Nanoparticles derived from proteins offer a smart material for the design of a new generation of anticancer therapies. In this thesis we describe a variety of novel photodynamic therapy (PDT), drug delivery, and imaging agents that have been combined to produce theranostic systems. The initial research focus was to produce a protein nanocage of Listeria innocua DNA binding protein from starved cells (LiDps) presenting the Gaussia princeps luciferase enzyme (GLuc) on its exterior (GLuc-LiDps) together with the Zn (II)-Protoporphyrin IX (ZnPP) photosensitiser that has been covalently attached to the protein surface. This system operates as a PDT based on Bioluminescence Resonance Energy Transfer (BRET). The cytotoxic effect of GLuc-LiDps-ZnPP nanoparticles in the presence of its substrate (coelenterazine) was tested against two types of breast cancer cell lines: SKBR3, MDA-MB-231, and also the MRC5 non-cancerous cell line, by means of an MTT assay. The results indicated that GLuc-LiDps-ZnPP nanoparticles plus coelenterazine could inhibit the growth and the migration of SKBR3 cells, out of those studied. Downregulation of the Bcl-2 and Mcl-1 anti-apoptotic proteins was also observed after treatment with GLuc-LiDps ZnPP-mediated PDT, suggesting the SKBR3 cells may be undergoing apoptosis. Furthermore, both flow cytometry analysis and confocal microscopy images demonstrated that the GLuc-LiDps-ZnPP appeared to be preferentially internalised in SKBR3 and MDA-MB-231 cell lines without uptake in the MRC5 cell line. Reactive Oxygen Species (ROS) levels were significantly increased in SKBR3 cells compared to MDA-MB-231 cells in the presence of this PDT agent. Similarly, a C-terminal mini Singlet Oxygen Generator (miniSOG) photosensitiser was conjugated to LiDps and human apoferritin (HuAft). Moreover HuAft was fused with the ZHER2:342 Affibody (Afb) targeting peptide to form Afb-HuAft. This 6.7 kDa Afb protein has a high binding affinity for the Human Epidermal Growth Factor receptor (HER2), which is overexpressed on the surface of a number of tumour cells especially breast cancers. Both miniSOG-LiDps and miniSOG-HuAft fusion proteins were successfully expressed and purified and their subunits were self-assembled to form GLuc-LiDps:miniSOG-LiDps and Afb-HuAft:miniSOG-HuAft hybrid chimeric cages. Additionally GLuc was directly fused with miniSOG supplemented with LTVSPWY targeting peptide to produce a novel (GLuc-miniSOG-LTVSPWY) for targeting of HER2 overexpressed cancer cells, potentially applicable for PDT. Further study is required in order to thoroughly characterise the GLuc-miniSOG-LTVSPWY and the hybrid cages as well as determine their respective cytotoxicity. In the second objective, lead sulfide quantum dots (PbS QDs) were utilised as a drug delivery system and an imaging agent. The PbS QDs were capped with a cancer cell targeting agent (mutated Afb (Afb2C)) to form Afb2C-PbS QDs. This construct was further modified by conjugation with ZnPP to produce ZnPP-Afb2C-PbS QDs. The cytotoxic effect of Afb2C-PbS and ZnPP-Afb2C-PbS QDs were studied in vitro using SKBR3 (HER2 positive) and MDA-MB-231 (HER2 negative) cell lines. The results indicate that both types of PbS QDs display anti-proliferative activity against SKBR3 cells through inducement of cancer cell apoptosis and/or necrosis. This was observed from Pre-G1 phase arrest and an increase in cell population in late apoptosis and necrosis. These results may contribute to the development of cancer treatment using of nanoparticles derived from protein.

