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Issues of the Cholinesterase Inhibition by the Selected Organophosphorous Pesticides In Vitro
Lázenská, Helena ; Opletalová, Veronika (advisor) ; Kuča, Kamil (referee)
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...
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Biological activity of plant metabolites III. Influence of alkaloids from Chelidonium majus L. on acetylcholinesterase
Kubincová, Dagmar ; Opletal, Lubomír (advisor) ; Kuča, Kamil (referee)
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...
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Synthesis of precursors for biologically active lactones IV.
Bémová, Hana ; Opletalová, Veronika (advisor) ; Kuča, Kamil (referee)
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...
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Electrosynthesis of Delta-Valero Lactones
Pochopová, Lenka ; Kubíček, Vladimír (advisor) ; Kuča, Kamil (referee)
Charles University in Prague Faculty of Pharmacy in Hradec Králové Department of pharmaceutical chemistry Candidate: Mgr. Pochopová Lenka Consultant: Ing. Kubíček Vladimír, CSc. Tittle of Thesis: ,,ELECTROSYNTHESIS OF DELTA-VALERO LACTONES" In this project electrolysis has been used as a method for a synthesis of new compounds. The main aim is cathodic reduction of the carbon-chlorine bond in a series of aldol trichloroacetates. Formed carbanion would form a 6 member ring by attaching the carbonyl group. Two structures can be formed: α, α-dichloro-β-hydroxy-δ-lactones and α, β-unsaturated-α-chloro -δ-lactones from the starting esters using electrolysis. The whole process was performed at controlled potential in a separated cell, on mercury pool as a cathode and platinum as an anode in an argon atmosphere with dichloromethane/tetraethylamonium system. During the process two electrons have been passed. The synthesized lactones are completely new compounds. In this report also electrochemical and spectroscopy properties of those compounds have been characterized to provide for their other use.
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Effects of antidepressants and depressive disorders on mitochondrial functions
Hroudová, Jana ; Fišar, Zdeněk (advisor) ; Martásek, Pavel (referee) ; Kuča, Kamil (referee)
Mood disorders are serious diseases. Nevertheless, their pathophysiology is not sufficiently clarified. Biological markers that would facilitate the diagnosis or successful prediction of pharmacotherapy are still being sought. The aim of the study was to find out whether mitochondrial functions are affected by antidepressants, mood stabilizers and depression. Our research is based on recent hypotheses of mood disorders, the advanced monoamine hypothesis, the neurotrophic hypothesis, and the mitochondrial dysfunction hypothesis. We assume that impaired function of mitochondria leads to neuronal damage and can be related to the origin of mood disorders. Effects of antidepressants and mood stabilizers on mitochondrial functions can be related to their therapeutic or side effects. In vitro effects of pharmacologically different antidepressants and mood stabilizers on the activities of mitochondrial enzymes were measured in mitochondria isolated from pig brains (in vitro model). Activity of monoamine oxidase (MAO) isoforms was determined radiochemically, activities of other mitochondrial enzymes were measured spectrophotometrically. Overall activity of the system of oxidative phosphorylation was measured electrochemically using high- resolution respirometry. Methods were modified to measure the same...
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Study of Antiproliferative effects of Novel Iron Chelators
Potůčková, Eliška ; Šimůnek, Tomáš (advisor) ; Boušová, Iva (referee) ; Kuča, Kamil (referee)
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...
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