This thesis describes the isolation and structure elucidation of secondary metabolites formed in static culture from a number of endophytic Xylariaceous fungi. Four Xylaria endophytes isolated from a palm tree in Thailand were surface cultured on an aqueous malt extract-glucose medium. They all produced cytochalasin D, coriloxin, (S)-mellein and (3R,4R)-4-hydroxymellein as the main secondary metabolites suggesting that the four endophytes could be the same species. The endophytic fungus A116 produced cytochalasin D as the main secondary metabolite. Another non-endophytic fungus B315, produced cytochalasin D, (R)-mellein, a mixture of two isomers of 4-hydroxymellein and phloroglucinol. X.62, an endophytic fungus, produced 19,20-epoxycytochalasin C from the mycelium as the main secondary metabolite. The fungus Engleromyces sinensis produced engleromycin acetate as the main secondary metabolite. Fungus X. polymorpha produced (3E)-4-(3'-acetyl-2',6'-dihydroxy-5'-methylphenyl)-2-methoxybut-3-enoic acid.

Title: Light platinum group metal complexes with a ferrocene N-phosphinoamide Author: Bc. Michal Navrátil Department: Department of Inorganic chemistry Supervisor: prof. RNDr. Petr Štěpnička, Ph.D., DSc. Abstract: The aim of this diploma thesis is the preparation of coordination compounds containing ferrocene phosphinoamide FcCONHPPh2 (1, Fc = ferrocenyl), whose preparation was discussed in author's bachelor thesis. Twelve new complexes are described in this thesis, including their characterisation by NMR and infrared spectroscopy, elemental analysis and mass spectrometry. In all cases the crystal structure was determined. Two palladium(II), one rhodium(III) and one ruthenium(II) precursors were used for the preparation of the complexes with each providing three new compounds. A reaction of the precursors with phosphinoamide 1 yielded complexes, in which the phosphinoamide was P-coordinated. This compound was a precursor for the other two complexes. The first one was obtained by a reaction with silver(I) perchlorate producing a cationic P,O-chelate. The other was obtained by a reaction with potassium tert-butoxide resulting in neutral P,O- chelate. All twelve complexes were prepared with optimised yields. Keywords: ferrocene, amides, phosphines, palladium, rhodium, ruthenium, structure elucidation.

Structural and mechanistic complexities of copper-dioxygen systems have attracted much attention in the field of bioinorganic chemistry, both in model systems and trapped protein intermediates. The research presented herein is focused on model and naturally occurring metallopeptide systems, from its design to catalysis. Copper is used as the coordinating metal ion, with cobalt and zinc as probes for metal binding. The bioinorganic chemistry of copper proteins and its coordination and spectroscopic properties are briefly discussed in chapter 1. The next two chapters are centered on the de novo design of a minimalistic metallopeptide system with an amino acid sequence of RHHPPHHE. Structural characterization of the peptide by means of CD and NMR spectroscopy techniques are presented in chapter 2, suggesting a characteristic beta-turn structure in its apo and di-metal bound form. The designed metallopeptide exhibits catecholase activity, which is presented in chapter 3. The data suggest the presence of two mononuclear copper active sites, exhibiting specificity towards the oxidation of catecholamine substrates. Similarly, the catecholase activity has been previously observed in copper complexes of Alzheimer's disease related peptide beta-amyloid, exhibiting metal-centered redox chemistry. The metallo-(beta-amyloid); complexes are the hallmark Alzheimer's disease and have been attributed to the generation of reactive oxygen species causing oxidative stress. Thus, inhibition of the observed oxidative activities was investigated. Probing the role of phosphate moieties in various compounds as potential inhibitors against the induced oxidative stress is presented in chapter 4. The phosphate analogs of the studied compounds exhibit more pronounced potency, where mutation of the beta amyloid peptide at Arg-5 and Lys-16 give insight into the interactions of the side chains of Arg and Lys with the phosphate moiety. 31P NMR relaxation studies further support the binding/interaction of phosphate with the Cu(II)-(beta-amyloid); complexes. The correlation of phosphate moiety binding/activity will allow for the design of more potent inhibitors toward the Cu(II)-(beta-amyloid); induced oxidative stress.