The pharmaceutical business is driven by innovation and new technologies. In order to improve the overall efficiency, the modern R&D organisations nowadays have integrated problem-solving techniques in their innovation process. This thesis aims to explore and analyse the application of TRIZ technique in the problem-solving process in the medical device sector of the pharmaceutical industry. The findings of the literature review indicate that TRIZ can effectively guide the problem-finding process with a tool kit that can recognise patterns and regularities based on the past solutions in a knowledgebase. The results suggest that such systematic approach is more effective than the conventional methods of trial-and-error. This study conducted a survey amongst the innovative medical device departments of various pharmaceutical companies in the Rhine-Main region in Germany and provided contemporary data on the application of problem-solving tools, especially TRIZ, in those institutions. As a result, the survey data also delivered some possible criteria for technical solutions of medical devices which were subsequently discussed and finalised with a group of experienced experts (expert panel). The next step of the study was organised as a 2x2 experiment. During the experiments, two groups of experienced practitioners were asked to improve the design of two sample medical devices, alternatively using TRIZ and brainstorming. The efficacy of TRIZ application was analysed both in terms of the quality of the technical solutions and that of the group work. The SYMLOG Adjective Rating Form method initiated by Bales was used for the assessment of the group work. The results of the experiment indicate that the impact of the problem-solving tools is influenced by the type of innovation problem. For the analysis of such influences, this research makes a contribution to knowledge by proposing a 2-dimensional framework to capture the problem types. In addition, a TRIZ procedure for the technical innovations of medical devices was developed based on the model of Su et al. Due to the sensitive protection of intellectual property in the pharmaceutical industry, field studies of R&D processes in large pharmaceutical firms are limited in the public literature. This work provides valuable insights into this business sector, especially in respect of application of problem-solving tools and how those tools may potentially improve the outcomes of the R&D activities in the pharmaceutical industry.

Polymer-based drug delivery systems have fantastic potential in chemotherapy as they can reduce drug side effects, help in patient compliance and provide targeting. Nanoprecipitation is used to encapsulate small drug molecules into polymer nanoparticles to form a drug delivery system. A major obstacle in polymer-based drug delivery systems reaching the clinic is their inability to load sufficient drug molecules. Little is known about the processes involved in the encapsulation of drug molecules into these delivery systems. An insight into the processes that govern the formation of these particles and encapsulation of small drug molecules within them is therefore desirable. We used molecular dynamics to model nanoprecipitation by simulating the dispersion of an acetone drop, containing polymer, into water containing drug. To allow sufficient dispersion of acetone a large amount of water is required, thus coarse-graining becomes mandatory. However, we maintain accuracy for our polymer-drug interactions by using a multiscale force field. Atomistic polymer and drug molecules contain coarse-grain virtual sites which facilitate interactions with the coarse-grain solvent molecules. We also employed fully atomistic reference simulations via resolution transformation to optimise our multiscale force field. This thesis details the theory and design behind this model of nanoprecipitation including how other techniques produced inferior results. Initial simulations with our multiscale model matched an experimental trend and were shown to be accurate relative to atomistic reference simulations. We also analysed a fully atomistic simulation of nanoprecipitation that took several months to complete. This atomistic simulation was used as a reference to update the multiscale force field. The updated force field improved on some aspects of the simulation but there are still areas that need improvement. Insight from the simulations provides an understanding of the experimental results and trends. The transferability of the model should help in designing more efficient polymer-based drug delivery systems in the future. We conclude with future work on modelling polymer-based drug delivery systems including alternate methods to gain understanding of not only drug incorporation but also drug release.

