Nováková D.: Biological activity of secondary plants metabolites I. Alkaloids of Narcissus jonquilla L. Charles University in Prague, Faculty of Pharmacy in Hradec Králové, Department of Pharmaceutical Botany and Ecology, Hradec Králové 2015, pp. 70. The aim of the diploma thesis was a preparation of alkaloid extracts to identification of alkaloid patterns and measure cholinesterase inhibitory activity. This activity is useful for treating Alzheimer's disease. Alkaloid extracts of seven Narcissus jonquilla L. (Amaryllidaceae) varieties (Sundial, Sundisc, Sweetness, Waterperry, Simplex, Twinkling Yellow, Yazz) were studied with respect to their acetylcholinesterase (HuAChE) and butyrylcholinesterase (HuBuChE) inhibitory activity and alkaloid patterns. Twenty-three alkaloids were determined by GC/MS, and ten of them identified from their mass spectra and retention times. All samples exhibited content of galanthamine, most samples contained lycorine and tazettine. Promising HuAChE inhibition activity was demonstrated by Narcissus jonquilla L. cv. Waterperry with IC50 values of 6.53 ± 0.88 μg/mL. The strongest inhibitory activity against HuBuChE was detected in extract from Narcissus jonquilla L. cv. Sundisc with IC50 value of 5.09 ± 0.64 μg/mL. Keywords: Alzheimer's disease, Amaryllidaceae, Narcissus,...

Panchartková, M.: Biological aktivity of secondary plants metabolites III. Alkaloids of Narcissus tazetta L., Diploma thesis, Charles University in Prague, Faculty of Pharmacy in Hradec Králové, Department of Pharmaceutical Botany and Ecology, Hradec Králové, 2015, 67s. Plants from the Amaryllidaceae family contain alkaloids that have multiple biological effects. There is described antiviral, antitumor, antibacterial, antimalarial and anti-fungal effect. Activity against human cholinesterases is important too. The aim of this thesis was to prepare seven alkaloid extracts of individual cultivars of the plants Narcissus and then to realize their GC/MS analysis. Thanks to this analysis, several different structural types of alkaloids from the Amaryllidaceae family were identified. The most frequently identified alkaloids were homolycorine, lycorine, tazettine and galanthamine type. This was followed by the measuring of biological activity against human acetylcholinesterase (HuAChE) and butyrylcholinesterase (HuBuChE). The highest inhibitory activity IC50 identified the alkaloidal extract of the Narcissus jonquilla cv. New baby with the values in relation to HuAChE 13,78 ± 1,48 and in relation to HuBuChE 96,12 ± 9,55. The main reason was probably the highest content of galanthamine of all cultivars,...

Jílek L.: The biological activity of secondary plants metabolites II. Alkaloids of Narcissus jonquilla L. Charles University in Prague, Faculty of Pharmacy in Hradec Králové, Department of Pharmaceutical Botany and Ecology, Hradec Králové 2015, pp. 74. The aim of the diploma thesis was a preparation of alkaloid extracts to identification of alkaloid patterns and measure cholinesterase inhibitory activity. This activity is useful for treating Alzheimer's disease. Alkaloid extracts of seven Narcissus jonquilla L. (Amaryllidaceae) varieties (Bella Estrella, Bell Song, Fruit Cup, Hill Star, Chit Chat, Martinette, Dick Sickel) were studied with respect to their acetylcholinesterase (HuAChE) and butyrylcholinesterase (HuBuChE) inhibitory activity and alkaloid patterns. Thirteen different alkaloids were identified from their mass spectra and retention times. All samples exhibited content of tazettine, most samples contained lycoramine and galanthamine. Promising HuAChE inhibition activity was demonstrated by Narcissus jonquilla L. cv. Bell Song with IC50 values of 6,19 ± 0,85 μg/mL. The strongest inhibitory activity against HuBuChE was detected in extract from Narcissus jonquilla L. cv. Bella Estrella with IC50 value of 18,39 ± 1,51 μg/mL. Keywords: Alzheimer's disease, Amaryllidaceae, Narcissus, GC/MS,...

