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Evaluation of the pharmacological properties of new potential drugs for Alzheimer's disease
Čechová, Pavlína ; Šorf, Aleš (advisor) ; Urbánková Rathouská, Jana (referee)
Charles University Faculty of Pharmacy in Hradec Králové Department of Social and Clinical Pharmacy Student: Pavlína Čechová Supervisor: PharmDr. Aleš Šorf, Ph.D. Consultant: mjr. PharmDr. Vendula Hepnarová, Ph.D. Title of diploma thesis: Assessment of pharmacological properties of new potencial drugs for Alzheimer's disease treatment Alzheimer's disease is a neurodegenerative disease with a progressive and irreversible course. The cause of AD is still not exactly known. There are currently four drugs available that have been approved by the State Institute for Drug Control. Three of them act as cholinesterase inhibitors (donepezil, rivastigmine, galantamine) and the fourth one as a N-methyl-D-aspartate (NMDA) receptor antagonist (memantine). Unfortunately, these drugs do not affect the course of the disease and provide only temporary symptomatic relief. An important area of current research is MTDLs (Multi-Target Directed Ligands), which act on multiple target structures related to the pathophysiology of the disease. These agents represent a promising therapeutic strategy. The aim of the study was to measure the ability to inhibit human acetylcholinesterase (AChE) and butyrylcholinesterase (BuChE) of 58 new potential MTDLs derived from biphenyl and benzhydryl using Ellman's method, as well as...
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In vitro characterization of novel potential modulators of cholinesterases
Táborská, Lenka ; Šorf, Aleš (advisor) ; Ťupová, Lenka (referee)
Charles University Faculty of Pharmacy in Hradec Králové Department of Social and Clinical Pharmacy Candidate: Lenka Táborská Supervisor: PharmDr. Aleš Šorf, Ph.D. Consultant: RNDr. Ľubica Múčková, Ph.D. Title of diploma thesis: In vitro characterization of novel potential modulators of cholinesterases This thesis focuses on investigating the interaction of newly synthesized potential drugs with human cholinesterases (ChE), particularly acetylcholinesterase (AChE) and butyrylcholinesterase (BChE). The main objective is to assess the inhibitory potential of these molecules against recombinant human forms of AChE and BChE. Furthermore, the cytotoxicity of selected compounds is examined using a colorimetric method on the SH-SY5Y cell line employing MTT tetrazolium salt. The theoretical part of the thesis delves into the structure and functions of cholinesterases, especially AChE and BChE, as well as the cognitive aspects of neurodegenerative diseases such as Alzheimer disease (AD), Parkinson's disease (PD), amyotrophic lateral sclerosis (ALS), and multiple system atrophy (MSA). Additionally, it addresses the toxic effects of organophosphates, which can inhibit the AChE enzyme, along with methods for determining cholinesterase activity. The practical part focuses on experimentally determining the...
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Synthesis and evaluation of potential drugs for neurodegenerative diseases
Foltýnová, Eva ; Krátký, Martin (advisor) ; Vinšová, Jarmila (referee)
The topic of this thesis is the synthesis of oxadiazole derivatives and their precursors as potential therapeutics for neurodegenerative diseases, and their evaluation primarily as potential inhibitors of cholinesterases (CHE). The theoretical part of this thesis focuses on Alzheimer's disease, particularly its pathophysiology, diagnosis, and treatment. It also discusses CHE inhibitors that are already clinically used, as well as new structures with potential for future clinical use. Additionally, it addresses the oxadiazole fragment, its chemical and biological properties. In the experimental work, derivatives of 1,3,4- and 1,2,4-oxadiazoles were synthesized, which differed not only in the type of oxadiazole but also in their substitution. These substances were prepared by the cyclization of precursors, which were also prepared and tested. For 1,3,4- oxadiazoles, besides their inhibitory activity against acetylcholinesterase (AChE) and butyrylcholinesterase (BuChE), their potential antimycobacterial, antibacterial, and antifungal activities were tested. For 1,2,4-oxadiazoles, so far, only their inhibitory activity against AChE and BuChE has been tested. A total of twenty-two substances were synthesized, and the yields of these syntheses ranged from 21-97%. The compound with the lowest IC50 value...