Development of novel biodegradable polymers from renewable resources has attracted attention due to the limitations associated with polymers obtained from petroleum resources. The objective of the work presented in this thesis was to develop various novel biodegradable amphiphilic block copolymers from commercially available sustainable feedstocks for drug delivery applications. Synthesis was performed using a reported method under mild reaction conditions. Renewable δ-decalactone was chosen as a key monomer to synthesise novel amphiphilic block copolymers via ROP using PEG as initiator. A diblock (i.e. mPEG-b-PDL) and a triblock (i.e. PDL-b-PEG-b-PDL) copolymer of poly(decalactone) (PDL) was synthesised and purified successfully. Additionally, a novel triblock copolymer (i.e. mPEG-b-PDL-b-PPDL) was synthesised using ω-pentadecalactone as monomer and mPEG-b-PDL as initiator via ROP to generate a copolymer with different physical properties. Further, a di-block copolymer of ε-caprolactone (i.e. mPEG-b-PCL) was synthesised for comparative studies with novel block copolymers. Micelles of synthesised block copolymers were fabricated using a reported nanoprecipitation method. Micelles fabricated from these novel block copolymers were of sizes <200nm and possessed low critical micelle concentration (CMC) values. Curcumin and Amphotericin B were successfully encapsulated in the novel block copolymer micelles via nanoprecipitation method. The results obtained from curcumin loading and release studies suggested that these novel PDL block copolymers could perform in similar fashion when compared with poly(caprolactone) (PCL) block copolymer micelles. However, in subsequent study micelle of mPEG-b-PDL gave high loading content compared to mPEG-b-PCL micelles when amphotericin B was used as a drug. Further, a preliminary in vitro degradation study of mPEG-b-PDL micelles was performed and the results proposed that the ester linkage of PDL chain were susceptible to hydrolytic degradation in physiological condition. Additionally, in vitro cytotoxicity studies performed on HCT-116 human colon cancer cells revealed that the novel mPEG-b-PDL micelles have similar toxicity profiles when compared to the well-established mPEG-b-PCL micelles. Ligand mediated targeting efficiency of novel diblock copolymer micelles was also studied for potential future applications in cancer therapy. Amphiphilic block copolymers using PEG and PDL were synthesised via click chemistry to generate functionalised block copolymers. Folic acid and rhodamine B were used as targeting ligand and tracker dye respectively. Mixed micelles fabricated from functionalised block copolymers (i.e. FA-PEG-b-PDL, RhB-PEG-b-PDL and mPEG-b-PDL) were tested on folate receptor positive (MCF-7 FR+ve) and folate receptor negative (A549 FR-ve) human cancer cell lines for receptor mediated endocytosis. The acquired confocal images demonstrated the nonspecific uptake of the PEG-b-PDL micelles formulations (targeted and non-targeted) in both cell lines selected in current study. The results obtained from this thesis study suggested that the synthesised novel PDL block copolymer micelles have potential to act as a novel drug delivery system. However, further studies have been proposed to explore the possible applications of these renewable block copolymers.

CoordenaÃÃo de AperfeiÃoamento de Pessoal de NÃvel Superior / A depressÃo à um dos mais prevalentes transtornos psiquiÃtricos. Os principais sintomas clÃnicos da depressÃo sÃo humor deprimido, anorexia, anedonia, reduÃÃo da atividade locomotora. Hà evidÃncias acumuladas de que a depressÃo pode se desenvolver em resposta à ativaÃÃo do sistema imune inato, sendo caracterizada por uma resposta inflamatÃria com aumento da produÃÃo de interleucina IL-1&#946;, IL-6, TNF-&#945; e outras. Com base nas propriedades anti-inflamatÃrias e antioxidantes da doxiciclina e supondo que esta droga apresenta menos efeitos colaterais e um melhor perfil farmacocinÃtico, em comparaÃÃo com a minociclina, a hipÃtese de que esta droga pode apresentar efeitos antidepressivos, utilizando o modelo de depressÃo induzido por lipopolissacarÃdeo (LPS) foi estudada. Para determinar as alteraÃÃes de comportamento, camundongos Swiss machos foram submetidos aos testes de campo aberto e nado forÃado. Para avaliar a capacidade da doxiciclina em prevenir ou reverter o comportamento tipo-depressivo induzido pela administraÃÃo sistÃmica de LPS, esta foi administrada nas doses de 25 ou 50 mg/kg, i.p. 30 min antes de LPS (prÃ-LPS) ou 1,5 e 23,5 horas apÃs a LPS (pÃs-LPS). A imipramina foi utilizada como antidepressivo padrÃo nas mesmas condiÃÃes de tempo. Em ambas as situaÃÃes, prevenÃÃo (prÃ-LPS) e tratamento (pÃs-LPS), o comportamento dos animais foi avaliado 24 horas apÃs a administraÃÃo de LPS, um perÃodo conhecido pela ocorrÃncia de um comportamento tipo-depressivo. Os nÃveis de citocinas (IL-1&#946; e TNF-&#945;) e nitrito foram avaliados no sangue (plasma) e as Ãreas cerebrais: cÃrtex prÃ-frontal (PFC), hipocampo (HC) e corpo estriado (ST). A administraÃÃo de LPS, 0,5 mg/kg aumentou significativamente o tempo de imobilidade em comparaÃÃo com os animais controle, enquanto que a doxiciclina, nas doses de 25 e 50 mg/kg e imipramina (10 mg/kg) foram capazes de prevenir e reverter a imobilidade induzida pelo LPS. A doxiciclina e imipramina, quando administrados prà e pÃs-LPS reduziram significativamente o tempo de imobilidade, mostrando um efeito antidepressivo. Em relaÃÃo a citocina IL-1&#946;, seus nÃveis foram diminuÃdos, enquanto os nÃveis de TNF-&#945; nÃo foram alterados significativamente. A doxiciclina e imipramina, preveniram e reverteram a diminuiÃÃo dos nÃveis de nitrito induzido por LPS. Com base nos resultados do presente estudo, avaliando o uso da doxiciclina, sugere-se que este antimicrobiano possa atuar como um antidepressivo. / Current evidences support inflammation, oxidative and nitrogen stress, as well as brain-derived neu-rotrophic factor (BDNF) signaling mechanisms as important in depression pathophysiology. Tetracycline antibiotics have anti-inflammatory and antioxidant properties. Preliminary evidence indicates