|
|
Utilization of mass spectrometry for analysis of biologically active compounds
Kaliba, David
This PhD thesis provides a commented set of four publications. These publications are focused on capillary electrophoresis, liquid chromatography, and UV/Vis spectrometry used to study complexes of rhenium with aromatic ligands. The methods of mass spectrometry with soft ionization techniques, 1 H and 13 C nuclear magnetic resonance, and infrared spectrometry were used for structural characterization of the individual complexes. The complexes were synthetized in reactions of the rhenium precursor tetrabutylammonium tetrachlorooxorhenate with the corresponding ligand under both aerobic and anaerobic conditions. In the course of the research, it was revealed that the prepared complexes (with Re in the oxidation number +V and +VI) are unstable and their oxidation numbers change to another more stable form (Re+VII ). Sub-projects which were successfully implemented during the research were as follows: Design and successful realization of the process of synthesis of selected rhenium complexes with aromatic ligands 1,2-dihydroxybenzene, 1,2,3-trihydroxybenzene, and 2,3-dihydroxynaphthalene in reactions with and without air access, and their structural characterization by ESI-MS, APPI- MS, LDI-MS, ESI-MS/MS, NMR, and IR. ESI-MS SRM and UV/Vis time studies of the behaviour of primary rhenium complexes...
|
|
|
Utilization of mass spectrometry for analysis of biologically active compounds
Kaliba, David ; Jelínek, Ivan (advisor) ; Pacáková, Věra (referee) ; Smrček, Stanislav (referee)
This PhD thesis provides a commented set of four publications. These publications are focused on capillary electrophoresis, liquid chromatography, and UV/Vis spectrometry used to study complexes of rhenium with aromatic ligands. The methods of mass spectrometry with soft ionization techniques, 1 H and 13 C nuclear magnetic resonance, and infrared spectrometry were used for structural characterization of the individual complexes. The complexes were synthetized in reactions of the rhenium precursor tetrabutylammonium tetrachlorooxorhenate with the corresponding ligand under both aerobic and anaerobic conditions. In the course of the research, it was revealed that the prepared complexes (with Re in the oxidation number +V and +VI) are unstable and their oxidation numbers change to another more stable form (Re+VII ). Sub-projects which were successfully implemented during the research were as follows: Design and successful realization of the process of synthesis of selected rhenium complexes with aromatic ligands 1,2-dihydroxybenzene, 1,2,3-trihydroxybenzene, and 2,3-dihydroxynaphthalene in reactions with and without air access, and their structural characterization by ESI-MS, APPI- MS, LDI-MS, ESI-MS/MS, NMR, and IR. ESI-MS SRM and UV/Vis time studies of the behaviour of primary rhenium complexes...
|
|
| |
|
|
The study of properties of anticancer drugs ellipticine, etoposide and doxorubicin in the forms of nanocarriers
Lengálová, Alžběta ; Stiborová, Marie (advisor) ; Martínková, Markéta (referee)
Currently available anticancer therapies are inadequate and spur demand for improved technologies. Among others, the utilization of nanocarriers for anticancer drug delivery has shown great potential in cancer treatment. Nanocarriers can improve the therapeutic efficiency of the drugs with minimization of the undesirable side effects. To evaluate potential application of this technology, two forms of nanocarriers have been studied: multi-walled carbon nanotubes (MWCNTs) and apoferritin. The aim of this study was to determine, whether given cytostatics (ellipticine, etoposide and doxorubicin) are bound to these nanotransporters and how are they released from them, especially depending on pH. Since the pH of the tumor cells is lower than the pH of healthy cells it would be preferred that the drugs would release from nanocarriers at the lower pH while at the physiological pH the release of the drug would be eliminated. The results found show that ellipticine is actually released from its MWCNT- and apoferrtin-encapsulated form at acidic pH (5.0), while at pH 7.4 its interaction with nanocarriers is stable. Ellipticine released from MWCNT is activated by microsomal enzymes to reactive metabolites (13- hydroxyellipticine and 12-hydroxyellipticine) forming DNA adducts. The results indicate that both...
|
|
|
Novel Electrochemical Biosensor for the Detection of DNA Damage Caused by Chemical Carcinogens
Blašková, Marta ; Vyskočil, Vlastimil (advisor) ; Zima, Jiří (referee)
Presented Bachelor Thesis is focused on the development and utilization of a simple and inexpensive electrochemical DNA biosensor for the detection of DNA damage caused by chemical carcinogens. A glassy carbon electrode (GCE), having several advantages such as broad potential window and well-renewable surface, was used for its preparation. A low- molecular-weight DNA isolated from salmon sperm was used. The initial part of the work is devoted to the optimization of the biosensor preparation and to its characterization, which was performed using several electrochemical techniques - cyclic voltammetry (CV), square-wave voltammetry (SWV) and electrochemical impedance spectroscopy (EIS). The influence of the DNA immobilization type on the quality of the biosensor response was also investigated. Two approaches were tested: (i) spontaneous adsorption of DNA by leaving the DNA stock solution to dry on the GCE surface and (ii) adsorptive accumulation of DNA on the GCE surface from the solution (selected as the optimum one). The second part of this Thesis deals with the detection of DNA damage by various chemical carcinogens. Four model substances were tested: flutamide (an antiandrogen antitumor drug), 4-nitro-3-(trifluoromethyl)aniline (NTMA; a metabolite of flutamide), 2-aminoanthracene (a genotoxic...
|
|
|
Inhibition of Thymidine Phosphorylase
Zákoucká, Eva ; Brynda, Jiří (advisor) ; Obšil, Tomáš (referee)
2. Abstract Thymidine phosphorylase (TPase), also known as gliostatin or Platelet-derived endothelial cell growth factor (PD-ECGF), is an enzyme with an important role in the nucleoside metabolism and is also involved in degradation and recycling of DNA. TPase catalyzes the reversible phosphorolysis of pyrimidine 2'-deoxynucleosides to 2-deoxy-D-ribose-1- phosphate and their respective bases, as well as the transfer of the deoxyribosyl moiety from one pyrimidine base to another. Thymidine phosphorylase is a therapeutic target of great importance because of its participation in angiogenesis especially in solid tumors of various tissues. Therefore, TPase stimulates tumor growth and progression, as well as metastasis. In addition to this, TPase inhibits apoptosis, particularly of tumor cells and causes degradation of several antiviral and anticancer drugs. Apart from the carcinoma tissues, thymidine phosphorylase is overexpressed in various other tissues affected by disorders characterized by proliferation of blood vessels including psoriasis, rheumatoid arthritis and atherosclerosis. Inhibiting the activity of TPase selectively in the tissues affected by the diseases listed above would be of great therapeutic significance. Therefore, many inhibitors, mainly substrate analogues, have been designed based on the...
|
|
|
Raman spectroscopy of biologically active species and antitumor drugs
Třeštíková, Liběna ; Vetterl,, Vladimír (referee) ; Vrána, Oldřich (advisor)
SERRS spektra of biological materials are very komplex, because they consist of signals from all molecules present in cells. In this text are presented SERRS spektra of antitumor drugs and its komplex with DNA. Experimental are rated on doxorubicin and another antitumor druha and on study of theirs potential by treatment for tumors. Doxorubicin belong to clase antracycline antibiotics and is used for stop of tumor cells reproduction. Scientists found still new ways, new drugs. SERRS is one of possibilities for study of this drugs and theirs interaction with DNA.
|
guest ::