Charles University Faculty of Pharmacy in Hradec Králové Department of Pharmacology & Toxicology Student: Brigita Mányová Supervisor: PharmDr. Marie Vopršalová, CSc. Supervisor-specialist: PharmDr. Vendula Hepnarová, Ph.D. Title of diploma thesis: Development of insecticides inhibiting acetylcholineseterase Pest insects cause problems and damage all over the world. They are vectors of serious diseases such as malaria, dengue fever, yellow fever, Zika virus disease and chikungunya disease. They cause huge damage to agricultural crops and are annoying to everyday life in living spaces. The aim of this diploma thesis was in vitro testing of selected compounds from the group of bis-isoquinoline and bispyridinium acetylcholinesterase inhibitors as potential insecticides. Another goal was also to create relationships between structure and effect. The ability of these compounds to inhibit both human (hAChE) and fly acetylcholinesterase (MdAChE) was evaluated. The modified Ellman spectrophotometric method was used. The half inhibitory concentration (IC50) values were obtained for both enzymes and the selectivity indexes (SI) were then calculated. Compounds having IC50s in micromolar or nanomolar range and exhibiting selectivity for MdAChE were most desirable. During the testing of these inhibitors, three...

Charles University Faculty of Pharmacy in Hradec Králové Department of Pharmacology and Toxicology Student: Ondrej Tomáš Supervisor: PharmDr. Marie Vopršalová, CSc. Consultant supervisor: mjr. PharmDr. Vendula Hepnarová, Ph.D. Title of diploma thesis: Evaluation of newly prepared insecticides in vitro Malaria is a widespread infection and one of the most dangerous diseases transmitted by insects. It threatens lives of millions of people all around the world, thus its regulation is necessary. Most common malaria vectors are mosquitoes of genus Anopheles. Novel structures of insecticides with selective inhibition of mosquito acetylcholinesterase are subjects of research, with an intention to deal with this problem. The aim of this work was to test six newly prepared succinimide derivatives with insecticidal potential. The ability of these compounds to inhibit Anopheles gambiae mosquito (AgAChE) and human acetylcholinesterase (hAChE) was evaluated. Leading structures of these compounds were also tested to find relations between chemical structure and biological activity. The modified Ellman's method was used to obtain the half maximal inhibitory concentration (IC50) values for both enzymes. Tested substances were able to inhibit only hAChE and none of them displayed activity against AgAChE. Compound...

Charles University in Prague Faculty of Pharmacy in Hradec Králové Department of Pharmacology and Toxicology Candidate: Natália Kuzmiaková Supervisor: PharmDr. Marie Vopršalová, CSc . Consultant: mjr. PharmDr. Vendula Hepnarová, Ph.D. Title of diploma thesis: In vivo evaluation of the efficacy of the novel reactivator against tabun. This study tackles the problem of irreversible inhibition of acetylcholinesterase (AChE). This enzyme degrades neurotransmitter acetylcholine (ACh), wich ensures transmisson of nerve impulses in central nervous system and in periphery. Organophospates (OP) are substances that cause irreversible blocade of AChE and that susubsequently leads to accumulation of AChE in synapses and inducing of muscarinic and nicotinic effects for life threatening condition. Oximic nature reactivators shown to this day the gratest potencial in inhibiting OP bond with AChE. Because reactivation abilities of to date synthesided oxime are not sufficient, new reactivators are being researched. The doal of my work was to test the potencial to reactivate AChe one of them (precisely oxime K 870). The method i used was colorimetric Ellman method modified by Bajgar, where the activity of AChE after reactivation was measured by absorbance in brain, diaphragm and blood of modeled orgamisms. The...

Charles University Faculty of Pharmacy in Hradec Králové Department of Pharmacology and Toxicology Student: Ondrej Tomáš Supervisor: PharmDr. Marie Vopršalová, CSc. Consultant supervisor: mjr. PharmDr. Vendula Hepnarová, Ph.D. Title of diploma thesis: Evaluation of newly prepared insecticides in vitro Malaria is a widespread infection and one of the most dangerous diseases transmitted by insects. It threatens lives of millions of people all around the world, thus its regulation is necessary. Most common malaria vectors are mosquitoes of genus Anopheles. Novel structures of insecticides with selective inhibition of mosquito acetylcholinesterase are subjects of research, with an intention to deal with this problem. The aim of this work was to test six newly prepared succinimide derivatives with insecticidal potential. The ability of these compounds to inhibit Anopheles gambiae mosquito (AgAChE) and human acetylcholinesterase (hAChE) was evaluated. Leading structures of these compounds were also tested to find relations between chemical structure and biological activity. The modified Ellman's method was used to obtain the half maximal inhibitory concentration (IC50) values for both enzymes. Tested substances were able to inhibit only hAChE and none of them displayed activity against AgAChE. Compound...