that minocycline has antidepressant properties. Doxycycline (DOXY) has favorable pharmacokinetic and safety profiles when compared to other tetracycline congeners. The antidepressant activity of DOXY has not been adequately investigated. This study evaluated the effects of DOXY (25 and 50 mg/kg, i.p.) on LPS-induced (0.5 mg/kg, i.p.) depressive-like behavior. Doxycycline was administered 30 min before LPS (pre-LPS) or 1.5 and 23.5 h following LPS (post-LPS) administration in mice. LPS-treated animals pre-sented an increase in immobility time in the forced swimming test (FST) when compared to controls 24 h after endotoxin administration. Similarly to imipramine (IMI-10 mg/kg, i.p.), DOXY at both doses pre-vented and reversed LPS-induced alterations in the FST. IL-1b content was increased 24 h after LPS administration in striatum, hippocampus and prefrontal cortex. IMI and DOXY prevented and reversed LPS-induced increase in IL-1b. IMI and DOXY also prevented and reversed LPS-induced alterations in nitrite content and oxidative stress parameters (lipid peroxidation and reduced glutathione levels). Both DOXY and IMI prevented LPS-induced decrease in hippocampal BDNF levels. Taken together, our results demonstrate that DOXY is comparable to IMI in effectively ameliorate LPS-induced depressive-like behavior, providing a rationale for testing DOXYâs antidepressant efficacy in humans.

Studies on electronic patient medication record (ePMR) systems that are used in community pharmacy in England have focused primarily on the ability of these systems to highlight potentially hazardous co-prescriptions and prevent clinical hazards and harm to patients. As such, there is a scarcity of literature on the use of ePMR systems, other safety functionality of the systems and user responses to alerts. This thesis aims to fill this gap by examining the functionality of ePMR systems used in community pharmacy in England, focusing on safety features and alerts. This research was conducted in England between July 2010 and July 2013. Evidence for the effectiveness of safety features and alerts in ePMR systems during the dispensing process was evaluated through a systematic review of the literature. Stakeholder perspectives of ePMR systems’ functionality were then obtained through qualitative interviews. The performance of ePMR systems licensed for the electronic prescription service (release 2) in the community pharmacy setting were then assessed using a simulated observational testing approach. Ethnographically informed observations in community pharmacies were subsequently used to study how community pharmacy professionals use ePMR systems and manage alerts in practice. The systematic review included five studies - three randomised controlled trials and two before-after studies, with drug-drug interaction (1), drug-laboratory (2), drug-pregnancy (1) and drug-age (1) alerts. The review revealed that ePMR systems in conjunction with embedded safety features are effective in picking up clinical hazards at the point of dispensing. However, there are problems of false alerts and inconsistencies in alert management. Empirical findings indicate that there are significant issues with the way ePMR systems and alerts are designed and used. Thirty participants took part in the qualitative interviews - community pharmacy professionals (13), health care policy makers (5), legal practitioners specialising in pharmacy (3), ePMR systems’ software vendors (4) and ePMR systems’ software knowledgebase creators (5). Participants attributed alert ineffectiveness in community pharmacy practice to factors such as lack of harmonisation of alert severity levels in systems, poor alert design, over-presentation of alerts and absence of management advice in alerts. Five unique ePMR systems were evaluated in eight participating pharmacies with a sixth ePMR system assessed in a demonstration setting. The systems’ performance was variable and sub-optimal. The ethnographically informed observations took place in the eight pharmacies where system assessment was conducted. The observations revealed that the current design of ePMR systems and presentation of alerts are limiting the quality of support provided to pharmacists and their support staff. This research is part of a growing body of work on the functionality of ePMR systems, their safety features and alerts indicating that ePMR systems require improvements if they are to effectively support patient safety and consistently deliver better patient outcomes. The findings highlight the need to incorporate patient context into alerting to increase alert relevance. In addition, system vendors need to make use of the evidence in the literature to design effective ePMR systems, alerts and user interfaces. An authoritative body should set the minimum specifications for ePMR systems and alerts, and identify the critical alerts that pharmacy professionals should evaluate at the point of dispensing. Additionally, training of pharmacy professionals in health information and communication technology is required to improve patient safety. This should cover areas such as informatics, human factors, safety culture, clinical decision-making, alert management, risk management and communication. Many of the findings are likely to be relevant to similar medication record systems in ambulatory pharmacies around the world; however, further work is required to understand fully the extent of the issues identified in this research.

