project 1: the use of 5-asa as a positive control in the dss ... projects 2013-14.pdfproject 2: the...

Project 1: The use of 5-ASA as a positive control in the DSS model of colitis: a North American experience

Name of Principal Supervisor Assist. Prof. Frankish Subject area Pharmacology Non-lab based project

5-Aminosalisylic acid (5-ASA) is a drug commonly used to treat patients with mild to moderate ulcerative colitis (UC). One of the most commonly used preclinical animal models used to establish efficacy of novel drugs for UC is the dextran sulfate sodium (DSS) murine colitis model. The student will be given the methodologies and the raw data from a study in the DSS model of colitis, using 5-ASA as a positive control, undertaken by a contract research organisation (CRO) based in North America. The aim of the project will be to analyse and present the data and draw conclusions.

Project 2: The use of 5-ASA as a positive control in the DSS model of colitis: a European experience


5-Aminosalisylic acid (5-ASA) is a drug commonly used to treat patients with mild to moderate ulcerative colitis (UC). One of the most commonly used preclinical animal models used to establish efficacy of novel drugs for UC is the dextran sulfate sodium (DSS) murine colitis model. The student will be given the methodologies and the raw data from a study in the DSS model of colitis, using 5-ASA as a positive control, undertaken by a contract research organisation (CRO) based in Europe. The aim of the project will be to analyse and present the data and draw conclusions.

Project 3: Treatment successes and failures in IBD: what are the odds, the costs and potential returns


Inflammatory bowel disease is a therapeutic area of considerable unmet clinical need. While 5-ASA and steroids are often used in mild to moderate disease and immunosuppressants and TNFa blockers may be resorted to in moderate to severe cases. The market segment is considerable and there have been many potential new entries into the field, with varying degrees of success. The aim of the project is to survey the literature and follow the various new potential therapies from the research lab through clinical trials to either the market place or obscurity. The student may then establish the success rate of new therapies and the cost of failure and may comment on the rationale of drug development in this area.

Project 4: Biodistribution and measurement of iron oxide nanoparticles using Magnetic Resonance Imaging

Name of Principal Supervisor Prof. Radomski Co-Supervisor Oliviero Gobbo Subject area Pharmacology Lab based project

Nanostructures are poised to play a crucial role in medicine by serving as carriers for drugs and imaging agents that will selectively bind to targets on diseased cells and tissues. In this context, it is very important to develop tools to evaluate and detect these therapeutic agents in pre-clinical development and in clinical trials. The EU-funded MULTIFUN consortium, including our group, develops multifunctionalised iron oxide nanoparticles (IONPS) combining diagnostic and therapeutic features against breast and pancreatic cancer. Although originally developed as liver-specific contrast agents, IONPS are widely used as drug carriers/imaging agents because of their strong magnetic properties. Therefore, the fate of IONPS in the body (biodistribution) can be determined using Magnetic Resonance Imaging (MRI). The aims of this study are to analyse the levels and biodistribution of IONPS and to establish a MRI protocol for the quantification of IONPs in vivo in rodents and ex-vivo in larger animals.

Project 5: Hepatic tryptophan dioxygenase – a target for antidepressant activity?

Name of Principal Supervisor Assoc. Prof. Harkin Co-Supervisor Eoin Sherwin/Brian Talbot Subject area Pharmacology Lab based project

The kynurenine pathway (KP) is the major pathway responsible for the metabolism of tryptophan in the central nervous system. The rate limiting enzymes in this pathway are indoleamine 2,3 dioxygnease (IDO) which is induced by inflammatory cytokines and tryptophan 2,3 dioxygenase (TDO). As tryptophan is required for the biosynthesis of the neurotransmitter serotonin which plays an important role in the regulation of mood, the induction of IDO and increased metabolism of tryptophan via the KP represents a mechanism linking inflammation to reduced serotonin availability and depressed mood. Whilst a considerable amount of work has been undertaken on IDO in our laboratory to date, we have not performed any experiments on a related enzyme in the liver, hepatic tryptophan 2,3 dioxygenase (TDO) which also leads to activation of the KP. The aim of this research will be to develop assays to determine the expression and activity of TDO and determine if changes to the expression and/or activity of TDO influence tryptophan metabolism, production of KP metabolites and serotonin biosynthesis of relevance to the pathophysiology of depressive illness. Assays will be performed on tissues harvested from animal experiments which simulate depression in the first instance. Clinical samples may also be tested. The aims of this project are to (1) determine TDO expression in hepatic samples obtained from laboratory experiments (2) to establish an enzyme assay for the quantification of changes in TDO activity i.e. catalytic conversion of substrate to product and (3) to develop HPLC assays for the quantification of tryptophan, serotonin and KP metabolites including kynurenine and kynurenic acid in tissue samples. Such analytes will be useful to confirm changes in expression or activity reported. The main techniques in use will be quantitative PCR, spectroscopic determination of enzyme activity and application of HPLC coupled to UV and fluorescence detection for metabolite analyses. The student will gain insight into the development and validation of a potential biomarker test for the clinic. References: Miller AH, Maletic V, Raison CL. (2009) Inflammation and its discontents: the role of cytokines in the pathophysiology of major depression. Biol Psychiatry 165(9):732-41. Rafice SA, Chauhan N, Efimov I, Basran J, Raven EL. (2009) Oxidation of L-tryptophan in biology: a comparison between tryptophan 2,3-dioxygenase and indoleamine 2,3-dioxygenase. Biochem Soc Trans. 37(Pt 2):408-12. Dantzer R, O'Connor JC, Freund GG, Johnson RW, Kelley KW. (2008) From inflammation to sickness and depression: when the immune system subjugates the brain. Nat Rev Neurosci. 9(1):46-56.

Project 6: Assessing the neuroprotective role of noradrenaline in combating Parkinson’s disease

Name of Principal Supervisor Assoc. Prof. Harkin Co-Supervisor Justin Yssel Subject area Pharmacology Lab based project

Evidence indicates that inflammation is a major contributor to loss of dopaminergic (DA) neurons and the consequent motor dysfunction in Parkinson’s Disease (PD). DA neuronal loss in the substantia nigra (SN) of PD patients is accompanied by activation of microglia, resulting in production of neurotoxic substances. Consequently, therapeutic approaches that inhibit microglial activation and inflammatory mediator production have the propensity to prevent degeneration of the nigrostriatal DA tract. Noradrenaline (NA) is a neurotransmitter that elicits anti-inflammatory and neuroprotective actions in the brain. The locus coeruleus (LC) is the primary source of NA cell bodies and their axons innervate almost all parts of the brain including the SN and striatum, and LC cell numbers are reduced by approximately 60% at autopsy in PD patients. Consequently it has been suggested that loss of LC NA neurons could be a significant contributor to the progression of PD, and that pharmacotherapies aimed at restoring normal NA tone are likely to have therapeutic potential. In this experiment we will examine the impact of lesioning the NA system using the neurotoxin DSP4 to determine if the noradrenergic loss that ensues will exacerbate the neuroinflammation and loss of DA neurons in the striatum that occurs following administration of the inflammagen LPS into the substantia nigra. Techniques employed - HPLC analysis of dopamine concentrations - Behavioural testing (amphetamine rotation test) to verify the dopaminergic lesion - Immunohistochemical staining of TH neurons and activated microglia Key references

-adrenoceptors enhances neuronal complexity in primary cortical neurons. PhD thesis, Trinity College Dublin. Hertz et al., (2004). Astrocytic adrenoceptors: a major drug target in neurological and psychiatric disorders? Curr Drug Targets CNS Neurol Disord. 3(3):239-67. Madrigal et al., (2007). Neuroprotective actions of noradrenaline: effects on glutathione synthesis and activation of peroxisome proliferator activated receptor delta. J Neurochem. 103(5):2092-101.

