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,...

Author: Petra Nekolná Title: Alkaloids of family Amaryllidaceae: genus Lycoris Diploma thesis Charles University in Prague, Faculty of Pharmacy in Hradec Králové, Department of Pharmaceutical Botany and Ecology 2015, 79 p. The aim of this diploma thesis was to summarize the findings about alkaloids which were isolated from Lycoris plants of Amaryllidaceae family. It contains a botanical characteristics of species of genus Lycoris which were studied phytochemically, a file of alkaloids which were isolated from these plants and findings about the biological activity of these compounds. Within the genus Lycoris 11 species were studied phytochemically and 118 alkaloids were isolated from these plants. Alkaloids which were isolated from Lycoris plants are divided in several structural groups. The lycorine-, homolycorine-, crinine-, galanthamine- and pancratistatine-type alkaloids occur the most numerously. Anticancer, acetylcholinesterase-inhibitory and antimalarial activity of the alkaloids were described. The biological activity of alkaloids is connected with their structure. The most significant anticancer activity was observed in alkaloids from lycorine-, crinine- and pancratistatine-type. Acetylcholinesterase-inhibitory activity was pronounced the most in galanthamine-type alkaloids. The most...

Charles University in Prague Faculty of Pharmacy in Hradec Králové Department of Pharmaceutical Botany and Ecology Candidate: Zuzana Kavková Supervisor: doc. Ing. Lucie Cahlíková, Ph.D. Title of diploma thesis: Alkaloids of family Amaryllidaceae and their analogues as potential drugs The object of this diploma thesis was to prepare derivatives of alkaloids of Amaryllidaceae family and to deal with their biological activity. These alkaloids are famous for their antibacterial, antiinfectives, antifungal, antimalarial and inhibitory activity against AChE, BuChE and POP and also for cytotoxic effect against cell lines. In the current studies about anticancer activity it was found that the most active alkaloids are Amaryllidaceae alkaloids of these types: lycorine, crinane and pancratistatine. Their biological activity relates closely with their structure. The changes of different parts of the structure can explain the relationship between structure and activity, and also the importance of their organization which is necessary for starting the activity. Based on this finding were for the experiments chosen alkaloids like haemanthamine, haemanthidine and lycorine. An eleven derivatives were prepared and identified mostly by GC-MS and NMR. These derivatives were tested on a wide spectrum of tumor lines....

Charles University in Prague, Faculty of Pharmacy in Hradci Králové Department of Pharmaceutical Botany and Ecology Lucie Gábrlová Diploma thesis: Alkaloids of family Amaryllidaceae and their biological activity The aim of this diploma thesis was to compile assigned fraction of alkaloidal extract obtained from Chlidanthus fragrans and isolation of at least two alkaloids for testing their biological aktivity. For the processing of the extract and isolation of the alkaloids contained in it was used preparative TLC. The obtained substances were then undegone structure analysis, specifically, there were used EI-MS and NMR methods. Based on the results obtained were isolated substances identified and prepared for screening of their biological activity, which was not part of this diploma thesis anymore. The isolated alkaloids were identified as 6α-hydroxybuphanidrine, crinamidine and crinine. 6α-Hydroxybuphanidrine failed to get enough for biological testing. Inhibitory activity of crinamidine and crinine against erythrocyte AChE and serum BuChE was quite weak. Equally, neither of alkaloids showed stronger ability to inhibit the POP, where for crinamidine was determined IC50 = 0,790 ± 0,062 mM and for crinine IC50 = 1,473 ± 0,122 mM. The measurment of cytotoxic activity has been carried out only with...