Adéla Dolejšová Testing the efficacy of potential therapeutics for Alzheimer,s disease Charles University in Prague, Faculty of Pharmacy in Hradec Kralove Pharmacy The aim of this thesis was to find out whether already implemented Ellman's method, which is used to analyse irreversible inhibitors AChE, is going to be a suitable technique for measuring reversible inhibitors. Furthermore, the efficiency of newly synthesized AChE inhibitors was established. These inhibitors will be used for treating AD or as prophylaxis against neural paralytic substances. Moreover, it was compared affect the efficacy of AChE inhibitors after intramuscular and intraperitoneal administration. The measurements were done in vivo on potkan species Wistar. In the first experiment standard AChE (tacrin, 7-MEOTA, donepezil, rivastigmin) inhibitors were applied to verify the method. In the second experiment the newly synthesizes AChE (K 298, K 344, K 474) inhibitors were medicine. The results confirmed the effectiveness of the tested method on commonly used inhibitors such as donepezil and rivastigmin. Out of the newly synthesized K 298, K 344 and K 474 inhibitors none was proven to have any significant inhibiting activity. Key words: acetylcholinesterase, acetylcholinesterase inhibitors, Alzheimer disease, cholinergic system,...

Bouz L.: Biological activity of secondary plants metabolites V. Alkaloids of Vinca minor L. Diploma thesis, Charles University, Faculty of Pharmacy in Hradec Králové, Department of Pharmaceutical Botany and Ecology, Hradec Králové 2016. Summary extract obtained from dried aerial parts of Vinca minor L. was separated by column chromatography with petrol, chloroform and ethanol to 531 fractions. By further separation of fractions, following preparative thin layer chromatography and crystallization 2 alkaloidal compounds marked LB-2 and LB-3 were isolated. These compounds were identified by GC/MS, 1 H- and 13 C-NMR spectroscopy and by use of physical-chemical methods. The structure of compounds were elucidated as indole alkaloids (+)-vincaminoreine (LB-2) and (+)-vincamine (LB-3). Both substances were tested for their inhibition activity against human cholinesterases. (+)-Vincamine didn't exhibited in comparison with standard drugs (galanthamine IC50 AChE: 1,710 ± 0,065 µM, IC50 BChE: 42,30 ± 1,30 µM; huperzine A IC50 AChE: 0,033 ± 0,001 µM) any inhibition activity. On the other hand (+)-vincaminoreine exhibited fairly strong selective inhibition of BChE (IC50 = 8,71 ± 0,49 µM) with no inhibition of AChE. Key words: Vinca minor L., indole alkaloids, vincamine, vincaminoreine, Alzheimer disease,...

Charles University in Prague, Faculty of Pharmacy in Hradec Králové Department of Pharmaceutical Botany and Ecology Candidate: Bc. Blanka Kaluzsná Supervisor: Prof. RNDr. Lubomír Opletal, CSc. Title of Diploma Thesis: Biological activity of plant metabolites XXIII. alkaloids of Corydalis cava (l.) Schweigg. & Körte and their effect on acetylcholinesterase and butyrylcholinesterase An ether extract which contained tertiary basic alkaloids was prepared from the dry bulb of Corydalis cava (L.) SCHWEIGG. & KÖRTE. A subfaction labeled 2/B was obtained by column chromatography and crystallization. Three alkaloids were isolated by preparative TLC and they were identified by GC/MS and 1 H a 13 C NMR analyzes as (+)-canadine, (+)-tetrahydropalmatine and domestine. Isolated alkaloids were tested by spectrophotometric Ellman method for inhibitory activity against human erythrocyte acetylcholinesterase (HuAChE) and plasma butyrylcholinesterase (HuBuChE). In comparison with the refernce substances, from these isolated alkaloids only (+)-canadine showed an relatively high inhibitory activity against HuAChE (12,4 µM). (+)-Tetrahydropalmatine and (+)-domestine had no significant inhibitory activity in this direction, against both HuAChE and HuBuChE. Key words: Corydalis cava, (+)-canadine, (+)-tetrahydropalmatine,...