Project 7: Astrocytic beta2-adrenoceptors as a pharmacological target to promote neurite outgrowth

Name of Principal Supervisor Assoc. Prof. Harkin Co-Supervisor Eimear O’Neill Subject area Pharmacology Lab based project

We have discovered that the neurotransmitter noradrenaline (NA) stimulates neurite outgrowth in primary cortical neurons by an action on astrocytic beta2-adrenoceptors . Moreover, recent data for our laboratory indicate that NA acting via glial cells can protect against the ability of the inflammatory cytokines IL-1beta and TNF-alpha to reduce neurite outgrowth. These are important findings as elucidating mechanisms to stimulate neurite outgrowth or to combat reductions in neurite outgrowth in the CNS induced by neurotoxic stimuli is an important objective in the context of providing the next generation of treatments for neurodegenerative disorders. Following on from our recent observations the primary aim of this project is to determine if stimulation of astrocytic beta2-adrenoceptors with NA and or selective agonists (such as formoterol, salmeterol and salbutamol that are used in the treatment of asthma) can combat neurotoxicity induced by a number of disease-relevant stimuli including amyloid-beta, glutamate and the inflammatory cytokines (IL-1beta and TNF-alpha). Techniques employed - Cell culture - Immunostaining - Analysis of neurite outgrowth/apoptosis - Pharmacological manipulations Key references

-adrenoceptors enhances neuronal complexity in primary cortical neurons. PhD thesis, Department of Physiology, Trinity College Dublin. Hertz et al., (2004). Astrocytic adrenoceptors: a major drug target in neurological and psychiatric disorders? Curr Drug Targets CNS Neurol Disord. 3(3):239-67. Madrigal et al., (2007). Neuroprotective actions of noradrenaline: effects on glutathione synthesis and activation of peroxisome proliferator activated receptor delta. J Neurochem. 103(5):2092-101

Project 8: Detection of resistance to the antibiotic clarithromycin in Helicobacter pylori infection

Name of Principal Supervisor Asst. Prof. Smith Subject area Pharmacology Lab based project

The bacterium Helicobacter pylori infects the stomach of approximately half of the world’s population. Most infected individuals have no symptoms, but infection may cause gastritis or stomach ulcers, and increases the risk of developing stomach cancer. Treatment is recommended in all symptomatic patients. For first time treatment of infection the recommended triple therapy consists of a proton pump inhibitor (to supress acid secretion in the stomach), and a combination of 2 antibiotics from amoxicillin, clarithromycin and metronidazole. However, the success rate of this treatment has decreased in recent years, mainly due to the rapid emergence of antibiotic resistant strains of H. pylori. This project aims to detect clarithromycin resistant strains of H. pylori in patients attending Tallaght Hospital. We have already begun collecting samples from H. pylori-infected patients. DNA will be isolated from these samples and examined for resistance-associated mutations using molecular biology methods. The results from this project will contribute to our knowledge on the rate of clarithromycin resistance in the local community and may guide doctors in their future choice of therapy for patients with clarithromycin resistant infections.

Project 9: Detection of resistance to the antibiotic levofloxacin in Helicobacter pylori infection

Name of Principal Supervisor Asst. Prof. Smith Subject area Pharmacology Lab based project

The bacterium Helicobacter pylori infects the stomach of approximately half of the world’s population. Most infected individuals have no symptoms, but infection may cause gastritis or stomach ulcers, and increases the risk of developing stomach cancer. Treatment is recommended in all symptomatic patients. For first time treatment of infection, the recommended triple therapy consists of a proton pump inhibitor (to supress acid secretion in the stomach) and a combination of 2 antibiotics from amoxicillin, clarithromycin and metronidazole. However, the success rate of this treatment has decreased in recent years, mainly due to the rapid emergence of antibiotic resistant strains of H. pylori. When first line therapy fails, levofloxacin is prescribed in the rescue therapy regimen, but levofloxacin resistance is also on the increase. This project aims to detect levofloxacin resistant strains of H. pylori in patients attending Tallaght Hospital. We have already begun collecting samples from H. pylori-infected patients. DNA will be isolated from these samples and examined for resistance-associated mutations using molecular biology methods. The results from this project will contribute to our knowledge on the rate of levofloxacin resistance in the local community and may guide doctors in their future choice of therapy for patients with multi-resistant infections.

Project 10: Measuring the effect of the nitric oxide donor DETA-NONOate on the activity of MMP-9 in Caco-2 cells

Name of Principal Supervisor Asst. Prof. Medina Martin Co-Supervisor Shane O’Sullivan Subject area Pharmacology Lab based project

Background: Matrix Metalloproteinase-9 is known to be upregulated in various pathological conditions including cancer and inflammatory bowel disease. An increase in circulating levels of the enzyme is associated with poor prognosis and so its selective inhibition remains the focus of much research. Several studies have reported on the influence of nitric oxide donors on MMP-9 activity. Aim: The project aims to discover the effect of DETA-NONOate at various concentrations on the activity of MMP-9 in Caco-2 cells. Methods: MMP-9 is induced in Caco-2 cells using the pro-inflammatory cytokines TNF-α and Il-1β. Various concentrations of DETA-NONOate will be co-incubated with the cytokines and the effect on MMP-9 activity will be measured by gelatin zymography of the conditioned media. This project will contribute to the uncovering of the complex relationship between nitric oxide and MMP-9 in inflammation. The student will gain an understanding and practical skills on in vitro work, protein assays, electrophoresis and gelatine zymographies.

Project 11: Comparing the effects of a barbiturate-nitrate hybrid compound and its two component parts on the activity of MMP-9 in a rat colon


Background: MMP-9 is known to be upregualated in inflammatory bowel disease and is associated with disruption of the epithelial barrier and progression of inflammation. Inhibition of this protease enzyme may aid in reducing the severity of the disease. Previous studies from our group have identified a series of barbiturate-nitrate compounds as potential therapeutics in MMP-9 inhibition in IBD. Aims: To evaluate and compare the effects of a barbiturate-dinitrate hybrid compound and its component parts; a non-nitrate analogue and a dinitrate salt on MMP-9 activity on the colon in a rat model of colitis. Methods: Colon samples from rats that have been exposed to dextran sulphate sodium (DSS) for five days and treated with one of the three compounds of interest are lysed. The samples will be normalised for protein concentration and the MMP-9 activity is to measure by gelatin zymography. The project aims to clarify the contribution towards MMP-9 inhibition of the various moieties of the barbiturate-dinitrate compound. The student will gain an understanding and practical skills on in vitro work, protein assays, electrophoresis and gelatine zymographies

Project 12: To discover the effect of a barbiturate-dinitrate compound on the inflammation in a rat model of colitis using rat inflammation array cards


Background: Inflammatory bowel disease (IBD) is a complex, chronic relapsing condition with inflammation and remodelling of the gastrointestinal tract. Dextran sulphate sodium (DSS) induced colitis in rats is bears many of the hallmarks of human IBD and is used to study the disease. Previous studies from our group have demonstrated the protective effects of a barbiturate-dinitrate compound in this model of colitis. Aims: This project aims to discover the effect of a barbiturate dinitrate on the expression of 90 genes known to be involved in inflammation and discover and to gain an understanding of the mechanism of the anti-inflammatory effect of the compound. Methods: mRNA isolated from the colons of rats that have been exposed to dextran sulphate sodium (DSS) and treated with the barbiturate-dinitrate compound is converted to cDNA. The cDNA is loaded into the into the inflammation array cards and the profile of the expression of inflammatory genes will be analysed. The project will elucidate the mechanism of action of the anti-inflammatory effects of the barbiturate-dinitrate compound. The student will gain an understanding and practical skills on in vitro work, RNA isolation and PCR.

Project 13: Analyzing mucin-nanoparticles interaction using QCM-D

Name of Principal Supervisor Asst. Prof. Santos Martinez Co-Supervisor Asst. Prof. Tajber Subject area Pharmacology Lab based project

During the last few years the potential use of engineered nanoparticles (NPs) as drug delivery systems has prompted lots of interest in studying interactions of NPs with living cells and their environment. Quartz Crystal Microbalance with Dissipation monitoring technology (QCM-D) is a useful analytical tool for measuring nanoparticle interactions with different surfaces and substrates. With this technique the viscoelastic properties and mass of the layer attached to the surface of a quartz crystal can be measured with nanogram sensitivity (www.q-sense.com). In order to be considered as drug delivery systems, NPs designed for the oral delivery have to overcome the mucus barrier that covers the epithelial surface in the gastrointestinal system. In this project interactions of NPs loaded with insulin designed for oral delivery (hyaluronic acid/protamine nanoparticles) with gastric mucin will be studied using QCM-D. These experiments will provide information about interactions of NPs intended for the oral delivery of insulin with gastric mucin layers and therefore relevant information for the optimization and the design of these NPs. The student will gain an understanding and practical skills on the production and characterization of nanoparticles; use of QCM-D technology as in vitro model for studying the interaction of NPs and mucin barrier and analysis and modeling of the data using this device.

http://www.q-sense.com/

Project 14: Looking into Silver Nanoparticles-Platelet interactions

Name of Principal Supervisor Asst. Prof. Santos Martinez Subject area Pharmacology Lab based project

During the last few years engineered nanoparticles (NPs) have become very attractive for their use in the medical field. Although nanoparticles can enter the body through various portals; their systemic distribution depends on interactions with blood including blood platelets. It has been demonstrated that NPs can activate platelets in vitro and induce vascular thrombosis in vivo. Therefore, the study of NP-platelet interactions is essential for the design and further use of platelet-biocompatible NPs. Nanotechnology has been applied in dental practice with the development of silver nanoparticles (Ag NPs). In fact, silver compounds and NPs are being used for their antimicrobial activity and as endodontic cements or dental implants. Regardless of the effectiveness that Ag NPs has shown in dental practice, the use of these NPs remains as a controversial area of research with respect to their potential toxicity. The general objective of this project will be to investigate and describe Ag NPs-platelet interactions. The student will gain an understanding and practical skills on the preparation and isolation of human platelets and in the study of platelet function using flow cytometry and in vitro models for the measurement of platelet aggregation.

