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Study of azaphthalocyanines as fluorescence quenchers in oligodeoxynucleotide probes
Demuth, Jiří ; Nováková, Veronika (advisor) ; Hocek, Michal (referee) ; Vávrová, Kateřina (referee)
Charles University, Faculty of Pharmacy in Hradec Kralove Department: Department of Pharmaceutical Chemistry and Pharmaceutical Analysis Author: Mgr. Jiří Demuth Supervisor: doc. PharmDr. Veronika Nováková, Ph.D. Consultant: prof. PharmDr. Petr Zimčík, Ph.D. Doctoral Thesis: Study of azaphthalocyanines suitable for quenching of fluorescence in oligodeoxynucleotide probes Phthalocyanines (Pcs) and azaphthalocyanines (AzaPcs) are planar macrocyclic compounds. These synthetic compounds are derived from naturally occurring dyes - porphyrins. Pcs and AzaPcs have very interesting photophysical properties due to their extensive 18 -electron conjugated system. They are used in electrotechnics, material industry or in medicinal applications. Octaalkylamino substituted tetrapyrazinoporphyrazines (TPyzPzs), which belong to AzaPc family, can quench fluorescence of other molecules due to the ultrafast intramolecular charge transfer. This thesis studies this quenching ability of TPyzPzs with the aim to develop efficient fluorescence probes. Such probes could find application in a variety of experiments across scientific disciplines i.e. - biochemistry, forensic or genetic analysis. Synthetic part of dissertation thesis is a continuation of previous investigation of TPyzPzs in our research group. It describes...
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Comparison of Photophysical Properties of Different Types of Azaphthalocyanines
Reimerová, Petra ; Nováková, Veronika (advisor) ; Kubíček, Vladimír (referee)
Charles Univerzity in Prague, Faculty of Pharmacy in Hradec Králové Department: Department of Biophysics and Physical Chemistry Candidate: Petra Reimerová Supervisor: PharmDr. Veronika Nováková, Ph.D. Title of Thesis: Comparison of Photophysical Properties of Different Types of Azaphthalocyanines Phthalocyanines and their aza-analogues azaphthalocyanines are organic dyes with planar macrocyclic structure and metal cation coordinated in the center. They can be used in many applications - for example as sensors, dyes or sensitizers in photodynamic therapy. The aim of this study was to compare photophysical properties (absorption, fluorescence, singlet oxygen production), solubility, aggregation and acid-base properties of several molecules 1-7 that differ in the type of macrocyclic core (see Figure below). The strength of absorption expressed as an extinction coefficient increased with the bathochromic shift of the Q-band. The position of the Q band depended on the number of conjugated double bonds and on the position of the peripheral substituents. Isosteric exchange of benzene rings to pyrazine rings shifted Q-band to shorter wavelength. Quantum yields of singlet oxygen (Φ∆) and quantum yields of fluorescence (ΦF) were determined in pyridine and THF by comparative method with ZnPc as a reference....
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Synthesis and study of ceramides with deuterated acyl chain
Hudská, Klára ; Vávrová, Kateřina (advisor) ; Nováková, Veronika (referee)
Charles University in Prague Faculty of Pharmacy in Hradec Králové Department of Inorganic and Organic Chemistry Candidate: Klára Hudská Supervisor: Doc. PharmDr. Kateřina Vávrová, Ph.D. Title of Diploma Thesis: Synthesis and study of ceramides with deuterated acyl chain Ceramides are a complex group of lipids belonging to sphingolipids. They are the most important component in stratum corneum and play an essential role in protecting the skin from excessive water loss and other environmental damages. Despite the fact that ceramides are a subject of intensive research, the exact effect of their structure on barrier skin function has not been fully explained. The aim of this diploma thesis was to synthesize a series of ceramides derived from sphingosin with deuterated acyl chains with chain lenght 4C, 16C and 24C and their study using Fourier transform infrared spectroscopy to understand the behaviour of ceramides in SC and their structure-activity relationships. The study of the synthesized ceramides using ATR-FTIR spectroscopy revealed that, at skin temperature, lipid chains are arranged in orthorhombic subcell, mostly with all-trans conformation. While the CerNS24 prefers extended conformation with its acyl associated with the acyl of LA and cholesterol associated with the sphingosin chain in...
