The Medicines Use Review (MUR) service was introduced in the UK in 2005 to improve patients' knowledge and use of medicines. The service, in essence, engages the patient and the pharmacist in a structured, private conversation about the patient's medicines. Pharmacies are permitted to deliver a limited number of annual MURs yet for a number of years service provision remained low. During the period of this investigation, the service attracted substantial controversy. In 2008 the UK Government called for improvements to be made to the 'quality' of service provision, with measurement of tangible patient outcomes a key concern. This thesis set out to investigate the potential value of the MUR service. First, using discourse analysis, this thesis considered the social construction of the MUR through written marketing material and its potential impact on uptake of the service, making suggestions for future situations. Next, based on a retrospective cross-sectional audit of MUR records, a practical tool for selecting patients who might benefit from an MUR consultation was developed and explained. Auditing MUR records was suggested by others as one way of tackling questions around service 'quality'. However, the cross-sectional audit suggested that such records were inadequate for assessing service quality and it is argued that quality measures should be based on the achievement of intended service outcomes. Finally, and relating to patient outcomes, this thesis includes a qualitative investigation of patients' MUR experiences, particularly patient satisfaction, as a measure of quality. In the absence of existing patient satisfaction questionnaires measuring the true dynamics of the MUR interaction, a novel conceptual framework for measuring patient satisfaction with this service was developed and is put forward. The results contained herein can contribute to the development of an intervention for measuring the benefits of the MUR versus usual care in terms of patient outcomes.

Paclitaxel is an important chemotherapeutic agent and is used in the treatment of many solid tumours including ovarian cancer, NSCLC, and breast cancer, with further applications under evaluation in clinical trials. In current practice, the dosage of paclitaxel is calculated according to body-surface area (BSA). However the scientific validity of individualised BSA-based dosing has been doubted for many years. In the case of paclitaxel, alternative dosing strategies such as flat-fixed dosing and dose-banding have been considered. However, prior to the studies reported in this thesis, no evidence was available to support the pharmaceutical and clinical implications of such strategies. This thesis includes a literature review, outlining the role of chemotherapy in the treatment of cancer and a detailed appraisal of paclitaxel from both clinical and pharmaceutical prospective. Robust stability data on ready-to-use preparations are required to support the inclusion of medicines in dose-banding (D-B) schemes. Studies on the physical and chemical stability of paclitaxel infusions are described of drug concentrations relevant to D-B. A clinical and pharmacokinetic study is described which was designed to assess the clinical effect of paclitaxel D-B, and to compare D-B with individualised BSA-based dosing and flat-fixed dosing schemes. This study used area under the plasma concentration-time curve (AUC) to assess effect of dose schedule on exposure of tissues to the drug. This was considered an appropriate surrogate for therapeutic effect and toxicity. Validated methods for the processing and analysis of plasma samples were a pre-requisite for this study and the development and validation of these methods are described in Chapter 3 of this thesis. External factors precluded implementation of the clinical-pharmacokinetic study and therefore a novel ‘ex vivo’ pharmacokinetic model was designed to simulate the clinical PK study. This laboratory simulation was developed and scaled-down from ‘in vivo’ data. This ‘ex vivo’ study suggested there was no significant difference between the D-B dosing and the individualised BSA-based dosing as well as between the flat-fixed dosing and the individualised dosing of paclitaxel on the basis of likely exposure of the tissues to the drug.

The tachykinins are the largest known peptide family containing the highly conserved carboxyl terminal motif FXGLM-NH[sub]2, in which the mammalian tachykinin family consists of Substance P (SP), Neurokinin A (NKA), NKB and Endokinin B (EKB). The tachykinins mediate their effects through three G protein-coupled receptors, neurokinin (NK) 1, 2 and 3 in mammals. Each of the tachykinins can ligand bind with each of the receptors though with varying affinities, and both SP and EKB appear to exhibit a similar pattern of potency, with preference to the NK[sub]1 receptor. Many studies have reported EKB to elicit similar effects to SP; this raises the questions as to why there are two peptides with apparently similar functions. The aim of the present study was to establish whether there are any distinct roles between SP and EKB in the model cell line U251 MG, looking at their effects on growth, cell signalling and their expression patterns after stress-related conditions. In addition their expression patterns and promoter regions were analysed using bioinformatics tools. SP and EKB affected growth of U251 MG cells through the NK[sub]1 receptor. Additionally both peptides elicited activation of the PKC-cRaf-MEK1/2-ERK1/2- p90RSK pathway. Activation of this pathway triggered mRNA expression of MMP- 9. ICAM-1 expression was affected by SP through the NK[sub]1 receptor, though EKB would appear to reduce signalling similar to NK[sub]1 receptor antagonism suggesting EKB may block ICAM-1 signalling pathway. Bioinformatic analysis of expression patterns and promoter regions revealed the tachykinin gene (TAC) 1 may play a role in development and immune system processes more than other peptide genes. TAC4 had a high proportion of stress-related transcription factors, and was the only gene whose expression is predicted not to be restricted to the CNS, which may explain its peripheral expression. Additionally the tachykinin receptor gene (T ACR) 1 has been implicated in cell cycle control, which is in agreement with NK[sub]1 receptors' ability to affect proliferation in U251 MG cells as described above. Expression analysis of the tachykinin peptide and receptor genes in U251 MG cells under stress-related conditions (serum starvation, hypoxia and oxidative stress) revealed TAC4 to be involved with responses to oxidative stress, along with short isoform of NK[sub]1. TACR2 and TACR3 expression were up-regulated during serum starvation and hypoxic conditions, implicating them in adapting to these conditions. This study has revealed many differences between SP and novel endokinin peptides, and has revealed potential distinct roles for TAC4 from TAC1, with TAC4 implicated in a major role during oxidative stress. These tachykinins may play a major role in both cancer progression and strokes or traumatic brain injury (TBI). Targeting these tachykinins may well provide a novel treatment option for both of these conditions.