Dočekalová L., Iron-chelating activity of selected alkaloids III, Diploma thesis, Charles University in Prague, Faculty of Pharmacy in Hradec Králové, Department of Pharmaceutical botanic and ecology, Hradec Králové, 2016, 60 pp. Iron is important trace element necessary for human organism. When excess it develops free radicals and structures of organism are harmed. It can lead to organ failure. Excess is solved with chelators, the most used is deferoxamine. But his terapeutic regimen is disadvantageous and it has a lot of side effects. That's reason for searching for substances with better features. This paper focuses on studium of iron-chelating activity of selected alkaloids from family Amaryllidaceae. Concretely, it's galanthamine, undulatine, buphanisine, caranine, 1-O- acetylbulbisine, homolycorine, tazettine, galanthamine, chlidanthine, ambelline, haemanthamine, haemanthidine, hamayne, 9-O-demethylgalanthine and lycoramine. As standard for comparison of activity was used deferoxamine. Activity was measured by spectrophotometer. As indicator was used ferrozine. Chelatation was measured for ferrous ionts and for total iron, where hydroxylamine as reduction agent was used. Ferrous ionts was measured also with pH alteration. Highest activity reached deferoxamine in all executed measurements. In...

Production of secondary metabolites in the explant cultures of Trichocereus pachanoi (Cactaceae) Lucie Urbanová Diploma thesis Charles University in Prague, Faculty of Pharmacy in Hradec Králové, Pharmacy Key words: Trichocereus pachanoi, the explant cultures, mescaline, secondary metabolites, prodrugs, elicitation, cinnamic acid, sodium cinnamate, tryptophan, ozone, methylene blue The aim of this thesis was the research of effect of selected precursors and elicitors for secondary metabolite production of mescaline in explant cultures Trichocereus pachanoi. Suspension cultures were cultivated on the Murashige and Skoog medium with addition of the growth regulators 2,4-dichlorophenoxyacetic acid and 6-benzylaminopurine. To observe changes in production these precursors: cinnamic acid, sodium cinnamate and tryptophan, and these elicitors: ozone and methylene blue, were selected. Precursors were prepared and used in concentrations of 50, 100 and 500 mg·l-1. Ozone was added in two time slots, 5 and 10 seconds. Methylene blue has been used in concentrations of 1, 10 and 100 mg·l-1. Cultivation with added compounds lasted for 24 and 168 hours. The content of mescaline was determined by HPLC analysis. A statistically significant effect on the production of mescaline in vitro culture was proved only with...

Since prehistoric times medicinal plants have been an invaluable source of medicinal preparations to mankind due to their respective chemical constituents. The family of the Mesembryanthemaceae is almost entirely endemic to southern Africa, and the subfamily Mesembryanthemoideae is a key source of previous reports of plants with traditional uses, both medicinal and culinary. In this thesis, phytochemical investigations were performed on a representative sample of the Mesembryanthemaceae genera and species, with emphasis on the subfamily Mesembryanthemoideae. The aims of this study were to record all ethnobotanical information on the medicinal and culinary uses of the Mesembryanthemaceae, and to isolate and identify alkaloids from selected taxa (especially those of medicinal interest) with specific emphasis on mesembrane alkaloids A review on the structural classification, biosynthesis and biological properties of the mesembrane alkaloids is presented in this dissertation. Experimentally, extracts of the 67 plants encompassing 15 genera and approximately 26 species which were investigated were prepared using one of two alkaloid extraction procedures, following which the extracts were screened for the presence of alkaloids by thin-layer chromatography. The extracts in which alkaloids were detected were analysed further using gas chromatography/mass spectrometry. Where possible, pure alkaloids were isolated from the crude alkaloid extracts and the pure compounds characterised using 1H and 13C nuclear magnetic resonance spectroscopy. Six of the mesembrane alkaloids as well as the tyrosine-derived non-mesembrane alkaloid hordenine were found to be distributed amongst the various genera and species of the Mesembryanthemoideae. The mesembrane alkaloids detected include mesembrine, mesembrenone, mesembrenol, mesembranol, 4’-Omethylsceletenone and 4,5-dihydro-4’-O-methylsceletenone. Within the genus Sceletium mesembrine, mesembrenone, mesembrenol, mesembranol and hordenine were identified. Mesembrine, mesembrenone, mesembranol and hordenine were also detected in Aridaria. The sceletenone derivatives 4’-Omethylsceletenone and 4,5-dihydro-4’-O-methylsceletenone as well as hordenine were identified as the major alkaloid constituents of the genus Aptenia. No alkaloids were detected in the species of Brownanthus and Prenia investigated in this study. Amongst the genera Mesembryanthemum, Phyllobolus and Psilocaulon, a random distribution of the alkaloid hordenine was observed. During the course of this study, the isolation and characterisation of 4’-O-methylsceletenone and 4,5-dihydro-4’-O-methylsceletenone from both Aptenia cordifolia and Aptenia lancifolia, as well as the non-mesembrane alkaloid hordenine from a number of species, was achieved. The two mesembrane alkaloids have been prepared synthetically, but this is the first report of the presence of the compounds in a natural source. / Prof. F.R. van Heerden

