The area of systems biology evolved in an attempt to introduce mathematical systems theory principles in biology. Although we believe that all biological processes are essentially chemical reactions, describing those using precise mathematical rules is not easy, primarily due to the complexity and enormity of biological systems. Here we introduce a formal approach for modeling biological dynamical relationships and diseases such as cancer. The immediate motivation behind this research is the urgency to find a practicable cure of cancer, the emperor of all maladies. Unlike other deadly endemic diseases such as plague, dengue and AIDS, cancer is characteristically heterogenic and hence requires a closer look into the genesis of the disease. The actual cause of cancer lies within our physiology. The process of cell division holds the clue to unravel the mysteries surrounding this disease. In normal scenario, all control mechanisms work in tandem and cell divides only when the division is required, for instance, to heal a wound platelet derived growth factor triggers cell division. The control mechanism is tightly regulated by several biochemical interactions commonly known as signal transduction pathways. However, from mathematical point of view, these pathways are marginal in nature and unable to cope with the multi-variability of a heterogenic disease like cancer. The present research is possibly one first attempt towards unraveling the mysteries surrounding the dynamics of a proliferating cell. A novel yet simple methodology is developed to bring all the marginal knowledge of the signaling pathways together to form the simplest mathematical abstract known as the Boolean Network. The malfunctioning in the cell by genetic mutations is formally modeled as stuck-at faults in the underlying Network. Finally a mathematical methodology is discovered to optimally find out the possible best combination drug therapy which can drive the cell from an undesirable condition of proliferation to a desirable condition of quiescence or apoptosis. Although, the complete biological validation was beyond the scope of the current research, the process of in-vitro validation has been already initiated by our collaborators. Once validated, this research will lead to a bright future in the field on personalized cancer therapy.

Le cancer est la principale cause de mortalité au Canada. Les taxanes (e.g. le paclitaxel et le docétaxel (DCTX)) constituent des remèdes efficaces contre une série de tumeurs solides telles que les cancers du sein, du poumon et de l’ovaire. Par ailleurs, des acides nucléiques (e.g. les oligonucléotides antisens (AON) ou les petits ARN interférents (siRNAs)), capables de supprimer sélectivement certains oncogènes impliqués dans la carcinogénèse, sont actuellement étudiés pour traiter une large gamme de cancers. Bien que l’activité des taxanes et des acides nucléiques soit bien établie sur des modèles humains et/ou animaux, plusieurs aspects physico-chimiques et cliniques restent encore à améliorer. Leur solubilité limitée (pour les taxanes), leur dégradation rapide dans le sang (pour les acides nucléiques), leur élimination précoce, leur absence de sélectivité et leur toxicité envers les tissus sains sont les principaux facteurs limitant leur efficacité. C’est pourquoi de nombreux efforts ont porté sur l’élaboration de systèmes de vectorisation ciblés à base de polymères, dans le but de surmonter les problèmes associés aux thérapies actuelles. Dans cette thèse, deux types de micelles polymères ont été développés pour la vectorisation de DCTX et d’acides nucléiques. D’une part, des micelles de poly(oxyde d’éthylène)-bloc-poly(oxyde de butylène/styrène) ont été étudiées pour la première fois pour solubiliser le DCTX et le protéger de l’hydrolyse. Ces polymères se sont révélés moins toxiques que le surfactant utilisé commercialement pour solubiliser le DCTX (i.e. polysorbate 80) et ont permis une libération prolongée du principe actif. D’autre part, deux systèmes différents de micelles polyioniques (PICM) ont été mis au point pour la vectorisation d’acides nucléiques. De nouveaux conjugués de poly(éthylène glycol) (PEG)-oligonucléotide ont été proposés pour la protection et la libération contrôlée d’AON. Lorsque ces conjugués ont été formulés avec des dendrimères de poly(amidoamine) (PAMAM), des complexes de taille homogène ont été