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Charles University in Prague Faculty of Pharmacy in Hradec Králové Department of Biochemical Sciences Candidate: Mgr. Eliška Potůčková Supervisor: Doc. PharmDr. Tomáš Šimůnek, Ph.D. Title of Doctoral Thesis: Study of antiproliferative effects of novel iron chelators Recent studies demonstrate that iron chelators possess marked potential as anticancer agents and also as substances protecting sensitive tissues against oxidative stress. This thesis is dealing mainly with design and synthesis of new antiproliferative iron chelators based on the structure of salicylaldehyde isonicotinoyl hydrazone (SIH) and with a characterization of their pharmacological and toxicological properties. Antiproliferative effects of these substances were studied on human breast adenocarcinoma cell line and human leukemic cell line and the non-specific toxicity on neonatal rat cardiac tissue-derived cells. During these studies some novel structure- activity relationships have been found. Furthermore, the suitability of simultaneous administration of iron chelators with antineoplastic agents used in breast cancer treatment was studied. In this project the anthracycline antibiotic, doxorubicin, and, in particular, the estrogen receptor antagonist, tamoxifen, were identified as the most suitable agents for potential treatment...

Charles University in Prague, Faculty of Pharmacy in Hradec Králové Department of Pharmaceutical Botany and Ecology Candidate: Mgr. Jakub Chlebek Supervisor: Doc. RNDr. Lubomír Opletal, CSc. Title of Doctoral Thesis: Study of biological activity of isolated alkaloids from Corydalis cava (Fumariaceae). Key words: Corydalis cava, isoquinoline alkaloids, DPPH, AChE, BuChE, BACE1, IMER. Tubers of Corydalis cava were selected on the basis of bio-guided spectrophotometric Ellman's method as a source of isoquinoline alkaloids for study of their selected biological activities. The tubers of Corydalis cava were extracted with ethanol and the mixture of summary tertiary alkaloids (fraction A) was fractionated in silica gel chromatography column using step gradient elution with hexane, chloroform and ethanol. Repeated column chromatography, preparative TLC and crystallization led to the isolation of fifteen isoquinoline alkaloids. The chemical structures of isolated compounds were determined on the basis of spectroscopic techniques and by comparison with literature data. Isolated alkaloids were tested on ability to inhibit AChE, serum BuChE (IC50), BACE1 and for its free-radical scavenging activity (EC50). The cholinesterase inhibitory activity was determined in vitro by modified spectrophotometric Ellman's...

Charles University in Prague, Faculty of Pharmacy in Hradec Králové Department of Department of Pharmaceutical Chemistry and Drug Control Candidate Mgr. Veronika Nováková Supervisor doc. PharmDr. Petr Zimčík, Ph.D. Title of Doctoral Thesis Synthesis and study of properties of aza-analogues of phthalocyanines Phthalocyanines (Pc) and their azaanalogues azaphthalocyanines (Azapc) are planar macrocyclic compounds with significant absorption band at 620-720 nm finding application in the areas such as photodynamic therapy, chemical catalysis and solar cells. Photophysical (fluorescence) and photochemical (singlet oxygen production) parameters are important in above mentioned applications. Peripheral substituents of Pc/AzaPc play key role in this case. Alkylsulfanyl and alkyloxy derivatives exerted high fluorescence quantum yields (ΦF) as well as quantum yields of singlet oxygen (ΦΔ). On the other hand, alkylamino substitution led to almost zero values of both ΦF and Φ∆. These derivatives have recently found new application as dark quenchers in DNA hybridization probes. The first part of this thesis demonstrated that intramolecular charge transfer (ICT) is responsible for zero values of quantum yields of alkylamino AzaPc and that the lone electron pair of peripheral nitrogen is a donor for ICT in AzaPc....

Charles University in Prague, Faculty of Pharmacy in Hradec Kralove Department of Pharmaceutical Chemistry and Drug Control SYNTHESIS OF PRECURSORS FOR BIOLOGICALLY ACTIVE LACTONES IV. Rigorous Thesis Hana Piskacova A high width of biological activity of natural or synthetic unsaturated five membered lactones from the family of furan-2(5H)-ones, is the main reason why to synthesize new substances of this structure. The introductory part of the thesis deals with antineoplastic natural compounds of this type. Except of antitumour activity some unsaturated five membered lactones exhibit antifungal, antiviral, antibacterial effects, or inhibit the synthesis of cholesterol, for example. The experimental project is an extension of my diploma thesis concerned with the synthesis of precursors for lactones. These precursors - methyl (E)- and (Z)-2-bromo-5- (subst.)arylpent-2-en-4-ynoates - were prepared by Sonogashira couplings. The parent substances for couplings were arylethynes (1-ethynyl-4-(methoxymethoxy)benzen, 1-ethynyl-2-nitrobenzene, 3-ethynylaniline, 2-ethynylpyridine) and methylesters of dihalogenated prop-2-enoic acid. The dominant products of the couplings were β-monoalkynylated esters, apart from them side products of homocouplings were obtained. Reactions of E-methylesters of dihalogenated...

