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Alkaloids of Narcissus 'Dutch Master '(Amaryllidaceae) and their biological activity. I.
Vacková, Lucie ; Chlebek, Jakub (advisor) ; Hošťálková, Anna (referee)
Vacková, L.: Alkaloids Narcissus 'Dutch Master' (Amaryllidaceae) and their biological activity. I. Diploma thesis, Charles University in Prague, Faculty of Pharmacy in Hradec Králové, Department of Pharmaceutical Botany and Ecology, Hradec Králové 2016. From a selected fraction ND-6, which was obtained by column chromatography of an alkaloid extract of Narcissus 'Dutch Master' (preparation of the alkaloid extract and column chromatography was performed by Mgr. Daniela Hulcová within her doctoral thesis), lycorine alakloid O-acetylpluviin was isolated using preparative TLC. Its structure was determined on the basis of MS, NMR analysis, and optical rotation, the obtained data were compared with the literature. The isolated alkaloid was tested on its possibility to inhibit human acetylcholinesterase and butyrylcholinesterase. The activity was expressed as IC 50 values (IC50 AChE = 648.03 ± 53.95 μM, IC50 BChE = 602.50 ± 48.50 μM) and compared with IC50 values of galanthamine, huperzine A and physostigmine. O-acetylpluviine showed a very low inhibitory cholinesterase activity, and so, the alkaloid does not seem to be a suitable cholinesterase inhibitor for potential use in the treatment of Alzheimer's disease. Keywords: Narcissus 'Dutch Master', Amaryllidaceae, lycorine alkaloids, Alzheimer's disease,...
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Alkaloids of Narcissus'Dutch Master ' (Amaryllidaceae) and their biological activity. II.
Dvořáková, Zdeňka ; Chlebek, Jakub (advisor) ; Ločárek, Miroslav (referee)
Dvořáková Zdeňka: Alkaloids of Narcissus 'Dutch Master' (Amaryllidaceae) and their biological activity II. Diploma thesis 2016, Charles University in Prague, Faculty of Pharmacy in Hradec Králové, Department of Pharmaceutical Botany and Ecology. The content of this work was isolation of compounds from the selected fraction ND-6 obtained by column chromatography of Narcissus 'Dutch Master' alkaloid extract. Preparation of extract and its column chromatography was performed by Mrg. Daniela Hulcová as part of her doctoral studies. By the means preparative TLC was from fraction ND- 6 homolycorine type alkaloid (+)-O-methyllycorenine gained. Its structure was determined on the basis NMR, GC-MS analysis and optical rotation. The obtained data were compared with facts in known literature. By the isolated alkaloid was determined its cholinesterase inhibitory activity against acetylcholinesterase and butyrylcholinesterase. Its inhibitory activity was expressed as IC50 (M) and compared with known standards galanthamine, physostigmine, and Huperzine A. This alkaloid is inactive against cholinesterase (IC50 AChE > 1000 M, IC50 BChE > 1000 M). On the basis of gained results, we can evaluate this alkaloid from the point of view of cholinesterase inhibition as potentially unusable in the treatment of AD. Key...
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Study of biological activity of alkaloids isolated from Fumaria officinalis L. (Fumariaceae) II.
Malý, Lukáš ; Chlebek, Jakub (advisor) ; Opletal, Lubomír (referee)
Malý, L.: Study of biological activity of alkaloids isolated from Fumaria officinalis L. (Fumariaceae) II. Diploma Thesis, Charles University in Prague, Faculty of Pharmacy in Hradec Králové, Department of Pharmaceutical Botany and Ecology, Hradec Králové 2014, 49 pp. Obtained diethylether extract of Fumaria officinalis L. was separated to fractions in column chromatography with petrol, chloroform and ethanol. Preparative TLC and crystalisation led to isolation of five alkaloids from fraction. Alkaloids were identified by GC-MS and NMR specters, optical rotation and melting point as protopine, cryptopine, (-)-fumaricine, (+)-fumariline and (+)-parfumidine. Isolated alkaloids were tested for their inhibition activity towards acetyl- and butyrylcholinesterase and towards prolyloligopeptidase. Activities were compared with standards. Natural inhibitor galanthamine showed IC50 AChE 1.710 ± 0.065 µM, IC50 BuChE 42.30 ± 1.30 µM. Best inhibition activity showed protopine (IC50 AChE 345.4 ± 24 µM, IC50 BuChE 239.6 ± 22.3 µM) and cryptopine (IC50 AChE 477.71 ± 47.33 µM, IC50 BuChE 270.82 ± 39.12 µM). The highest prolyloligopeptidase inhibition activity showed (+)-parfumidine with IC50 POP 99.2 µM, which was more active than used natural inhibitor baicaline (IC50 POP 605.9 ± 0.021 µM). Synthetic POP...
