No description available.

With the incidence of Alzheimer’s disease (AD) growing worldwide and in Canada along with the growing economic and social burdens, the need for more effective therapies becomes of great importance. Since the discovery of AD, a number of proposed theories have arisen to explain the pathophysiology including the i) cholinergic theory, ii) oxidative stress pathways, and iii) metal ion imbalance. The major class of drug therapies to treat AD are cholinesterase inhibitors; however, the “one drug, one target” approach has not proven fruitful and generally becomes ineffective in later stages of disease progression. In this project, we synthesized a library of 1,2,3,4-tetrahydroacridine derivatives (10a-d, 11a-e, 12a-e, and 13a-f) as potential agents to target the cholinergic and oxidative stress pathways of AD. Chapter I provides background information on the role of AChE and BuChE enzymes in AD. Furthermore, this chapter describes the neurotoxicity of reactive oxygen species (ROS) and metals in AD. Chapter II provides a summary of project hypothesis and rationale. Chapter III describes the synthetic details regarding the synthesis of target small molecules. It further describes the principles involved in carrying out biological evaluation such as AChE and BuChE inhibition, antioxidant properties via DPPH stable radical scavenging, iron chelation capacity using ferrozine and in vitro cell viability data in neuroblastoma cells. Chapter IV describes the SAR details on ChE inhibition, antioxidant activities, iron chelation and cell viability profiles and molecular modeling details. A brief conclusion and future directions are included in Chapter V and the final section, Chapter VI provides experimental details for synthetic chemistry including analytical data of synthesized compounds and protocols for biological evaluations. This study identified novel tetrahydroacridine derivatives with nanomolar inhibition of both human AChE and human BuChE enzymes that were more potent relative to the reference agent tacrine. Compound 10d[N-(3,4-dimethoxybenzyl)-1,2,3,4-tetrahydroacridin-9-amine] was identified as a potent inhibitor of BuChE (IC50 = 24.0 nM) and compound 13c [6-chloro-N-(pyridine- 2-ylmethyl)-1,2,3,4-tetrahydroacridin-9-amine] was identified as a potent inhibitor of AChE (IC50 = 95.0 nM) with good inhibition of BuChE (IC50 = 1.61 μM) whereas compound 11e [6-chloro-N-(3,4-dimethoxybenzyl)-1,2,3,4-tetrahydroacridin-9-amine] was identified with an optimum combination of dual AChE and BuChE inhibition (AChE IC50 = 0.9 μM; BuChE IC50= 1.4 μM). In conclusion, our studies provide new insight into the design and development of novel tetrahydroacridine derivatives to target multiple pathological routes of AD.

