Drug discovery is reliant on new developments in natural product chemistry as well as advances in chemical synthesis. The interconnectivity and interdependence of natural and synthetic investigation in drug discovery is evident. The chemical exploration reported herein elaborates the relationship between natural product chemistry and chemical synthesis. Of particular interest are chemicals from organisms residing in less accessible environments, particularly Antarctica and endophytic microbial communities. Degradation via reductive ozonolysis of palmerolide A, a macrocyclic polyketide isolated from the Antarctic tunicate Synoicum adareanum, and subsequent synthetic preparation of the resulting polyols (1,2,6-hexanetriol and 1,2,3,6-hexanetetraol) led to a revision in the absolute configuration of the bioactive natural product (7R, 10R, 11R to 7S, 10S, 11S). A partial synthesis of palmerolide A (C3-14) was completed using Grubb’s 2nd generation catalyst to couple fragments formed using the previously developed methodology from the degradation study. Isolation of indole-pyrimidine containing alkaloids meridianins A, B, C, and E from the Antarctic tunicate Synoicum sp. prompted a synthetic investigation of psammopemmin A, a related alkaloid from the Antarctic sponge Psammopemma sp. resulting in reassignment of the structure of psammopemmin A to that of meridianin A. Both meridianin A and psammopemmin A were synthesized through a Suzuki coupling of the same 4-indolol nucleophile to the apposite pyrimidine electrophile. Several synthetic 3-pyrimidylindole analogs were also prepared and investigated for central nervous system, antimalarial, and cytotoxic activity. Chemical investigation of extracts from mangrove fungal endophytes that displayed antimalarial properties in vitro resulted in the isolation of several potent but cytotoxic and cytostatic compounds: cytochalasin D, roridin E, and 12,13-deoxyroridin E.

published_or_final_version / Chemistry / Doctoral / Doctor of Philosophy

published_or_final_version / Chemistry / Doctoral / Doctor of Philosophy

The use of homogeneous organometallic catalysis for otherwise challenging chemical transformations is a concept that has gained significant interest in recent decades, providing access to a variety of useful chemical products. The catalytic reactivity of transition metals and non-reactive ancillary ligands that bind to the metal center has played an important role in such methods, with notable breakthroughs being Nobel Prize-winning reactions (palladium-catalyzed C-C cross-coupling, 2010). The research compiled in the thesis further develops the themes of ligand design and catalytic applications currently studied in the Stradiotto group. Key ideas throughout the thesis are to establish an understanding of the palladium/Mor-DalPhos catalyst system in ammonia arylation with respect to mechanism and substrate scope, and to expand the reactivity profile of the DalPhos ligand set to more challenging C-N and related cross-coupling processes. The first section describes an examination of the [Pd(cinnamyl)Cl] dimer/Mor-DalPhos catalyst system in C-N cross-coupling employing ammonia to better understand the catalyst formation process and to provide a guide for the development of precatalysts for otherwise challenging room-temperature ammonia monoarylations. Oxidative addition complex [(Mor-DalPhos)Pd(Ph)Cl] proved to be the optimal catalyst for arylation of ammonia at room temperature using aryl halides and tosylates. In the second section, ammonia cross-coupling was extended by applying it in the construction of indole frameworks, for the first time, which gave access to NH-indoles directly from ortho-alkynylbromoarenes. The Pd/JosiPhos was the superior catalyst system in comparison to Pd/Mor-DalPhos for this reaction and further stoichiometric studies revealed the reasons for this may be that the bulky arylalkyne ligand induces loss of ammonia from (Mor-DalPhos)Pd catalytic intermediates, and that catalyst inhibition by the alkyne substrate through irreversible metal binding is also a possible factor prior to the oxidative addition step. The reactivity profile of the DalPhos ligand set was successfully expanded in the third section of the thesis to palladium-catalyzed aminocarbonylation of aryl bromides using a pyridine-derived DalPhos variant (Pyr-DalPhos). Several different aryl and some heteroaryl bromides were accommodated in the coupling reaction with ammonia and carbon monoxide as reagents, providing aryl amide products in synthetically useful yields. The methodology described in the final thesis section demonstrated the use of Mor-DalPhos and [Pd(cinnamyl)Cl] dimer mixtures for gaining access to the first examples of ketone alpha-arylation employing aryl methanesulfonates (mesylates) and expanding the scope of amination reactions involving these non-halide aryl electrophiles to primary alkyl amines for the first time. These transformations featured acetone and methylamine as coupling partners, both of which can be difficult substrates to monoarylate but were found to be coupled with ease in this chemistry.