Project 15: Investigating mechanisms of sepsis-associated thrombocytopenia and thrombosis

Name of Principal Supervisor Asst. Prof. Santos Martinez Subject area Pharmacology Lab based project

Sepsis is defined as a potentially life-threatening complication secondary to a systemic inflammatory response caused by an infection. Severe sepsis accounts for 25% of all admissions to Intensive Care Units and the mortality rate from septic shock is around 30% in developed countries. The inflammatory responses associated to severe sepsis can lead to platelet activation and therefore, platelet consumption and thrombocytopenia (low counting in the number of platelets). The development of thrombocytopenia at any time of the septic process is also associated with an increased mortality in those patients. Matrix metalloproteinases (MMPs) play an important role in platelet activation and are very strongly linked to inflammation. The general objective of this project is to investigate the relationship between sepsis, thrombocytopenia and MMP levels. For this purpose MMP levels will be measured in plasma from healthy donors and septic patients (with and without thrombocytopenia). During this project the student will gain a deeper understanding on the pathophysiology of thrombocytopenia in patients with sepsis and practical skills on some laboratory techniques (protein assay, western blotting and/or gelating zymography).

Project 16: Developing 3D breast cancer cell models to help elucidate resistance to targeted anti-cancer drugs

Name of Principal Supervisor Assoc. Prof. O’Driscoll Co-Supervisor Susan Breslin Subject area Pharmacology Lab based project

Background: Traditionally, in vitro cell culture involves growing cells flat in a monolayer (2D) on cell culture plastic. However, it has been suggested that this method of cell growth does not mimic the complex cell-cell and cell-extracellular matrix signalling that occurs in the in vivo environment. The use of 3D cell culture, which produces multicellular spheroids, has been proposed as an in vitro method that would better replicate the in vivo situation. HER2-overexpression occurs in 20-25% of breast cancer cases and is commonly treated with HER2-targeting drugs. While development of targeted therapy has greatly improved the outcome for HER2-positive patients, issues due to acquired resistance remain problematic. Neratinib is an irreversible EGFR, HER2 and HER4 tyrosine kinase inhibitor currently in phase III clinical trials for breast cancer treatment. We have developed 2 novel neratinib-resistant (NR) breast cancer cell line variants to serve as in vitro models of drug-resistance. We are currently using these models to investigate the effects and mechanisms of resistance to neratinib. Aim: As part of our research into drug resistance, this project will require the nominated student to culture monolayer and 3D spheroid cell culture samples (both drug-sensitive and drug-resistant cell variants) and to prepare these samples for SEM imaging in order to capture the difference in morphology of the cells within the two different culture methods. Techniques involved: Cell culture – monolayer and 3D; scanning electron microscope imaging.

Project 17: Evaluation of the influence of exosomes in vivo in priming the pre-metastatic niche for breast cancer metastasis

Name of Principal Supervisor Assoc. Prof. O’Driscoll Co-Supervisor Keith O’Brien Subject area Pharmacology Lab based project

Background: Exosomes are membrane-bound nanoparticles (30-120nm) which are naturally released for a number of cell types. From analysis of isogenic breast cancer cell lines and their exosomes, our studies have shown that once released into extracellular space, such exosomes are involved in cell-to-cell communication, subsequently increasing the aggression of triple-negative breast cancer (TNBC). We have previously identified a panel of microRNAs (miRNAs) within these exosomes which may be of clinical/therapeutic value. One particular miRNA, miR-134, has been identified and evaluated in silico and in vivo, determining that miR-134 is downregulated in breast cancers, apparently functioning as a tumour suppressor, with reduced expression correlating to increased aggression. We have also shown that miR-134 exerts its effects by blocking STAT5B translation, subsequently reducing HSP90 transcription in the nucleus and thus reducing cellular aggressiveness. Recently, in vivo analysis has been performed on Balb/c nude mice using these exosomes to determine if they can prepare the pre-metastatic niche for breast cancer metastasis. Our results suggest that the more aggressive exosomes hone towards organs commonly effected in breast cancer metastasis. Preliminary in vivo/ex vivo results have determined the exosomes influence the miRNA and protein expression in the mouse brain. From this, we hypothesise that TNBC-derived exosomes may support TNBC metastasis by priming/initiating organs (inc. the brain) for subsequent tumour establishment and growth. Further work is now underway to analyse the effects of these exosomes on miRNA and protein expression in other typical metastatic regions, including: lymph nodes, lungs, bone, and liver, as well as spleen, kidneys and heart. Human serum samples, representing a number of breast cancer subtypes, are also being analysed to establish the clinical relevance of these findings. Aim: The aim of this project is to further analyse the effects that TNBC exosomes have in vivo. This will include: (i) assessing, by qPCR, effects of TNBC exosomes on levels of numerous miRNA expressed in murine brain, lymph nodes, lungs, spleen, bone, liver, kidneys and heart (ii) determining, by western blotting, effects of TNBC exosomes on the protein expression in the same organs (ii) investigating, by confocal microscopy, more precisely the sites of exosome distribution in murine organs; (iv) histological investigation and visualisation of the exact regions of protein expression alterations in mice organs. Techniques involved: Ex vivo analysis, real time PCR, miRNA isolation, protein extraction from cells and tissues, western blotting, immunohistochemistry, data analysis and interpretation, bioinformatics, Excel; GraphPad.

Project 18: Investigating amino acid regulation of mTORC1 activity in NK cells.

Name of Principal Supervisor Assist. Prof. Finlay Subject area Pharmacology/Biotechnology Lab based project

mTORC1 is a serine/threonine kinase which has a crucial role in directing immune responses such that targeting mTORC1 signalling is an attractive approach for a range of immune related diseases, from inflammatory disorders to cancer and autoimmune conditions. However, our understanding of mTORC1 regulation and function in many immune subsets is incomplete. An in depth knowledge of how mTORC1 is controlled in immune cells and how mTORC1 controls the function of these cells will facilitate the development of novel immunotherapies. Research in non-immune cells has established complex regulation of mTORC1 activity. Thus mTORC1 senses the cellular microenvironment for nutrients and growth factors while also monitoring the cellular energy status. To understand how mTORC1 signalling might be manipulated for therapeutic benefit is is important to establish whether mTORC1 activity is subject to similar regulation in immune cells. NK cells have cytolytic functions and are important for the direct killing of virally infected cells and malignant tumour cells. It is likely that amino acid concentrations can become limiting at sites of disease such as within the tumour microenvironment. This project will investigate whether mTORC1 activity is regulated by amino acid availability in NK cells. The amino acids required to maintain mTORC1 activity can differ depending on the cell type. This project will establish exact amino acid requirements for mTORC1 activity in NK cells. NKL cells (an NK cell line) will be cultured in IL-2 with and without amino acids and with a range of amino acid combinations. Cells will then be lysed and mTORC1 activity monitored by western blot analysis for the mTORC1 substrates p70 S6 ribosomal kinase (S6K) and 4E binding protein 1 (4EBP1). Likely outcome: This project will determine which amino acids are required for mTORC1 activity in NK cells and will thus make a significant contribution to our understanding of mTORC1 signalling in this important immune cell subset.

Project 19: Investigating glucose regulation of mTORC1 activity in NK cells.


mTORC1 is a serine/threonine kinase which has a crucial role in directing immune responses such that targeting mTORC1 signalling is an attractive approach for a range of immune related diseases, from inflammatory disorders to cancer and autoimmune conditions. However, our understanding of mTORC1 regulation and function in many immune subsets is incomplete. An in depth knowledge of how mTORC1 is controlled in immune cells and how mTORC1 controls the function of these cells will facilitate the development of novel immunotherapies. Research in non-immune cells has established complex regulation of mTORC1 activity. Thus mTORC1 senses the cellular microenvironment for nutrients and growth factors while also monitoring the cellular energy status. To understand how mTORC1 signalling might be manipulated for therapeutic benefit is is important to establish whether mTORC1 activity is subject to similar regulation in immune cells. NK cells have cytolytic functions and are important for the direct killing of virally infected cells and malignant tumour cells. It is likely that glucose concentrations can become limiting at sites of disease such as within the tumour microenvironment. This project will investigate whether mTORC1 activity is regulated by limiting glucose availability in NK cells. NKL cells (an NK cell line) will be cultured in IL-2 and a range of glucose concentrations. Cells will then be lysed and mTORC1 activity monitored by western blot analysis for the mTORC1 substrates p70 S6 ribosomal kinase (S6K) and 4E binding protein 1 (4EBP1). The AMP dependent kinase (AMPK) is responsible for turning mTORC1 off when energy levels are depleted. Therefore, the activity of AMPK will also be monitored using western blot analysis. Likely outcome: This project will determine whether mTORC1 is a glucose sensor in NK cells and will thus make a significant contribution to our understanding of mTORC1 signalling in this important immune cell subset.

