Charles University Faculty of Pharmacy in Hradec Králové Department of Pharmaceutical Botany Candidat: Jana Faberová Supervisor: doc. Ing. Kateřina Macáková, Ph.D. Title of Thesis: Interaction of chalcones with transition metals I. Iron is a biogenic metal element with unpaired d-electrons that plays a key role in the survival of unicellular organisms, plants and mammals. The electron shell of the iron atom is able to easily bind and release electrons and thus contribute to the proper function of the organism. Iron is necessary to ensure the transport of respiratory gases, cell differentiation and the functioning of some specific enzymes. However, despite the clearly positive effect of iron on human health, inappropriately low or high level of iron in the body can cause the development many diseases. Chalcones are plant polyphenols belonging to the family of flavonoids, secondary metabolites of plants. The term chalcone generally refers to a chemical structure with an ,-unsaturated ketone system based on 1,3-diphenylprop-2-en-1-on. These substances have a wide spectrum of biological activities and in the right amount can be good for human health. The aim of this diploma thesis was to monitor the iron-chelating and iron-reducting activity of selected chalcones: isoliquiritigenin, licochalcon A,...

Charles University in Prague Faculty of Phramacy in Hradec Králové Department of Pharmaceutical Botany Candidate: Martina Šulcová Supervisor: Assoc. Prof. Ing. Kateřina Macáková, Ph.D. Title of the diploma thesis: Interaction of chalcones with transition metals II. Copper is one of the essencial elements in human body which is involved in the functioning of many enzymes and metabolic pathways. Although it appears in trace amounts in the body, its functions are irreplaceable. However, the ion level must be kept in balance by the various mechanisms of the body. If there is an excess or deficiency of copper in the body, this condition can lead to the development of health issues. Chalcones are substances belonging to the group of flavonoids. Their polyphenolic structure does not have a closed ring C. Studies describe their anticancer, anti-inflammatory, antiviral, antidiabetic, antibacterial, antioxidant, immunosuppressive and other properties. Chosen substances in the group of dihydrochalcones (naringin dihydrochalcone, neohesperidin dihydrochalcone, phloretin, phlorizin) and chalcones (isoliquiritigenin, licochalcone A) were tested for their chelation and reduction activity of the copper ions in this diploma thesis. The activity was measured at different pH environments (7.5; 6.8; 5.5 and 4.5) and...

Charles University Faculty of Pharmacy in Hradec Králové Department of Pharmacology & Toxicology Student: Zuzana Sobolová Supervisor: Prof. Přemysl Mladěnka, Pharm.D., Ph.D. Consultant: Marcel Hrubša, MS.c. Title of diploma thesis: The impact of synthetic chelators from the group of 4-acylpyrazole-5-ones on alcohol dehydrogenase Inhibitors of alcohol dehydrogenase play a key role in the treatment of poisoning caused by methanol, ethanol, ethylene glycol, and their toxic metabolites. Such treatment can therefore be considered as life-saving. In clinical practice, a single substance, fomepizole, is used. This thesis is aimed at investigation of the effect of eleven experimental chelators from the 4- acylpyrazol-5-ones group on equine and yeast alcohol dehydrogenases, and the possible definition of structural features that would be the most suitable for interaction with this enzyme. Due to the ability of these substances to chelate zinc ions, it is believed that they could inhibit an enzyme that has a zinc molecule in the structure. The measurement itself is based on principle of increase in absorbance during the enzymatic reaction, which is caused by the formation of NADH. This measurement is relatively simple, efficient and, due to used kinetic method, also less susceptible to be affected by...

Charles University Faculty of Pharmacy in Hradec Králové Department of Pharmacology & Toxicology Student: Karolína Pelnářová Supervisor: Prof. Přemysl Mladěnka, PharmD., Ph.D. Consultant: Václav Tvrdý, MSc.; Marcel Hrubša, MSc. Title of diploma thesis: Searching of effective cobalt chelators - 8-hydroxyquinolines Cobalt is one of the essential trace elements present in the human body. It forms a part of the organometallic complex of vitamin B12, which is essential for many physiological functions. Both overload and lack of cobalt in the body is associated with pathological conditions. Manifestations of deficiency can lead up to pernicious anemia or hypofunction of the thyroid gland. Cobalt intoxication can occur in an industrial environment, such as by cobalt metal dust during heavy metal processing, or by its release due to corrosion from orthopedic prostheses. Systemic toxicity is manifested by a number of endocrine, cardiovascular and neurological symptoms. The main aim of this diploma thesis was to find effective chelators of cobalt from the group of substances derived from 8-hydroxyquinoline. In vitro spectrophotometric measurement was used to determine the degree of chelation. The effect of chelation was also monitored ex vivo in rat erythrocytes. All tested chelators - 8-hydroxyquinoline,...

