The ¹HNMR spectra of all possible structural isomers of mono and dihydroxy substituted naphthalenes are measured in trifluoracetic acid and trifluormethanesulfonic acid as well as in their deuterated derivatives. These spectra indicate different protonation/deuteration positions in the compounds studied. Whereas with the relative weak TFA the OH group as most basic group of the substrates is protonated, with the more acidic TFS also aromatic positions with lower basicity are protonated. However, some of these position were indicated only by using deutertated acids. To quantify the degree of proton/deuterium exchange at the aromatic rings, dioxane as proton standard was used. For the OH-protonation a complex between the appropriate naphthol and the acid used, in which a quick proton exchange can be occur, is assumed. Furthermore, the formation of corresponding trifluoroacetates or triflates by reaction of the naphthols with the corresponding acids has been indicated. In some cases, in which the ring positions of protonation are not definitely clear, 2D NOESYNMR experiments have been performed.

No description available.

The use of enzymes as catalysts for chemical transformations has emerged as a “greener” alternative to traditional organic synthesis. An issue to solve though, is that enzymes are designed by nature to catalyze reactions in a living cell and therefore, in many cases, do not meet the requirements of a suitable biocatalyst. By mimicking Darwinian evolution these problems can be addressed in vitro by different types of directed evolution strategies. α-Hydroxy aldehydes and α-hydroxy ketones are important building blocks in the synthesis of natural products, fine chemicals and pharmaceuticals. In this thesis, two alcohol dehydrogenases, FucO and ADH-A, have been studied. Their potentials to serve as useful biocatalysts for the production of these classes of molecules have been investigated, and shown to be good. FucO for its strict regiospecificity towards primary alcohols and that it strongly prefers the S-enantiomer of diol substrates. ADH-A for its regiospecificity towards secondary alcohols, its enantioselectivity and that is has the ability to use a wide variety of bulky substrates. The kinetic mechanisms of these enzymes were investigated using pre-steady state kinetics, product inhibition, kinetic isotope effects and solvent viscosity effects, and in both cases, the rate limiting steps were pin-pointed to conformational changes occurring at the enzyme-nucleotide complex state. These characterizations provide an important foundation for further studies on these two enzymes.   FucO is specialized for activity with small aliphatic substrates but is virtually inactive with aryl-substituted compounds. By the use of iterative saturation mutagenesis, FucO was re-engineered and several enzyme variants active with S-3-phenylpropane-1,2-diol and phenylacetaldehyde were obtained. It was shown that these variants capability to act on larger substrates are mainly due to an enlargement of the active site cavity. Furthermore, several amino acids which are important for catalysis and specificity were identified. Phe254 interacts with aryl-substituted substrates through π-π stacking and may be essential for activity with these larger substrates. One mutation caused a loss in the interactions made between the enzyme and the nucleotide and thereby enhanced the turnover number for the preferred substrate

Edible potatoes originated in the Andes and were brought to Europe in the 16th century. Their introduction spurred both the European population growth and economic development. Being the world's fourth-largest food crop, potatoes continue to shape the global economy and world history. Glycoalkaloids are natural insect deterrents generated by potatoes, and are known for their toxic effects as well as potential medicinal utilities. Demissidine, the aglycone of the primary glycoalkaloids, represents one major Solanum alkaloid. Its unique indolizidine framework presents a challenging synthetic target in organic chemistry. Our synthesis of demissidine starts from readily available epiandrosterone and takes advantage of a Lewis acid-mediated fragmentation of a γ-silyloxy-β-hydroxy-α-diazoester; the D-ring of a diazo ester derivative of epiandrosterone was efficiently ruptured to provide an aldehyde tethered ynoate product. In combination with a subsequent azomethine ylide 1,3-dipolar cycloaddition and a transition metal catalyzed oxidation/reduction, the core indolizidine framework of demissidine was successfully prepared in a stereoselective manner. In addition, the syntheses of two amino acids, 5-methylenepipecolic acid and (5S)-5-methylpipecolic acid were explored; they are used for the installation of the α-oriented C25 methyl group on demissidine. The successful preparation of demissidine was supported by NMR analysis of the synthetic compound in comparison with a natural sample. As an efficient and stereoselective synthesis, our efforts toward demissidine illuminate a strategy to indolizidine frameworks that could be applied in the preparation of other polycyclic amine natural products.