From the roots of the African plant Bulbine frutescens (Asphodelaceae), two unprecedented novel dimeric phenylanthraquinones, named joziknipholones A and B, possessing axial and centrochirality, were isolated, together with six known compounds. Structural elucidation of the new metabolites was achieved by spectroscopic and chiroptical methods, by reductive cleavage of the central bond between the monomeric phenylanthraquinone and -anthrone portions with sodium dithionite, and by quantum chemical CD calculations. Based on the recently revised absolute axial configuration of the parent phenylanthraquinones, knipholone and knipholone anthrone, the new dimers were attributed to possess the P-configuration (i.e., with the acetyl portions below the anthraquinone plane) at both axes in the case of joziknipholone A, whereas in joziknipholone B, the knipholone part was found to be M-configured. Joziknipholones A and B are active against the chloroquine resistant strain K1 of the malaria pathogen, Plasmodium falciparum, and show moderate activity against murine leukemic lymphoma L5178y cells.

A new synthetic route towards methylene-spaced hybrid phosphinoferrocene ligands was developed, making use of a phosphinoferrocene betaine 3-(N,N,N-{[1'-(diphenylphosphino)- ferrocenyl]methyl}dimethylammonium)propane-1-sulfonate (2), that is readily accessible from 1-(diphenylphosphino)-1'-(N,N-dimethylaminomethyl)ferrocene (XIX) via treatment with 1,3-propanesultone. The betaine 2 was reacted with sodium alkylsulfinates (alkyl = methyl, phenyl, 4-tolyl) to afford the corresponding sulfones 4. The coordination chemistry of [1'-(diphenylphosphino)ferrocenyl]methyl(methyl)sulfone (4a) was investigated in its zinc(II) complexes. Surprisingly, only insoluble mixed-metal coordination polymers of the general formula [MZnX3(4a)(MeOH)]n (M/X = Na/Br, K/Br, Na/I) and six-centre complex [Li2Zn2Br6(4a)2(MeOH)4(H2O)] could be obtained. Two isomeric homologues of 1'-(diphenylphosphino)ferrocene-1-carboxylic acid (Hdpf, II), viz. 1'-(diphenylphosphino)ferrocenylacetic acid (HIIa) and 1'- [(diphenylphosphino)methyl]ferrocene-1-carboxylic acid (HIIb), were prepared and their coordination preferences were assessed in their palladium(II) complexes. The palladium(II) complexes were prepared by reacting various (acetylacetonate)palladium(II) precursors with these acids with concomitant release of acetylacetone....

In our previous publication of Thiele cage diols, we describe the first pinacol rearrangement to occur over a C(sp3)–C(sp3). Two mechanisms were initially proposed: a concerted mechanism and a stepwise mechanism, proceeding through a carbocation intermediate. Interestingly, the rearrangement provides only a single diastereomer. The aim of this thesis is to investigate the nature of the reaction by measuring the relative rates of reaction with varying substituents that either stabilize or impede the formation of carbocations. From the relative rates, we can narrow in on whether the mechanism is stepwise or concerted, based on substituent effects, and determine how the reaction may provide a single diastereomer. In Chapter 1, the pinacol rearrangement is introduced, and each analogue that followed after, including stereoselective pinacol rearrangements and a series of different semipinacol rearrangements that provide useful synthetic pathways for chemists with desirable stereochemical outcomes. In Chapter 2, we describe the isolation and characterization of two analogues of a key side product. The structure of each analogue was determined through a series of spectroscopic techniques including 1H-1H COSY, HSQC, HMBC, and 1D-selective gradient NOE NMR. From the solved structures, we proposed a possible mechanism to describe their formation during the main rearrangement reaction – one that shares a carbocation intermediate with the stepwise mechanism already proposed. In Chapter 3, we prepared five Thiele cage diol analogues with aryl substituents with different electronic properties: two substituents that stabilize carbocation intermediates (p-OCH3 and p-CH3), two that destabilize (p-F and 3,5-diOCH3), and the base tetraphenyl Thiele cage diol. We measured the rates of reaction of each diol with p-toluenesulfonic acid at 25°C, 35°C, 45°C, and 52°C via variable temperature quantitative 1H-NMR over time. From the rates of reaction, we found that diols with carbocation-stabilizing aryl substituents reacted faster than the destabilizing analogues, providing evidence that the rearrangement proceeds through a carbocation intermediate. We also found that the diols with electron-deficient aryl substituents showed an increase in the entropy of activation with increasing electron-deficiency in the aryl groups, suggesting an associative pathway for the electron-deficient substituents toward the rearrangement product. Considering the pathway proposed for the side product, we propose an updated stepwise mechanism. Based on computational studies and a previously-isolated X-ray crystal structure, we determined that the diastereoselectivity of the reaction was facilitated by favourable π-π stacking interactions between two aryl substituents. The interaction of the aryl groups twists the geometry of the molecules, placing the migrating aryl substituent in the ideal position for the rearrangement to occur stereoselectively. / Graduate / 2023-10-31

Metaphosphimates are possible precursors for oxonitridophosphates. A new polymorph of trimetaphosphimic acid, in fact an oxonium trimetaphosphimate monohydrate, crystallizes in the monoclinic space group P21/c. It exhibits coin-roll-like stacks of H2(PO2NH)3 􀀀 anions in a rectangular pattern, whereas these form a hexagonal arrangement in the known polymorph. Ring conformation analysis reveals a state between boat and twist conformations, closer to the boat conformation. Temperaturedependent powder diffraction shows decomposition upon heating to ca. 100 °C.