No description available.

No description available.

This study was undertaken to increase knowledge of the mechanisms of inter- and intracellular signalling in the gastrointestinal tract. Specific aims were: to use cell lines to elucidate factors affecting growth of gastric cells, to investigate the distribution and aspects of function of isoforms of protein kinase C in a gastric cell line and in the rat gastrointestinal tract and to determine the presence and regulation of nitric oxide synthase in gastrointestinal tissues from the rat and in cell lines. The gastric cancer cell line HGT-1 was used to investigate control of growth. Increases in cell number were found to be dependent on the seeding density of the cells. In cells plated at low density insulin, epidermal growth factor and gastrin all increased cell number. Gastrin produced a bell-shaped dose response curve with a maximum activity at 5nM. No effect of gastrin was apparent in cells plated at high density. and isoforms of protein kinase C were found, by immunoblotting procedures, to be widespread in the gastrointestinal tract of the rat, but protein kinase C was confined to the gastric mucosa and gastrointestinal smooth muscle. HGT-1 cells contained protein kinase C and but or were not detected. Preincubation of HGT-1 cells for 24h with 1M phorbol-12,13-dibutyrate down-regulated protein kinase C but not . The inhibition by the activator of protein kinase C, 12-O-tetradecanoylphorbol 13-acetate (TPA) of the histamine-stimulated increase in cAMP in HGT-1 cells was down regulated by phorbol-12,13-dibutyrate. Inhibition of histamine-stimulation of adenylate cyclase by TPA was Ca2+-dependent and inhibited by the addition of an antibody to protein kinase C . A role for protein kinase C in modulating the effect of histamine on adenylate cyclase in HGT-1 cells is suggested.

Despite having been described by the then (2003) Chief Pharmaceutical Officer for England as ·probably the biggest untapped resource for health improvement", the development of the public health function of community pharmacists has been limited. However, devolution of healthcare budgets has led 10 differential rates of development of the public health function in each administration of the UK (England, Scotland, Wales and Northern Ireland). This is measured and reflected upon in this thesis. Two large-scale surveys were conducted, one of key strategic personnel (Directors of Public Health and Chief Pharmacists) in Primary Care Organisations (PCOs) and one of practicing community pharmacists. This research highlights the fact that community pharmacists have developed an individualistic, service-based approach to their engagement with public health that is contrary to the more collective approach adopted by the wider public health movement. The study measures the scope and level of health-improving services through community pharmacy across the UK and shows that the nature of the pharmacy contractor (independent, multiple etc.) may impact on the range and nature of services provided. Survey data also suggest that attitudes towards pharmacy involvement in the public health agenda vary markedly between Directors of Public Health, PCO Chief Pharmacists, and community pharmacists. Furthermore, within the community pharmacist population, attitudes are affected by a wide range of factors including the nature of employment (owner, employee, self-employed) and the type of employing pharmacy (independent, multiple etc.). Implications for policy and areas for further research aimed at maximising the public health function of community pharmacists are suggested.