Student Number ; 9901384G - MPharm dissertation - School of Pharmacy and Pharmacology - Faculty of Health Sciences / A lyophilized polymeric wafer system was formulated for the provision of rapid drug release in the oramucosal region. Lyophilization produced a porous sponge-like matrix which allowed simulated saliva to be rapidly imbibed into the hydrophilic structure. This surge of simulated saliva resulted in rapid disintegration of the wafer. Hydroxypropyl cellulose (HPC) was selected as the polymeric platform based on its low gelation potential. Other excipients incorporated into the system were lactose and mannitol as diluents, and glycine as a collapse protectant. A Face Centred Central Composite Design was chosen to establish the significant effects of the independent formulation variables on the physicochemical and physicomechanical properties of the wafer. The formulation variables investigated were, HPC concentration, type of diluent (lactose, mannitol or mixture), concentration of diluent, quantity of glycine and fill volume. An analysis of these variables elucidated the influential factors that may be controlled to form an ‘ideal’ wafer. The concentration of HPC significantly affected the disintegration rate (p=0.003), influx of simulated saliva (p=0.011) and friability (p=0.023). The quantity of diluent present in the system also had significant effect on matrix tolerance (p=0.029) and friability (p=0.032). Statistical optimization was undertaken using stepwise forward and backward regression, and Artificial Neural Networks to predict the ideal combination of the independent variables that would produce an ideal formulation. This wafer was required to produce a matrix disintegration of 3.33%/s, friability of 0.1% loss and maximum matrix resilience. Formulations manufactured with and without model drug, diphenhydramine hydrochloride, reflected no significant differences in their physicomechanical and physicochemical properties. In an attempt to expand the scope of this technology, a preliminary investigation was undertaken to develop a prolonged release wafer system. This was successfully achieved trough the application of crosslinking technology. It was possible to achieve drug released over a period of 6 hours.

A thesis submitted in fulfilment of the requirements for the degree Magister Technologiae (Chemistry) in the Faculty of Applied Sciences, Department of Chemistry, Cape Peninsula University of Technology, 2008 / Green et al. attempted to synthesize linear naphthopyranquinones from a naphthyl dioxolane using a TiCl4 as a catalyst. They managed to synthesise an angular naphthopyran as well as a linear naphthopyran in low yield. They showed that reducing the steric strain at position 1 of the naphthyl dioxolane afforded a low percentage yield of the linear naphthopyran plus an angular one. This thesis describes the synthesis of linear naphthopyrans with an improved percentage yield using TiCl4 as a catalyst. This was achieved by placing a OMe group of less steric hinderance at position 1 and a Br atom at position 4 of a naphthyl dioxolane. The OMe group at position 1 was to allow isomerisation to occur at position 2, and the Br atom was to inhibit isomerisation at position 4, thereby inhibiting the formation of the angular naphthopyran.

