This study focused on biochemical and structural characterisation of two lipid binding proteins: human phosphatidylinositol transfer protein (PITP) Nir2 and American hookworm fatty acid and retinol binding protein (FAR) Na-FAR-1. Nir2 is a large multi-domain PITP that has recently been implicated in phosphoinositide signalling, where it was demonstrated to regulate phosphatidylinositol-4,5-bisphosphate [PI(4,5)P2] homeostasis. Nir2 acts by reciprocally transporting phosphatidylinositol (PI) and phosphatidic acid (PA) between the plasma membrane (PM) and the endoplasmic reticulum (ER), which allows PI(4,5)P2 to be re-synthesised at the PM. Upon cell stimulation, Nir2 translocates to the ER-PM contact sites and is believed to associate with the PM by binding to PA via its C-terminal LNS2 domain. Due to the proposed role of LNS2-PA binding in Nir2 targeting, a detailed investigation of the binding mechanism is desirable, which could help to reveal more details about Nir2 function in the cell. Expression screening of the difficult-to-express Nir2 LNS2 domain yielded a highly-expressed construct that was employed for characterisation of LNS2-PA binding. The data suggested that Nir2 LNS2 binds PA in a specific manner, interacts with both the polar and apolar regions of PA and might associate with the membrane via both hydrophobic and polar interactions. Although the structure of the LNS2 domain could not be determined, several assays are proposed for the identification of LNS2-PA interaction inhibitors that could be used as tool compounds in the investigation of Nir2 and its homologs. Na-FAR-1 is a small lipid binding protein secreted by the human hookworm Necator americanus that infects hundreds of millions of people globally. Na-FAR-1 is known to bind a range of lipid ligands including fatty acids, retinoids and phospholipids, and was proposed to play a role in parasite-host interactions by facilitating nutrient uptake or sequestering lipid signalling molecules in the host tissues. The structure of Na-FAR-1 has been determined previously, but the molecular details of ligand binding by Na-FAR-1 remained unclear. In this study, the high-resolution structure of Na-FAR-1 in complex with its natural ligand oleic acid was determined, and the ligand binding sites were mapped. Furthermore, phospholipid binding by Na-FAR-1 was investigated, and resonance assignment of Na-FAR-1 in complex with PA was carried out, which can be used to obtain the structure of the complex. In addition, Na-FAR-1’s interaction with lysophosphatidic acid was demonstrated in vitro. As lysophosphatidic acid is a mediator of inflammation, the interaction might have important biological implications if it also occurs in vivo.

Palladium-catalysis is extremely important in fine chemical synthesis. This thesis looks at the development of new palladium-catalysed carbon–carbon bond formation reactions, with particular attention to forming new bonds to sp3 carbons. The opening chapter of this thesis gives an overview of current methods for palladium-catalysed heterocyclisation, and the methods for incorporating further functionalisation into this process, then focuses on the optimisation and expansion of a new palladium-catalysed carboallylation reaction. The reaction mechanism was demonstrated via a deuterium-labelling study, confirming that the reaction proceeds through an isohypsic mechanism. Chapter 2 begins with a summary of palladium-catalysed isohypsic reactions, and the introduction of the isohypsic–redox sequence. New results are presented on the expansion of this isohypsic–redox sequence to include the oxyallylation–Heck-coupling, and work on the aminoallylation–Grubbs–Wacker oxidation. Chapter 3 commences with an introduction to MIDA boronates, describing their useful properties along with some uses, particularly in step-wise synthesis. The development of a new palladium-catalysed allylation of MIDA boronates is then detailed. Using MIDA boronates to form a new bond to an sp3 carbon for the first time, this was applicable to a range of allyl halides as well as a large number of MIDA boronates containing a range of functionality. Formation of a new sp3–sp3 carbon–carbon bond was explored, as well as an enantioselective allylation. The application of the allylation was demonstrated in the development of a new palladium-catalysed synthesis of Ibuprofen. Experimental procedures and data are summarised in Chapter 4. An appendix containing NMR spectra for new compounds is attached.

