Charles University Faculty of Pharmacy in Hradec Králové Department of Pharmacognosy Candidate: Iva Valachová Supervisor: Pharm.Dr. Daniela Hulcová, Ph.D. Title of diploma thesis: Derivatives of Amaryllidaceae alkaloids as potential drugs in the treatment of tumor diseases Twelve aromatic derivatives were prepared. The substitutions were performed on the hydroxyl group at C-3. This was acylated with differently substituted benzoyl chlorides affording the corresponding esters. Structural identification was performed by 1D- and 2D- NMR, ESI-HRMS spectroscopic techniques and optical rotation measurement. The yield of all reactions was more than 60 %. After converting these derivatives to hydrochlorides, their biological activity was tested. The Amaryllidaceae alkaloid vittaine itself exhibits cytotoxic activity and therefore its derivatives have been investigated in this respect. It has been tested against 9 cancerous and 1 non-cancerous cell lines. 3-O-(4-Chloro-3-nitrobenzoyl)vittatine showed the highest cytotoxic activity, unfortunatelly, it does not selectively affect only cancerous cells. Conversely, 3-O-(2-naphthoyl)vittatine has a selective effect on the HT-29 cancerous cell line (colorectal carcinoma) with a viability value of 32 ± 3 %, potentially being the subject of further studies of cytotoxicity...

Charles University Faculty of Pharmacy in Hradec Králové Department of Pharmaceutical Botany Candidate: Dušan Zelina Supervisor: doc. Ing. Lucie Cahlíková, Ph.D. Title of diploma thesis: Derivates of Amaryllidaceae alkaloids and their biological activity: Derivates of tazettine II Plants of family Amaryllidaceae are a rich source of alkaloids that are characterized by a broad biological activity. Perhaps the world's best-known member of this family has become galanthamine, which, because of its potent inhibitory potential, has found a place in the treatment of Alzheimer's disease. Based on the conducted studies, the alkaloid tazettine has been chosen for this thesis. The main purpose was to prepare more active derivatives of tazettine and to test the biological activity for potential use in the treatment of cancer and Alzheimer's disease. Ten aromatic esters of tazettine were prepared. Reaction yields ranged from 33,5-69,3 %. Derivatives were identified by MS, NMR and optical rotation. Prepared compounds were tested for their inhibitory potential against human cholinesterases - AChE and BuChE. Unfortunately, it has not been possible to prepare derivatives, which exhibit significant activity in inhibiting AChE or BuChE. Anti-tumor activity of the derivates has been tested on the panel of selected...

Muchová, A.: Antiplatelet activity of alkaloids and its potential use in the Alzheimer's disease therapy. Diploma thesis, Charles University in Prague, Faculty of Pharmacy in Hradec Králové, Department of Pharmaceutical Botany, Hradec Králové, 2020. According to recent studies, the pathophysiology of Alzheimer's disease, as a neurodegenerative disease, has been implicated in platelets and the associated haemostasis disorder, the release of inflammatory cells, which may result in the facilitation of amyloid plaque formation. Alkaloids as alkaline nitrogen compounds, which have many effects on humans or animals. In connection with this issue, researchers are investigating the antiplatelet effect of many structural types of alkaloids. The isoquinoline alkaloids of the families Lauraceae, Annonaceae, Piperaceae, Magnoliaceae and Papaveraceae are perpective for the study. In addition to their antiplatelet effects, they also have an antioxidant and inhibitory effect on acetylcholinesterase, which is also used in Alzheimer's disease therapy. Another promising substance is the pyrrolidinoindoline alkaloid Psm2, which was isolated from the plant Selaginella moellendorffii Hieron. It has an antiplatelet effect higher than the reference substance acetylsalicylic acid and is also associated with a lower risk...

Valová D.: Vinca minor L. alkaloids and their biological activity III. Diploma thesis, Charles University, Faculty of Pharmacy in Hradec Králové, Department of Pharmaceutical Botany, Hradec Králové, 2020, 75 p. Alzheimer's disease (AD) is a progressive, neurodegenerative disorder and it's the most common form of dementia. Because we're still not able to treat the cause of disease, searching for a new substance is relevant. This thesis is focused on isolation of alkaloids from a Vinca minor L. alkaloidal extract as a potential drug. The preparation and column chromatography of the summary extract were performed by Ing. Miroslav Ločárek as a part of his doctoral studies. Subsequent preparative TLC led to the isolation of three compounds. The chemical structures of the isolated alkaloids were elucidated by means of optical rotation, NMR and MS analyses and by comparison of the obtained data with those in the literature. One of the compounds was determined as(-)-vincine, other two compounds have not been isolated yet. Isolated compounds were also tested for their biological activity. Vincine, DV-1 a DV-3 were tested for their ability to inhibit acetylcholinesterase (AChE) and butyrylcholinesterase (BuChE). Additionally, vincine and DV-3 were also tested for their inhibitory activity on prolyl...

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.