Amphibian alkaloids are attractive targets for synthesis due to their biological activity. An important class of amphibian alkaloids is the 2,5-disubstituted pyrrolidine-based family of compounds. There are many synthetic approaches for the preparation of the trans-2,5- disubstituted pyrrolidines, but methods for the construction of the cis-2,5-pyrrolidines are limited. Therefore, it was desired to develop an enantioselective approach for the preparation of cis-2,5-disubsituted pyrrolidines. (+)-Tropin-2-one derived from cocaine was used as starting material to exploit the inherent stereochemistry for construction of the cis-pyrrolidine ring. This permitted the unequivocal assignment of the absolute configuration of the target pyrrolidine. The structurally simple pyrrolidine alkaloid, 225H, was selected as a target to develop a general synthetic approach. The enantioselective synthesis of 225H was achieved in nine steps and good overall yield. The search for potent cannabinoid receptor partial agonist ligands as potential marijuana addiction therapeutic agents has led to an investigation of the synthesis of diaryl ether hybrid analogues of BAY 59-3074. A series of 2-(3-alkyl-5-hydroxyphenoxy)-6- (trifluoromethyl)benzonitriles, 3-(2-cyano-3-(trifluoromethyl)phenoxy)phenylalkanoates, and (3- (benzyloxy)phenoxy)-6-(trifluoromethyl)benzonitriles were synthesized and evaluated in vitro for CB1 affinity. The olivetol diaryl ether analogue was the most potent ligand of the alkyl series, but the diaryl ester analogues exhibited modest affinity for CB1 receptors. The most potent compound of the series was the 2-(3-(benzyloxy)phenoxy)-6- (trifluoromethyl)benzonitrile.

This thesis describes the development of new thermal and oxidative radical cyclization methodologies and their application to the total syntheses of alkaloids, particularly to bridged diketopiperazine (DKP) alkaloids. A practical solvent free approach to diverse DKPs and quinazolines is described. The methodology proceeds by thermal silica gel mediated deprotection of the Boc protecting group and intramolecular condensation of the resulting free dipeptides and tripeptides. It was applied to the total syntheses of alkaloids glyantrypine and ardeemin. A major part of the thesis concerns with the discovery and applications of novel diketopiperazine derived alkoxyamines. Their propensity to undergo facile thermal C-O bond homolysis to generate captodative DKP radicals and persistent TEMPO radical allowed using them as radical surrogates. The methodology takes advantage of the persistent radical effect (PRE). The methodology based on PRE was applied in an asymmetric approach to the alkaloid asperparaline C. An asymmetric synthesis of a very advanced precursor to asperparaline C, 8- oxoasperparaline C, was accomplished in 11 steps and 15% overall yield. The key steps of the synthesis include a direct oxidative cyclization of DKPs, regioselective furan dearomatization with singlet oxygen and a reductive...