Ink-jet printing is a precise and versatile technique that accurately deposits small volumes of solutions (pico litres) in specific locations. Recently inkjet printing has attracted increasing attention in the pharmaceutical industry because of its ability to deliver low adjustable doses, variable drug release profiles and drug combinations suitable for the paradigm of personalised medicines. The significant growth in the aging population and the rise in the number of patients suffering from multiple chronic diseases are the key drivers. The current traditional tablet compression methods are largely limited in terms of flexibility and complexity of dosage form. There is a need for new innovative technologies that can produce bespoke medicines in a relatively cheap and efficient manner at the point of care. 3D inkjet printing (3DIJP) provides a platform with the potential to address the above need. This thesis investigates the capability of 3DIJP as a tool for manufacturing solid dosage forms. In chapter 3, a piezoelectric drop on demand printer was used. The chapter focuses on two solvent based inkjet printing methods. In the first solvent based method, excipients including hydroxypropyl methylcellulose (HPMC), poly (vinyl pyrrolidone) (PVP) and Eudragit RL were investigated for printability. PVP (K10) which showed the best printability behaviour was loaded with digoxin or carbamazepine (CBZ) and printed to obtain films. In the second solvent based method, a solution containing CBZ dissolved in a mixture of of polyethylene glycol diacrylate (PEGDA) and with poly(caprolactone dimethyl acrylate) (PCLDMA) was printed and polymerised in situ using ultraviolet light to form films. The printed drug loaded films were investigated using time of flight secondary ion mass spectroscopy (ToF SIMS), atomic force microscopy (AFM), scanning electron microscopy (SEM) and differential scanning microscopy (DSC). PVP formulations were homogeneous, with no evidence of crystallisation PEGDA/PCLDA/CBZAFM images showed a clear phase separation at the micron scale and no drug was detected at the surface. In this chapter, the production of adjustable doses was also evaluatedusing UV-VIS spectrophotometry. In chapters 4 and 5, a solvent-free hot-melt 3D inkjet printing method suitable for manufacturing solid dosage forms was developed. Excipients including beeswax, carnuba wax, gelucire 44/14 and trimyristin were examined for printability. Beeswax a naturally derived and FDA approved material showed the best printability behaviour and was selected as the drug carrier. Traditional circular shaped tablets and cylindrical implants loaded with 5% w/w fenofibrate were successfully fabricated. The printed tablets and implants were well-defined, smooth surfaced and with no apparent defects. The architecture of the tablets was investigated using 3D micro X-ray computed tomography (μCT), revealing well defined and ordered honeycomb channels in the interior of the tablets. The distribution of the drug was evaluated at the macro scale level using DSC and at the micro scale level using ToF - SIMS and Raman spectroscopy. The drug was homogenously distributed within the drug carrier (beeswax matrix ) at the microscale level. At the micron scale level, the drug was heterogeneously distributed. ToF - SIMS studies also revealed that the drug was depleted from the upper most top surfaces. Production of solid dosage forms with intricate and adaptable geometries was demonstrated by printing honeycomb architecture tablets with predetermined variable cell diameters. The diamater of the honeycomb cells was varied, in order to achieve controlled variable drug release profiles. The ablity to control drug release was only applicable above an established critical cell diameter of 0.5 mm. An analytical model describing Fickian diffusion from a slab geometry was developed to allow for the prediction of drug release from the honeycomb tablets. The predicted drug release profiles varied slightly from the experimental data, but the trends for the two data set were identical. For both data sets the rate of drug release increased with increase in the surface area to volume ratio. The findings and the developments demonstrated in this thesis provide an insight into the potential application of 3DIJP as a tool for manufacturing solid dosage forms with bespoke properties for controlled drug release but also highlights some limitations.