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Determination of Lipophilicity of Drugs by HPLC
Pleváková, Magdaléna ; Kubíček, Vladimír (advisor) ; Nováková, Veronika (referee)
3 ABSTRACT Charles University in Prague, Faculty of Pharmacy in Hradec Králové Department of biophysics and physical chemistry Author: Magdaléna Pleváková Supervisor: Ing. Vladimír Kubíček, CSc. Thesis title: Determination of Lipophilicity of Drugs by HPLC In this thesis a RP-HPLC method for fast and reliable determination of lipophilicity was proposed and tested. Stationary phase was selected by using hydrophobic substraction model. Capacity factors of the chosen substances were initially measured on Zorbax ECLIPSE XDB C18 250x4,6 mm, 5µm column, which exhibits almost identical retention characteristics as the column used for this purpose until now. Then the capacity factors of the same substances were determined by using Zorbax ECLIPSE XDB-C18 50x4,6 mm, 1,8µm column that was selected to reduce retention times significantly. A group of newly synthesised drugs based on structure of pyrazine served as samples for the measurement. The reproducibility of the capacity factor values determined using both columns was compared and the independence of the capacity factor on the mobile phase flow was confirmed. The capacity factors of two homologous series and a group of benzimidazols were consequently determined on Zorbax ECLIPSE XDB-C18 50x4,6 mm, 1,8µm column using various compositions of mobile phases. Several...
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Syntheses of azaphthalocyanine fluorescence sensors with the improved selectivity towards desired metal cations
Uhlířová, Štěpánka ; Nováková, Veronika (advisor) ; Zimčík, Petr (referee)
Charles University, Faculty of Pharmacy in Hradec Králové Department Department of Biophysics and Physical Chemistry Candidate Štěpánka Uhlířová Supervisor doc. PharmDr. Veronika Nováková, Ph.D. Title of thesis Syntheses of azaphthalocyanine fluorescence sensors with the improved selectivity towards desired metal cations Azaphthalocyanines (AzaPc) bearing aza-crowns on the periphery may be used as fluorescence sensors for metal cations. Emission in the red part of the spectrum is a great advantage of these compounds because such light is not absorbed by endogenous chromophores. Previous projects served mostly as a proof of concept where sensitivity of AzaPc sensors to a series of alkali and alkaline earth metal cations was shown. The aim of this diploma thesis is focused on the improvement the selectivity of a recognition moiety for particular cations. This can be achieved by the attachment of lariat ethers or crown ethers close to aza-crown moiety. Firstly, appropriate precursors, i.e. 5,6-disubstituted pyrazine-2,3-dicarbonitriles, were prepared by nucleophilic substitution. Then, cyclotetramerization of two different precursors (A and B) using a template method with zinc acetate in a high boiling solvent provided a statistical mixture of AzaPc congeners. Desired unsymmetrical AzaPc of ABBB type was...
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Synthesis of "Chemical Tags" and their application for selective protein labelling
Prokešová, Kristína ; Miletín, Miroslav (advisor) ; Nováková, Veronika (referee)
Charles University in Prague Faculty of Pharmacy in Hradec Králové Department of Pharmaceutical Chemistry and Drug Control Student: Kristína Prokešová Supervisor: doc. PharmDr. Miroslav Miletín, PhD. Consultant: Dr. Richard Wombacher Title: Synthesis of "Chemical Tags" and their application for selective protein labelling This diploma thesis is aimed on the synthesis and application of chemical tags. In theoretical part, protein labelling in general is discussed and fluorescent proteins, as routinely used technique for protein tracking, are shortly presented. The greatest attention is dedicated to chemical tags which consist of genetically encoded protein or peptide tag fused to the protein of interest (POI) and from a small fluorescent molecule which labels the POI-tag fusion. Particular representatives with their advantageous and disadvantageous properties are mentioned and super-resolution microscopy and calcium imaging, as applications of chemical tags, are explained. Experimental part is divided into chemical synthesis and biological methods. In synthetic part, four precursors of chemical tags have been prepared - two precursors of TMP-tag small molecule and two precursors of a chemical calcium dye. These precursors can be connected to create a new TMP-tag applicable for calcium imaging. One...
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Synthesis and study of 6-((7-nitrobenzo[c][1,2,5]oxadiazol-4-yl)amino)hexanoic acid derivatives as transdermal permeation enhancers
Kučera, Ondřej ; Vávrová, Kateřina (advisor) ; Nováková, Veronika (referee)
Ondřej Kučera Charles University in Prague, Faculty of Pharmacy in Hradec Králové Synthesis and study of 6-((7-nitrobenzo[c][1,2,5]oxadiazol-4-yl)amino)hexanoic acid derivatives as transdermal permeation enhancers Transdermal permeation enhancers are used to increase absorption of drugs through skin or, more importantly, through the stratum corneum, which is the uppermost layer of the skin. Mechanism of action of enhancers is not fully understood. In general the most active enhancers consist of hydrophilic and hydrophobic parts. In this work, we prepared and studied fluorescent permeation enhancers. Fluorescent dye NBD (7-nitrobenzo[c][1,2,5]oxadiazol) is fairly hydrophilic, so we used it as a hydrophilic head of potential enhancers based on dodecyl 6- (dimethylamino)hexanoate (DDAK). Such fluorescent enhancers could help us understand more about the mechanism of action. It enables determination of this enhancer and imaging of its penetration pathways in the skin. We synthesized three esters of 6-((7-nitrobenzo[c][1,2,5]oxadiazol-4-yl)amino)-hexanoic acid (NBD-acid) with ester-linked C8-C12 alkyls. 6-Aminocaproic acid reacted with 4-chloro- NBD and then with a series of alcohols to give us different lengths of alkyl chains. We applied these enhancers to human skin in Franz diffusion cells using two...