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Cytotoxic and cholinesterase inhibitory activity of extracts from selected species of the Centaurea L. genus
Faschingbauer, Jakub ; Chlebek, Jakub (advisor) ; Šafratová, Marcela (referee)
Faschingbauer J.: Cytotoxic and cholinesterase inhibitory activity of extracts from selected species of the Centaurea L. genus. Diploma thesis, Charles University, Faculty of Pharmacy in Hradec Králové, Department of Pharmaceutical Botany, Hradec Králové, 2019. During the screening of biologically active secondary metabolites of plants carried out at the Department of Pharmaceutical Botany FAF UK, selected taxa of the genus Centaurea (Asteraceae) were investigated. This study is focused on a basic phytohemical research of extracts prepared from Centaurea cyanus, Centaurea jacea, Centaurea scabiosa, Centaurea pseudophrygia, Centuarea stoebe, Centaurea solstitialis a Centaurea benedicta. Extracts were prepared for evidence of the proof reactions of TLC and MS analysis (EI, ESI) to clarify a potential presence of alkaloids. EtOAc and ethanol extracts were evaluated for potential inhibitory activity against human erythrocyte acetylcholinesterase (AChE) and plasma butyrylcholinesterase (BChE) and cytotoxicity against selected 9 tumor lines. C. cyanus alkaloid extract had interesting cholinesterase activity which selectively inhibited BChE (IC50 BChE = 22.62 ± 3.62 μg / ml, IC50 AChE = 221.50 ± 44.56 g / ml). Other EtOAc extracts of selected Centaurea species were considered inactive (IC50 > 100 μg/ml)....
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Magnetic particles as a reversible carrier for enzyme electrodes
Janíček, Zdeněk ; Žeravík,, Jiří (referee) ; Skládal,, Petr (advisor)
Master’s thesis contains information about the enzymes, biosensors, enzyme biosensors and magnetic particles. Cholinesterasa (ChE) is the generally name for the two related enzymes, mutually differing appearance and biological functions. Acetylcholinesterasa (AChE) is necessary for the transmission of nervous excitement. AChE is located at cholinergic synapses, where is the hydrolysis of the neurotransmitter acetylcholine and this termination by the nervous excitement. Butyrylcholinesterasa (BChE) is related to AChE and it is present in serum. Experimental part was focused on electrochemical biosensors with cholinesterase enzyme using magnetic particles for reversible reconstruction layer. Cholinesterase immobilization was carried out on the magnetic particles by covalent binding after glutaraldehyde activated. The measurement of activity take place with acetylthiocholin as a substrate in a flow system, magnetic particles were captured before the platinum electrode and produced by thiocholin is detected amperometric. The aim was to measure inhibition of AChE. Significant inhibition ChE is caused by certain drugs and pesticides based on organophosphates. There was tested by inhibition of AChE pesticide dichlorvos. Emphasis was on finding possible experimental conditions for the creation of the automated procedure to measure the AChE activity, which was based on auto-injector and pumps of Gilson, together with a 735 programme for the definition of the working configuration, preparation of measurement scripts and implementation of application methods.
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Electrochemical biosensors with spatially separated enzymatic and detection parts for selective analysis in flow-through arrangement
Tvorynska, Sofiia ; Barek, Jiří (advisor) ; Labuda, Ján (referee) ; Korecká, Lucie (referee)
This dissertation thesis presents the newly developed four highly reusable, stable as well as simple, and cost-effective electrochemical (bi)enzymatic biosensors for the selective and reliable determination of choline, acetylcholine, uric acid, and L-lactic acid in flow injection analysis. All biosensors are based on the concept of the spatial separation of the biorecognition part from detection one and amperometric monitoring of the enzymatically consumed oxygen via its four-electron reduction at the highly negative detection potential. In this way, the design of the biosensors includes an easily replaceable enzymatic mini-reactor(s) connected upstream to the flow cell that contains the appropriate silver amalgam-based transducer. The enzymatic mini-reactor based on choline oxidase, uricase, or lactate oxidase was used for choline, uric acid, or L-lactic acid biosensors, respectively. The acetylcholine bienzymatic biosensor includes the consequently connected choline oxidase- and acetylcholinesterase-based mini-reactors. The first part of this thesis focuses on the construction of two different silver amalgam-based electrodes. Specifically, this section discusses the fabrication of a silver solid amalgam electrode covered by mercury film operating in a wall-jet cell and also highlights the...
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Alkaloids of Papaver rhoeas L. (Papaveraceae) and their biological activity related to Alzheimerʼs disease VI.
Ludvik, Matěj ; Chlebek, Jakub (advisor) ; Šafratová, Marcela (referee)
Ludvik M., Papaver rhoeas L. (Papaveraceae) alkaloids and their activity related to Alzheimer disease VI., diploma thesis, Charles University, Faculty of Pharmacy in Hradec Králové 2023, number of pages 69. This diploma thesis was focused on alkaloids from combined fractions 13-15 of diethylether extract from Papaver rhoeas L. The methods used for extraction of the alcaloids include flash chromatography and preparative thin-layer chromatography. The structures of isolated compounds were determined by using mass spectrometric and spectrophotometric methods (GC-MS, NMR and optical rotation). Alkaloids were identified as galanthamine and N,N-dimethyltryptamine. Subsequently, N,N-dimethyltryptamine was tested for its biological activity against human cholinesterases, specifically acetylcholinesterase and butyrylcholinesterase. Galanthamine was not tested on ability to inhibit cholinesterase, since the compound has been already tested on this activity at the Department of Pharmacognosy and Pharmaceutical Botany. N,N-dimethyltryptamine showed mild inhibitory activity against butyrylcholinesterase (IC50 = 44,14 ± 1,76) and no activity against acetylcholinesterase (IC50 = >100). Despite its mild activity towards butyrylcholinesterase, N,N-dimethyltryptamine can't be used in therapy of Alzheimer's disease,...