Patients with Alzheimer disease often report presence of neuropsychiatric symptoms even before the presence of cognitive decline itself. The most reported ones are anxiety, depression, emotional dysregulation, apathy and agitation. The aim of this work was to investigate the presence of anxiety- and depression-like behaviour, and eventually dysregulation of social behaviour and emotional memory impairment via behavioural approach on the TgF344-AD (tg) rat model. Results show that tg rats display anxiety-like behaviour in several different tests and parameters. Tg rats of the age of 10 months spent more time around the wall and less in the middle of the arena. Surprisingly, this anxiety-like phenotype has not been demonstrated in the older (14 months) rats. Tg rats spent also less time peeking out from closed arms and looking down from open arms. Moreover, tg rats displayed anxiety-like behaviour in all observed parameters in the Social interaction test. Social deficit expressed as a smaller investment of time into the anogenital and non- anogenital exploration and following of counterparts. In the Forced swim test, tg rats did not spend more time immobile, thus depression-like phenotype has not been demonstrated in these animals. Moreover, 10 months old tg rats spent surprisingly less time...

Polycystic kidney disease is a severe genetic disease occurring in both adult and pediatric patients. The basic characteristic of this disease is the development and progressive enlargement of renal cysts gradually replacing functional kidney tissue. This leads to renal failure in many patients. However, renal cysts may also occur in a number of other diseases, including multisystem syndromes. This complicates differential diagnosis in some patients. In our study, we first focused on the diagnosis and characterization of genotypic-phenotypic relationships in patients with polycystic disease arising in childhood, later we extended our study to adult patients and patients with unclear clinical diagnosis. At the same time, we expanded the portfolio of analyzed disorders to a number of diseases in which the phenotype of polycystic kidneys may occur, and noncystic diseases as well. During our project, massive parallel sequencing was used to analyze 149 patients - 128 with cystic and 21 with noncystic clinically diagnosed nephropathies. At the same time, the findings were verified by Sanger sequencing in 176 relatives of our probands. Mutation detection reached 59% in cystic patients, and 43% in non-cystic patients, respectively. In many patients, molecular genetic analysis revealed a different etiology...

This dissertation is a collection of a total of seven publications that deal with eye movement disorders in patients with basal ganglia disorders. We obtained normative data for videooculography in healthy individuals. We have described the eye movement evolution during a human life such as the increase of latency, movements become hypometric and antisaccadic error rate increases. We have shown that sex and education do not affect the eye movements. Our study highlighted the asymmetry in the eye movement performance. As the first, we studied the vergence in patients with Parkinson's disease (PN) using videooculography (VOG). We devised and defined a paradigm for this examination and saw that in patients with PN there is a prolonged latency and hypometry of divergence. In patients with ephedrone induced parkinsonism (EP), we were the first who examined eye movements and found that it was possible to identify between this toxic Parkinson's syndrome and PN on the basis of a videooculography. In EP patients, we described velocity decsrease and hypometry in horizontal saccades, prolonged latency in horizontal saccades, and higher error rate in the antisacadic task. Behavioral disorder in REM sleep (RBD) as a prodromal stage of PN leads to impaired eye movement. In the evaluation with PN patients, we...

Huntington's disease is an autosomal dominant neurodegenerative disorder caused by an extended (≥36) CAG repeat in the huntingtin gene. Its hallmark is brain athrophy, but huntingtin is widely deposited in all tissues of the body, most notably in the brain and testes. Its pathogenic effect is conditioned by the formation of cytotoxic forms of aggregates and fragments, which occur in both brain and peripheral tissues. Testicular atrophy has been demonstrated in postmortem samples from human patients with Huntington's disease and in transgenic mouse models. We investigated reproductive decline in a large animal model of Huntington's disease. A transgenic (tgHD) minipig model was created by inserting a lentiviral vector into the genome of a pig. Vector contained a truncated form of the N terminal part of huntingtin gene. Boars of this transgenic line showed a reduced ability to produce offspring from 13 months of age. We confirmed apoptosis of seminiferous epithelial cells and Sertoli cells, and a production of morphologically damaged spermatozoa, which were unable to efficiently fertilize the oocyte under in vitro conditions. We found a reduction of mitochondrial metabolism parameters in the sperm of tgHD boars. These changes were not dependent on the age of the boars., It is directly related to the...

Introduction: The thesis "Alzheimer's disease and department of intensive care" deals with the specifics and difficulties of nursing care for patients with this chronic and degenerative disease in intensive care units (ICUs). The number of patients with Alzheimer's disease is constantly growing and caring for them in such specific wards is extremely demanding. The purpose of this thesis was to map the most problematic areas in the care provided and propose their solution. Methods and results: The empirical part presents the results of a conducted qualitative research which took place in the form of interviews with 10 general nurses who have experience with such patients. Following a word-for-word transcription of the obtained interviews, the results are summarized in a final discussion. They show that when caring for a patient with dementia, general nurses see the main problems in his decreased ability to cooperate, behavioral changes, which often appear as a result of environment change, and difficult communication. They consider the patient's confusion, his negativism, and the inability to find the right words to be communication barriers. This results in enormous demands on the caring staff's patience. The interviewed general nurses however approach patients with dementia generally positively...