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Cationic surface active compounds and evaluation of its properties
Pekařová, Kateřina ; Nováková, Veronika (advisor) ; Kuchařová, Monika (referee)
Charles University Faculty of Pharmacy in Hradec Králové Department of Biophysics and Physical Chemistry Consultant: PharmDr. Jan Marek, Ph.D. Supervisor: doc. PharmDr. Veronika Nováková, Ph.D. Student: Kateřina Pekařová Title of Thesis: Cationic surface active compounds and evaluation of its properties Quaternary ammonium salts are used as surfactants in many branches of the food, pharmaceutical and chemical industries. The basic properties of these substances were studied in the thesis. The critical micellar concentration of the homologous series of five substances was determined using the conductometric method. The theory of declining critical micellar concentration with the increasing length of the side alkyl chain has been verified. The hydrolytic activity of two commonly used cationic surfactants benzalconium chloride and benzoxonium chloride for the decomposition of the organophosphorus pesticide fenitrothion was tested. For these two substances, the effect of pH and temperature changes was evaluated. The influence of tenside concentration was also monitored. Benzoxonium chloride at 50 řC and pH 11was found as the most effective hydrolytic catalyst for the decomposition of the organophosphorus pesticide fenitrothione.
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Preparation of Substituted Arylsulfanyl Azaphthalocyanine
Filandrová, Tereza ; Nováková, Veronika (advisor) ; Miletín, Miroslav (referee)
Charles Univerzity in Prague, Faculty of Pharmacy in Hradec Králové Department of Department of Pharmaceutical Chemistry and Drug Control Candidate Mgr. Tereza Filandrová Consultant PharmDr. Veronika Nováková, PhD. Title of Thesis Preparation of substituted arylsulfanyl azaphthalocyanine The aim of this thesis was to prepare a series of symmetrical azaphthalocyanines (AzaPc) with arylsulfanyls on the periphery with different bulky substituents in the ortho positions (hydrogen, methyl, isopropyl) or a functional group in the para position (-Br). These AzaPc were prepared by cyclotetramerization reactions of 5,6-disubstituted pyrazine-2,3-dicarbonitrile namely 5,6-bis(phenylsulfanyl)pyrazine-2,3-dicarbonitrile, 5,6-bis((2,6-dimethylphenyl)sulfanyl)pyrazine-2,3-dicarbonitrile and 5,6-bis-((2,6- diisopropylphenyl)sulfanyl)pyrazine-2,3-dicarbonitrile. These precursors were prepared by nucleophilic substitution of 5,6-dichloropyrazine-2,3-dicarbonitrile with commercially available thiophenol and 2,6-dimethylbenzenthiol or synthesized 2,6- diisopropylbenzentiol.The latter one had to be prepared by the Newman-Kwart rearrangement. Several cyclotetramerization methods were compared resulting thus in magnesium, zinc and metal free AzaPcs. Properties of prepared AzaPcs were also studied and compared...
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The influence of anesthesia on the degree of DNA oxidative damage
Zubáňová, Veronika ; Kuchařová, Monika (advisor) ; Nováková, Veronika (referee)
Background: Oxidative damage is one of the most frequent types of cell components damage leading to oxidation of lipids, proteins and the molecule of DNA. As a consequence, there is a higher occurrence of several pathologies such as atherosclerosis, neurodegenerative diseases, cancer; or diabetes. In our study, influence of whole body anesthesia during minor surgery on the level of DNA damage was examined using comet assay technique. Methods: The basic principle of this method is fixing the cells (lymphocytes) in agarose, their lysis for the removal of membranes, incubation with the specific enzymes and electrophoresis of the released cell nuclei. During the electrophoresis, free low-molecular weight and negatively charged fragments of DNA move towards anode which causes the formation of the typical comet cell shape. Finally, the gels are stained by ethidium bromide (DNA intercalating dye) and visualized. Results: We have observed single strand breakages (SSBs) and, with the use of modified assay using specific enzymes for detection of specific lesions, also oxidized purines and pyrimidines. The extent of DNA damage as determined by the intensity of the tail of the comet was quantified using LUCIA Comet Assay (Laboratory Imaging, Czech Republic) software for image analysis. The results were used...
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