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Interaction of chosen natural compounds with farnesoid X receptor
Trojanová, Eliška ; Pávek, Petr (advisor) ; Cahlíková, Lucie (referee)
Charles University in Prague Faculty of Pharmacy in Hradec Králové Department of Pharmacology & Toxicology Student: Mgr. Eliška Trojanová Supervisor: Prof. PharmDr. Petr Pávek, Ph.D. Title of rigorous thesis: Interaction of chosen natural compounds with farnesoid X receptor Metabolism of many substances taken in the diet, formed in the body and administered in the form of pharmaceuticals and dietary supplements may be influenced by the specific structures called the nuclear receptors. The focus of this work is farnesoid X receptor (FXR) and its interaction with selected natural ingredients, specifically anthocyanidins peonidin, petunidin, pelargonidin, malvidin, cyanidin and delphinidin and chosen alkaloids from Fumaria officinalis. For these substances has been previously observed and described in different studies the positive influence of the development and progression of atherosclerosis, metabolism of triglycerides, bile acids secretion, total and low density lipoprotein (LDL) cholesterol, glucose metabolism and also the influence on the insulin signaling pathway. Given the critical role of FXR in these processes, these compounds have been selected for the examination as the potential agonists of FXR. Substances were tested using TR-FRET LanthaScreen TM farnesoid X receptor co-activator analysis. We...
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Individualized pharmacokinetically guided dosage adjustment of fluorouracil using population pharmacokinetic modeling
Čechlovský, David ; Vlček, Jiří (advisor) ; Pávek, Petr (referee)
vii ABSTRACT David Čechlovský‡ , Stefanie Kraff*, Ulrich Jaehde* *Institute of Pharmacy, Clinical Pharmacy, University of Bonn, Germany ‡ Department of social and clinical pharmacy, Faculty of pharmacy in Hradec Králové, Charles University in Prague Title of diploma thesis: Individualized pharmacokinetically guided dosage adjustment of fluorouracil (5-FU) using population pharmacokinetic modelling. Objectives: To develop a method to increase the efficacy and tolerability of fluorouracil (5-FU) with pharmacokinetically-guided dose adjustment based on a target AUC. Methods: Blood samples were collected from 90 patients with the diagnosis of colorectal carcinoma treated with fluorouracil (5-FU) administered at various infusion durations. Several versions of compartmental pharmacokinetic models were fitted to the plasma concentration data, using nonlinear mixed effect modelling (NONMEM). Different error models were evaluated. The potential effect of patient covariates was evaluated using a stepwise method. Model evaluation was performed by using the bootstrap method. Results: The one-compartment linear model was chosen as a base model as it was successful in fitting to the data collected..The final model contained Additive Residual Error. A covariate BSA>CL and IIV on CL were significantly correlated to the...
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In vitro and in silico evaluation of biological activity of new compounds
Želazková, Jana ; Pávek, Petr (advisor) ; Matoušková, Petra (referee)
The cytotoxicity against HepG2 cell line has been determined in the MTS and LDH assay for the six coumarins, some of which have been determined in previous study to have platelet antiaggregatory activities and to act as cyklooxygenase I inhibitors. Three of the coumarins did not decrease HepG2 cells viability at 30μM concentration after 48h of exposure. None of the other three coumarins caused decrease in viability under 79% under identical conditions. Further, for all the coumarins, the important physicochemical properties determining the behaviour in biological systems have been calculated using the ADMET Predictor. The software has predicted four coumarins to have suitable drug-like properties (zero global risk) and the other two with the lowest risk in the ADMET Risk scale range. These two coumarins were identical with the coumarins showing certain cytotoxicity in the in vitro assays. Cytotoxicity in vitro has been also determined for the representative of the flavones' derivatives group the 3-hydroxyflavone. The 3-hydroxyflavone led to the decrease of HepG2 cells viability observed in MTS assay up to 80%, whereas no cytotoxicity has been observed in LDH assay. This may be explain by the cytostatic and not cytotoxic effect of 3-hydroxyflavone. Thus, the flavone derivative shall be further investigated....