This work presents three new classes of 6-oxoverdazyl radicals. Each of these classes of radicals bears a different substituent in the 3-position of the verdazyl ring. These classes include N-heteroaromatic monoverdazyls, oligopyridine diradicals, and ferrocenyl-based verdazyl radicals and diradicals. With the exception of the ferrocenyl-based radicals, these verdazyl radicals have been designed to serve as ligands and the direct metal-radical interactions have been explored. The ferrocenyl verdazyls have been designed to investigate the indirect interactions between iron(II) and the covalently linked verdazyl radical. All verdazyl radicals and precursors were fully characterized and the metal-radical magnetic interactions were investigated where structural characterization was available. A series of bidentate verdazyl radical ligands were prepared and the metal-radical magnetic interactions have been investigated. The magnetic susceptibility data for the octahedral complexes indicates that cobalt(II) couples ferromagnetically (Jco_vd = +95 cm-l) and iron(II) antiferromagnetically (JFe_vd = -66 cm-1) to the verdazyl radical. The nature of these interactions appears to be dictated by orbital symmetry and is consistent with previously reported nickel and manganese verdazyl complexes. This work also demonstrates that imidizole-based verdazyl radicals are effective ligands in tetrahedral copper(I) complexes. Oligopyridine-based diradicals have been designed as ligands, but decompose in solution preventing metal complexation. An attempt was made to construct grids as a higher order molecular structure. To this end, a diradical ligand with the topology necessary to form discrete grid architectures was synthesized, but has not yet demonstrated the ability to coordinate to metal ions. Other monoverdazyl radicals that are symmetrically substituted in the N1 and N5 positions were also investigated as potential grid forming ligands. A series of ferrocenyl verdazyl and methylated ferrocenyl verdazyls were prepared to investigate the electronic and magnetic interactions between the ferrocene and verdazyl electrophores. As evidenced by UV-Vis and electrochemical solution measurements, the two electrophores exhibit mutual electronic perturbations. In the case of the methylated ferrocene derivatives the degree and pattern of methylation appears to have a regiospecific influence on the verdazyl electrochemistry. In the solid state Mössbauer data is consistent with a Fe2+ ground state and there is no evidence of Fe3+ at or below room temperature. Weak antiferromagnetic behaviour (|J| < -13 cm-1) was observed within and between pairs of ferrocenyl monoverdazyls in the solid state. In order to investigate the interactions between radicals separated by an organometallic spacer, a ferrocenyl verdazyl diradical was prepared. The ferrocene diradical demonstrated significant differences between the solid state and solution phase. While the ferrocene diradical and ferrocene monoverdazyl exhibited similar solution electronic properties, the magnetic properties were vastly different. In solution the spins associated with the diradical appeared to be weakly coupled, but in the solid state the diradical has been characterized as a strongly coupled antiferromagnetic π-dimer. This is the first example of a verdazyl π-dimer. The π-dimer appears to be diamagnetic with the lower limit of exchange estimated at Jinter ≈ 2000 cm 1.

Supramolecular systems are held together by non-covalent forces, and include systems involving the interaction of small molecules with biomolecules such as DNA and proteins. The inherent reversibility of supramolecular systems means that dynamic processes are important for many of the functions achieved. The first objective of this work was to develop methodology to study the dynamics and binding mechanism of small molecules with DNA. Though there is a great deal known about the thermodynamics of molecules binding to DNA there is much less known about the binding dynamics. Aminoxanthones were chosen as appropriate guest molecules, and their photophysics were examined in a number of solvents, showing that both the singlet and triplet excited states are strongly affected by solvent polarity, with the excited state energies decreasing in polar solvents. Laser flash photolysis experiments for quenching of the triplet excited state by nitrite anions in the absence and presence of DNA allowed a residence time of microseconds to be estimated for these guests. These experiments also showed that this methodology is not widely applicable when studying the binding dynamics of small molecules with DNA, and that other fast kinetic techniques are necessary. Laser temperature jump experiments allow measurement of dynamics in supramolecular systems, while avoiding the problems encountered in laser flash photolysis experiments. A custom-built system was developed as an adaptation of systems described in the literature, with the main difference being a laser optimized for the excitation of water, and the ability to operate across a wide dynamic range. The laser and the detection systems for absorbance and fluorescence were successfully implemented and a number of artifacts were eliminated. The expected temperature jump was obtained and signals were detected by both fluorescence and absorption; the signal-to-noise ratio still needs improvement before systematic studies can be carried out. The second objective of this work was to study bimolecular reactions in proteins. Using biomolecules to effectively modulate reactivity in bimolecular reactions requires knowledge of the reaction mechanism. When the dimerization of 2-anthracenecarboxylate (AC) takes place in serum albumin proteins enantiomeric excess (ee) is obtained for the chiral products. There are a number of binding sites for AC in these proteins and it has been shown that a balance between strength of binding and mobility of the reactants is essential to achieve high ee's. In human serum albumin remarkable ee's of more than 80% are achieved. It is important to differentiate between reactant molecules bound to different binding sites so that the binding sites where reaction preferentially forms one enantiomer can be identified. Steady-state and time-resolved fluorescence studies identified two types of 2-anthracenecarboxylate bound to HSA: one that has a short lifetime and is very protected from the bulk aqueous solution, and one that has a longer lifetime, but is exposed to the bulk solution. Further work using binding site inhibitors allowed identification of a third type of molecule that has a longer lifetime, but is very protected from the bulk solution, which is likely the species responsible for the majority of the observed ee.