Project 20: Investigating growth factor regulation of mTORC1 activity in NK cells.


mTORC1 is a serine/threonine kinase which has a crucial role in directing immune responses such that targeting mTORC1 signalling is an attractive approach for a range of immune related diseases, from inflammatory disorders to cancer and autoimmune conditions. However, our understanding of mTORC1 regulation and function in many immune subsets is incomplete. An in depth knowledge of how mTORC1 is controlled in immune cells and how mTORC1 controls the function of these cells will facilitate the development of novel immunotherapies. Research in non-immune cells has established complex regulation of mTORC1 activity. Thus mTORC1 senses the cellular microenvironment for nutrients and growth factors while also monitoring the cellular energy status. To understand how mTORC1 signalling might be manipulated for therapeutic benefit is is important to establish whether mTORC1 activity is subject to similar regulation in immune cells. In most cell types PI3-kinae and PKB signaling are required for growth factor activation of mTORC1. However, our research reveals that this is not the case in certain T cell subsets. As PI3-kinase and PKB inhibitors are in clinical trials for cancer therapy it is important to undersand the impact that inhibiting these pathways will have on the immune system. This project will investigate whether PI3-kinase and PKB are required for IL2 activation of mTORC1 in NK cells. NK cells have cytolytic functions and are important for the direct killing of virally infected cells and malignant tumour cells. NKL cells (an NK cell line) will be cultured in IL-2 +/- various inhibitors of PI3-kinase and PKB signalling. Cells will then be lysed and mTORC1 activity monitored by western blot analysis for the mTORC1 substrates p70 S6 ribosomal kinase (S6K) and 4E binding protein 1 (4EBP1). PKB activity will also be monitored by western blot analysis. Likely outcome: This project will determine whether PI3-kinase and PKB are required for mTORC1 activity in NK cells and will thus make a significant contribution to our understanding of mTORC1 signalling in this important immune cell subset.

Project 21: Does inhalable insulin cause or promote lung cancer?

Name of Principal Supervisor Assoc. Prof. Ehrhardt Subject area Biopharmaceutics/Pharmacology Non-lab based project

Inhalable insulin was available from September 2006 to October 2007 as a new method of delivering insulin, a drug used in the treatment of diabetes, to the body. The first such product to be marketed was Exubera®, a powdered form of recombinant human insulin, delivered through an inhaler into the lungs where it is absorbed. A systematic review concluded that inhaled insulin "appears to be as effective, but no better than injected short-acting insulin. The additional cost is so much more that it is unlikely to be cost-effective." In October 2007, Pfizer announced that it would be discontinuing the production and sale of Exubera due to poor sales. Several other companies were developing inhaled forms of the drug to reduce the need for daily injections among diabetics. After the withdrawal of Exubera work on most other inhalable formulation was discontinued, therefore, all currently available insulin formulations are administered by subcutaneous or intravenous injection. In April 2008, Pfizer announced in its "Dear Doctor" letter that Exubera may have been associated with lung cancer: of the 4,740 patients who used Exubera in clinical trials, six have developed lung cancer as of April 2008, compared to only one of the 4,292 patients in the placebo group. The association was not statistically significant, and Pfizer maintained in its letter that "Exubera remains a safe and effective medication." In a letter 18 June 2008, Pfizer informed UK doctors of the above mentioned six cases, noting that they all had a prior history of cigarette smoking and that they were planning to investigate further the "observed imbalance in diagnosed lung cases" with an international observational trial. This international observational trial was never conducted. The aim of this project, therefore, is conducting a thorough literature review on the insulin-cancer relationship with particular emphasis on lung malignancies.

Project 22: Characterisation of NBD-MTMA uptake in NCI-H441 human bronchiolar epithelial cells

Name of Principal Supervisor Assoc. Prof. Ehrhardt Co-Supervisor Sergey Zaichik Subject area Biopharmaceutics/Pharmacology Lab based project

Drug transporter effects on pulmonary PK have been hypothesised for several years now and a number of in vitro studies presented data consistent with this notion. Transporters may alter airway residence times of drugs, modulate access of drugs to intracellular targets and submucosal lung tissue, and potentially influence drug absorption profiles into and from the systemic circulation. Very recent studies conducted in experimental animal models and human volunteers, for the first time, report findings that support the idea that drug-transporter interactions are indeed a clinical reality. Particularly, the family of ATP-binding-cassette (ABC) transporters - to which many efflux pumps belong – has been associated with increased airway retention time of inhaled bronchodilators, i.e., β-agonists and anti-muscarinics. Consequently, inhibition of these transporters by drugs such as verapamil, which is frequently used by patients suffering from co-morbidities like hypertension, has profound implications on local drug concentrations at the target receptors, but also systemic exposure and side effects. In the same context, it is important to confirm if our currently available in vitro models are powerful enough to predict in vivo findings. This project is aiming to functionally characterise organic cation transporter (OCTs) using a novel fluorescent substrate, N,N,N-trimethyl-2-[methyl(7-nitrobenzo[c][l,2,5]oxadiazol-4-yl)amino]ethanaminium iodide (NBD-MTMA).

Project 23: Multicomponent solid state systems of lidocaine

Name of Principal Supervisor Assist. Prof. Tajber Subject area Pharmaceutics Lab based project

Lidocaine (LD) is a local anesthetic and an antiarrhythmic agent, used mainly as a hydrochloride salt. LD hydrochloride has also very interesting physicochemical properties e.g. is a glass-forming pharmaceutical that is an ionic conductor. We have recently published on the synthesis of LD hemisuccinate, which has similar physical properties to LD hydrochloride. The nature of the counterion, i.e. the acid component of LD salt, has also a direct impact on the skin permeation properties of LD, as it is hypothesised that the drug is transported via ion-pair mechanism. The aim of this project is to investigate the potential of LD to form multicomponent systems (e.g. salts) with acidic molecules, especially with dicarboxylic acids, and to study the ion-pair behaviour of such multicomponent systems in water. The student will learn a range of preformulation techniques such as differential scanning calorimetry (DSC), X-ray diffraction (XRD) and well as infrared (IR) and UV-Vis spectroscopies. Selected references:

Serajuddin AT. Salt formation to improve drug solubility, Adv Drug Deliv Rev. 2007, 59(7), 603-616.

Wojnarowska Z et al. Decoupling of conductivity relaxation from structural relaxation in protic ionic liquids and general properties, Phys Chem Chem Phys. 2013, 15(23), 9205-9211.

Project 24: In vitro stability studies of insulin –free and -loaded nanoparticles in the presence of simulated gastrointestinal (GI) fluids

Name of Principal Supervisor Assist. Prof. Tajber Co-Supervisor Svenja Sladek Subject area Pharmaceutics Lab based project

Peptide encapsulation in nanoparticles (NPs) is one of the many strategies which have been established to e.g. enhance absorption of insulin via oral delivery. We have developed a new NP system, which has shown promising properties as carrier of insulin. This novel formulation comprises natural polymers (hyaluronic acid or alginic acid) and protamine (as a cell penetrating peptide). It is expected that the polymer will protect insulin against proteolytic enzymes and an acidic gastric environment. However, the nanoparticulate formulations must also have an adequate physical stability (no aggregation, no dissolution) in GI fluids, which is challenging considering inherent instability of NPs. The aim of this project is to assess the physical stability of the various NPs (with and without insulin) in simulated gastrointestinal fluids (SGIFs). The student will have an opportunity to learn how to prepare empty and peptide loaded nanoparticles and evaluate their physical stability by measuring the particle size and surface potential as well as estimate the degree of peptide leakage. Proposed plan of work: 1st week Literature review of publications relevant to the project. Researching info on the composition of SGIFs and best practice guidelines for physical stability testing of NPs. Planning the experiments for the rest of project, preparation of experimental protocols and stock solutions. 2nd week Learning the techniques of carrier and peptide loaded NPs preparation and characterisation. Stability studies of NP carriers (NPs with no insulin): estimation of particle size, size distribution and surface potential. Estimation of degree of NP aggregation based on the intensity of scattered light. 3rd week Stability studies of insulin-loaded NPs (as week 2). Examination of peptide leakage by HPLC. Critical evaluation of the data obtained and presentation of the write-up plan. 4th week Writing up the project. Selected references:

Umerska A. et al. Exploring the assembly process and properties of novel crosslinker-free hyaluronate-based polyelectrolyte complex nanocarriers. Int J Pharm., 2012, 436(1-2):75-87.

Roger E. et al. The gastrointestinal stability of lipid nanocapsules. Int J Pharm., 2009, 379(2), 260-265.