Organoclays modelling the Athabasca oil sands were prepared in heptane and toluene showing indole loading occurring exclusively on the external surface of the clay, via a multilayer adsorption mechanism. Solvent adsorption was minimal. Vermicular microstructures, similar to natural kaolinite were formed. Isotherms were constructed and fitted to the BET equation, giving monolayer quantities (9.28mg) that matched well to the theoretical amount calculated from surface area measurements (8.87mg). Dispersing the organoclays in isopropanol and in toluene left a monolayer equivalent. Using cellulose as a competitive desorption agent in asphaltene based organoclay dispersions achieved complete disaggregation of the dispersed organoclay stacks.13 C CP - MAS NMR, showed up to a 25% increase in desorption for aliphatic and up to 40% increase in desorption for aromatic functionalities of the loaded organic matter. Investigation of other saccharides and modified celluloses as competitive agents is recommended for future work.

Perophoramidine 1 is a halogenated natural product which contains two contiguous quaternary centres within its structure. In this thesis, approaches towards the synthesis of perophoramidine are described. In particular, the synthesis of the tetracyclic core structure and the formation of the quaternary centres have been examined. In Chapter 1, the natural product perophoramidine 1 is introduced and its isolation, structure and biological activity is discussed. The structurally related communesin family of natural products are also introduced before the literature published on both the biosynthesis and laboratory synthesis of perophoramidine 1, is reviewed. Finally the Westwood group's approach towards the synthesis of perophoramidine 1 is introduced with a summary of non-halogenated model system investigations previously carried out within the group being provided. Chapter 2 describes studies towards the synthesis of an appropriately halogenated indolo[2,3-b]quinoline core structure of perophoramidine 1. This then allowed methodology previously developed within the group on model system substrates to be applied to the formation of the first of the two quaternary centres required for the synthesis of perophoramidine 1. Chapter 3 describes the attempted formation of the second quaternary centre using an ester alkylation approach. After initial studies failed to generate the desired quaternary centre, non-halogenated model system studies were carried out in an attempt to develop an alternative approach. In Chapter 4, model system studies were continued with cyclic ether compounds investigated as potential intermediates towards the synthesis of perophoramidine 1. The results obtained in this chapter provided a novel route to the formation of the second quaternary centre and led to a redesigned approach towards perophoramidine 1 being developed. In Chapter 5, this redesigned approach was applied to the halogenated intermediates synthesised in Chapter 1. This led to the formation of the first halogenated intermediate synthesised within the group which contained the two contiguous quaternary centres required for the synthesis of perophoramidine 1.

Magister Scientiae - MSc (Biotechnology) / Endophytic bacteria are known to have an endosymbiotic relationship with plants and provide them with many beneficial properties. These bacteria stimulate plant hormones, provide protection from pathogens and increase nutrient availability in the environment. In this study some of these potential growth factors were tested. Endophytic bacteria have the potential to be of great value for the increase of crop production. They offer a variety of processes that aid in plant growth promotion in an ecofriendly manner. The use of endophytic bacteria provides a cheaper and cleaner approach compared to industrial made fertilizers. They also have potential uses in bioremediation to clean the environment polluted by industrial processes. Endophytes were isolated and showed significant growth improvement. Each isolate displayed different morphologies. Isolates were tested for classical growth promotion mechanisms such as the ability to solubilize phosphate, Indole-3-acetic acid and siderophore production. Inductively Coupled Plasma Optical Emission Spectrometry was performed to measure the effect of the isolates on the plants nutrient profile. The isolates were then tested again while the plants were under heavy metal stress to determine if they were still capable of growth promotion. The plants were then assayed for cell death using Evans blue and biomass was measured to determine the effect of vanadium stress. Inductively Coupled Plasma Optical Emission Spectrometry was performed again to assess the change in nutrient profile while under vanadium stress. / 2021-08-31