Introdução: A acidemia 3-hidroxi-3-metilglutárica é causada pela deficiência da 3-hidroxi-3-metil-glutaril-CoA-liase, uma enzima do metabolismo da leucina, levando ao acúmulo, especialmente, do ácido 3-hidroxi-3-metilglutárico nos tecidos. Estudos sugerem que o estresse oxidativo pode contribuir para os danos neurológicos observados em algumas acidúrias orgânicas. Objetivo: Avaliar parâmetros de estresse oxidativo em pacientes com acidúria 3-hidroxi-3-metilglutárica antes e após o tratamento. Materiais e Métodos: Amostras de sangue e urina foram coletadas de pacientes no momento do diagnóstico e após tratamento com dieta com restrição de proteínas e suplementação de L-carnitina (100mg/kg/dia) e de controles. O TBA, um subproduto final da peroxidação lipídica, foi medido no plasma. A determinação do teor de carbonilas e de grupos sulfidrila, marcadores de dano oxidativo a proteínas, foi realizada no plasma. Para avaliar na urina a oxidação de proteínas, os níveis de di-tirosina foram medidos por autofluorescência. O ensaio da capacidade antioxidante urinária foi realizado utilizando um kit comercial. Os níveis de carnitina livre e isovalerilcarnitina foram analisados em amostras de sangue por espectrometria de massas em tandem usando o método de monitorização de reação múltipla (MRM). A concentração de proteínas foi determinada pelo método de biureto em amostras de plasma usando um kit comercial. Resultados e Discussão: Os resultados demonstraram um aumento significativo nos níveis de isovalerilcarnitina em sangue total, das concentrações plasmáticas de malondialdeído e urinárias de di-tirosina, além de uma redução significativa da capacidade antioxidante urinária e dos níveis sanguíneos de carnitina livre nos pacientes no momento do diagnóstico em relação aos controles. Verificou-se uma diminuição nas concentrações do malondialdeído plasmático e da di-tirosina na urina dos pacientes tratados, o que sugere um efeito de proteção do tratamento sobre a peroxidação de lípidos e do dano oxidativo a proteínas, bem como uma normalização dos níveis de L-carnitina durante o tratamento. Conclusões: Esses resultados permitem sugerir que o estresse oxidativo ocorre em pacientes com acidemia 3-hidroxi-3-metilglutárica e que o tratamento com a dieta restrita de proteína e suplementada com L-carnitina pode oferecer proteção contra o dano oxidativo a biomoléculas. / Introduction: The 3-hydroxy-3-methylglutaric acidemia is caused by the deficiency of 3-hydroxy-3-methyl-glutaryl-CoA lyase, an enzyme of leucine metabolism, leading to accumulation of 3-hydroxy-3-methylglutaric acid in tissues. Studies have suggested that oxidative stress may contribute to the neurological damage observed in some organic acidurias. Objective: Evaluate oxidative stress parameters in patients with 3-hydroxy-3-methylglutaric aciduria patiets before and after treatment. Materials and Methods: Blood and urine samples were collected from patients at diagnosis and after treatment with restricted protein diet and supplemented with L-carnitine (100mg/kg/dia) and from controls. TBA , an end subproduct of lipid peroxidation, was measured in plasma. Determination of carbonyl and sulphydryl content, biomarkers of oxidative damage to proteins, was done in plasma. To assess urine protein oxidation, levels of di-tyrosine were measured by autofluorescence. The assay of antioxidant urinary capacity was performed using a commercial kit. The levels of free carnitine and isovalerylcarnitine were analyzed in blood samples by tandem mass spectrometry using the method of multiple reaction monitoring (MRM). Protein content was determined by the biuret method for plasma samples using a commercial kit. Results and Discussion: The results demonstrated a significant increase of total blood isovalerylcarnitine, malondialdehyde plasma concentrations and di-tyrosine urinary levels and a significant reduction of the urinary antioxidant capacity and free-carnitine blood levels in pacients at diagnosis compared to controls. It was verified a decrease in plasma malondialdehyde concentrations and urinary di-tyrosine levels in treated patients, suggesting a protective effect of the treatment on lipid peroxidation and protein oxidative damage, as well as a normalization of L-carnitine levels during treatment. Conclusions: These results allow to suggest that oxidative stress occurs in 3-hydroxy-3-methyl-glutaryl-CoA lyase deficient patients and treatment with restricted protein diet and L-carnitine may offer protection against oxidative damage.

No description available.