In this project the photodegradation of 3,5-diamino-6-chloro-N-(2-(methylamino)ethyl)pyrazine-2-carboxamide was studied. A hypothetical degradation pattern for the compound was proposed and the aim of the project was to study the formed secondary photodegradants and to, if possible, structure elucidate some of these compounds. In order to do this, the parent compound was photodegraded in two steps, where a primary photodegradant was isolated using semi-preparative supercritical fluid chromatography (SFC) and then further degraded into the secondary photodegradants. The photodegradation was first carried out in aqueous solution, where the parent compound was irradiated in UV-A light of 300-400 nm. This resulted in a primary photodegradant with a molecular ion of m/z = 227, where the chloride in position 6 of the pyrazine group had been replaced by a hydroxyl group. During the large scale photodegradation, prior to the preparative purification, the yield of primary photodegradant was very low due to the photodegradation being dependent on both sample volume and concentration and due to the primary photodegradant also being unstable in aqueous solution at room temperature. Due to the above mentioned difficulties the parent compound was photodegraded in methanol instead of water in order to avoid the freeze-drying process where a lot of the primary photodegradant was lost. This resulted in a primary photodegradant with a molecular ion of m/z = 241, where the chloride had been replaced by a methoxy group instead of a hydroxyl group. This compound was more stable which allowed workup by rotary evaporation, instead of freeze-drying, before the preparative purification. This primary photodegradant was isolated using semi-preparative SFC on a Viridis® BEH Prep OBD TM column (250 x 30 mm, 5 µm) and a Luna HILIC column (250 x 30 mm, 5 µm) with MeOH/NH3 100/1 v/v as organic modifier. About 1.2 mg material was isolated and further photodegradation tests in ordinary water and 18O-water were conducted. Some secondary photodegradants were observed in LC-MS analyses, and their element compositions were proposed by accurate mass results. Fundamental structures for these compounds were proposed. Further structural investigational analyses are needed for confirmation in the future.

Title: Homologous ferrocene phosphines Author: Bc. Petr Vosáhlo Department: Department of Inorganic Chemistry Supervisor: prof. RNDr. Petr Štěpnička, Ph.D., DSc. Abstract: This thesis describes the synthesis and coordination behaviour of ferrocene diphosphines derived from 1,1'-bis(diphenylphosphino)ferrocene (dppf). Dppf is one of the most succesful ferrocene ligands, which can be used in various metal-catalyzed reactions. This study aimed to prepare analogous ligands with one dialkylphosphino substituent and one (diphenylphosphino)methyl group. These ligands were oxidized by KSeCN to afford the corresponding phosphinoselenides. The phosphinoselenides were used to assess sigma-donor abilities by measuring the coupling constant 1 JSeP via 31 P NMR spectroscopy. Lastly, the coordination behaviour of these ligands in palladium complexes was studied. The homologous ligands usually formed a mixtures containing a chelate complex and dimeric species with trans- coordinated ligands. Key words: ferrocene, phosphines, homologous ligands, palladium(II) complexes, structure elucidation.