A considerable amount of effort has been directed to develop porous materials as drug delivery systems (DDSs) – one of the most promising emerging applications in healthcare, as most anticancer therapeutics have toxic dose dependence due to a lack of tumour selectivity – as their hierarchical porosity can be used to store and release challenging drugs. Among them, Metal-Organic Frameworks (MOFs) – emerging hybrid, highly porous crystalline structures – offer several advantages compared to other available DDS, as they combine desirable features from both organic (biocompatibility, e.g. porous polymers) and inorganic (high loadings, e.g. mesoporous silica) porous materials. MOFs are highly amenable to functionalisation, meaning fine control over their physical properties can be achieved, and thus they have experienced tremendous development during the past decade in many applications. Despite surface engineering being advantageous for diverse fields – in biomedicine, it can both improve stability and dispersion, and provide the possibility of targeted carriers, decreasing the immune system recognition – surface functionalization of MOFs is underdeveloped. The multiple synthetic steps – synthesis, drug loading and surface modification – and the lack of orthogonality between them hinder their industrial manufacturing as DDSs. This thesis focuses on the development of surface functionalisation protocols of Zirconium MOFs, particularly UiO-66, a Zr-terephthalate MOF, the study of their cell internalisation fate and routes and the correlation with their therapeutic activity. During Chapter 1, an introduction to the use of DDSs in anticancer therapy, followed by examples of the most relevant MOFs from a coordination chemistry point of view, is given, in which zirconium MOFs and their synthesis are highlighted. Particular focus is given to the coordination modulation process, in which monodentate modulators are introduced to the MOFs synthesis to compete with the multidentate linkers during nucleation, enhancing properties such as porosity through the induction of defects. Then, the most relevant examples of surface functionalization of Zr MOFs for drug delivery are discussed with respect to the effects on properties such as colloidal dispersion in aqueous solvents, physiological stability, and drug release kinetics. In Chapter 2 different functionalised modulators (i.e p-functionalised benzoic acids, folic acid or biotin) are introduced to UiO-66 synthesis to obtain surface-functionalised UiO-66 with the appropriate size for drug delivery by one-pot synthesis. Full characterisation of the materials shows them to be remarkably porous due to the defects formed when modulators attach to available zirconium positions in the pores and on the surfaces of the MOFs. Furthermore, the use of a carboxylate-containing anticancer metabolic target (dichloroacetic acid, DCA) as a modulator of UiO-66 synthesis is explored, and co-modulated samples, in which both DCA and functionalised modulators are introduced to UiO-66 synthesis, are synthesised and fully characterised, resulting in drug-containing (ca. 20% w/w) surface-functionalised MOFs by one pot syntheses. Importantly, DCA modulation induces a high number of defects, and consequently highly charged nanoparticles which are colloidally stable in aqueous solvents. Particle size control in the DCA modulated synthesis of the UiO family of isoreticular MOFs – including UiO-66 and its bromo, amino and nitro derivatives, and extended structures Zr-Naphthalenedicarboxylate (NDC) and Zr-Biphenyldicarboxylate (BPDC) – is achieved, obtaining ca. 100 nm particles of UiO-66 derivatives and microcrystals of Zr-NDC and Zr-BPDC when ZrCl4 is the metal precursor, and mesoporous < 20 nm UiO-66 derivatives and ca. 200 nm Zr-NDC and Zr-BPDC when ZrOCl2 is used as the metal precursor. The high porosity of the DCA modulated samples, due to DCA attachment to the inner and outer surface at defect sites, allows the loading of a second drug, the well-known anticancer drug 5-fluorouracil (5-FU), into the pores of the isoreticular MOFs to create dual DDSs. Different postsynthetic modes of surface coating, based in both coordination and covalent chemistry, are studied during Chapter 3. The functionalities of the p-functionalised benzoic acid modulators, introduced to UiO-66 structure during Chapter 2, are used to covalently attach short-chain alkanes and long-chain polymers to UiO-66 surface through copper-catalysed azide-alkyne cycloaddition. Exhaustive characterisation confirms that the attachment occurs through covalent chemistry and not through surface adhesion or electrostatic forces. Folic acid and biotin, which are introduced to UiO-66 surface as synthetic modulators during Chapter 2, are also introduced to UiO-66 surface postsynthetically. Colloidal dispersion and stability towards phosphates are investigated and compared to bare MOFs, in order to gain insights into the effect of both surface chemistry and mode of attachment on physical properties. A comprehensive overview of in vitro