Pancreatic cancer is the fourth main cancer in the western world. Currently the only chemotherapy available clinically is gemcitabine. However, gemcitabine treatment only proves effective in 23.8% of patients. Nano-structures (< 120 nm) are capable of entering the highly permeable blood capillaries which supply the rapidly growing tumours. Once inside the capillaries they accumulate and are retained in the tumour as a result of the poor lymphatic drainage. This allows for a deeper tissue penetration which is otherwise difficult to achieve. Hybrid nanoparticles with an iron oxide core covered by gold shell (HNPs) have shown great potential for anti-cancer therapies. The magnetic iron oxide cores and the surface plasmon resonance (SPR) properties of the gold surface provide the HNPs with the capabilities of diagnostic imaging and drug delivery, making them true theranostic agents. A novel prodrug of gemcitabine has been developed by a regioselective coupling of gemcitabine and lipoic acid, itself a potent antioxidant. Gemcitabine-N-lipoate (GL) was obtained in a one-pot synthesis and the optimum conditions for the reaction were established. GL prodrug loading on to the HNPs surface was confirmed and the release profile of gemcitabine from the GL-HNPs formulation was studied at pH 3.6, 5.6 and 7.4 utilising different temperature conditions (20, 37, 44 °C) using RPMI serum free media under sink conditions. The data showed the stability of the formulation at pH 7.4, 20 °C while the optimum release conditions for gemcitabine from the GL-HNPs formulation were at pH 5.6, 44 °C with the highest release of 41.1% recorded after 24 hrs. III Preliminary in vitro MTT assay together with the drug uptake study show the superior inhibitory effect of the GL-HNPs formulation on the cell viability over gemcitabine after 24 hrs which indicates faster uptake of the formulation, however the overall effect of gemcitabine is greater after 48 hrs which is mainly due to the slow release of gemcitabine from the formulation. The behaviour of the GL-HNPs formulation as a drug delivery system shows a great potential for the system to act as a theranostic tool and to overcome the significant drawbacks associated with gemcitabine.

A novel method for monitoring of pharmaceutically relevant reactions using PADI-MS has been developed in this study. The visible non-thermal plasma plume from a coaxial helium gas flow non-thermal RF plasma was optimised using a range of samples. PADI-MS was found to be a powerful analytical technique for pharmaceutically relevant solid and liquid samples and can readily be adapted for use in reaction monitoring. This study has determined that PADI-MS is fast, easy to set up and requires little or no sample preparation. PADI-MS has significant advantages over other techniques as it is faster, more sensitive and more convenient and suitable for investigation of complex molecules and mixtures. However, two limitations of PADI-MS are that the plasma can affect the sample surface chemistry upon exposure and quantification is not always trivial. A number of other analytical methods were used in conjunction with PADI-MS: TLC, HPLC, FTIR and Raman spectroscopy. Chapter 3 investigated PADI-MS and Raman analysis of paracetamol tablet as model for pharmaceutically relevant solids. Raman microscopy was used to develop an understanding of how the plasma affects the sample surface. The plasma effect was also studied by examining changes in PADI-MS spectra. In Chapter 4, PADI-MS acquisition settings were improved by adding water vapour to the outer He flow gas and increasing the plasma power to 8 W, optimising the analysis of mixture solutions from TLC plates. Although molecules could be identified with optimum sensitivity after separation, this may not be necessary, unless the highest possible sensitivity is required. Chapter 5 deals with PADI-MS for direct analysis of pharmaceutical compounds in solid and liquid forms from glass slides and cotton swabs. PADI-MS was determined to besuitable for analysis of pharmaceutical tablets from solutions via both substrates, with distinct advantages for the latter. The final Chapter studied PADI-MS for the monitoring of imine formation as model pharmaceutical reaction. Both FTIR and PADI-MS were successfully used, but the latter is faster and more versatile. TLC and HPLC could not be used for this reaction. PADI-MS was successfully used for this reaction using cotton swabs without preparation or pre-concentration of sample solutions.