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Nuclear factor kappa B and inflammatory reaction
Klabačková, Sáva ; Jílek, Petr (advisor) ; Pávek, Petr (referee)
Sáva Klabačková Nuclear factor kappa B and inflammatory reaction Bachelor thesis Charles University in Prague Faculty of Pharmacy in Hradec Králové Medical laboratory technician Background: Proposed of LNA probe-based PCR systems for study of NF-B family-related molecules will make possible a comparison of differences between activation of immune systems of groups of germ-free and conventional piglets. This requires: a purification of total RNA and synthesis of cDNA, PCR, relative quantifications of transcriptions of NF-B-related molecules and a statistical evaluation of possible differences between piglet groups. Methods: 1) Germfree and conventional piglets, 2) a proposal of PCR systems with LNA probes for genes of NF-B family and related molecules, 3) a column-based isolation of total RNA from ileum, 4) spectrophotometric evaluation of RNA purity and concentration, 5) a reverse transcription, 6) real-time PCR, 7) an electrophoretic confirmation of amplicons sizes, 8) a relative mRNA expression normalized against reference genes. Results: The LNA probes-based PCR systems for RelA-85, RelB-1, cRel-63, NF-B1-63, NF-B2-21, NFBIA-27, Bcl3-9, IKK1-7, IKK2-3, NEMO-27 of predicted amplicons molecular sizes. Differences in gene transcriptions between germ-free and conventional piglets were found,...
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Chelation ability of experimentally used iron chelators
Morkusová, Michaela ; Mladěnka, Přemysl (advisor) ; Pávek, Petr (referee)
v angličtině Charles University in Prague Faculty of Pharmacy in Hradec Králové Department of Pharmacology & Toxicology Student: Michela Morkusová Supervisor: Přemysl Mladěnka, Pharm.D., Ph.D. Title of diploma thesis: Chelation ability of experimentally used iron chelators Iron is a vital element, playing many important functions in the body. Its presence in too low or too high quantities is harmful. The body has no excretion route for iron, and in certain circumstances, excess iron can accumulate in body tissues. Iron is present in the body in excess particularly in individuals undergoing frequent blood transfusions or having genetically based diseases associated with iron overload (hemochromatosis). Iron excess is medically treated by administration of iron chelators. Such substances can find wider therapeutic use as well, e.g. in individuals with acute myocardial infarction or with tumours. This Thesis was aimed at determining the stoichiometric ratio in the complexes of two experimental chelators, salicylaldehyde isonicotinoyl hydrazone (SIH) and ethylenediaminetetraacetic acid (EDTA), with iron(II) and iron(III) ions at four different pathophysiologically significant pH levels (4.5, 5.5, 6.8 and 7.5). Spectrophotometry was used as the analytical method. In the processing stage, various new...
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Interaction studies of MEK inhibitors and Cytochrome P450 3A4
Urban, Michal ; Pávek, Petr (advisor) ; Nová, Alice (referee)
Charles University in Prague Faculty of Pharmacy in Hradec Králové Department of Pharmacology & Toxicology Student: Mgr. Michal Urban Supervisor: Doc. PharmDr. Petr Pávek, Ph.D. Title of rigorous thesis: Interaction studies of MEK inhibitors and Cytochrome P450 3A4 Objective of our work was to introduce a new method for detection of CYP3A4 inhibition effect of potentially therapeutical compounds. CYP3A4 is the main biotransformation enzyme of 50% current drugs in clinical use and its inhibition may lead to the plasma levels increase of other coadministered drugs which may result in various drug- drug interactions and potentially life threatening adverse effects. Potential of a drug candidate to cause severe pharmacokinetic interactions may be a reason of the clinical trials suspension or withdrawal of a medicinal product from the market and therefore it is essential to determine pharmacokinetic profile of drug candidates at early stages of the development. In addition, we focused on study of interaction potential of MEK1/2 inhibitors PD0325901, PD184352 and U0126 known to inhibit Ras/Raf/MEK/ERK pathway playing an important role in the oncogenesis of various malignancies. Due to the fact that some of these MEK1/2 inhibitors have already entered clinical trials it is absolutely crucial to study...