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Biological aktivity of secondary plants metabolites V. Alkaloids of Vinca minor L.
Bouz, Lukáš ; Opletal, Lubomír (advisor) ; Cahlíková, Lucie (referee)
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,...
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Study of biological activity of alkaloids isolated from Fumaria officinalis L. (Fumariaceae) I.
Kostelník, Jan ; Chlebek, Jakub (advisor) ; Cahlíková, Lucie (referee)
Kostelník, J.: Study of biological activity of alkaloids isolated from Fumaria officinalis L. (Fumariaceae) I. Diploma thesis, Charles University in Prague,Faculty of Pharmacy in Hradec Králové, Department of Pharmaceutical Botany and Ecology, Hradec Králové 2014, 63 p. The aim of this study was to isolate alkaloids from joined fraction no. 55-67 (A2) obtained from the total alkaloid fraction of extract of Fumaria officinalis L. (Fumariaceae) plant. Using chromatography methods three alkaloids were isolated and then identified by structural analysis (GC-MS, NMR). Three alkaloids were isolated by using common chromagografic methods and then identified by structural analyses optical rotation and melting point as (-)-O- methylfumarophycine, (-)-sinactine a (-)-stylopine. Inhibitory activity of isolated alkaloids was assessed against human erythrocyte acetylcholinesterase, human butyrylcholineesterase and prolyl oligopeptidase. The results were expressed as IC50 values ((-)-stylopine: IC50 AChE and IC50 BuChE > 1000 μM, IC50 POP > 1000 mM; (-)-O-methylfumarophycine: IC50 AChE = 963.10 ± 135.98 µM, IC50 BuChE = 1771.0 ± 380.94 µM, IC50 POP - unmeasured; (-)-sinactine IC50 AChE = 632.0 ± 68.12 µM, IC50 BuChE = 8154.3 ± 981.42 µM, IC50 POP = IC50 POP = 52.9 ± 1.8 µM). None of alkaloids isolated showed...
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Study of biological activity of alkaloids isolated from Fumaria officinalis L. (Fumariaceae) II.
Malý, Lukáš ; Chlebek, Jakub (advisor) ; Opletal, Lubomír (referee)
Malý, L.: Study of biological activity of alkaloids isolated from Fumaria officinalis L. (Fumariaceae) II. Diploma Thesis, Charles University in Prague, Faculty of Pharmacy in Hradec Králové, Department of Pharmaceutical Botany and Ecology, Hradec Králové 2014, 49 pp. Obtained diethylether extract of Fumaria officinalis L. was separated to fractions in column chromatography with petrol, chloroform and ethanol. Preparative TLC and crystalisation led to isolation of five alkaloids from fraction. Alkaloids were identified by GC-MS and NMR specters, optical rotation and melting point as protopine, cryptopine, (-)-fumaricine, (+)-fumariline and (+)-parfumidine. Isolated alkaloids were tested for their inhibition activity towards acetyl- and butyrylcholinesterase and towards prolyloligopeptidase. Activities were compared with standards. Natural inhibitor galanthamine showed IC50 AChE 1.710 ± 0.065 µM, IC50 BuChE 42.30 ± 1.30 µM. Best inhibition activity showed protopine (IC50 AChE 345.4 ± 24 µM, IC50 BuChE 239.6 ± 22.3 µM) and cryptopine (IC50 AChE 477.71 ± 47.33 µM, IC50 BuChE 270.82 ± 39.12 µM). The highest prolyloligopeptidase inhibition activity showed (+)-parfumidine with IC50 POP 99.2 µM, which was more active than used natural inhibitor baicaline (IC50 POP 605.9 ± 0.021 µM). Synthetic POP...