Ovarian cancer is the 5th leading cause of cancer-related death. The disease responds initially to treatment which is most often surgical cytoreduction followed by chemotherapy. The primary response rates to chemotherapy are approximately 80%. Unfortunately, most patients relapse and eventually tumours become refractory to frontline therapy. The lack of widely effective therapies at this points leads to a low 5-year survival. Therefore, new therapeutic agents or treatment strategies are required. It has been reported previously that gain of function mutations of p53 which upregulate the mevalonate pathway in breast cancer. TP53 is commonly altered in high grade serous ovarian cancer which might suggest that the mevalonate pathway may also be deregulated in ovarian cancer. The result reported in this thesis indicate that p53 upregulate the expression of key enzymes of the mevalonate pathway in ovarian cancer cell lines. In particular, it was found that 3-Hydroxy-3-Methylglutaryl-CoA Reductase (HMGCR), the rate limiting enzymes of the pathway, geranylgeranyl transferase-I (GGTI), GGTII and farnesyltransferase are upregulated in number of ovarian cancer cell lines. These observations suggest that pharmaceutical inhibition of the mevalonate pathway may be a promising therapeutic approach. Pitavastatin, a member of statin family of HMGCR inhibitors, has been found to have significant activity against ovarian cancer cells and induce regression of ovarian cancer xenografts in mice in previously published result from our laboratory. Although repurposing statins for use in oncology is an attractive strategy, there are legitimate concerns about the potential for the drug to cause myopathy. Therefore, other pharmacological agents which inhibit the mevalonate pathway were evaluated to test the hypothesis that dual inhibition of the mevalonate pathway would synergistically cause ovarian cancer cell death. Bisphosphonates, such as zolendronic acid, are inhibitors of farnesyl diphosphate synthase. Zolendronic acid, and to lesser extends risedronate, potentiated the activity of pitavastatin in several assays assessing the growth and viability of ovarian cancer cells. In contrast, the geranylgeranyl transferase I inhibitor, GGTI-2133, antagonised the activity of pitavastatin. Similarly, knockdown of either GGTI-β or GGTII-β by RNAi failed to potentiate the activity of pitavastatin. However, combined knockdown of both geranylgeranyl transferases potentiated the activity of pitavastatin. To identify further drugs which could interact synergistically with pitavastatin, a library of 100 off-patent drugs was screened in combination with pitavastatin in cell growth assays. Several compounds were identified which potentiated the activity of pitavastatin and/or had notable activity as single agents. The most striking hit from this screen was prednisolone, a synthetic glucocorticoid. Subsequent studies confirmed the synergistic interaction between prednisolone and pitavastatin in several cell growth and viability assays. To evaluate the mechanism underlying this synergistic interaction, publically-available expression data were interrogated to identify mevalonate pathway enzymes whose expression was regulated by prednisolone. The effect of these candidate genes was then tested in ovarian cancer cells and levels of HMGCR, farnesyl diphosphate synthase and geranylgeranyl transferase II were found to be reduced. These data suggest that drug combinations inhibiting multiple points in the mevalonate pathway may increase the therapeutic window for pitavastatin and offer a potential treatment for ovarian cancer.

Background: The Royal Stoke Pharmacy Workforce Calculator (RSPWC) was developed to meet a local need of identifying clinical pharmacy staffing levels, not described nationally. This study demonstrates the validity and transferability of the RSPWC and its application to other settings. Methods: A two-round Delphi consensus study was conducted to confirm the activity standard (tasks, times and frequencies) for clinical pharmacy services. An operator evaluation was undertaken to demonstrate the reliability of the tool by multiple operators and a series of qualitative interviews explored the utility of the tool in different settings. These research strands ran concurrently from April 2016 to December 2016. Results: Participants from 21 sites across the UK were recruited, including district general hospitals (nine), teaching hospitals (eleven) and one mental health trust. A wide range of staffing levels, across all staff groups, was reported. Consensus was achieved for 68% of components of the algorithm that drives the RSPWC. For a further 21% of components, ‘nationally-representative’ figures were identified from the data. Eleven percent of the components, those relating to elements dependent on individual patient responses to medicines, failed to achieve agreement. A pragmatic approach was taken in the derivation, from study data, of these activity frequencies for typical patient groups. Content validity of the tool was demonstrated. The ‘operator evaluation’ demonstrated reliability in its use by different operators. The application of this tool to a variety of settings was identified through the qualitative data. Discussion: The results of the study demonstrate the validity, transferability and utility of the RSPWC. They capture, for the first time, a consensus on the required service components for the delivery of pharmaceutical care, across multiple hospital sites nationally in the UK. Through this study a clinical pharmacy workforce calculator for acute hospital settings has been developed and validated.