Submitted by Irene Nascimento (irene.kessia@ufpe.br) on 2017-05-04T19:42:55Z No. of bitstreams: 2 license_rdf: 1232 bytes, checksum: 66e71c371cc565284e70f40736c94386 (MD5) TESE.ELIZABETH.ALMEIDA.LAFAYETTE..pdf: 5409552 bytes, checksum: e75a96e9227604d98bcc02d9668c2c4a (MD5) / Made available in DSpace on 2017-05-04T19:42:55Z (GMT). No. of bitstreams: 2 license_rdf: 1232 bytes, checksum: 66e71c371cc565284e70f40736c94386 (MD5) TESE.ELIZABETH.ALMEIDA.LAFAYETTE..pdf: 5409552 bytes, checksum: e75a96e9227604d98bcc02d9668c2c4a (MD5) Previous issue date: 2016-05-27 / FACEPE / O DNA é um significativo receptor celular, onde muitos compostos químicos exercem seus efeitos antitumorais. A ligação de drogas ao DNA pode afetar a sua transcrição e a expressão da informação genética nas células, influenciando assim, na inibição do crescimento de células antitumorais, que é a base da concepção de fármacos mais eficazes. Moléculas com núcleos indois e acridínicos são biologicamente ativas e possuem atividades como antitumoral, antituberculose, anti-inflamatória e antiasmática; associado não apenas a interação com o DNA, mas também com proteínas, especialmente algumas enzimas-chaves na proliferação celular, as topoisomerases. Com base nestas informações, este trabalho teve como objetivo a síntese de onze novos derivados indólicos com anéis 3-amino-2-tioxo-tiazolidin-4-ona, 2-tioxo-tiazolidin-4-ona, tiazolidin-4-ona e 2-tioxo-imidazolidin-4-ona condensados como cadeias laterais e a síntese de oito novos derivados acridínicos com diferentes tiosemicarbazidas condensadas. Além disso, objetivou-se verificar a atividade antitumoral, investigar a capacidade de interação o DNA e atividade antitopoisomerase dos novos derivados obtidos. Os derivados foram sintetizados com êxito, com ponto de fusão em faixa de pureza e com estruturas elucidadas e comprovadas por espectrometria de massas (MS), infravermelho (IV), técnicas espectroscópicas unidimensionais de RMN1H e RMN13C, e algumas técnicas espectroscópicas bidimensionais, como COSY1H-1H e HSQC1H-13C. A avaliação antitumoral foi realizada com diferentes linhagens de células cancerígenas, por meio do ensaio antiproliferativo MTT e Sulforrodamina B. A análise da ligação ao DNA foi conduzido através da espectroscopia de absorção e da fluorescência utilizando o brometo de etídio (BE) como sonda fluorescente. Os derivados testados exibiram mudanças em suas propriedades espectroscópicas após a interação com ctDNA (DNA de timo bovino), com efeitos hipocrômico e hipercrômico, além de alterações na forma com desvio para a região do vermelho e do azul. Na série das acridinas testadas, o composto mais ativo no teste antiproliferativo foi o derivado não substituido (LT.26) na porção tiossemicarbazona, e o derivado com o substituinte cloro (LT.27) mostrou-se o mais eficiente na ligação com o DNA. Dentre os derivados indólicos, o composto com o grupamento amino livre e não substituído no anel indólico (SE.01) se destacou com eficácia nas linhagens de leucemias testadas e com o alto valor da constante de ligação ao DNA, Kb de 5.69 x 104, aliado a maior supressão da fluorescência, Ksv = 1.81 x 104, assegurando a participação do grupamento amino na interação com o DNA. O derivado indólico SE.05 apresentou destaque importante como compostos promissor, com atividade antitumoral para uma linhagem de mama testada, por apresentar potência superior ao controle positivo, a doxorrubicina. O derivado bis-indólico (TE.04) sintetizado, apresentou também eficácia nas linhagens de câncer testadas, entretanto com menor capacidade de ligação ao DNA. O derivado indólico SE.01 e TE.04 destaque foram conduzidos a uma avaliação da