The PMR spectra of the two isomers of pyridine-2-aldehyde=2'-pyridyl hydrazone in carbon tetrachloride, dimethyl sulphoxide, and benzene solutions have been studied. Specific associations between PAPHY molecules and solvent molecules have been proposed to account for the observed solvent shifts. Dilution studies have shown that there are PAPHY-PAPHY molecular associations in solution. In the E-isomer these are of the n-donor type involving the lone pair electrons of the ring nitrogen atoms, whereas the Z-isomer association is a dipole-dipole interaction involving the aldehyde ring of different PAPHY molecules. Evidence from long range coupling shows that the uncomplexed E and Z-PAPHY molecules possess a different configuration to the complexed molecules. The chemical shift changes on isomerisation of the E to Z-isomer have been measured and those for the aldehyde ring are consistent with the withdrawal of charge from this ring via the nitrogen atom. Metal complexes of the two isomers of PAPHY have been prepared and their PMR spectra studied. The chemical shifts changes on complex formation have been interpreted in terms of the variation in magnetic anistropy of the ring nitrogen atoms in the ligand molecule and the degree of back donation of d-electrons from the metal atom.

Spin-orbit coupling and the related effects of A-doubling and spin-splitting have been well known to spectroscopists for some considerable time. The importance of these phenomena stems from the advent of radioastronomy and the study of the interstellar medium. Identification of the molecules, and the molecular transitions, in the interstellar dust clouds is necessary for an understanding of the cooling process by which these clouds can contract to form new stars.

No description available.

Simulations were used to assist in both the optimization and experimental support of polymer-supported immobilized homogeneous catalysts. This work is a starting point for using molecular modeling to assist in the design of immobilized homogeneous catalysts, where the broader impact is the use of such catalysts which offer high reactivity and selectivity while also providing improved separability and recyclability over heterogeneous catalysts. ROMP poly(norbornene) was examined because it was hypothesized that one of its isomeric configurations might have a helical conformation like vinylic PNB. Alpha shapes were used to determine the accessibility of these polymers with an approximated catalyst group attached to the backbone. The polymer size, reactant size, catalyst size, and linker length were all varied. The simulations were validated by reproducing the expected trends of a random coil for accessibility across the range of the varied properties. Structural analysis of the final conformations showed that these structures were all random coils. It was found that the assumption that the backbone cyclopentane ring was a non-rotatable bond was invalid, which was most likely the largest contributing factor in the lack of a helical structure. It was also found that increasing the size of the virtual catalyst group caused this polymer to have a regions with a local helical conformation. The backbone cyclopentane ring of ROMP PNB was stiffened by adding a dicarboximide group to the ring. The simulation results showed that the TR configuration produced a broad helical conformation. This helix is broad, so its radius of gyration is indistinguishable from that of an equivalent random coil with less than 100 repeat units. Additionally, accessibility did not properly capture this structural difference, but that was mainly because these simulations were pre-optimized for accessibility by having a long linker length and relatively small polymer dimensions. Co(III)salen catalysts were simulated to determine a way to use simulations to optimize polymer supports for these catalysts. The supports examined were an oligomer synthesized by Jacobsen, poly(cyclooctene) polymerized as a macrocycle, and PCO polymerized as a straight chain polymer. The MMFF94 force field was extended to accommodate cobalt terms based on the ESFF force field, X-ray diffraction structures, and ab initio quantum calculations. In order to compare the supports, the individual catalyst efficiency and the overall catalyst efficiency were combined into a "reaction score." The results showed that the PCO macrocycle was the optimal support in the range of 3-5 repeat units, which was consistent with experimental work.