Background: Anticoagulation therapy for patients with atrial fibrillation (AF) remains a national challenge. A decision support tool (DST) was developed to assist healthcare professionals (HCPs) in the appropriate prescribing of anticoagulants in patients with AF. This thesis aimed to evaluate the utility of the DST and associated patient decision aid (PDA) for anticoagulant decision making in clinical practice. Methods: This study involved a series of sequential stages in the evaluation of the DST. Semi-structured interviews were conducted with forty-seven HCPs to explore their perceptions of anticoagulation prescribing decision. Using a vignette, the perspective of HCPs on the potential utility of the DST and associated PDA were explored using both semi-structured interviews and questionnaires. Second interviews were conducted approximately eight weeks from the initial contact to explore HCPs’ perspectives on the actual utility from implementing the DST and associated PDA in routine clinical practice. The perspectives of a group of AF patients’ who had experienced the DA during consultation were explored using semi-structured interviews and questionnaires. Results: Qualitative themes elicited during initial contact revealed that anticoagulants prescribing decision can be suboptimal. Findings from the pre-intervention evaluation showed that the DST has potential to improve the quality of anticoagulants decision process. Findings from post-intervention evaluation demonstrated improvements in anticoagulants decision-making in clinical practice. Findings from fourteen patients revealed that the DA was effective in facilitating a quality decision that was informed and consistent with personal values and expectations. Conclusions: This study demonstrated the positive impact the DST can have on the quality of anticoagulants decision-making in clinical practice and provides a unique contribution to the existing CDSS research. The ever-increasing demand for a quality decision-making process in clinical practice is a fertile environment for clinicians and policymakers to consider the potential impact that the DST and associated PDA can offer.

Virtual patients (VPs) are routinely used in the training of medicine and nursing professionals but uptake into pharmacy has been slower. The pharmacy pre-registration training year takes place in the workplace and a disparity in the perceptions of support provided and the pre-registration examination pass rates has been established between the training sectors. This programme of work aimed to evaluate the effectiveness of virtual patients (VPs) at supporting pre-registration training when compared to a non-interactive (NI) learning tool. Following institutional ethical approval, a mixed methods approach was adopted to evaluate the VP technology. A purposive sample of 165 pre-registration trainees (2014-2015) who were completing their training in a UK-based community or hospital pharmacy were recruited. Participants were randomly stratified to receive three VP or NI case studies. Knowledge surrounding the case studies was assessed using a quasi-experimental evaluation and thoughts on the two learning tools were obtained and compared via questionnaires and semi-structured telephone interviews. Quantitative data was analysed using descriptive and inferential statistics and qualitative data was analysed using content analysis (questionnaire) and framework analysis (interviews).No significant differences in knowledge improvement between pre-registration trainees in the VP and NI groups were obtained. Significant improvements in knowledge were found between the sectors of training for the three case studies. Pre-registration trainees reported that the VP enabled them to apply their learning and engage in experiential learning. The VP case studies were associated with greater satisfaction and were reported to provide a more realistic, interactive and enjoyable learning experience. Pre-registration trainee’s perspectives of the VP technology as a learning tool were more favourable regarding the development of real-life complex skills and aspects of learning, which provides a remit for further evaluation of the technology in undergraduate and postgraduate pharmacy training.