studies of cellular internalisation of zirconium MOFs is given in Chapter 4, focussing on the relevance of the endocytosis internalisation routes, which are strictly correlated with therapeutic efficacy. The postsynthetic surface functionalisation protocols investigated in Chapter 3 are applied to analogous calcein-loaded UiO-66 samples. Calcein is a fluorescent molecule not able to efficiently cross the cell membrane by itself, and hence serves as an in ideal probe of MOFs cellular internalisation. Its release from bare and poly(ethylene glycol) coated UiO-66 into phosphate buffered saline at pH 7.4 and 5.5, in order to simulate extracellular and intracellular conditions, is found to be pH responsive (more pronounced at 5.5) for all MOFs, but an ideal decrease in calcein release at pH 7.4 occurs only for PEGylated MOFs. Internalisation of calcein-loaded MOFs by HeLa cervical cancer cells is studied by fluorescence assisted cell sorting, highlighting the effects of surface chemistry on endocytosis efficiencies and internalisation mechanisms. A discussion of in vitro studies into anticancer drug delivery from Zr MOFs is provided in Chapter 5, alongside a summary of the therapeutic effects of DCA and approaches to enhance its anticancer efficacy. Experimental assessment of the in vitro anticancer performance towards MCF-7 breast cancer cells of the DCA-containing MOFs of the UiO family of different sizes (ca. 100 nm and <20 nm), synthesised by coordination modulation during Chapter 2, is given. The effect of dual-drug containing MOFs (DCA and 5-FU) is also examined, to investigate the possible synergic effect of the drug combination. Then, the cytoxicity of bare and surface functionalised, DCA-loaded and empty UiO-66 MOFs is studied at first upon incubation with HeLa cells, for which the cellular routes of internalisation were elucidated in Chapter 4. The most promising MOFs are then tested for selective anticancer activity against a series of cancerous and healthy cells lines, and their macrophage uptake and ROS production is also analysed, to determine the effect of surface functionalization. The selective anticancer cytotoxicity of folate-coated MOFs is attributed to a combination of cancer cell targeting and optimal cell internalisation routes. To summarise, the one-pot synthesis of drug-loaded, surface functionalised UiO-66 has been successfully performed, resulting in porous, crystalline MOFs with the appropriate size for drug delivery. The use of a carboxylate-containing anticancer metabolic target as a modulator has been explored for the UiO family of isoreticular MOFs, resulting in well-dispersed nanoMOFs with enhanced anticancer activity, into which a second drug can be loaded, enabling the creation of dual DDSs. / A series of postsynthetic surface modifications are performed, enabling the study of the MOF’s properties (colloidal dispersion, physiological stability and biocompatibility) with respect to their surface chemistry and coating mode, but more importantly providing valuable insights into correlations between surface chemistry, routes of cellular internalisation and therapeutic effect.

The formation and subsequent decomposition of iron and molybdenum carboxylate precursors in a molten solvent made of the corresponding carboxylic acid was investigated for the purposes of iron molybdate catalyst production. These catalysts were tested for their ability to partially oxidise methanol to formaldehyde using gaseous oxygen. The use of oxalic acid was tested first and was found to be successful in production of iron molybdate forming highly active catalysts. These samples were found to be highly sensitive to Fe:Mo ratios with the best in terms of formaldehyde yield were 1:2.2 and 1:1.7 due to the superior mixing of iron and molybdenum on the surface and in the bulk content of the catalyst. Other Fe:Mo ratios tested caused non-homogeneous mixing of the iron and molybdenum resulting in lower formaldehyde yields overall. The oxalate method was found to be highly sensitive to water additions into the oxalate mixture with the formation of large quantities of COx selective Fe2O3 with small water additions, large additions caused a more coprecipitation approach to be observed. The use of malonic acid was tested and was found to outperform the oxalic acid examples. This was due to superior mixing of the iron and molybdenum causing a highly homogeneous composition. Similar Fe:Mo study found that a range of ratios(1:1.5, 1:1.9, 1:2.2 and 1:3.0) achieved very high formaldehyde yields. A calcination study showed that the malonate method was highly sensitive to changes in calcination conditions with both composition and performance altered dependent on conditions. Alterations of iron precursor using the malonate method found significant changes in catalyst composition depending on the anion used. Chloride and nitrate were found to be the best achieving very high yields. Acetate, oxalate, sulphate and phosphate were found to produce less homogeneous samples which negatively affected catalytic performance.