A química de produtos naturais sempre foi uma fonte importante de novas substâncias e de substâncias bioativas. No mundo moderno, o homem utiliza os produtos naturais para diversos fins: corantes, edulcorantes, essências, defensivos agrícolas e principalmente medicamentos. Com o desenvolvimento das técnicas de isolamento de substâncias, cresceu a necessidade de organizar as informações obtidas e também a criação de meios para a identificação mais rápida das substâncias isoladas. Esta foi uma das necessidades que fez surgir a Quimioinformática. Quimioinformática é uma disciplina que utiliza os métodos da informática para organizar dados químicos, analisar estes dados e gerar novas informações a partir destes dados. Esta ferramenta tem sido utilizada com sucesso em procura por novas drogas (QSAR/QSPR), elucidação estrutural automatizada de substâncias orgânicas e em cálculos e previsão de propriedades físico-químicas de diversas moléculas. Os objetivos do presente trabalho foram o estudo fitoquímico de espécies dos gêneros Dimerostemma e Ichthyothere com o intuito de isolar novas substâncias e o desenvolvimento de técnicas envolvendo quimioinformática com o intuito de auxiliar a elucidação estrutural de produtos naturais. Realizou-se a técnica de microamstragem de tricomas glandulares de diversas espécies pertencentes a gêneros da tribo Heliantheae (Viguiera, Tithonia, Dimerostemma). Através da microamostragem foi possível identificar diversas substâncias presentes nos tricomas glandulares das espécies analisadas. Das duas espécies de Dimerostemma investigadas (D. brasilianum e D. rotundifolium) foi possível identificar dois germacrolidos e dois eudesmanolidos, enquanto que de Ichthyothere terminalis foi possível a identificação de dois melampolidos, todos eles lactonas sesquiterpênicas. Foram treinadas redes neurais artificiais para a realização da identificação dos esqueletos carbônicos de determinadas substâncias a partir dos dados obtidos através dos espectros de RMN 13C, sendo que os resultados obtidos podem ser considerados satisfatórios. Foi desenvolvido um software para efetuar a identificação automática de substâncias através da comparação com uma biblioteca de padrões que possui dados cromatográficos de 51 lactonas sesquiterpênicas. Esse software, chamado de NAPROSYS, também é capaz de fazer comparação de dados de RMN de amostra com dados de RMN presentes em uma biblioteca de dados, tornando possível a identificação imediata de substâncias presentes na biblioteca e também auxiliar a elucidação estrutural de substâncias que não estão nela presentes. Para testar a eficiência do NAPROSYS, o programa foi utilizado com sucesso para identificar LSTs através da microamostragem de tricomas glandulares. A eficiência do NAPROSYS em identificar dados de RMN de substâncias presentes na biblioteca foi testada com substâncias isoladas do gênero Tithonia e Viguiera que possuem substâncias bem descritas na literatura e já isoladas no nosso laboratório, sendo que os resultados apresentados foram excelentes. Criou-se também dois modelos de redes neurais para prever tempos de retenção de lactonas sesquiterpênicas em cromatografia líquida (QSRR) com o objetivo de melhorar o desempenho do NAPROSYS em análises de dados cromatográficos. Os resultados para este caso, embora coerentes, precisam ser melhorados. Neste trabalho concluimos que o uso das técnicas clássicas juntamente com as novas técnicas de Quimoinformática pode se tornar uma ferramenta muito eficaz para a elucidação estrutural e busca de substâncias com determinadas propriedades químicas ou mesmo na bioprospecção de novas substâncias bioativas. / Natural products chemistry has always been an important source for new andbioactive compounds. In modern world, mankind uses natural products to do many tasks: colouring, as essences, as agricultural defensives and many as medicines. Within the development of compound isolation techniques, the need for information organisation has grown. The need for quickly identification of isolated compounds has also grown. This was one of the necessities that made Chemoinformatics emerge. Chemoinformatics is a discipline that uses informatics as a tool to organise, analise and to generate new knowledge from chemical data. This tool has been used with success in automate structure elucidation, drug development (QSAR/QSPR) and to predict chemical-physical data of many molecules. The aims of the present work were the phytochemical study of species of the genera Dimerostemma and Ichthyothere to isolate new compounds, and the development of chemoinformatics techniques to aid natural products structure elucidation. The glandular trichome microsampling was made for diverse species of genera from the tribe Heliantheae (Viguiera, Tithonia, Dimerostemma). Many compounds were identified through glandular trichome microsampling. Two germacrolides and two eudesmanolides were identified from Dimerostemma species (D. brasilianum and D. episcopale), while from Ichthyothere terminalis two melampolides were identified, all of them being sesquiterpene lactones. Artificial Neural Networks were trained to make skeleton identification from data obtained from 13C NMR and the obtained results can be considered satisfactory. A software was developed to make automatic compound identification through the comparation with a compound library that possesses data from 51 STLs. This software is called NAPROSYS is also able to compare the NMR data of the sample with the NMR data stored into a compound library, making the imediate identification of compounds present into library possible and also help the structure elucidation of unknown compounds. To test NAPROSYS\' efficience to identify NMR data of compunds sored into the library was made with compounds isolated from species of Tithonia and Viguiera genera, because these genera has well describe compounds in the literature and that has been isolated in our laboratory, and the obtained results are excellent. Two Artificial Neural Network models were created to predict the retention time of sesquiterpene lactones in liquid cromatography (QSRR) with the aim of improve NAPROSYS performance in cromatographic data analysis. The results for this case, although coherent, can be improved. The conclusion of this work is that the use of classical techniques with the new techniques of chemoinformatics can be a very efficient tool to make structure elucidation, search for compounds with certain chemical properties and even the search for new bioactive compounds.