Project 25: Amorphous solid dispersions prepared by spray drying

Name of Principal Supervisor Prof. Healy Co-Supervisor Claire Brennan Subject area Pharmaceutics/Pharmaceutical Technology Lab based project

Background: The therapeutic application of a high proportion of new chemical entities in the drug discovery pipeline is limited by their low solubility. By many estimates up to 40 per cent of new chemical entities (NCEs) discovered by the pharmaceutical industry today are poorly soluble or lipophilic compounds. A strategy to facilitate oral delivery of such substances is the formulation of amorphous solid dispersions, comprising an active pharmaceutical ingredient combined with a water-soluble polymer to produce a single-phase amorphous mixture. Drugs in the amorphous state have a higher apparent solubility and faster dissolution than their crystalline counterparts but are usually less physically and chemically stable. Co-formulation with a polymer is a strategy used to physically stabilise the amorphous drug form. Aim: The aim of the current study is to co-spray dry carbamazepine, which is a BCS Class II compound, with three different polymers - Kollidon VA 64, HPMC AS LF and PVP K17 - at a range of different drug/polymer weight ratios and to characterise the solid dispersions produced. The objective is to determine the ease of processing of the various composite systems and to evaluate how the product characteristics vary depending on solid dispersion composition. Methods: Spray drying will be undertaken using a Büchi B-290 Mini Spray Dryer. The solid-state characteristics of raw materials and co-spray dried systems will be assessed using powder X-ray diffraction, FTIR and thermal analysis. Thermal analysis using differential scanning calorimetry (DSC) will be directed at evaluating the glass transition temperature of the amorphous phase, as an indicator of the physical stability of the amorphous form in the solid state.

Project 26: Complementary and alternative medicine: Patterns of use in the republic of Ireland.

Name of Principal Supervisor Assoc. Prof. Sheridan Subject area Pharmacognosy Non-lab based project

Background: There has been an increasing use of complementary and alternative medicine in Ireland over the last number of years. This is paralleling a trend in the North of Ireland that has led to the proposed plans for patients to be able to access CAM through GP’s and Health Boards. Despite this trend in Northern Ireland there are no figures available in the republic for CAM use in The Republic of Ireland. This project: This project involves carrying out an online survey (using Survey monkey) looking at a variety of factors influencing the use of CAM therapies. The survey will be anonymous. Questions will be designed to identify the aspects of CAM most used by respondents and to reveal the motivations behind its use. Outcomes: Upon completion of project, the candidate will have enhanced their knowledge of CAM (an integral part of the S. Sophister curriculum (PH4003)), data acquisition, analysis and interpretation.

Project 27: Synthesis of small chemical fragments for incorporation into novel anti-inflammatory drugs

Name of Principal Supervisor Assoc. Prof. Sheridan Subject area Pharmacognosy Lab based project

Background: As part of an ongoing drug discovery programme we continue the quest to identify potential new medicines from the natural world and also by the synthesis of new molecules incorporating nature’s bioactive designs. This project involves the development of an approach to the synthesis of a number of small chemical fragments which are destined to be incorporated into novel chemical scaffolds. This project: The candidate will carry out a number of simple synthetic chemical steps including the addition and removal of protecting groups under atmospheric and inert atmospheres, and the coupling of small chemical components using aldol type reactions. The student will carry out chromatographic separations and purifications and will use advanced spectroscopic techniques to characterise the products of their reactions. Outcomes: Upon completion of project, the candidate will have enhanced their knowledge of medicinal chemistry; will have developed laboratory skills related to synthesis and purification of organic compounds. Will have applied their knowledge of spectroscopy to characterisation of novel molecules.

Project 28: Isolation of cannabidiol from Hemp

Name of Principal Supervisor Assist. Prof. Walsh Subject area Pharmacognosy Lab based project

The purpose of this project is to develop a novel, student friendly method for the isolation of cannabidiol from Hemp. Part of the project will also involve a detailed assignment of its structure using IR, NMR and HRMS. The student will also be expected to conduct a critical review of the pharmacological data published in support of the potential use of this substance in the treatment of cancer and psychiatric conditions. References Carroll AM, Kavanagh DJ, McGovern FP, Reilly JW and Walsh JJ. Nature’s Chiral Catalyst and Anti-Malarial: Isolation and Structure Elucidation of Cinchonine and Quinine from Cinchona calisaya. 2012, 89, 1578-1581. Walsh EL, Ashe S. and Walsh JJ Nature’s Migraine Treatment: Isolation and Structure Elucidation of parthenolide from Tanacetum parthenium Journal of Chemical Education, J. Chem. Educ. 2012, 89, 134–137. Mohd Nazri M, Samat FD, Kavanagh PV and Walsh JJ Nature’s Cholesterol Lowering Drug: Isolation and Structure Elucidation of Lovastatin from Red Yeast Rice-containing Dietary Supplements J. Chem. Educ. 2012, 89, 138–140. Halpin CM, Reilly C, and Walsh JJ Nature’s Anti-Alzheimer’s Drug: Isolation and Structure Elucidation of Galantamine from Leucojum aestivum, Journal of Chemical Education, 87 , (11), 2010, p1242 – 1243. Doyle AM, Reilly J, Murphy N, Kavanagh PV, O'Brien JE, Walsh MS, Walsh JJ Nature's sedative: Isolation and structural elucidation of valtrate from Centranthus ruber, Journal of Chemical Education, 81, (10), 2004, p1486 – 1487.

Project 29: Isolation of berberine and hydrastine from Hydrastis canadensis (Golden seal).


The purpose of this project is to develop an elegant method for the isolation of berberine and hydrastine from Hydrastis canadensis. The second part of the project will involve the use of modern spectroscopic techniques to confirm the identity of the compounds isolated. The project will also involve a critical assessment of the pharmacological and clinical data supporting the use of berberine for the treatment of diabetes. References Carroll AM, Kavanagh DJ, McGovern FP, Reilly JW and Walsh JJ. Nature’s Chiral Catalyst and Anti-Malarial: Isolation and Structure Elucidation of Cinchonine and Quinine from Cinchona calisaya. 2012, 89, 1578-1581. Walsh EL, Ashe S. and Walsh JJ Nature’s Migraine Treatment: Isolation and Structure Elucidation of parthenolide from Tanacetum parthenium Journal of Chemical Education, J. Chem. Educ. 2012, 89, 134–137. Mohd Nazri M, Samat FD, Kavanagh PV and Walsh JJ Nature’s Cholesterol Lowering Drug: Isolation and Structure Elucidation of Lovastatin from Red Yeast Rice-containing Dietary Supplements J. Chem. Educ. 2012, 89, 138–140. Halpin CM, Reilly C, and Walsh JJ Nature’s Anti-Alzheimer’s Drug: Isolation and Structure Elucidation of Galantamine from Leucojum aestivum, Journal of Chemical Education, 87 , (11), 2010, p1242 – 1243. Doyle AM, Reilly J, Murphy N, Kavanagh PV, O'Brien JE, Walsh MS, Walsh JJ Nature's sedative: Isolation and structural elucidation of valtrate from Centranthus ruber, Journal of Chemical Education, 81, (10), 2004, p1486 – 1487.

Project 30: Isolation of benzoyl cinnamate and benzoyl benzoate from Balsam Peru.


Balsam Peru is contained in several preparations for the treatment of treatment of severe and acute discomfort or pain associated with internal and external haemorrhoids. The purpose of this project is to develop an elegant method for the isolation of its purported active constituents, benzoyl cinnamate, benzoyl benzoate and cinnamic acid and to study the chemistry by which these substances may be transformed into other derivatives of pharmaceutical importance. Selected References Carroll AM, Kavanagh DJ, McGovern FP, Reilly JW and Walsh JJ. Nature’s Chiral Catalyst and Anti-Malarial: Isolation and Structure Elucidation of Cinchonine and Quinine from Cinchona calisaya. 2012, 89, 1578-1581. Walsh EL, Ashe S. and Walsh JJ Nature’s Migraine Treatment: Isolation and Structure Elucidation of parthenolide from Tanacetum parthenium Journal of Chemical Education, J. Chem. Educ. 2012, 89, 134–137. Mohd Nazri M, Samat FD, Kavanagh PV and Walsh JJ Nature’s Cholesterol Lowering Drug: Isolation and Structure Elucidation of Lovastatin from Red Yeast Rice-containing Dietary Supplements J. Chem. Educ. 2012, 89, 138–140. Halpin CM, Reilly C, and Walsh JJ Nature’s Anti-Alzheimer’s Drug: Isolation and Structure Elucidation of Galantamine from Leucojum aestivum, Journal of Chemical Education, 87 , (11), 2010, p1242 – 1243. Doyle AM, Reilly J, Murphy N, Kavanagh PV, O'Brien JE, Walsh MS, Walsh JJ Nature's sedative: Isolation and structural elucidation of valtrate from Centranthus ruber, Journal of Chemical Education, 81, (10), 2004, p1486 – 1487.