Fluorinated compounds are extremely desirable in several fields of the chemical industry, such as pharmaceuticals, materials and agrochemicals. Consequently, the development of novel methods of fluorination represents a major challenge in contemporary chemistry. Although several breakthroughs have been achieved in this area in recent years, the development of an active, stable, reusable and truly heterogeneous catalyst, able to perform selective C(sp3)-F formation, has not yet been achieved. Accordingly, this thesis explores the applicability of heterogeneous catalysts, to perform novel and selective C(sp3)-F bond formations. Different strategies to achieve C(sp3)-F bond synthesis were explored, including: i) fluorination of preactivated substrates (e.g. carboxylic acids), and ii) direct C(sp3)-H fluorination. Firstly, the applicability of heterogeneous materials, such as AgxO/TiO2 and pure TiO2, were investigated for decarboxylative fluorination reactions (R-COOH → R-F), with (photofluorination) and without the employment of light sources. Following an investigation of the catalytic performances of AgxO/TiO2 and pure TiO2 for fluorination of pre-activated substrates, an alternative route, whereby the applicability of heterogeneous materials for direct C(sp3)-H fluorination, was explored. Although more elegant and desirable, direct C(sp3)-H fluorination represents an immense challenge, mainly due to the scarce substrate reactivity, which typically results in a requirement of harsher reaction conditions. Due to the challenges associated with activation of C(sp3)-H bonds, two different systems to achieve direct C(sp3)-H fluorination were investigated. Firstly, the viability of benzylic fluorinations was explored, as these substrates represent excellent model substrates with slightly activated C-H bonds. Secondly, substrates possessing less active C(sp3)-H bonds, such as cyclooctane, were investigated for direct alkane fluorination.

The Ab peptide is a key molecule in the development of Alzheimer’s Disease - Ab peptides form toxic aggregates in the brain. Density functional theory (DFT), Parametric Model 7 (PM7) and ligand-Field Molecular Mechanics (LFMM) methods have been used to model the interactions of a series of potential therapeutic PtII(ligand) complexes with various fragments of the Ab peptide. LFMM calculations with the AMBER forcefield were used to generate conformations of PtII-Ab6-14 via LowMode MD and results validated against BHandH. While LFMM showed insufficient agreement with DFT, the semi-empirical PM7 method displayed strong geometric and energetic agreement and was used to predict coordination preference and stable conformations of PtII(bipy) and PtII(phen) complexes. These species are shown to restrict the conformational freedom of Ab and the coordination of PtII(phen) is shown to be in agreement with experimental data. Studies of additional PtII(ligand) complexes in this manner revealed distinct preferences in metal binding mode for each of the complexes studied, with varied His Nd and His Ne coordination observed. Analysis of peptide conformations using Ramachandran plots and the STRIDE algorithm indicate that coordination of the PtII(ligand) complexes disrupts existing peptide structure in a ligand-specific fashion, interrupting or translating the turn structure, suggesting that controlling peptide structure and behaviour may be achieved via ligand design. Ligand-field molecular dynamics (LFMD) simulations of PtII(phen) -Ab16 and -Ab42 are compared to those of the metal-free peptides to investigate the influence of the PtII complex on the structure and properties of the peptide. Simulations of Ab16 and PtII(phen)-Ab16 revealed that PtII coordination does not drastically alter peptide size, but increases the occurrence of 3,10-helical conformations while disrupting hydrogen bond and salt bridge networks. Simulation data also highlights the prevalence of p-p stacking interactions between residues Phe4 and His13 with the phenanthroline ligand. Similarly, microsecond timescale simulations of PtII(phen)-Ab42 and Ab42 illustrate profound effects of PtII coordination on peptide structure; while Ab is shown to adopt collapsed conformations, PtII-Ab42 systems assume extended structures. PtII coordination also induces large changes in peptide secondary structure, particularly an increase in helical character throughout the central hydrophobic region of the peptide, considered a potential route to preventing formation of Ab fibrils. The results detailed here provide insight into the coordination of these complexes to the peptide and present a new understanding of the effects of PtII complexes on Ab.