The results of total knee arthroplasty are inferior in younger patients. The challenge today is therefore to develop designs and concepts that will last at least 25 years. This thesis has evaluated the fixation to bone of modern designs of knee prostheses uring RSA analysis. Coating implant surfaces with hydroxy-apatite have proven to enhance fixation to bone. Addition of screws for fixation of the tibial component enhances the fixation, but has negative side effects such as osteolysis around the screws, in turn leading to a higher risk of component loosening. The magnitude and pattern of migration was studied in a randomized study of uncemented tibial implants coated with hydroxy-apatite with and without additional screw fixation in patients younger than 65 years. The uncemented implants migrated initially more than the cemented implants that constituted the control group. Both uncemented groups stabilized at 3 monthes with no further migration, while the cemented implants showed a continuous migration up to the 2 year follow-up, indicating continuous bone resorption at the implant-bone interface, a fact that might lead to an increased risk of late implant loosening. This may not be a problem in older patients, but may have consequences for long-term fixation in younger patients. There was no difference between the two uncemented groups indicating that screws do not improve fixation. Hydroxy-apatite coated knee implants might be well suited for younger patients. Mobile bearing total knee arthroplasty theoretically uncouples the forces at the implant-bone interface, thus improving fixation of the implant to bone. The magnitude and pattern of migration of a cemented mobile bearing knee arthroplasty and a fixed bearing total knee arthroplasty was compared in a randomized study. The results showed that mobile bearings did not improve fixation. Trabecular metal, a new material recently introduced for total knee arthroplasty, has several theoretical advantages. Trabecular metal tibial implants were evaluated in a randomized study in patients younger than 60 years. The implants displayed the typical migration pattern for uncemented implants with greater migration initially followed by early stabilization. The majority of the trabecular metal implants subsided into the bone with no lift-off. Lift-off has the potential of exposing the interface to joint fluid with the potential risk of bone resorption and late loosening, and is commonly seen in metal-backed implants. The finding of absence of lift-off is regarded beneficial for uncemented fixation. Trabecular metal tibial implants might be suited for younger patients. The optimal mode of fixation of the femoral component is yet to be established. Comparing cemented femoral components with uncemented femoral components in a randomized study in patients younger than 60 years revealed no differences of the magnitude or the pattern of migration. Uncemented femoral component seems equally as good as cemented components in younger patients.

The thesis entitled “Total synthesis of bio-active natural products microcarpalide, synargentolide A, jaspine B and anamarine.” demonstrates the utility of chiral pool tartaric acid as the source in the synthesis of bio-active natural products. The thesis was divided into four sections. Section I of the thesis deals with the enantiodivergent synthesis of microcarpalide from tartaric acid. Microcarpalide is a 10-membered lactone of polyketide origin isolated from the fermentation broths of an unidentified endophytic fungi, found to be weekly cytotoxic to mammalian cells and acts as a microfilament discrupting agent. Stereoselective approach for the synthesis of ()-microcarpalide is described from D- and L-tartaric acids, while enantiodivergent approach for the synthesis of both enantiomers is described from L-tartaric acid using ring closing metathesis as the Scheme 2: Enantiodivergent total synthesis of microcarpalide. In section II of the thesis, stereoselective synthesis of synargentolide A is described. Synargentolide A is a polyhydroxy -lactone, isolated from Syncolostemon argenteus, which was founf to exhibit cytotoxic and antitumor properties. Stereoselective synthesis of synargentolide A was accomplished, starting from L-tartaric acid employing, Keck and Brown allylations and ring closing metathesis, as the key steps. Scheme 3: Stereoselective total synthesis of ()-synargentolide A. Section III of the thesis deals with the synthesis of ()-jaspine B. Pachastrissamine (jaspine B), is an anhydrophytoshingosine derivative, isolated from marine sponges Pachastrissa and Jaspis speces. Pachastrissamine was shown to exhibit cytotoxicity (IC 50 0.01 g/mL) against P388, A549, HT29, and MEL28 cell lines. Enantioselective synthesis of jaspine B is accomplished from L-tartaric acid employing, Keck allylation, acid mediated formation of tetrahydrofuran, and olefin cross metathesis as the key reactions. In section IV of the thesis, enantioselective synthesis of ()-anamarine is described. Anamarine is a polyhydroxy -lactone isolated from the flowers and leaves of Peruvian hyptis, possessing cytotoxicity against human tumor cell lines. Enantioselective synthesis of -anamarine is accomplishedelaboration of hitherto unknown -keto phosphonate derived from tartaric acid amide. In an appendix for the thesis, enantiodivergent synthesis for 4-siloxy-pent-2-enone was described. The usefulness of asymmetric aldol reaction is exemplifiedin this section. hydroxy amide synthesized from crotonaldehyde is suitably elaborated to the diene which on RCM yielded 4-silyloxycyclopent-2-enone. Further synthetic modification of this compound afforded the other enantiomer. Scheme 6: Enantiodivergent synthesis of hydroxy cyclopentenones. (For structural formula pl the abstract pdf file)