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Alkaloids of Papaver rhoeas L. (Papaveraceae) and their biological activity related to Alzheimerʼs disease V.
Dušánková, Zita ; Chlebek, Jakub (advisor) ; Hošťálková, Anna (referee)
Dušánková Z.: Alkaloids of Papaver rhoeas L. (Papaveraceae) and their biological activity related to Alzheimerʼs diesease V.; Diploma thesis, Charles University, Faculty of Pharmacy in Hradec Králové, Department of Pharmacognozy and Pharmaceutical Botany, Hradec Králové 2023, pages 72. The aim of this diploma thesis was focused on isolation of alkaloids from a combined fraction 7-8 of an alkaloidal diethyl ether (Et2O) extract of aerial parts of Papaver rhoeas L. (Papaveraceae). Column chromatography (flash chromatography), planar chromatography (preparative TLC) and crystallization were used for the isolation of alkaloids. The structures of isolated substances were elucidated using NMR, GC-MS methods and optical rotation. Isolated alkaloids were identified as ZD-1, (+)-3,4-dehydrotheaspirone, (±)-juziphine, (+)-lirinidine and protopine. Alkaloids were tested for biological activity against enzymes (acetylcholinesterase, butyrylcolinesterase and prolyl oligopeptidase). (+)-3,4-Dehydrotheaspirone, (±)-juziphine, and protopine did not show significant inhibitory activities against cholinesterases (IC50 values >100 µM). Protopine was also inactive against prolyl oligopeptidase (IC50 >1000 µM ). (+)-Lirinidine showed a moderate inhibitory activity against BuChE (IC50 value of 23,45 ± 0,55 µM), towards...
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Synthesis and evaluation of dual acting potential drugs for neurodegenerative diseases
Miffek, Dominik ; Krátký, Martin (advisor) ; Vinšová, Jarmila (referee)
Charles University, Faculty of Pharmacy in Hradec Králové Department of Organic and Bioorganic Chemistry Author: Dominik Miffek Supervisor: doc. PharmDr. Mgr. Martin Krátký, Ph.D. Title of diploma thesis: Synthesis and evaluation of dual acting potential drugs for neurodegenerative diseases Alzheimer's disease (AD) and Parkinson's disease (PD) are neurodegenerative diseases that mainly affect the elderly population. The incidence of these diseases is still increasing worldwide, and currently, there is no cure that can permanently treat patients. Current medications mainly aim to slow the progression of these diseases. The goal is to improve the accessibility of treatment, prevent progression, or completely cure patients suffering from AD and PD. New potential drugs are also developing in this direction, which could exhibit a satisfactory effect in inhibiting cholinesterases. Acetylcholinesterase (ACh) and butyrylcholinesterase (BuChE) play a significant role in the pathogenesis of these diseases. Particularly in Alzheimer's disease (AD), there is a decrease in acetylcholine as a neurotransmitter that ensures the homeostasis and proper functioning of the central nervous system (CNS). In AD treatment, acetylcholinesterase inhibitors (AChEIs) such as rivastigmine, galantamine, and donepezil are used...
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Vinca minor L. alkaloids and their biological activity (inhibition of human cholinesterases) V.
Vašíčková, Alžběta ; Chlebek, Jakub (advisor) ; Hošťálková, Anna (referee)
Vašíčková A.: Alkaloids of Vinca minor L. and their biological activity (inhibition of human cholinesterases) V. Diploma thesis, Charles University, Faculty of Pharmacy in Hradec Králové, Department of Pharmaceutical Botany, Hradec Králové 2020. From the selected fraction VM 215-258 were isolated 3 alkaloids by flash chromatography followed, preparative thin layer chromatography and crystallization. Their structure was determined by mass spectroscopy, NMR and optical rotation, and the obtained data were compared with those in literature. Strictamine and minovincinine belong to alkaloids previously isolated, the alkaloid AV-3 has not been isolated yet. These alkaloids were tested for their ability to inhibit human cholinesterases and their inhibitory activity was compared to standards galanthamine and physostigmine. Compound AV-3 showed mild inhibitory activity against BChE (IC50 μM > 86.3 ± 2.3), other alkaloids were considered inactive, their IC50 values against cholinesterases were > 100 μM. Key words: Vinca minor L. (Apocynaceae), vinca alkaloids, minovincinine, strictamine, Alzheimer's disease, acetylcholinesterase, butyrylcholinesterase.
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