Organic electrosynthesis is recognised as a green enabling methodology to perform reactions in an efficient and straightforward way. Electrons are used as the reagent to form anion and cation radical species from neutral organic molecules achieving oxidations and reductions by replacing toxic and dangerous reagents. Within this field, the use of microreactors in continuous flow is also concurrent with electrochemistry because of its convenient advantages over batch, such as: i) low loading or no supporting electrolyte at all, due to the small distance between electrodes, providing significant advantages in downstream processing; ii) high electrode surface-to-reactor volume ratio; iii) short residence time; iv) improved mixing effect. In this thesis, a novel easy-to-machine flow electrochemical reactor has been designed and fabricated. This reactor can be made either of aluminium or polymers, if additive manufacturing employed. The efficiency of the reactor has been shown in the electrosynthesis of nitrogen-containing heterocycles. The products obtained have been further functionalised in a single two-step flow system connected to the electrochemical reactor: For the electrochemical synthesis of benzothiazoles from N-arylthioamides in flow a catalyst- and supporting electrolyte-free method has been developed. A library of benzothiazoles was synthesised only in the presence of solvent and electricity: Finally, a new synthetic method for the electrochemical formation of benzoxazoles from easily accessible and inexpensive resorcinol and nitriles was discovered: In conclusion, flow electrosynthesis has shown to be a promising tool for electroorganic synthesis, improving the outcome of standard batch cells.

Hydro(solvo)thermal synthesis was used to synthesise a range of oxide, oxyhydroxide and hydroxide materials. Their structures were characterised by a wide variety of diffraction, spectroscopic, and other analytical techniques. Many of the oxides were screened for potential applications in catalysis. High energy X-ray diffraction was used to follow in situ the crystallisation of a cobalt gallium oxide prepared from metallic gallium in ethanolamine. Two transient metastable layered double hydroxide phases were observed prior to the formation of the spinel product when a solvent of 1:1 water:ethanolamine mixture was used. Photocatalytic studies showed that the spinel had activity as a water oxidation catalyst. A different cobalt gallium oxide spinel was prepared by solvothermal reaction using pre-formed γ-Ga2O3 and a solution of Co2+. Its metastable structure was characterised by a variety of techniques including analysis of Bragg and diffuse neutron scattering data. The material was investigated for three-way catalysis. The polymorphism of Ga2-xAlxO3 was investigated and the structures of materials characterised. The solvothermal synthesis of the defect spinel γ-Ga2-xAlxO3 in 2-propanol was optimised for 240 °C with up to 90% of the Ga replaced by Al. A novel oxyhydroxide Ga5-xAlxO7(OH) was prepared by solvothermal reaction in 1,4-butanediol. Solid-state MAS NMR was used to determine the coordination of the metals in these materials. All the materials are found to be metastable, transforming into thermodynamically stable polymorphs above 1400 °C. Ga2O3 and Ga2-xAlxO3 polymorphs were tested as palladium supports in the diesel oxidation reaction and the semi-hydrogenation of acetylene. A novel oxyhydroxide, Ga2.52V2.48O7.31(OH)0.69, was prepared by reaction of Ga metal and Na3VO4 in a 1:1 ethanolamine:water mixture. The structure of this material was investigated by neutron diffraction and found to isostructural with the minerals tohdite and nolanite. The material is metastable, dehydrating around 300 °C to form Ga2.52V2.48O8, and then decomposing above 500 °C. The material was found to show good activity as a catalyst for oxidative propane dehydrogenation. A new chromium substituted γ-Ga2O3 has been prepared and characterised. This material was prepared by solvothermal reaction in 1,4-butanediol and found to be very poorly crystalline. The spinel is stable to 900 °C before phase separating into its respective binary oxides. The hydrothermal reaction of RhCl3·3H2O with AO2 or A(OH)2 (A = Ca, Sr, Ba) in either NaOH or KOH at 200 °C yielded a range of hydroxides. Ca3Rh2(OH)12 and Sr3Rh2(OH)12 are hydrogarnets which can be dehydrated to oxides on heating. A new hydroxide in the system Ca-Na-Rh-OH was synthesised. Reaction with barium salts in NaOH yielded BaNaRh(OH)6 a new hydroxide. The structure of this material was solved by single crystal diffraction and found to contain isolated Rh-OH octahedra, two crystallographically different 8 coordinate Na sites and a 10 coordinate Ba site.