obtenus. Ces PICM ont permis de prolonger la libération de l’AON et de le protéger efficacement contre la dégradation enzymatique. De plus, des polymères de poly(oxyde d’éthylène)-bloc-poly(méthacrylate de propyle-co-acide méthacrylique) ont été incorporés afin de conférer des propriétés acido-sensibles aux PICM. Dans ces micelles, formées de ce dernier polymère formulé avec le dendrimère PAMAM, des oligonucléotides (AON et siRNA) ciblant l’oncogène Bcl-2 ont été encapsulés. L’internalisation cellulaire fut assurée par un fragment d’anticorps monoclonal (Fab’) situé à l’extrémité de la couronne de PEG. Après l’internalisation cellulaire et la protonation des unités d’acide méthacrylique sous l’effet de l’acidification des endosomes, les micelles se sont affranchies de leur couronne. Elles ont ainsi exposé leur cœur composé d’acide nucléique et de dendrimère PAMAM, qui possède une charge positive et des propriétés endosomolytiques. En effet, ces PICM acido-sensibles ciblées ont permis d’augmenter la biodisponibilité des acides nucléiques vectorisés et se sont avérées plus efficaces pour silencer l’oncoprotéine Bcl-2 que les micelles non ciblées ou que le dendrimère de PAMAM commercial seul. Finalement, les nanovecteurs polymères présentés dans cette thèse se révèlent être des systèmes prometteurs pour la vectorisation des anticancéreux et des acides nucléiques. / Cancer is considered as the leading cause of premature death in Canada. Taxanes (e.g. paclitaxel and docetaxel (DCTX)) are effective against a range of solid tumors including breast, lung, and ovarian malignancies. In addition, nucleic acids (e.g. antisense oligonucleotides (AON) and short interfering RNA (siRNA)) which are capable of selectively suppressing oncogenes involved in carcinogenesis are currently being investigated for the treatment of a wide variety of cancers. Although the activity of taxanes and nucleic acid drugs is well-established in human and/or animal models, several physicochemical and clinical issues still need to be addressed. Low aqueous solubility (i.e. taxanes), rapid degradation in the blood (i.e. nucleic acids), fast clearance, non-selectivity and toxicity to normal tissues are limiting factors to their effectiveness. Hence, many efforts have been focused on developing targeted polymeric delivery systems to overcome the problems associated with the current therapies. In this thesis, two types of polymeric micelles have been developed for the delivery of DCTX and nucleic acids. On the one hand, poly(ethylene oxide)-block-poly(butylene oxide/styrene oxide) micelles were tested for the first time to solubilize and protect DCTX from hydrolytic degradation. The polymers showed less toxicity than the surfactant used commercially to dissolve DCTX (i.e. polysorbate 80) and released the drug in a sustained fashion. On the other hand, two different systems of polyion complex micelles (PICM) were developed for the sustained release and intracellular delivery of nucleic acids. Novel poly(ethylene glycol) (PEG)-oligonucleotide conjugates were assessed to protect AON against degradation and release them in a sustained manner. When these conjugates were mixed with poly(amidoamine) (PAMAM) dendrimers, monodisperse PICM were formed. These PICM further slowed down AON release and significantly protected it against enzymatic degradation. In addition, the incorporation of poly(ethylene oxide)-block-poly(propyl methacrylate-co-methacrylic acid) was exploited to impart pH-sensitivity to PAMAM-based PICM. This system was composed of the previous copolymer mixed with PAMAM dendrimer. Such PICM were loaded with AON or siRNA targeting the Bcl-2 oncogene. Micelles uptake by the cancer cells was mediated by a monoclonal antibody fragment (i.e. Fab') positioned at the extremity of the PEG corona. Upon cellular uptake and protonation of the methacrylic acid units in the acidic endosomal environment, the micelles lost their corona, thereby exposing their positively-charged endosomolytic PAMAM/nucleic acid core. The targeted, pH-sensitive PICM were found to increase the intracellular bioavailability of the entrapped nucleic acids and knock down the Bcl-2 oncoprotein more than either non-targeted micelles or commercial PAMAM dendrimers. The polymeric nanocarriers reported in this thesis appear to be promising vehicles for the delivery of anticancer drugs and nucleic acids.