Negli ultimi anni si è assistito ad un miglioramento della qualità di vita dei piccoli animali che, oltre ad aumentarne l'aspettativa di vita, ha determinato un aumento della frequenza di patologie associate all'età medio-avanzata, quali le patologie renali croniche. Il presente studio si fonda sulla necessità, sempre più sentita nella pratica clinica veterinaria, di poter fornire al proprietario del paziente affetto da CKD, una serie di parametri che, oltre a fungere da target terapeutico, possano aiutare a comprenderne la prognosi. Lo studio ha valutato una popolazione di cani affetti da CKD e ne ha seguito o ricostruito il follow-up, per tutto il periodo di sopravvivenza fino al momento dell’exitus. Di tali soggetti sono stati raccolti dati relativi ad anamnesi, esame clinico, misurazione della pressione arteriosa, diagnostica per immagini, esami ematochimici, analisi delle urine ed eventuale esame istologico renale. È stato possibile individuare alcuni importanti fattori prognostici per la sopravvivenza in pazienti con CKD. Oltre a fattori ben noti in letteratura, come ad esempio elevati valori di creatinina e fosforo, o la presenza di proteinuria, è stato possibile anche evidenziare il ruolo prognostico negativo di alcuni parametri meno noti, ed in particolare delle proteine di fase acuta positive e negative, e del rapporto albumina/globuline. Una possibile spiegazione del valore prognostico di tali parametri risiede nel ruolo prognostico negativo dell’infiammazione nel paziente con CKD: tale ruolo è stato suggerito e dimostrato nell’uomo e avrebbe alla base numerosi possibili meccanismi (sviluppo di anemia, complicazioni gastroenteriche, neoplasie, etc.), ma dati analoghi sono mancanti in medicina veterinaria. Una seconda possibile spiegazione risiede nel fatto che potenzialmente i livelli delle proteine di fase acuta possono essere influenzati dalla presenza di proteinuria nel paziente con CKD e di conseguenza potrebbero essere una conferma di come la proteinuria influenzi negativamente l'outcome. / In recent years there has been an improvement in the quality of life of small animals. In addition of increasing life expectancy, this has led to an increase in the frequency of diseases associated with elderly, such as chronic kidney disease. The present study is based on the need, in the veterinary practice, to be able to provide the owner of a patient with CKD some parameters which can help to understand the prognosis, besides being useful as a therapeutic target. The study evaluated a population of dogs with CKD and followed these dogs or rebuilt the follow-up, until the exitus. We collected data on medical history, clinical examination, blood pressure measurement, diagnostic imaging, blood tests, urinalysis and, when possible, renal histology. It was possible to identify important prognostic factors for survival in patients with CKD. The prognostic role of high levels of creatinine and phosphorus, or the presence of proteinuria is well known in the literature. It was also possible to emphasize the negative prognostic role of some less known parameters, such as acute phase proteins and albumin to globulin ratio. One possible explanation of the prognostic value of these parameters lies in the negative prognostic role of inflammation in patients with CKD: this role has been suggested and demonstrated in humans and has many possible underlying mechanisms (development of anemia, gastrointestinal complications, cancer, etc.), but similar data are missing in veterinary medicine. A second possible explanation lies in the fact that the levels of acute phase proteins may be influenced by the presence of proteinuria in the patient with CKD and therefore may be a confirmation of how proteinuria negatively affects the outcome.