Annona crassiflora é uma espécie nativa do cerrado brasileiro, conhecida pelo uso popular como alimento e na etnofarmacologia para o tratamento de parasitas do couro cabeludo e de transtornos menstruais. Quimicamente, Annonaceae é conhecida pela produção de acetogeninas e alcaloides benzilisoquinolínicos, dentro dos quais o gênero Annona apresenta a maior diversidade de compostos aporfínicos. O objetivo central deste trabalho foi avaliar potenciais atividades biológicas para Annona crassiflora, relacionadas a sua composição alcaloídica. A análise do extrato foliar de A. crassiflora através de técnicas cromatográficas, espectrométricas e por ressonância magnética nuclear levou a detecção de oito compostos nitrogenados, sendo sete deles identificados: anonaina, annoretina, estefalagina e xilopina, já previamente descritos na espécie, litseglutina B e tetrahidropalmatrubina, identificados pela primeira vez no gênero, e um alcaloide aporfínico inédito proposto como crassiflorina, em homenagem a espécie. A partir das amostras obtidas, foram desenvolvidos diversos ensaios de atividades biológicas, sendo testado o potencial fitotóxico, a atividade antimicrobiana frente as bactérias (Escherichia coli, Pseudomonas aeruginosa e Bacillus subtilis), a inibição de atividade enzimática (acetilcolinesterase e transcriptase-reversa do HIV1) e a atividade antiparasitária (amastigotas e tripomastigotas de Trypanosoma cruzi). Somente dois dos alcaloides identificados, estefalagina e xilopina, foram isolados em quantidades suficientes para alguns bioensaios. De maneira geral, a estefalagina apresentou maior efeito inibitório no alongamento dos coleóptilos de trigo, além de melhores resultados nos ensaios de inibição de atividade enzimática (acetilcolinesterase e HIV1-RT). A xilopina foi mais eficiente nos ensaios fitotóxicos com braquiária (Urochloa decumbens) e com o parasita causador da doença de Chagas (Trypanossoma cruzi). Resultados interessantes com a fração enriquecida em alcaloides também foram obtidos para os ensaios fitotóxicos com espécies-alvo (Lactuca sativa e Lycopersicon esculentum) e antimicrobianos. Estes resultados reforçam o grande potencial biológico dos alcaloides produzidos por Annona crassiflora, sendo, a maioria das atividades descritas neste trabalho, inéditas para estas subtâncias / Annona crassiflora is a native species of Brazilian cerrado, used as food for local people and for treatment of scalp parasites and menstrual disorders in folk medicine. Chemically, Annonaceae is known for the production of acetogenins and benzylisoquinoline alkaloids, within which the Annona genus presents the greatest diversity of aporphine compounds. The aim of this work was to evaluate potential biological activities for Annona crassiflora, related to its alkaloid composition. The analysis of the foliar extract of A. crassiflora by chromatographic, spectrometric and nuclear magnetic resonance techniques led to the detection of eight nitrogen compounds. Seven of which were identified: anonaine, annoretine, stephalagine and xylopine, previously described in the species, litseglutine B and tetrahydropalmatrubine, as first report for the genus, and an unpublished aporphic alkaloid proposed as crassiflorine, in honor of the species. From the obtained samples, several biological assays were developed, being tested the phytotoxic potential, the antimicrobial activity against three bacteria (Escherichia coli, Pseudomonas aeruginosa and Bacillus subtilis), the inhibition of enzymatic activity (acetylcholinesterase and transcriptase-reverse of HIV1) and the antiparasitic activity (amastigotes and trypomastigotes of Trypanosoma cruzi). Only two of the identified alkaloids, stephalagine and xylopine, were isolated in sufficient amounts for some bioassays. In general, stephalagine had significant inhibitory effect on wheat coleoptile elongation, as well as strong results in inhibition of enzymatic activity (acetylcholinesterase and HIV1-RT). Xylopine was more efficient in phytotoxic trials with Urochloa decumbens, and with the parasite that causes Chagas disease (Trypanosoma cruzi). Interesting results with an alkaloid enriched fraction were also obtained for the phytotoxic assays with target species (Lactuca sativa and Lycopersicon esculentum) and antimicrobials. These results reinforce the great biological potential of the alkaloids produced by Annona crassiflora. As the best of our knowledge, most of the activities described in this work are unpublished for these compounds