Hepatocellular carcinoma accounts for 85% of primary liver cancer that are usually characterised by defective or ineffective apoptosis which is considered to be the main cause of cancer progression. In this study, Cytochrome-C which is a pro-apoptotic protein is combined with hybrid (iron oxide-gold) nanoparticles and triggers mitochondrial downstream apoptosis pathway in the tumour cells. The nanoparticle complex enables delivery of this difficult protein through the cell membrane. In this research, five different anticancer drugs (doxorubicin, oxaliplatin, paclitaxel, vincristine and vinblastine) were used against liver cancer and U937 cell lines to assess their IC50 values alone and to check their toxicity after their co-administration with cytochrome C hybrid formulation. These combinations resulted in an increase in the cytotoxicity of the used chemotherapeutic drugs and remarkable decrease in the amount needed to kill hepatic cancer cells. For that reason, Iron-gold hybrid nanoparticles offer a promising tool for cytochrome-c delivery into tumour cells and enhance the specific targeting of therapeutic particles to their site of action. The preliminary results reflected the increasing killing abilities of chemotherapeutic therapies when co-administered with cytochrome C hybrid formulation by targeting the natural killing mechanism inside the cells and activating its pathways. Subsequent to these results, further work was done in formulation of one platform therapeutic device with Polymeric amphiphiles hydrophilic poly(allylamine) polymer (PAA) grafting with 5-(4-chlorophenyl)-1,3,4-oxadiazole-2-thiol(oxadiazole, Ox). Paclitaxel (PTX) was selected as a hydrophobic drug model to check its water solubility behaviour after loading into PAA-HNP-C platform. These new devices showed a significant increase in drug uptake level and increase in PTX cytotoxicity against liver cancer cell lines. The data from this work showed a significant increase in the apoptosis activities of combining treatment anticancer agents (doxorubicin, paclitaxel, oxaliplatin, vinblastine and vincristine) and the hybrid formulation of the cytochrome-C within the liver cell lines, which leads to cellular death. Therefore, this combined method may give promise step for the future of liver cancer treatment regimes. In addition to, formulating the HNP-CYT-C and PTX into as active single platform for increasing the PTX cytotoxicity. More laboratory investigation is needed to check the activity of this formulation as a preparing step to further in vivo studies.

Thermo-responsive polymers are a class of smart polymers that respond to change in temperature. This property makes this type of polymers useful materials in a wide range of applications especially, in the field of drug delivery system. Polymers, which have primary amino groups, such as N-(3-aminopropyl) methacrylamide hydrochloride (APMA), have been used for post-polymerization modification reactions for the development of new materials for biomedical applications. Here I aim to fabricate an amphiphilic monomers composed of APMA by substituting (palmitoyl, dansyl, cholesteryl and oxadiazole) groups onto the primary amine in the APMA monomer. These monomers were then copolymerized with N-(2-hydroxypropyl)methacrylamide (HPMA) in six different monomer feed ratios in order to characterise the effect of hydrophobic pendants on the copolymer properties including lower critical solution temperature (LCST), solubility, drug loading and drug release. In this study drug loading and release properties of HPMA-co-APMA copolymers were studied by using four model hydrophobic drugs, propofol, griseofulvin, prednisolone and paclitaxel. High performance liquid chromatography (HPLC) measurements were performed in order to compare drug loading properties of the copolymer formulas. The most potent carrier candidate was loaded in order to carry out thermo-responsive release study. The results showed that all the copolymer formulations in this study possessed the ability to encapsulate practically poor-soluble drugs within their hydrophobic core. HPLC measurement has demonstrated that HPMA-co-(APMA-Oxadiazole 1%) (O-1) and HPMA-co-(APMA-Dansyl 2%) (D-2) have a higher drug solubilisation capacity than other copolymer formulations. In vitro release profiles of different model drugs of the optimal formulation of copolymers were investigated at four different temperatures. Unfortunately, these copolymers showed uncontrolled release of all of the loaded drugs. To set-up thermo-responsive copolymers for controlled drug release, approaches based on introducing poly ethylene glycol (PEG) block as a part of O-1 and D-2 amphiphilic copolymers. A significant enhancement in response to the change in temperature as the drug release across the membrane was seen when PEG was present for the three hydrophobic drugs models. Then hybrid nanoparticles (HNPs) were prepared and attached to O-1 as localised nano-heaters to accelerate drug release in responded to temperature change. Finally, the optimal formulation of (O-1)-b-(PEG) and (O-1)-b-(PEG)-HNPs loaded with paclitaxel (PTX) were tested for their cytotoxicity in vitro on BxPC-3 cells. In-vitro MTT assay results established the ability of (O-1)-b-(PEG)-PTX and (O-1)-b-(PEG)-HNPs-PTX novel formulations to accumulate and kill pancreatic cancer cells more effectively, compared to the free PTX.