Transition metal coordination complexes show great promise as novel therapeutic agents with new mechanisms of action, but their characterisation, and identification of their target sites present significant challenges. In this thesis a variety of new analytical methods is explored for the study of platinum, ruthenium, osmium and iridium anticancer complexes. High performance liquid chromatography (HPLC) was used to determine the relative hydrophobicity of a series of photoactivatable Pt(IV) diazido complexes of the general type trans,trans,trans-[Pt(N3)2(OH)2(R)(F11]. Interestingly the hydrophobicities did not follow trends based on literature Log P values of individual ligands and did not correlate with the cellular uptake or antiproliferative activity of the drugs. Other factors such as the quantum yield of the complex, and the type of DNA adducts appear to be more important for their efficacy. Chromatography and high-resolution mass spectrometry were used to study the formation of platinum adducts on DNA when the most active complex trans,trans,trans-[Pt(N3)2(OH)2(PYridine)2], 8 was irradiated in the presence of short single strand oligonucleotides 14 bases in length. Complex 8 was found to bind to the oligonucleotides as a {Pt(pyridine)2}2+ adduct. Modifying the wavelength of activation from UVA to 420 nm had no effect on the type of adduct formed, but the higher energy irradiation achieved maximum levels of DNA platination more quickly. Changing the sequence of the oligonucleotide suggested that the photoactivated form of 8 does not favour the formation of the 1,2-(GpG) bisadduct formed by cisplatin and other clinically approved platinum based drugs, but may form 1,3-(GpNpG) or 1,3-(ApNpG) adducts, as is the case with other trans-platinum complexes. Chiral chromatography using cellulose- and amylose-based stationary phases successfully separated the enantiomers of a series of organometallic 'piano stool' anticancer complexes. This appears to be the first successful separation of facially chiral Ru(II) arene complexes, the enantiomers of which were stable in solution for over 3 h. In contrast, separated cyclopentadienyl WI) complexes with chiral metal centres epimerized within 2 h in solution at ambient temperature. Under similar conditions the enantiomers of the Os(II) arene complex [Os(n6-p-cym)(4-(2-pyridylazo)-N,N-dimethylaniline)Ir remained stable, as did those of the ruthenium-based complex [Ru(9,10- diydrophenanthrene)(en)C1r. It was shown that it is possible to separate the diasteriomers of [Ru(t)6-para-cymene)(iminopyridine)I], that can also be resolved by crystallisationtechniques, and hence, decrease the time required to separate the enantiomers. This work will therefore allow exploration of the biological properties of some of these enantiomers A novel technique for the rapid irradiation and detection of light¬senstive species was developed. Photonic crystal fibers (PCFs) were coupled to a mass spectrometer using HPLC tubing and fittings. This continuous flow method of analysis was validated using the photaquation of cyanocobalamin. The PCF system was compared to the conventional cuvette-based approach. No significant difference in the species detected by MS could be found, but the PCF system had the advantage of requiring 20 times less sample (25 pL), and only 15 min of irradiation compared to 10 h by conventional methods. The new PCF-MS system was then used to study the interaction of the photoactivateable ruthenium-based drug [{(e-indan)RuC1}201-2,3-dPa2+ with a range of small molecules that acted as models for intracellular components, e.g. 5'GMP for DNA. The nucleobase binding properties were consistent with those previously reported with plasmid DNA by Magennis et al: a small amount of binding took place in the dark in view of the aquation of the mondentate leaving groups but this dramatically increased upon photoactivation and loss of the arene ligands. The complex was also found to bind to glutathione (GSH), which is known to detoxify metal-based drugs, an observation possibly explaining its poor anticancer activity.

The presence of organic salts as drug counter-ions and buffers in hydroxypropylmethylcellulose (HPMC) matrices is often overlooked. This study investigates their potential to influence polymer solution properties and matrix drug release kinetics. A homologous series of aliphatic organic salts influenced solution and matrix properties in rank order of hydrocarbon chain length. Monovalent salts containing 1to4 C-atoms had little effect on polymer surface activity, but lowered sol:gel transition temperatures (SGTT), and accelerated matrix drug release in comparison with a dextrose control. Divalent salts were more potent. These observations are consistent with Hofmeister effects in which anions restructure water in the polymer hydration sheath, induce 'salting-out' and suppressing particle swelling and matrix gel layer formation. Organic salts with StoB C-atoms increasingly influenced polymer surface activity, elevated SGTT, and retarded matrix drug release. This suggests these salts enhance HPMC hydration, possibly through interaction with hydrophobic regions. The effects of these salts on matrix drug release show that these ions impact on water:polymer interactions important to gel layer formation and diffusion barrier properties. HPMC matrices containing SOS and its homologues were also investigated. Turbidimetric, tensiometric and rheological studies supported a mechanism in which these surfactants solubilise HPMC at post-micellar concentrations. Incorporating 10% SOS into HPMC matrices was shown to increase the resistance of HPMC matrices to sucrose medium up to 2.0M, suggesting a role for surfactants in avoiding food solute effects. This study shows that organic salts incorporated in HPMC matrices have the potential to influence drug release in a rank order that reflects their modulation of the HPMC polymer hydration sheath in solution. SOS and its homologous series could retard drug release from HPMC matrices only when their critical aggregation concentration (CAC) was reached. However, it suggests this excipient may have uses as an excipient for improving HPMC matrix release performance.