Markéta Komlóová: Synthesis of bisquarternary asymmetrical reactivators of acetylcholinesterase Nine potential bisquaternary reactivators of AChE have been prepared and their reactivation activity against paraoxon or tabun-inhibited enzyme has been tested in vitro. Three reactivators of AChE proved to be suitable for further testing against paraoxon. One of these compounds also demonstrated satisfactory efficiency when reacting against tabun. The reactivation abilities of prepared compounds depend on structural factors such as the presence of oxime group, its location, the presence of quaternary nitrogen and other functional groups. Also four monoquaternary inhibitors have been prepared and their inhibitive activity tested in vitro.

The aim of present work was a comparison of effectiveness of oximes chosen in the experimental therapy of nerve agents intoxication (soman and russian VX). All experiments were performed in male Wistar rats. All oximes tested were administered intramuscularly either in a separate injection or in combination with atropine. The preparation of the brain parts for determination of acetylcholinesterase was performed in several time intervals after drugs tested administration. Antidotal potency of oxime HI-6 was superior in comparison with the rest of oximes tested.

QUESTIONS OF THE IN VITRO INHIBITION OF CHOLINESTERASES BY SELECTED ORGANOPHOSPHORUS PESTICIDES Helena Lázenská Department of Pharmaceutical Chemistry and Drug Control, Faculty of Pharmacy in Hradec Kralove, Charles University in Prague, Heyrovskeho 1203, 500 05, Hradec Kralove, Czech Republic. The aim of this study was to describe organophosphorus inhibitors paraoxone, DDVP and DFP from the aspect of the kinetics of their reaction with human erythrocyte acetylcholinesterase (AChE) and plasma butyrylcholinesterase (BuChE), to find IC50, and to test the in vitro potency of five selected oximes (pralidoxime, methoxime, trimedoxime, obidoxime and HI-6) to reactivate AChE and BuChE inhibited by three mentioned organophosphorus inhibitors. The inhibition of AChE and BuChE was performed by incubation with organophosphorus inhibitors at a convenient concentration and such time, that would result in about 90% activity of an enzyme. A solution of a reactivator at a final concentration 1 μM or 10 μM was added to an enzyme, after 10 min of reactivation, a solution of a substrate - acetylthiocholine-iodide or butyrylthiocholine-iodide was added, and the activity of an enzyme was measured by the spectrophotometric Ellman's method. The experiment was performed at 25 žC, pH 7,4 and in a 0,1 M phosphate buffer. According...

SOUHRN Spolu s ostatními diplomantkami (Šárkou Brožovou, Janou Nagyovou a Evou Vítkovou) jsem provedla extrakci 41,8 kg suché nati s kořeny. Primární extrakt jsme vyčistily filtrací a následným oddestilováním rozpouštědla na vakuové odparce. Dále jsme připravily sekvenčním postupem výtřepky s jednotlivými typy alkaloidů: šlo o dva diethyletherové výtřepky (získané po předchozí alkalizaci primárního extraktu uhličitanem sodným a hydroxidem draselným), které byly následně okyseleny kyselinou chlorovodíkovou, vzniklé kvartérní jodidy po přidání jodidu draselného vytřepány chloroformem a po následné alkalizaci extraktu amoniakem získány další kvartérní jodidy vytřepáním do směsi chloroform + ethanol 8,5:1,5. Já jsem se zabývala výtřepkem B, který obsahoval silně basické alkaloidy. Provedla jsem separaci výtřepku B pomocí sloupcové chromatografie. Zpracovávala jsem frakci 27- 46, jejíž odparek jsem po vysušení rozdělila na dvě části. První část (0,02 g) byla podrobena barevným reakcím s činidly, které prokázaly, že izolovaný alkaloid CH-M/B-1 je berberin.Dále jsem změřila teplotu tání Z druhé části odparku jsem získala 0,346 g žlutého prášku, který byl v závěru práce podroben testu vlivu látky na aktivitu AChE. Z naměřené hodnoty IC50= 0,221 (0,146- 0,228) μM je patrné, že látka CH-M/B-1 je vysoce aktivní v...