Background: Biological medicines are effective but expensive options for treating patients with chronic and life-threatening diseases unresponsive to conventional small molecule medicines. The importance of biosimilars as an alternative to expensive originator biologics in treatment regimens is increasing year by year. This thesis aimed to examine the prescribing pattern of current marketed biosimilars and understand the potential factors influencing their prescribing in UK. Methods: Examples of generic medicines entry were analysed and compared and contrasted with the entry of biosimilars in both primary and secondary care settings. Web-surveys were conducted with 234 healthcare professionals (HCPs) and 182 service users to investigate their knowledge and attitudes towards biosimilars. Face-to-face semi-structured interviews were then conducted with 26 HCPs and service users to further elucidate interviewee’s perceptions of biosimilars. Results: Findings from the market analyses showed that the penetration of a generic and/or a biosimilar is governed by the cost, the number of products (competitors) and chronicity of use when the delivery devices are the same. When delivery device or another factor influencing prescribers’ perceptions of patient benefit differed, biosimilar uptake was inhibited. The findings from web surveys and interviews showed that both HCPs and service users had a good knowledge and understanding of biosimilars and their importance for cost savings. Concerns about safety and efficacy of biosimilars during the process of switching to biosimilars were expressed by HCPs and services users, although the extent of this concern varied with the clinical discipline. Conclusion: The introduction of biosimilars is associated with considerable cost savings to the NHS. There are subtle differences between specialists’ views on biosimilars and different uptake patterns. Factors influencing prescribing biosimilar medicines were similar but more complicated than generic medicines. Cost is a key driver for the uptake of biosimilars only when patients related factors were justified or equal.

The bone graft procedure is the gold standard therapy for large bone defects and usually involves implanting autologous bone into the defect site. The need to find more patient-friendly alternatives to the bone graft procedure is a driving force behind recent advances in bone tissue engineering. Different synthetic and natural biomaterials are being investigated for their use in the repair of large bone defects. However, there is still a need for scaffold materials that are able to sinter in situ with the capability of delivering growth factors that promote the bone repair process with minimal invasion. In this study, our main aim was to develop a porous, protein-loaded microsphere delivery system with the capability of sintering in situ and with injectable potential. Poly (lactic-co-glycolic acid) (PLGA) was used for this formulation due to its biodegradability, biocompatibility, controllable mechanical properties and good processing capabilities. An optimised procedure for formulating porous and non-porous (protein-loaded) microspheres was established and the microspheres were extensively characterised. There was an inverse relationship between the molecular weight of the PLGA and both the protein release and compressive strength. An intermediate molecular weight (53 kDa) variety of PLGA was chosen for further work based on balancing the need for retaining sufficient compressive strength and a slower protein release profile. The burst release of protein was addressed by investigating various coatings. The combination of chitosan and PLGA to form composite PLGA/chitosan microspheres resulted in the desired reduction in the burst release. The protein-loaded and porous microspheres were combined as a paste and found to sinter at body temperature (37°C) into scaffolds. Whilst previous investigations have focused primarily on a single type of PLGA microsphere (with or without an additional biomaterial), in this study we combined both porous and protein-loaded microspheres into a single delivery system. Furthermore, successful sintering was confirmed when a suspension of the microspheres was injected through a 19 G needle. The biocompatibility and osteogenic potential of the scaffolds were investigated. The composite PLGA/chitosan scaffolds supported the growth of MG-63 osteosarcoma cells and a primary human stem cell line. Furthermore, the scaffolds also supported the osteogenesis of the stem cells, as demonstrated by the presence of the late protein marker of osteogenesis, osteocalcin, and positive von Kossa staining, which is indicative of mineralization. The composite PLGA/chitosan and porous microspheres combine both porosity and the ability to load and sustain the release of protein into one system. Moreover, their ability to sinter at body temperature when injected or applied as a paste demonstrates the dual functionality of the system. This represents a novel approach to delivering protein for tissue regeneration as presently, there has been no report of the combination of PLGA/chitosan microspheres with porous PLGA microspheres as a system that is able to sinter, both post-injection and when packed as a paste, at 37°C. Therefore, the formulation presented in this thesis shows potential for further in vitro and in vivo testing to determine its suitability for bone tissue engineering applications.