The mucosa of the small intestine is clearly able to discriminate specific chemical components of ingested meals to stimulate gastrointestinal feedback pathways and reduce further food intake. Luminal carbohydrates delay gastric emptying and initiate satiation, which are mediated by reflexes via the vagus nerve upon activation of vagal afferent endings in the mucosa. Nutrients activate these nerve fibres through intermediary epithelial cells, which release neuromediators upon transduction of luminal signals through the apical membrane. 5-hydroxytryptamine (5-HT) and glucagon-like peptide-1 (GLP-1) are released from enteroendocrine cells in response to luminal carbohydrates and both slow gastric emptying and inhibit food intake via vagal afferent pathways. The molecular mechanisms for carbohydrate detection and transduction leading to 5-HT and GLP-1 release are unknown. However molecules key to transduction of taste by receptor cells in the lingual epithelium are expressed in the gastrointestinal mucosa. The studies in this thesis aimed to investigate 1) the possibility that taste molecules expressed in the intestine form part of the carbohydrate sensing pathway that leads to 5-HT and GLP-1 release, which in turn activate mucosal vagal afferents and 2) to gauge any alterations in taste molecule expression that may relate to adaptation of carbohydrate-induced gastric motility reflexes that occurs in dietary and disease states. Firstly these studies show key taste molecules, including sweet taste receptors T1R2 and T1R3, the Gprotein gustducin (alpha-subunit Gαgust), and the taste transduction channel TRPM5, are expressed in the mouse gastrointestinal mucosa shown by RT-PCR and were further localised to individual epithelial ‘taste’ cells using immunohistochemistry. Quantification of transcript levels by real time RT-PCR revealed the proximal small intestine as the preferential site of sweet taste receptor expression along the gastrointestinal tract. This finding was also confirmed in humans using gastric and intestinal mucosal biopsies obtained at enteroscopy with significantly higher transcript expression levels in the small intestine compared to stomach. In the mouse, double label immunohistochemistry with Gα[subscript]gust antibody, as a marker of intestinal taste cells, was performed using lectin UEA-1, a marker of intestinal brush cells, and 5-HT or GLP-1 to link intestinal taste transduction to 5-HT and GLP-1 release. Results show Gα[subscript]gust is expressed within a subset of all three cell types in the small intestine but predominantly within UEA-1-expressing cells. Although Gα[subscript]gust, 5-HT and GLP-1 are largely expressed in mutually exclusive cells, within the jejunum a portion Gαgust positive cells coexpressed 5-HT or GLP-1. This Indicates a subpopulation of intestinal taste cells may be dedicated to carbohydrate-evoked gastrointestinal reflexes through 5-HT and GLP-1 mediated pathways, however, taste transduction within the small intestine appears to predominantly link to alternate mediators. After nutrient detection at the luminal surface, activation of mucosal afferents by 5-HT released from enterochromaffin cells is well documented, however although vagal afferents express GLP-1 receptors direct activation has not been demonstrated. For this purpose the effects of GLP-1 on gastrointestinal vagal afferents were investigated through single fibre recordings in in vitro tissue preparations. GLP-1 had no effect on the activity of mouse gastroesophageal vagal afferents but a rat duodenal preparation proved too problematic to be able to test GLP-1 specifically on duodenal vagal afferents. Altered gastric motility in response to carbohydrate meals due to prior dietary patterns and diabetes mellitus suggest adaptation in feedback mechanisms. Towards the second aim of this thesis taste molecule expression was quantified in fed and fasted mice by real time RT-PCR and revealed taste gene transcription is altered with the changing luminal environment, specifically transcription of taste genes was significantly decreased after feeding compared to the fasted state. Studies comparing expression in the duodenum of type 2 diabetics and non-diabetic controls show no significant difference in taste transcript levels between the two groups. However taste molecule expression was correlated to blood glucose levels in diabetics suggesting transcription of these signal molecules is adapted to both luminal and systemic carbohydrate levels. Findings in both the mouse and human gastrointestinal tract in terms of intestinal chemosensing are discussed. / http://proxy.library.adelaide.edu.au/login?url= http://library.adelaide.edu.au/cgi-bin/Pwebrecon.cgi?BBID=1363582 / Thesis (Ph.D.) -- University of Adelaide, School of Molecular and Biomedical Sciences, 2009