Project 31: Chemical Evaluation of caffeine containing pills from Head Shops

Name of Principal Supervisor Assist. Prof. Boylan Subject area Pharmacognosy Lab based project

Background: According to NACD (National Advisory Committee on Drugs) 2010 was a very busy year, marked by many challenges, not least responding to the threat of new and emerging psychoactive substances available through Head Shop outlets. In January 2010, at the request of the Minister with responsibility for the National Drugs Strategy, an expert and multi-disciplinary Research Advisory Group(RAG) was established by the NACD to carry out a review of new psychoactive substances and the outlets supplying them. On foot of the deliberations of the RAG, the NACD advised Government in February 2010 to bring a range of synthetic drugs under the control of the Misuse of Drugs Act1. This culminated with the closure of all head shops in Ireland. Although the shops are not there anymore, one can still acquire these new synthetic drugs from websites. Party pills, also known as "herbal highs", "pep pills" "dance pills" and "natural power", is a colloquialism for a type of recreational drugs that can be purchases in these websites. Their main ingredient was originally benzylpiperazine (BZP), but has now expanded to a wide range of compounds with a variety of effects, including caffeine which is a legal compound. Although BZP is banned in Ireland, several of the originally sold party pills are now reporting to contain caffeine and a blend of medicinal plants, which makes it legal to be sold in Ireland. Aims of this project: The idea of this project is to investigate the chemical nature of two of these caffeine containing party pills acquired online from one UK head shop. These are new products that are not listed by the Gardai’s National drugs Unit list.2 Methodology to be used: Extraction of the new drugs, analysis by different types of chromatography and final evaluation using spectroscopic techniques. Expected outcomes: Come up with the possible identity of any possible component other than caffeine in these new drugs in order to contribute with the Gardai up-to-date National Criminal Intelligence Unit’s list. References:

1. NACD Annual report 2010, May 2011, 36 pages. 2. Drug Names and Street Terminology – Ireland and Head Shop products and

Substances list. National Criminal Intelligence Unit, January 2011. 3. Narrowing of regional differences in the use of illegal drugs, News Release,

issued by the National Advisory Committee on Drugs,Tuesday June 19th, 2012.

Project 32: Quantification of Vitamin C in nutraceuticals – the case of Hyppophae rhamnoids L


Background: Hippophae rhamnoides, common sea-buckthorn, is a species of flowering plant in the family Elaeagnaceae, native to fixed dunes and sea cliffs in Europe and Asia. The fruits of sea buckthorn are used in a wide variety of products. Especially in France (southern Alps) sea buckthorn is commonly sold as fruit juice or as an ingredient in non-alcoholic and alcoholic mixed beverages. Other uses include the berries to be processed as fruit wine or into liquor as well as jam. Buckthorn tea is also made out of the fruits and originates from India. The fruits have very high vitamin C content, on average exceeding that of lemons and oranges. Recently two specimens of sea-buckthorn were collected one from Ireland and one from Poland. The fruits were cleaned and deseeded Aims of this project: The idea of this project is to quantify the vitamin C content in the two samples of Hippophae recently collected and ascertain whether there is any significant difference between the Irish and the Polish samples, which could have economic implication in terms of sourcing material to the food and cosmetic industries. Methodology to be used: Extraction of the plant material, partition with different solvents, quantification of the vitamin C content by both titration and HPLC. The vitamin C analysis as well as the determination of any other hydro soluble vitamin will be performed by thin layer chromatography with stationary phase gradient. Expected outcomes: Establishment of the quantity of vitamin C in the two specimens of sea buckthorn from two different countries, comparison with other sources of vitamin C and proving that deseeding the material leads to no loss of vitamin C. References:

1. Claudia Cimpoiu, Anamaria Hosu, Anitta Puscas. Journal of Chromatography A, 1223: 142-146, 2012.

2. Jie Zheng, Heikki Kallio, Kaisa Linderborg, Baoru Yang. Food Research International, 44: 2018-2026, 2011.

Project 33: Chemical Evaluation of sexual enhancer pills from Head Shops


Background: Products promising to enhance sexual performance have been promoted for over a century. They have always been characterized by wildly exaggerated claims and they usually lack evidence of safety or effectiveness. This exposes the consumer to unknown health hazards, as these products use ingredients that are of unknown therapeutic benefit. Marketing hype and high profits should never override the pharmacist's professional responsibility to demand proof of safety and efficacy in the recommended products. Head shops in the world are now selling this sexual enhancer pills together with their party pills with a very aggressive marketing strategy. They have products “designed” for men and products “designed” for women. They claim to contain a blend of medicinal plants containing Ginkgo, Muira Puama, Ginseng and Ashwaganda as the main core and Yohimbine for men and Dong Quai for women. Aims of this project: The idea of this project is to investigate the chemical nature of two of these sexual enhancer pills (one for women and one for men) acquired online from one UK head shop. These are new products that are not listed by the Gardai’s National drugs Unit list.2 Methodology to be used: Extraction of the new drugs, analysis by different types of chromatography and final evaluation using spectroscopic techniques. Expected outcomes: Come up with the possible identity of any possible component other than the plant blend in these new drugs in order to contribute with the Gardai up-to-date National Criminal Intelligence Unit’s list and also to determine whether the male product differs from the female’s one. References:

1. Hafez B, Hafez ES. Andropause: endocrinology, erectile dysfunction, and prostate pathophysiology. Arch Androl. 2004;50:45-68.

2. Drug Names and Street Terminology – Ireland and Head Shop products and Substances list. National Criminal Intelligence Unit, January 2011.

Project 34: Analysis of Dilute Adrenaline Samples Stored in Various Packaging Materials for Stability Testing using HPLC and LC-MS

Name of Principal Supervisor Assist. Prof. Sasse Co-Supervisor Peter O’Connell Subject area Pharmaceutical Chemistry Lab based project

The School of Pharmacy & Pharmaceutical Sciences is currently involved in an industry sponsored project to assess the suitability of a new polymer material (Barex™) as a potential replacement for glass and polypropylene in the packaging of parenteral pharmaceutical products. To date samples of dilute adrenaline for injection have been transferred to Barex™, borosilicate glass and polypropylene containers and stored under a selection of temperature and humidity controlled environments. The potency of the active ingredient has been assessed using a HPLC assay at regular time intervals. It has been observed that the concentration of adrenaline present in some container types and storage conditions has decreased over time. The aim of this project is to identify and quantify any impurities or degradants that may be present in each sample of adrenaline in order to establish the related substances profile of the product. This profile is important as it will allow us to evaluate the suitability of the chosen packaging material by comparing the range of impurities obtained in each packaging material and storage condition. The identity of each impurity will be confirmed using LC-MS by comparing the resulting mass spectra to all known impurities of adrenaline. Suitable methods will be chosen following a literature review of journal articles and current pharmacopoeial methods and an assessment of the compatibility of the chosen methods with equipment and materials available within the School prior to analysing the sample solutions. It is then planned to validate the method(s) according to industry guidelines (ICH) to ensure that the method is fit for purpose. The student will benefit from the close alignment of this project with PH4002 lectures.

Project 35: Optimisation and Validation of a HPLC Method for the Simultaneous Detection & Quantification of Budesonide and Formoterol Fumarate in a Commercial Dry Powder Inhaler

Name of Principal Supervisor Assist. Prof. Sasse Co-Supervisor Peter O’Connell Subject area Pharmaceutical Chemistry/Pharmaceutics Lab based project

The School of Pharmacy and Pharmaceutical Scienes, in collaboration with physicians at the RCSI and biomedical engineers located in the TBSI, is involved in a project to investigate the amount of drug product delivered to a patient via commonly available inhalers. Current research has demonstrated a noticeable effect of the velocity, duration and the relative humidity of the inhalation on the amount of drug product administered to the patient and it is intended to continue this research by investigating additional pharmaceutical products and drug delivery devices. This project is primarily concerned with developing, optimising and validating a High Performance Liquid Chromatography (HPLC) method to allow the detection and quantification of both active pharmaceutical ingredients in the Symbicort® Turbohaler® (Budesonide and Formoterol Fumarate). A suitable method will be identified from a review of existing literature and the method will be optimised for use with the chosen formulation as well as with the reagents and equipment available at the School to allow the detection of low levels of each component. The method will then be validated in accordance with industry guidelines for specificity, linearity, accuracy, precision, ruggedness, and robustness to demonstrate the suitability of the method for its intended purpose. Finally prior to submitting the method for use by our colleagues in the RCSI and TBSI some in-vitro experiments will be carried out to assess the particle size distribution of the inhaled drug products using both the Next Generation Impactor (NGI) and the Anderson Cascade Impactor (ACI) in order to troubleshoot any potential problems that may be encountered during routine analysis.

Project 36: The derivation of the eccentric adjacency index

Name of Principal Supervisor Assist. Prof. Quigley Subject area Pharmaceutical Chemistry Non-lab based project

The discriminating power of a relatively novel topological index, the eccentric adjacency index, is investigated. The derivation will be applied to a series of phenethylamines. The intercorrelation of this index with other indices and properties will be examined, in order to assess the appropriate combination for an SAR analysis. The Computational software suites employed will be CLogP, Molecular Modelling Pro and SPARTAN.