inibição da enzima topoisomerase I humana, porém mostraram-se ineficientes na inibição até a concentração de 50 μM analisada. Tais resultados mostram que o núcleo indol e acridínico, associados a heterocíclicos tiazolidinicos, imidazolidinicos e tiosemicarbazonas, são promissores como agentes antitumorais, com potencial capacidade de interação com o DNA. / DNA is a major cellular receptor, where many chemical compounds exercise their antitumor effects. The DNA binding drugs may affect its transcription and expression of genetic information in cells, thereby influencing, in inhibiting the growth of antitumor cells, which is the basis for design of more effective drugs. Molecules with indoles and acridines ring are biologically active and have activities such as anticancer, anti-tuberculosis, anti-inflammatory and anti-asthamatic; associated not only the interaction with DNA, but also to proteins, especially some key enzymes in cell proliferation, topoisomerases. Bases on this information, this study had as objective to the synthesis of eleven new indole derivatives with heterocyclic rings 3-amino-2-thioxo-thiazolidin-4-one, 2-thioxo-thiazolidin-4-one, thiazolidin-4-one and 2-thioxo-imidazolidin-4-one condensed as side chains; and synthesis of eight new derivatives acridine with different condensed thiosemicarbazides. Moreover, the objective was to determine the antitumor activity, investigate their ability to interact DNA and anti-topoisomerase activity. The derivative were synthesized successfully, with a melting range in purity and elucidated structures and proven by mass spectrometry (MS), infrared (IR), spectroscopic one-dimensional 1H NMR, 13C NMR, and some dimensional spectroscopic techniques such as COSY1H-1H and HSQC1H-13C. The antitumor evaluation was performed with different cancer cell lines, using the antiproliferative assay MTT and sulforrodamina B. Analysis of DNA binding was executed by absorption spectroscopy and fluorescence using ethidium bromide (EB) as a fluorescent probe. The derivatives tested exhibited changes in their spectroscopic properties, after interacting with ctDNA (DNA calf thymus), with hypochromic and hyperchromic effects and changes in form with deviation to the region of the red and blue. In the series of acridines tested, the most active compound in the antiproliferative test was the unsubstituted derivative (LT.26) in thiosemicarbazone moiety, and acridine derivative with the chloro substituent (LT.27) proved to be the most efficient in binding to DNA. Among the indole derivatives, the compound with the free amino group and unsubstituted in the indole ring (SE.01) was highlighted effectively tested on leucemia lines and the high value of the DNA binding constant, Kb of 5.69 x 104 ally to greater suppression of fluorescence, Ksv of 1.41 x 104, ensuring the participation of the amino group in the interaction with DNA. The indole derivative SE.05 presented significant attention as promising compounds with antitumor activity for a breast line tested by presenting power greater than the positivi control, doxorubicin. The bis-indole derivative synthesized (TE.04) also exhibit efficacy in cancer cell lines tested, however with lower DNA binding capacity. The indole derivatives SE.01 and TE.04 highlighted were conducted an evaluation of the inhibition of the enzyme topoisomerase I human, but proved to be inefficient in inhibition to the concentration of 50 μM analyzed. These results show that the indole and acridine nucleus, associated with thiazolidine and imidazolidine heterocyclic and thiosemicarbazones, are promising as antitumor agents with the potential ability to interact with DNA.