The mucosa of the small intestine is clearly able to discriminate specific chemical components of ingested meals to stimulate gastrointestinal feedback pathways and reduce further food intake. Luminal carbohydrates delay gastric emptying and initiate satiation, which are mediated by reflexes via the vagus nerve upon activation of vagal afferent endings in the mucosa. Nutrients activate these nerve fibres through intermediary epithelial cells, which release neuromediators upon transduction of luminal signals through the apical membrane. 5-hydroxytryptamine (5-HT) and glucagon-like peptide-1 (GLP-1) are released from enteroendocrine cells in response to luminal carbohydrates and both slow gastric emptying and inhibit food intake via vagal afferent pathways. The molecular mechanisms for carbohydrate detection and transduction leading to 5-HT and GLP-1 release are unknown. However molecules key to transduction of taste by receptor cells in the lingual epithelium are expressed in the gastrointestinal mucosa. The studies in this thesis aimed to investigate 1) the possibility that taste molecules expressed in the intestine form part of the carbohydrate sensing pathway that leads to 5-HT and GLP-1 release, which in turn activate mucosal vagal afferents and 2) to gauge any alterations in taste molecule expression that may relate to adaptation of carbohydrate-induced gastric motility reflexes that occurs in dietary and disease states. Firstly these studies show key taste molecules, including sweet taste receptors T1R2 and T1R3, the Gprotein gustducin (alpha-subunit Gαgust), and the taste transduction channel TRPM5, are expressed in the mouse gastrointestinal mucosa shown by RT-PCR and were further localised to individual epithelial ‘taste’ cells using immunohistochemistry. Quantification of transcript levels by real time RT-PCR revealed the proximal small intestine as the preferential site of sweet taste receptor expression along the gastrointestinal tract. This finding was also confirmed in humans using gastric and intestinal mucosal biopsies obtained at enteroscopy with significantly higher transcript expression levels in the small intestine compared to stomach. In the mouse, double label immunohistochemistry with Gα[subscript]gust antibody, as a marker of intestinal taste cells, was performed using lectin UEA-1, a marker of intestinal brush cells, and 5-HT or GLP-1 to link intestinal taste transduction to 5-HT and GLP-1 release. Results show Gα[subscript]gust is expressed within a subset of all three cell types in the small intestine but predominantly within UEA-1-expressing cells. Although Gα[subscript]gust, 5-HT and GLP-1 are largely expressed in mutually exclusive cells, within the jejunum a portion Gαgust positive cells coexpressed 5-HT or GLP-1. This Indicates a subpopulation of intestinal taste cells may be dedicated to carbohydrate-evoked gastrointestinal reflexes through 5-HT and GLP-1 mediated pathways, however, taste transduction within the small intestine appears to predominantly link to alternate mediators. After nutrient detection at the luminal surface, activation of mucosal afferents by 5-HT released from enterochromaffin cells is well documented, however although vagal afferents express GLP-1 receptors direct activation has not been demonstrated. For this purpose the effects of GLP-1 on gastrointestinal vagal afferents were investigated through single fibre recordings in in vitro tissue preparations. GLP-1 had no effect on the activity of mouse gastroesophageal vagal afferents but a rat duodenal preparation proved too problematic to be able to test GLP-1 specifically on duodenal vagal afferents. Altered gastric motility in response to carbohydrate meals due to prior dietary patterns and diabetes mellitus suggest adaptation in feedback mechanisms. Towards the second aim of this thesis taste molecule expression was quantified in fed and fasted mice by real time RT-PCR and revealed taste gene transcription is altered with the changing luminal environment, specifically transcription of taste genes was significantly decreased after feeding compared to the fasted state. Studies comparing expression in the duodenum of type 2 diabetics and non-diabetic controls show no significant difference in taste transcript levels between the two groups. However taste molecule expression was correlated to blood glucose levels in diabetics suggesting transcription of these signal molecules is adapted to both luminal and systemic carbohydrate levels. Findings in both the mouse and human gastrointestinal tract in terms of intestinal chemosensing are discussed. / http://proxy.library.adelaide.edu.au/login?url= http://library.adelaide.edu.au/cgi-bin/Pwebrecon.cgi?BBID=1363582 / Thesis (Ph.D.) -- University of Adelaide, School of Molecular and Biomedical Sciences, 2009