Project 37: In search of inhibitors of protein aggregation

Name of Principal Supervisor Assist. Prof. Gilmer Subject area Pharmaceutical Chemistry Lab based project

Protein aggregation is thought to contribute to several diseases collectively termed amyloidosis or amyloid diseases. These are especially prominent in the central nervous system including Alzheimer’s Disease, Parkinson’s, Huntington’s corea. Mis-folded proteins also aggregate intracellularly causing ER stress leading to an even wider range of pathophysiological contributions through effects on protein secretion. Protein aggregation is also a key mechanism of degradation of biopharmaceuticals leading to reduced potency, efficacy and increased side effects. Inhibitors of protein aggregation are highly desirable therefore for disease treatment or for stabilisation of biological products. Several compounds are known that have the ability to inhibit protein aggregation and/or reduce ER stress reducing associated problems. The bile acid ursodeoxycholic (UDCA) and its taurine conjugate (TUDCA) have these inhibitory properties and related protein chaperone activity (they may help proteins fold properly by stopping them aggregating in the unfolded state). We have noticed that several new derivatives of UDCA and TUDCA are potent inhibitors of thermally induced aggregation of bovine serum albumin (BSA). We would like to see if this is an effect specific to BSA. Therefore in this project we have selected another smaller protein –beta-lactoglobulin. The researcher will study the denaturation –aggregation of betalactoglobulin and generate some simple new derivatives of UDC A to test as inhibitors.

Project 38: In vitro metabolism studies on a candidate cancer chemopreventative agent

Name of Principal Supervisor Assist. Prof. Gilmer Subject area Pharmaceutical Chemistry Lab based project

Esophageal adenocarcinoma is a leading cause of cancer death. It is usually preceded by a condition known as Barrett’s esophagus in which the normal squamous epithelium is replaced by an intestinal phenotype. Bile acids present in gastroduodenal refluxate of GERD patients may be involved in this process. We and others are interested in the potential of ursodeoxycholic acid (UDCA) in this setting because its established clinical use in liver diseases is to some extent explained by an ability to block the pathophysiological effects of other bile acids. UDCA has independent anti-inflammatory effects mediated in part by the glucocorticoid receptor, which it weakly activates. We produced several compounds related to UDCA with much greater potency for GR activation. One of the UDCA analogs – a simple amide- was subsequently shown to be highly effective in a surgical rat model of GERD which produces a condition similar to BE leading to EA. The successful analog was significantly different in effect to UDCA. The compound has promise for the treatment and chemoprevention of disorders of the esophagus, colon and liver. We would like to know the extent to which the effects observed were due to the compound itself as opposed to UDCA which is likely to have been a metabolite in the rat study. Therefore, we propose to study the behaviour of the compound under conditions simulating those it will have encountered in the rat model. This includes rat intestinal microsomes, rat liver and rat serum. HPLC/HPLC MS will be used to estimate a half-life for the candidate drug in vitro and to help identify it byproducts.

Project 39: Design and chemistry of novel cytotoxic indazoles

Name of Principal Supervisor Prof. Meegan Co-Supervisor Tadhg Cotter Subject area Pharmaceutical Chemistry Lab based project

Both tubulin and the Hsp90 chaperone protein are recognised as important targets for new cancer chemotherapeutic compounds. A “first in class” dual tubulin/Hsp90 binding compound was recently discovered and identified by our research group. The lead compound identified from the study contains an indazole core structure. We have recently discovered interesting cytotoxic effects of indazoles associated with their tubulin and heat shock protein (HSP90) targeting activities. The indazole scaffold is found in many compounds displaying a wide range of biological activities including HIV protease inhibitors, kinase inhibitors, factor XIa inhibitors and human neutrophil elastase inhibitors. In this project, the use of the indazole ring as a scaffold for the design of bioactive compounds is investigated to gain access to novel therapeutic agents that possess vascular targeting and anticancer effects in tumour cells. The specific objective of this project is to prepare a small focused library of analogues of the lead indazole molecule and to evaluate these compounds as molecular probes for proteins which regulate cancer cell proliferation. The chemistry will focus primarily on the development of the structure of the indazole ring and will investigate methods for the introduction of aryl substitutents onto the C-3 position of the molecule. This approach will add greater functionality to the lead indazole compound with the objective of increasing the potency of the lead compound. The project will have elements of organic synthesis, purification (chromatography) and analytical chemistry (Mass Spectrometry, 1H and 13C Nuclear Magnetic Resonance spectroscopy, Infrared Spectroscopy). The biochemical evaluation of the synthesized compounds will be investigated to determine potency.

Project 40: Investigation of 1,4-dihydropyridines as multi-drug resistance reversal agents

Name of Principal Supervisor Prof. Meegan Co-Supervisor Conor O’Shea Subject area Pharmaceutical Chemistry Lab based project

Multidrug resistance(MDR) is considered to be one of the major reasons of failure of chemotherapy for the majority of cancer patients. There is much interest in the discovery of small molecule inhibitors of MDR. The 1,4-dihydropyridines (DHPs) were initially introduced as a class of calcium channel blockers, and are widely used in cardiovascular diseases and in the treatment of hypertension. It is now known that these calcium channel blockers can be effective as a new class of drug resistance reversers in cancer treatment. Many studies have been carried out to identify new DHPs for reversal of MDR while reducing their calcium channel blocking effects. A series of biologically active 1,4-dihydropyridine scaffold structures has been identified in our research group as lead structures from initial high throughput screening (HTS) with both interesting anti-proliferative and Pgp inhibitory properties. This project will investigate the identification of novel cancer chemotherapeutic agents and will focus on the synthesis, structural characterisation and evaluation of 1,4-dihydropyridine and related dihydropyridone compounds. The aims of this present research project are:

i. Synthesis of a small library of dihydropyridine and dihydropyridone compounds, based on the initial molecular scaffold structures identified as a lead structure from initial in silico screening.

ii. Spectroscopic characterisation to confirm identity of synthetic products

iii. In vitro biochemical evaluation of selected synthesised compounds in MCF-7 breast cancer cell line to confirm target of compound.

Optimisation studies of the initial scaffold will help to determine the structural requirements of the compounds at the ER and Pgp binding sites.

Project 41: Design of novel molecules to target Burkitt’s lymphoma

Name of Principal Supervisor Prof. Meegan Co-Supervisor Andrew Byrne Subject area Pharmaceutical Chemistry Lab based project

Burkitt’s lymphoma (BL) is a non-Hodgkin’s lymphoma (NHL), which manifests as tumours composed of small non cleaved B-cell lymphocytes. Cancers of the lymphatic cells (lymphomas) account for approximately 12% of malignant diseases worldwide and 80% of these cases are subdivided into non-Hodgkin’s lymphomas (NHL). As part of an ongoing research project we have sought to identify possible alternatives to the current clinical drugs used in the treatments for Burkitt’s lymphoma. We have recently discovered structural derivatives of the anti-depressant drug maprotiline which are effective against Burkitt’s lymphoma and related lymphomas and are potential new targets for the design of compounds with activity against Burkitt’s lymphoma . We have demonstrated that a series of these maprotiline related 9,10-ethanoanthracenes express significant antiproliferative effects in vitro in two BL cell lines: EBV- MUTU-I and the chemoresistant EBV+ DG75 lymphoma cell lines. The objective of this project is the design and evaluation of novel small molecules which target Burkitt’s lymphoma. Based on our previous studies, a small focussed library of structurally related anthracene compounds will be designed and characterised. Many of these compounds contained a classic nitrostyrene core which has also been previously shown to possess activity in the Burkitt cell lines. In this project the structures proposed for investigation are based on the anthracene and related triptycene scaffolds. These compounds will be synthesised via a number of established routes and each compound will be subsequently characterised by spectroscopic methods (1H and 13C NMR, IR and HRMS). The antiproliferative activities of the compounds are evaluated using two BL cell lines: EBV- MUTU-I and the chemoresistant EBV+ DG75 and a possible mechanism of action for the compounds will be investigated. The structure activity relationships for the series of products will be examined.

Project 42: Patient self-administration of medication in Tallaght Hospital – a descriptive survey of the frequency and the type of products administered.

Name of Principal Supervisor Assoc. Prof. Grimes Co-Supervisor Evelyn Deasy Subject area Clinical Pharmacy Non-lab based project

Objectives:

1. Identify what proportion of prescribed medication is self-administered. 2. Characterise what medications are being self-adminstered. 3. Assess suitability of the patient and medication for self-administration.