Many diseases are caused by pathogenic bacteria. A key example of this is sepsis, which is mostly caused by staphylococci and Gram-negative bacteria. In addition, the highly resistant ESKAPE pathogens are responsible for the majority of hospital acquired infections. In order to treat bacterial infections effectively, and to avoid promoting bacterial resistance against antibacterial drugs, the correct agents must be used, for which in turn the detection and identification of pathogenic strains is essential. This research aims to develop selective chromogenic culture media, by introducing new antibacterial agents for the improved selectivity and new chromogenic substrates for selective visualisation of certain bacterial strains. The intention of the major part of this work was to inhibit the growth of commensal bacteria in clinical samples, as they mask the growth of the infection-causing bacteria. New and known compounds were prepared for 3 evaluation as alanine racemase inhibitors. The compounds were tested on a range of clinical pathogenic and non-pathogenic bacterial strains. The molecules developed were based on the amino acid alanine and utilised bioisosteres and other replacements for the carboxylic acid moiety. Key compounds targeted included alanylmethanesulfonamide 27-L, 1-aminoethyl5-oxo-1,2,4-oxadiazole 33-L and 1-aminoethyltetrazole 32a-L. Each compound was tested initially as the alanyl-X dipeptide form. While most of the alanine bioisosteres were known structures, their novel peptide derivatives required synthetic development using both solution and solid phase techniques. The solid phase synthesis of several C-terminal 1aminoethyltetrazole peptides was successfully established by using 2-chlorotrityl chloride resin. The investigation of the antimicrobial activity of the synthesised compounds identified several clinically applicable selective inhibitors. These compounds were shown to provide differentiation between Salmonella and Escherichia coli, or enterococci and streptococci. This work also gave a useful comparison between the different alanine bioisosteres, and showed the importance of di- and oligopeptide permease systems in order to reach sufficient bacterial activity. The microbiological activity of 1- aminoethyltetrazole peptide derivatives was studied in more detail, due to their potential in clinical applications for the diagnosis of food poisoning. In other work, also directed towards the rapid and selective detection and identification of pathogenic bacteria in a clinical environment, new chromogenic substrates were prepared. Each of these compounds contained a chromogen with a phenoxazin-3-one scaffold linked to an amino acid residue. The purpose of the amino acid is to act as a unit recognised and cleaved by specific hydrolytic bacterial enzymes. Upon liberation, electronic differences between the conjugated and free forms of the chromogen resulted in the development of distinct colour changes, which provide the basis of 4 bacterial detection and identification. Synthetic methods have been developed for the efficient and economical production of this series of substrates. After preparation, these compounds were tested against a panel of clinically relevant bacteria. The aim of these substrates was to present an alternative substrate for (N-β-alanyl)-7-amino-1-pentylphenoxazine-3-one 86a, which is applied commercially in chromID® Pseudomonas aeruginosa chromogenic medium designed for the clinical detection of P.aeruginosa. The new substrates are designed to fully explore the chemical space of phenoxazinonebased chromogenic substrates, and to decrease the colour, as substrate 86a causes significant background colour in culture media. The future application of these substrates in chromogenic media resides in their potential to advance the identification of specific pathogenic bacteria and to thus facilitate the treatment of bacterial infections.

Amorphous solid dispersion (ASD) of a poorly soluble active pharmaceutical ingredient (API) in a polymeric matrix is a promising approach to increase the solubility, dissolution rate and hence bioavailability of the API. From an industrial point of view, spray-drying represents the main solvent evaporation process used for the manufacture of solid dispersions. The aim of the present PhD thesis was to evaluate the accuracy of: i) thermodynamic models (e.g. solubility parameter and Flory-Huggins theories), ii) standard screening methodologies (e.g. solvent casting and quench cooling) and iii) novel screening approaches for predicting the miscibility of binary spray-dried solid dispersions (SDSDs) so that the best performing API-polymer systems at adequate drug-loading (DL) can be selected. Two novel approaches for screening improvement, miniaturization and downscaling of regular spray-drying, were investigated and applied to API-polymer systems consisting of models drugs (Ibuprofen, Naproxen, Carbamazepine and Itraconazole) and seven polymers at four DLs. Screened samples were characterized using modulated differential scanning calorimetry (mDSC), X-ray powder diffraction (XRPD), thermo-gravimetric analysis (TGA) and scanning electron microscopy (SEM). Results obtained from miscibility and solid state characterization were compiled into principal components analysis (PCA) in order to qualitatively rank the screening approaches based upon their prediction accuracy. Non-sink dissolutions conditions with regard to the crystalline API were performed to assess the solubility enhancement and the extent of supersaturation of screened samples. The two proposed screening approaches were found to provide a greater accuracy than traditional screening methodologies to predict the miscibility of SDSDs. The limitations of theoretical models and standard screening methods tested are symptomatic of the gap existing between equilibrium solid solubility to kinetic miscibility as well as the importance of the preparation method with regard to ASD properties. Therefore, the main benefits of the novel approaches rely on their capacity to better reproduce the operating mode and process conditions of regular spray-dryer, while minimizing API needs for a production, significantly. The outcome of this work favours the downscaling approach due to its improved ability to ease the transfer from screening phases to manufacturing stage in the selection of adequate polymer and DL. In this regard, a novel three-stage decision protocol that implements spray-drying in a methodical small-scale approach for the development of ASDs during preclinical activities was developed and has successfully replaced all former practices in UCB projects.