The present investigation has involved a kinetic and mechanistic study of the thermal cyclization of 3-acetoxy-3-(2-pyridyl)-2-methylenepropanoate esters and related compounds to 2-substituted indolizines. Substrates for the kinetic study were prepared via the Baylis-Hillmann reaction of pyridine-2-carboxaldehydes with acrylate esters, acrylonitrile and methyl vinyl ketone. The resulting hydroxy compounds were then acetylated to afford the acetoxy derivatives, thermal cyclization of which gave the corresponding 2-substituted indolizines. The cyclization reactions was followed using 'H NMR spectroscopy and were shown to follow firstorder kinetics. The influence of the various substituents on the observed first-order rate constants has been examined and variable temperature studies have permitted evaluation of activation parameters for the formation of methyl indolizine-2-carboxylate and ethyl indolizine-2-carboxylate. An alternative route to 2-substituted indolizines via halogenated derivatives was explored and several halogenated 2-pyridyl derivatives were synthesised and their thermal cyclization to indolizines was attempted. Novel 5-methylindolizine-2-carboxamides were prepared as part of this investigation and dynamic NMR spectroscopy was used to study internal rotation about the amide N-CO bond in these compounds.

The crystalline compound resulting from thermal cyclization of the Baylis-Hillman product, methyl 3-hydroxy-2-methylene-3-(2-pyridyl)propanoate, has been identified as the indolizine derivative, methyl indolizine-2-carboxylate, and this approach involving the reaction of pyridine-2-carboxaldehydes and acrylate analogues has been established as a general route to 2-substituted indolizines. The ease of cyclization the Baylis-Hillman products to indolizines has been shown to increase by converting the hydroxy group to an acetoxy group, and a range of acetylated Baylis-Hillman products were prepared and cyc1ized to the corresponding 2-substituted indolizines, generally in good overall yield. In the reaction of pyridine-2-carboxaldehyde and methyl vinyl ketone, the intermediate cyclized readily and directly to the corresponding indolizine. One- and two-dimensional ¹H and ¹³C NMR analysis of the 2-substituted indolizine products has permitted complete assignment of all ¹H and ¹³C NMR signals, as well as the measurement of all coupling constants for these compounds. A kinetic and mechanistic study has been conducted on the Baylis-Hillman reaction using ¹H NMR spectroscopy. A range of substrates has been examined and the reaction has been found to be third-order overall. A mechanism involving an addition - elimination sequence is proposed, which fits the kinetic data and accounts for observed substituent effects. Reaction of N,N-dimethylacrylamide with pyridine-2-carboxaldehyde in the presence of the tertiary amine catalyst, DABCO, in chloroform, yielded an unexpected product which has been identified by single crystal X-ray diffraction analysis as 1-(2,2,2-trichloro-1-hydroxyethyl)pyridine. Attempted extension of the general indolizine route to the preparation of chromene systems by reacting salicylaldehyde with methyl acrylate in the presence of DABCO, also led to an unexpected, crystalline material, identified by single crystal X-ray diffraction analysis as the coumarin derivative, 3-[(2-formylphenoxy)methyl]coumarin.A series of chloroquine analogues have been prepared from indolizine-2-carboxylic acid, pyrrolo[I,2-a]quinoline-2-carboxylic acid and imidazo[I,2-a]pyridine-2-carboxylic acid by reaction with suitable amines in the presence of the coupling reagent 1, I' -carbonyldiimidazole. This route has been shown to be a vast improvement on earlier procedures and has provided access to both secondary and tertiary indolizine-2-carboxamides. A range of N,N-dialkylindolizine-2-carboxamides have been prepared by this route, and the influence of substituents on their N-CO rotational energy barriers has been determined using variable temperature ¹H and ¹³C NMR techniques. Intercalation with natural DNA by both chloroquine and the synthesized chloroquine analogues has been examined using UV spectrophotometry, and ¹H and ³¹P NMR spectroscopy. The pyrrolo[I,2-a]quinolines have been shown to be DNA intercalators with binding affinities similar to that of the known antimalarial intercalator, chloroquine. In a preliminary study the synthesis of a short oligonucleotide has been undertaken and changes have been observed in the ¹H and ³¹P NMR spectra of the oligonucleotide on addition of the intercalator, chloroquine.