Charles University Faculty of Pharmacy in Hradec Králové Department of Pharmaceutical Botany Candidate: Hana Němcová Supervisor: PharmDr. Jana Karlíčková, Ph.D. Title of Thesis: Comparison of interactions of equol and desmethylangolensin with iron and copper Iron and copper are essential trace elements, which are important for our body. Both elements have a significant effect on the correct function of organs and make part of many enzymes. They are able to accept or donate electrons - conversion between oxidized (Fe3+, Cu2+) and reduced (Fe2+, Cu+) forms. If these metals are excess in organism, they are accumulated in the cells and mediate the creation of free radicals, that destroy cell structures. This deficiency is treated with chelators, which facilitate the excretion of metals from the body. Isoflavonoids are polyfenolic substances, which can have antioxidant effects and they are involved in the scavening of free radicals. Isoflavonoids can have also a pro-oxidative effect, because they are able to reduce metal ions. In this study were tested interactions (chelation and reduction) between the metabolites of isoflavonoids (equol and desmethylangolensin) and ions of iron and copper. Both metabolites only weakly chelate metal ions, but they significantly reduce cupric ions. KEYWORDS: Iron,...

Charles University Faculty of Pharmacy in Hradec Králové Department of Pharmaceutical Botany Candidate: Zuzana Lomozová Supervisor: PharmDr. Jana Karlíčková, Ph.D. Title of Thesis: Interactions of tamarixetin and isorhamnetin with copper Keywords: tamarixetin, isorhamnetin, chelation, reduction, copper, bathocuproine, hematoxylin Copper is a biogenic trace element important for proper function of human organism. It is an essential part of several enzymes and is involved in metabolic processes in the body. Excess or lack of serum copper can lead to pathological conditions. Copper chelating agents are used to treat copper toxic effects. Flavonoids are polyphenolic substances belonging to secondary metabolites of various plants. They are part of the human diet and have a positive impact on our health. They exhibit antioxidant and anti-inflammatory effects and are able to chelate transient metals, especially iron and copper. Chelation therapy is currently used in Wilson's disease in which copper is overloaded. In the future, the chelating effects of flavonoids could be used to treat neurodegenerative diseases, cardiovascular diseases or cancer. In this diploma thesis, interactions of two flavonoids (namely tamarixetin and isorhamnetin) with copper ions in different buffers were tested....

Charles University Faculty of Pharmacy in Hradec Králové Department of Pharmaceutical Botany and Ecology Candidate: Kamila Zemanová Supervisor: PharmDr. Jana Karlíčková, Ph.D. Title of Thesis: Interaction of phenylpropionic acids with copper Copper is an essential trace element which is essential for our body. It has a significant effect on the correct functioning of important organs and it plays an important role in the transfer of electrons at the major enzymatic pathways as a prostetic group. On the other hand, excess or deficiency of copper in the human body can cause many diseases. Phenylpropionic acids are group of substances which can have antioxidant, anti-inflammatory and anticancer effects. They are involved in the scavenging of free radicals and reactive oxygen species. In this diploma thesis, I tested copper chelating activity of six phenylpropionic acids at different pHs by spectrofotometric methods using a hematoxylin and a bathocuproinedisulfonic acid disodium salt as an indicator. 3-(3,4-dihydroxyphenyl)propionic acid showed the highest chelating potential but only using a hematoxylin. KEYWORDS: Copper, Phenylpropionic acids, Antioxidants, Chelating activity, Hematoxylin, Bathocuproin

Charles University Faculty of Pharmacy in Hradec Králové Department of Pharmacology and Toxicology Candidate: Barbora Gorová Supervisor: Assoc Prof. Přemysl Mladěnka, Pharm.D., Ph.D. Title of thesis: Iron-chelating properties of fruit extracts of various elderberries Inorganic iron is the major food source of iron in humans. It plays role in many biochemical reactions. Thus, iron metabolism disorders can lead to different diseases associated with lack of iron or iron overload. One of the possible treatment modalities of the latter represents the administration of iron chelators. Elderberry, Sambucus nigra L. (Adoxaceae), has been used in traditional medicine. The fruits of elderberry are a rich source of cyanidin-based anthocyanins as the main component. There are important differences, both in chemical and physical properties between several cultivars of elderberry. Anthocyanins might interact with metals in the gastrointestinal tract by formation of chelates. However, data on metal interactions with anthocyanins are sparse. The main aim of this study was to perform the analysis of interaction of iron with elderberry fruit extracts as a rich and cheap source of anthocyanins with cyanidin as the aglycon. In this in vitro study ten elderberry fruit exctracts were tested for iron chelating activities...

Charles University Faculty of Pharmacy in Hradec Králové Department of Pharmaceutical Botany Candidate: Nikola Holotíková Supervisor: PharmDr. Jana Karlíčková, Ph.D. Title of Thesis: Can essential amino acids be considered as copper chelators? Copper is a trace element playing an important role in the human organism. Copper levels in the body have to be carefully controlled because disruption of copper homeostasis can cause oxidative damage which may lead to various pathologies. Chelation of copper can potentially be used as a therapeutic tool. Medical chelators should ideally mimic chelators, especially peptides and proteins which are important for regulation of copper homeostasis. Their structure is formed by amino acids linked by peptide bonds. For this reason, the study is focused on selected amino acids. Proteinogenic amino acids are classified into essential and nonessential. Essential amino acids must be obtained from the diet. The aim of this study was to compare the copper chelation and reduction activity of essential amino acids L-histidine and L-methionine, and nonessential amino acids L-cysteine, its dimer L-cystine, L-aspartic acid and L-glutamic acid. Also a L-cysteine-derived compound, N-acetylcysteine, was tested. All of them were tested using spectrophotometric methods at pHs which...