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Stanovení anthokyanidinů jako potenciálních ligandů lidského konstitutivního androstanového receptoru (hCAR) in vitro
Škoricová, Dagmar ; Pávek, Petr (advisor) ; Filipský, Tomáš (referee)
Charles University in Prague Faculty of Pharmacy in Hradec Králové Department of Pharmacology and Toxicology Candidate: Mgr. Dagmar Škoricová Supervisors: Doc. PharmDr. Petr Pávek, Ph.D. Title of rigorosum thesis: Investigating of anthocyanidins as the putative ligands of the human constitutive androstane receptor (hCAR) in vitro Anthocyanidins are natural flavonoid compounds occurring in various fruits and berries, which represent an important part of the normal healthy diet. In addition to their doubtless health benefits, recent studies have suggested that flavonoids are able to activate xenosensors and therefore regulate a wide range of xenobiotic enzymes involved in the biotransformation of drugs, which might be associated with the increased risk of drug-food interaction. Human constitutive androstane receptor (hCAR) is a xenosensor and a nuclear receptor, which regulates the activity of various hepatic and extrahepatic biotransformation enzymes, especially from the CYP superfamily. In this thesis we used the molecular biology methods - Gene reporter assay and Two- hybrid assay to examine the ability of 6 diverse anthocyanidins, cyanidin, delphinidin, malvidin, pelargonidin, peonidin and petunidin to activate the hCAR receptor in vitro. We found, that none of the examined anthocyanidins has...
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Combination treatment of tumour cell culture with miRNA mimetics and oncogene targeted siRNAs.
Šabouková, Martina ; Miletín, Miroslav (advisor) ; Pávek, Petr (referee)
During the last two decades, the attention of many scientists has been attracted by a newly discovered group of regulatory molecules - RNA interference. These RNA molecules are capable of very efficient gene silencing. The main pathways of this mechanism are used by two major types of small noncoding RNAs - microRNA (miRNA) and small interfering RNA (siRNA). These RNAs are a direct product of genes and are able to bind to mRNA molecules and influence their activity. Due to this knowledge, scientists started examining the effect of RNAi in cancer cells and tried to find a way how RNAi can be applied in therapy. The main topic of this thesis is the potential of miRNA replacement to harm cancer cell proliferation. In cancer cells, miRNA levels are downregulated or upregulated, depending on the nature of cancer and miRNA. Restoring levels of miRNAs can lead to elimination of cancer cell by apoptosis or stop cell in cell cycle arrest. miRNA replacement was used first to examine the function of a single miRNA in HeLa cancer cells. The change in the expression level of miRNA treated cells was verified by reverse transcriptase quantitative PCR and the proliferation inhibition was examined using proliferation assays. The most potent miRNAs inducing significant proliferation inhibition were examined. In the...
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Influence of biotransformation and transport to the effects of chemotherapy and prognosis of breast cancer
Hubáčková, Miluše ; Souček, Pavel (advisor) ; Pávek, Petr (referee) ; Janatová, Markéta (referee)
Influence of biotransformation and transport to the effects of chemotherapy and prognosis of breast cancer Abstract: Introduction: The aim of this study was to evaluate the influence of biotransformation enzymes and enzymes of multidrug resistance for the prognosis and effects of chemotherapy of breast cancer patients. The first part dealed with the genetic factors and the influence of genetic polymorphisms in selected biotransformation genes and in ABCB1 gene and their influence in the formation and prognosis of breast cancer. In the second part, the expression profiles of these genes were analyzed as potential prognostic and predictive properties of a treatment's outcome. Materials and methods: Polymorphisms of biotransformation genes were determined using real-time PCR and polymorphisms of ABCB1 gene were determined using NanoChip Assay, which was performed in our cooperated department. In both cases, the polymorphisms were detected in DNA obtained from the blood of patients. The expression of genes was determined using quantitative real-time PCR in paired tumor and adjacent non-tumor tissue of breast cancer samples. The expression of NQO1 and p53 protein was assesed by immunohistochemistry in the tumor breast tissues. Results: The frequencies of the studied SNPs did not differ from the previous...
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