Background: Breast cancer remains a prominent cause of mortality and morbidity in the female population. Despite advances in terms of novel compounds, exhibiting chemotherapeutic activity successful delivery remains challenging. Particulate-based system such as Nanostructured Lipid Carriers (NLCs) a new generation carrier allow encapsulation of drug and targeting of desired tissue. Resveratrol (RES) is a chemotherapeutic drug limited by its physiochemical properties (i.e. poor solubility and bioavailability), encapsulation into NLCs is potentially viable solution to the aforementioned issues. Aim: The main aim of the present work is to develop a nanostructured lipid carriers based drug delivery system of RES in order to overcome its physicochemical and pharmacokinetic limitations and impart suitable functionalities for targeting breast cancer cells. Methods: Box-Behnken experimental design (BBD) was used to understand the effect of three independent factors (amount of liquid lipid, amount of drug and surfactant concentration) and interactions between these factors for each of the six liquid lipids on the selected response variables particle size (PS), polydispersity index (PDI), zeta potential (ZP), drug encapsulation efficiency (%EE), and drug loading (%DL). The formulated resveratrol nanostructured lipid carriers (RES-NLCs) were subjected to a series of physicochemical characterization including particles size, zeta potential measurements, in-vitro drug release while the morphology of RES-NLCs was confirmed through various microscopic methods, differential scanning calorimetry, x ray diffraction and Fourier transfer infrared studies was carried out in order to understand the interactions between RES and the components of the formulation. Undertake stability studies for the formulated RES-NLCs, at various storage conditions in order to select the most stable formulation and take it for further functionalization with targeting ligands hyaluronic acid; HA-NLCs, folic acid; FA-NLCs and dual targeting system using both hyaluronic acid and folic acid; HAFANLCs, and optimization of ligand density and quantify the amount of amine groups present on the surface on RES-NLCs. The efficacy of RES-NLCs was demonstrated through various in-vitro cell lines studies carried on three breast cancer cell lines entailed: MCF-7, MCF-10A and MDAMB-231 cells. Finally, the permeability of the formulations was evaluated through Caco-2 monolayer and Caco2/HT29 co-culture cell lines in order to understand the intestinal barrier transport mechanism. Results: The employed Box Behnken design resulted in the formulation of particles which were <100nm in size, PDI <0.3, a negative surface charge (-24), high entrapment efficiency (91-99%) and drug loading of 3-4%, with tuneable characteristics within the design space. Differential scanning calorimetry and x-ray diffraction indicated amorphous nature of the drug in nanoparticles indicating its entrapment. Upon exposure to acidic medium (pH 1.2), <20% drug release was observed in 4 hours, with 100% drug release observed upon the increase of pH to 5 over a period of 24 hours. Upon stability testing of NLCs, nanoparticles formulated with GTO as a liquid lipid showed good stability and therefore was taken forward for further PEGylating and surface modification with three ligands; hyaluronic acid, folic acid and combination of both ligand in order to impart targetability for breast cancer cells. Surface modification drastically reduced drug release to < 2 % at pH 1.2, however at pH 5, drug release time plots indicated slower overall release from the surface modified NLCs. In-vitro cytotoxicity studies showed that RES-NLCs were effective against both non-TNBC; MCF-7 and TNBC; MDAMB-231 breast cancer cell lines. Dual ligand appended nanoparticles showed 2.7 folds higher toxicity in MCF-7 and 3.6 fold higher toxicity in MDAMB-231 cells as compared to RES-NLC-GTO-PEGS40 demonstrating its potential for treatment in the aggressive triple negative cancer. None of the RES-NLCs formulations containing different liquid lipids or their blanks were cytotoxic to healthy MCF-10A cells demonstrating safety of the formulations. All bare, PEGylated and surface modified RES-NLCs showed time dependent cellular uptake on both MCF-7 and MDAMB-231 cell lines. Surface modification lead to 3 fold increase in the cellular uptake confirming the targetability potential of the ligand appended formulations toward overexpressed receptors on the surface of both cancer MCF-7 and MDAMB-231 cells. Upon examination of endocytosis mechanisms when cells were treated with the formulations, it was noted that two mechanisms were prominent. Clathrin-mediated endocytosis was identified as the primary method of endocytosis for all formulations. However, specifically for surface modified NLCs receptor mediated endocytosis was also found to be responsible for uptake of nanoparticles. Bidirectional transport study demonstrated that the permeability was sensitive to the type of liquid lipid incorporated with the highest permeability exhibited for PGML based formulation, when studied using a Caco-2 monolayer. On comparison to Caco-2/HT29 co-culture free resveratrol showed higher permeability when compared to the formulated NLCs. Conclusion: The aforementioned research has demonstrated that resveratrol NLCs are a viable potential product for use in the treatment of breast cancer, exhibiting high versatility and specificity.