesis was to study the metabolism of the anticancer drug vandetanib. Vandetanib is a tyrosine kinase inhibitor, that has been used in Europe since 2012 for treatment of symptomatic or progressive medullary thyroid cancer. The kinetics of vandetanib oxidation by cytochromes P450 3A5 was studied in this thesis. Oxidation was investigated by two different systems. The first were recombinant cytochromes P450 3A5 expressed in baculovirus-transfected insect cells (SupersomesTM ) and the second were human recombinant cytochromes P450 3A5 expressed in E.coli cells (Bactosomes). Furthermore, the effect of NADPH:CYP reductase and cytochrome b5 on vandetanib oxidation was investigated. Both systems formed the demethylated metabolite of vandetanib, N-desmethylvandetanib, which was separated by HPLC. The study of enzyme kinetics of vandetanib oxidation by human CYP3A5R, 3A5BR, 3A5BLR in Bactosomes indicates that two vandetanib molecules can bind into the active site of the enzyme, resulting in more efficient oxidation. The results also indicate that not only NADPH: CYP reductase, but also cytochrome b5 affects vandetanib...
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