This body of work is concerned with the synthesis and characterisation of new inorganic clusters based on the [Mo2O2S2]2+ dimeric unit. By utilising simple chalcogenide building blocks, we have discovered several collections of new compounds and gained valuable insight into the self-assembly process that governs the formation of inorganic supramolecular materials. Three clusters were successfully discovered and characterised with the tellurite anion, TeO32-, as the only template- the first confirmation that this species is is able to act as a template in this system with no other templating agents present. The template was observed to form five different building blocks with the dimer, which then combined to form the full clusters. While the basic architecture is consistent with precedents that had already been set for this chemistry, none of the building blocks had been observed before. This template was found to have a remarkable symmetry lowering effect on the structures, with the first fully inorganic chiral polyoxothiometalate cluster a part of this set. When the squarate anion, C4O42-, a species already proven to template a vast array of structures in this system, was introduced to add an element of competition between the two templates, another new collection of four compounds was discovered. A valuable discovery here was that squarate was the preferred template, as shown by the fact that most of the building blocks from which these compounds were derived had been observed before in compounds where squarate was the only template present. Over the course of this investigation, it was observed that no variable has a greater influence over the reaction system than pH, and identifying the optimal pH ranges for each compound allowed for the creation of a rudimentary map of the parameter space that governs the self-assembly process With the selenite anion being established as a viable POTM template in previous work, it was also added to the squarate anion so it could compete in a similar manner to tellurite. In addition to this, mononuclear molybdate anions were added to the system as a third potential templating agent in an effort to increase the competition even further. The results of these studies were the discovery of five new compounds, two from the dual-templated reactions, and three compounds from reactions with three templates present. These compounds were based on a common library of three building blocks, one of which was newly discovered with these compounds.

The constant problems encountered in the fields of medicinal and pharmaceutical chemistry, especially those related to the side effects of drug candidates, have risen the concern of developing methods that can help us achieve the therapeutic effect without undesired properties. This thesis describes the development of two different Drug Delivery Systems based on the modification of natural occurring molecules. The first one is directed to the treatment of parasitic tropical diseases. The alteration of pantothenic acid with the introduction of a double bond has proved to increase the uptake of fluorescent labelled molecules in different model systems with low cytotoxicity. The concept of this Drug Delivery System relies on the necessity of the parasites to consume the host’s pantothenic acid for their own biological processes. Due to their inability to synthesise this vitamin along with the huge supply they need to survive, it was hypothesised that the increased uptake of CJ-15,801 would allow us to attach interesting molecules that could be selectively delivered into parasites. The second example of a Drug Delivery System presented in this work is based on peptides released by cells of the immune system. The so called Cationic Amphiphilic Peptides are released by an organism that is under the attack of potential pathogens. Due to their physicochemical properties, they can stop an infection by direct killing of microorganisms by different mechanisms. Either by the membrane disruption or internalisation and intracellular targeting, the presence of positively charged residues play a major role on the activity of these peptides. By substitution of the natural occurring lysine and arginine residues with a new class of phosphonium based amino acids, a new class of cationic amphiphilic peptides was synthesised. Fluorescent versions of these peptides have allowed us to investigate their properties. They are characterised by their ability to cross cellular membranes with relatively low toxicity compared to the natural occurring versions of the sequences and even though their direct antimicrobial activity is diminished they can be used as potential Cell Penetrating Peptides. Finally, due to the nature of the cation present in these new peptides, it is theorised that they can have certain selectivity to deliver drugs into mitochondria. Although further studies to prove this need to be done, an initial experiment is reported at the end of this work.