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Comparison of original Ellman's method and its modification using microplate reader
Morávková, Martina ; Macáková, Kateřina (advisor) ; Siatka, Tomáš (referee)
Morávková, Martina. Comparison of original Ellman's method and its modification using microplate reader. Diploma thesis 2015. Charles University in Prague, Faculty of Pharmacy in Hradec Králové, Department of Pharmaceutical Botany and Ecology. Supervisor: Ing. Kateřina Macáková, Ph.D. Key words: Ellman's method, acetylcholinesterase, butyrylcholinesterase, spectrophotometer, microplate reader. Ellman's method is often used procedure for measuring an inhibition of cholinesterases acetylcholinesterase (AChE) and butyrylcholinesterase (BuChE). The principle is based on hydrolysis of acetylthiocholiniodide and butyrylthiocholiniodide to give an appropriate acid and thiocholine. Amount of thiocholine is determined spectrophotometrically by reaction with 5,5'-dithiobis-2-nitrobenzoic acid. Results of measurement from two different devices (spectrophotometer and microplate reader) and at two different temperatures (room temperature and 37 řC) were compared in this thesis. Substances with different amount of inhibition of both enzymes were chosen: allocryptopine, californidine iodide, eserine, galanthamine, huperzine A and tazettine. For californidine iodide (AChE, BuChE), eserine (BuChE) and huperzine A (AChE) were detected statistically significant differences during measurement on different devices and at...
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Study of biological activity of isolated alkaloids from Argemone grandiflora (Papaveraceae)I.
Adamcová, Markéta ; Chlebek, Jakub (advisor) ; Opletal, Lubomír (referee)
Adamcová, M.: Study of biological activity of alkaloids isolated from Argemone grandiflora (Papaveraceae) I. Diploma thesis, Charles University in Prague, Faculty of Pharmacy in Hradec Králové, Department of Pharmaceutical Botany and Ecology, Hradec Králové 2015. The aim of this study was isolation of substances from total diethyl ether alkaloid extract of Argemone grandiflora Sweet, their identification and assessment of their inhibition activity towards acetylcholinesterase, butyrylcholinesterase and prolyl oligopeptidase. Using common chromatografic methods, four alkaloids were isolated, that was identified as (+)-laudanosine, protopine, (-)-argemonine a (-)-platycerine. These substances was tested for their inhibition activity IC50: (+)-laudanosine (IC50 AChE = 617,00 ± 46,55 μM, IC50 BuChE = 644,77 ± 55,52 μM, IC50 POP = not mesured yet); protopine (IC50 AChE = 229,98 ± 21,02 μM, IC50 BuChE = 208,87 ± 17,67 μM, IC50 POP ˃ 1000 μM); (-)-argemonine (IC50 AChE = 4677,75 ± 1241,08 μM, IC50 BuChE = 885,45 ± 119,50 μM, IC50 POP = 337 ± 83,1 μM); (-)-platycerine (IC50 AChE = 223,65 ± 19,61 μM, IC50 BuChE = 1651,25 ± 327,7 μM, IC50 POP = 687 ± 74 μM). In comparison with the standards galanthamine (IC50 AChE = 1,710 ± 0,065 μM, IC50 BuChE = 42,30 ± 1,30 μM) and huperzine A (IC50 AChE = 0,033 ± 0,001...
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Biological aktivity of secondary plants metabolites III. Alkaloids of Narcissus tazetta L.
Panchartková, Markéta ; Ločárek, Miroslav (advisor) ; Opletal, Lubomír (referee)
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,...
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Biological aktivity of secondary plants metabolites I. Alkaloids of Narcissus jonquilla L.
Nováková, Dana ; Ločárek, Miroslav (advisor) ; Chlebek, Jakub (referee)
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,...
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