Background: Self-administration of medication by hospital in-patients is a desirable practice for selected patients and medications in that it promotes timely and empowered medicines management by the patient. For self-administration to be successful it needs a supporting programme, policies and procedures. A recent continuous improvement project of self-administration of self-administration in ST Vincent’s University Hospital (SVUH) fine-tuned their approach which includes assessing the medicine for suitability, the patient for suitability, documenting patient agreement, observing patient medication administration for the first 24h. The SVUH project also increased the range of medication which could be self-administered by the patient. Self-administration of medication is not covered comprehensively by any medication management policy in Tallaght Hospital and there is no self-administration programme. In Tallaght, the majority of medications are administered by nurses to the patients, who are encouraged not to self-medicate. Certain medications e.g. inhalers, phosphate binders, are commonly self-administered and recorded as ‘self’ on the in-patient prescription chart. We would like to conduct a baseline survey to identify what proportion of medicines are self-administered, to characterise which medicines are self-administered and to assess patient and medicine suitability for self-administration. Out of scope for the current project:

Identifying self-administration by patients which is not captured on the prescription chart administration record

Project 43: Pharmacist competencies and pharmacy education

Name of Principal Supervisor Assist. Prof. Ryder Subject area Practice of Pharmacy Non-lab based project

There is an increasing move towards the specification and evaluation of ‘competencies’ as a means of ensuring practising pharmacists are in a position to provide services to an appropriate standard. The Pharmaceutical Society of Ireland has recently [August 2013] finalised and published a Core Competency Framework setting out standards of practice for pharmacists who wish to join or remain on the PSI’s register. It is clearly necessary for undergraduate pharmacy degree programmes to prepare students for professional practice by development of the specified competencies, and it is also important for Schools of Pharmacy to have a means of determining whether the required standards have been met by each student. While the Pharmaceutical Society of Ireland has developed accreditation standards for pharmacy degree programmes (the undergraduate bachelor’s degree, the one year internship programme, and most recently [August 2013] draft accreditation standards for potential future 5-year integrated M.Pharm degrees), these standards are not fully aligned with one another, nor with the Core Competency Framework. This project will entail:

- Exploring fulfilment of existing regulatory requirements by the School’s current B.Sc.(Pharm.) degree, both in terms of developing the required skills and in evaluating whether they have been attained;

- Preparation of a ‘gap analysis’ identifying additional components required for a 5 year integrated M.Pharm. programme;

- Determining students’ views on how well the existing B.Sc.(Pharm.) degree meets their needs.

- Preparing a tool to guide undergraduates to integrate material from multiple disciplines as they develop into competent pharmacists.

The project should therefore not only give the student a detailed insight into their future evaluation by the regulator as an intern and practising pharmacist, but will also provide the opportunity to influence future pharmacy education.

Project 44: Supporting the Safe Use of Insulin in St Vincent’s University Hospital

Name of Principal Supervisor Assist. Prof. Ryder Co-Supervisor Maeve Moran / Claire Keane Subject area Practice of Pharmacy/Clinical Pharmacy Non-lab based project

Insulin is a high alert medicine, and medication errors with insulin can cause serious harm to patients. There are many reasons why errors with insulin occur: Different insulin treatment regimens, dosages individualised to each patient, altering insulin requirements due to changed diet all contribute to the complexity of using insulin safely. In hospitals, multiple insulin products may be required on the ward to manage different patients. The current insulin formulations do not have tamper seals, making it difficult to identify insulin pens that have been used. The aim of this project is to assess if insulin dispensing and storage on wards in St Vincent’s University Hospital supports the safe use of insulin. The student will be required to visit each ward in the hospital, and review the storage and labelling of insulin in stock there. In addition they will complete two case studies on patients treated with insulin and describe the potential risks with insulin treatment, along with the actions taken to address those risks. The project will also provide an opportunity for the student to gain insight into hospital pharmacy practice and may be of benefit to those seeking to practice in this sector in the future.

Project 45: Benzodiazepine prescribing and its regulation

Name of Principal Supervisor Assist. Prof. Ryder Co-Supervisor Denis O’Driscoll (HSE) Subject area Practice of Pharmacy Non-lab based project

There is increasing evidence of increasing problem benzodiazepine use among young people in Dublin, and the HSE has identified a need to identify potential ways in which prescribed drugs are being diverted for illicit use. The project will entail the collection and analysis of pharmacy prescription data using methodology adapted from the French OSIAP (Ordonnances Suspectes, Indicateur d’Abus Possible) system, in the Dublin 12 Local Drug Task Force and Canal Community Local Drug Task Force areas, in association with the Chief Pharmacist (Addiction Services) of the HSE. The government is seeking to improve the prescribing of benzodiazepines and z-drugs through legislative change. The student will also audit existing prescriptions with a view to exploring how the proposed forthcoming amendment to the Misuse of Drugs regulations [currently in draft form, for consultation] may affect the prescribing and supply of benzodiazepines.

Project 46: Knowledge of alcohol and alcohol-drug interactions

Name of Principal Supervisor Assoc. Prof. Henman Subject area Practice of Pharmacy Non-lab based project

Alcohol, the most widely used social drug, produces a diverse range of effects on the body that vary with the dose and duration of use. Although there is quite a lot of evidence from North America there is relatively little from Europe on this topic and most of it, inevitably is observational in nature and requires careful interpretation. Alcohol-drug interactions are frequently derived from case studies. Consequently the assessment of their clinical significance and the robustness of the advice offered in reference sources may vary. The systematic appraisal of the evidence for these interactions would enable better advice for practitioners to be formulated. A review of the content of standard reference sources and a systematic review of the literature will be conducted to examine (a) the biological basis of the interaction, (b) the relationship between dose and duration of exposure, (c) the quality of the case studies, observational studies or other published evidence and (d) the structured interpretation of all that information into practical advice. The student will gain experience of the review and systematic analysis of case studies and observational studies and of the appraisal and translation of the results of the review into practical, evidence-based advice.

Project 47: Evaluation of Proton Pump Inhibitor Prescribing in the Older Patient

Name of Principal Supervisor Assoc. Prof. Henman Co-Supervisor Niamh McMahon Subject area Practice of Pharmacy Non-lab based project

Proton pump inhibitors (PPIs) are a widely utilised group of medicines (esomeprazole, omeprazole, lansoprazole, pantoprazole all rank within the top 20 most commonly prescribed medicines in the GMS list 2011). PPIs are generally well tolerated and many older patients are prescribed PPIs both for treatment and prophylaxis of gastric conditions (perhaps specify the licenced indications?). Recent evidence suggests overprescribing of PPIs, particularly in the older patient. Risks associated with high doses and long courses of treatment include increased risk of fractures, electrolyte disturbances (especially hypomagnesaemia), increased risk of Clostridium difficile infection and also of community and hospital acquired pneumonia. In addition to these risks, there are also significant cost implications for the overuse of this group of drugs. This project aims to evaluate appropriateness of PPI prescribing (including appropriate indication, dose and duration of treatment) within the Medicine of the Elderly (MedEl) Directorate patients in St.James’s hospital. Where inappropriate prescribing is identified, the patient will be assessed for evidence of adverse reactions as detailed above. The cost implications of inappropriate prescribing will also be evaluated. Inclusion criteria: Inpatients in MedEl wards Exclusion criteria: Intensive care, surgical patients, and medical patients on non-MedEl wards.

Project 48: Analysing and tracking in-patient glycaemic control

Name of Principal Supervisor Assoc. Prof. Henman Co-Supervisor Ciara Kirke, Tallaght Hospital Subject area Practice of Pharmacy/Clinical Pharmacy Non-lab based project

Project Aim/Research Hypothesis: To analyse factors influencing in-patient glycaemic control and set up to track glycaemic control over time, using the blood glucose Point of Care Testing (POCT) database. Two audits in 2010 showed poor glycaemic control in in-patients, despite considerable improvement efforts to address this between audits. An improvement programme is again trying to address these risks, beginning with perioperative/fasting patients. Literature Search: General, plus review of previous audits and write-ups. Data collection plan: Student liaises with POCT manager in laboratory to obtain Excel files of blood glucose data for the previous 2 months (in Excel files containing two days’ results at a time). Sampling: Full database for initial analysis. Run charts may be based on full data or a sample, as long as it can be used to provide meaningful tracking of glycaemic control over time. Inclusion criteria: All in-patients with measurements on the POCT database (adult medical and surgical, paediatric medical and surgical and psychiatry). Exclusion criteria: ICU, Theatre and day patients. Non-diabetic patients will not be excluded, as the large volumes of data mean that the influence of non-diabetic patients’ levels should be negligible and relatively constant over time. Data analysis/statistical methods to be employed: Analysis of the information to determine additional information for low and/or high blood glucose levels, e.g. day of week, time of day, ward, surgical/medical etc, will be carried out using Excel. Excel will also be used to build run charts or control charts (Statistical Process Control) to track levels over time. Anticipated outcome measures primary +/- secondary: Understanding of the factors associated with low and/or high blood glucose levels to inform priorities for the improvement programme and appropriate measures to track over time. Excel database set up to facilitate automatic building of run chart(s) once data is imported, which can be used to track progress with improvement programme.

project 1: the use of 5-asa as a positive control in the dss ... projects 2013-14.pdfproject 2: the...

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