Cotton has been a world-wide economic staple in textiles and oil production. There has been a concerted effort for cotton improvement to increase yield and quality to compete with non-natural man-made fibers. Unfortunately, cultivated cotton has limited genetic diversity; therefore finding new marketable traits within cultivated cotton has reached a plateau. To alleviate this problem, traditional breeding programs have been attempting to incorporate practical traits from wild relatives into cultivated lines. This incorporation has presented a new problem: uncultivated cotton hampered by photoperiodism. Traditionally, due to differing floral times, wild and cultivated cotton species were unable to be bred together in many commercial production areas world-wide. This worldwide breeding problem has inhibited new trait incorporation. Before favorable traits from undomesticated cotton could be integrated into cultivated elite lines using marker-assisted selection breeding, the markers associated with photoperiod independence needed to be discovered. In order to increase information about this debilitating trait, we set out to identify informative markers associated with photoperiodism. This study was segmented into four areas. First, we reviewed the history of cotton to highlight current problems in production. Next, we explored cotton’s floral development through a study of floral transition candidate genes. The third area was an in-depth analysis of Phytochrome C (previously linked to photoperiod independence in other crops). In the final area of study, we used Genotype-By-Sequencing (GBS), in a segregating population, was used to determine photoperiod independence associated with single nucleotide polymorphisms (SNPs). In short, this research reported SNP differences in thirty-eight candidate gene homologs within the flowering time network, including photoreceptors, light dependent transcripts, circadian clock regulators, and floral integrators. Also, our research linked other discrete SNP differences, in addition to those contained within candidate genes, to photoperiodicity within cotton. In conclusion, the SNP markers that our study found may be used in future marker assisted selection (MAS) breeding schemas to incorporate desirable traits into elite lines without the introgression of photoperiod sensitivity.

This thesis emphasizes the role of political risk in international business by analyzing the impact of political events on the valuation of firms. The guiding question is how governments interfere with the business interests of firms located in their own country as well as with the business interests of firms from other nations, as a consequence of the application of international sanctions. Therefore, the focus is on multi-country and multi-sector effects due to the occurrence of specific sanction events. The empirical methodology is the event-study approach, which analyzes stock market reactions to new information. The research objective is to detect abnormal stock returns across multiple markets and sectors, as a consequence of events related to the imposition of or threat of international sanctions. The empirical model of this thesis differentiates between risk-effects for firms located in the sender country (i.e., the origin of sanctions), for firms located in or specifically related to target countries (i.e., the receiver of sanctions) and firms located in third countries (i.e., countries not directly involved). There are three different cases analyzed: E.U. Economic Sanctions against African countries (2002-2005), the U.S. Steel Tariff (2002) and the Iran Sanctions Act (2007). The cases represent sanctions applied on the nationwide, sector- and firm-specific level. The event studies provide empirical evidence for the existence of political risk-effects due to sector-specific sanctions. Risk-effects are detected for firms in target countries and for firms in the sender country itself. The applied political risk framework describes how political risk affects multinational firm value and explains that it varies among firms. The impact of political risk on a firm's value depends on the risk exposure of a firm's individual business interests to it. This contributes a new perspective on political risk that emphasizes multinational and multi-sectoral effects and underlines that a specific political risk can be relevant for a variety of different international business interests. (author's abstract)

It is a major clinical problem that survival rates after out-of-hospital cardiac arrest have not markedly improved during the last decades, despite extensive research and the introduction of new interventions. However, recent studies have demonstrated promising treatments such as targeted temperature management (TTM) and methylene blue (MB). In our first study, we investigated the effect of MB administered during experi-mental cardiopulmonary resuscitation (CPR) in the setting of postponed hypother-mia in piglets. We set out to study if MB could compensate for a delay to establish targeted TTM. The study demonstrated that MB more than compensated for 30 min delay in induction of TTM. The effect of MB added to that of TTM. The second study examined the effects of TTM and S-PBN on the endothelin system and nitric oxide synthases (NOS) after prolonged CA in a porcine CPR mod-el. The study was designed to understand the cardioprotective mechanism of S-PBN and TTM by their influence on the endothelin system and NOS regulation. We veri-fied for the first time, that these two cardioprotective postresuscitative interventions activate endothelin-1 and its receptors concomitantly with eNOS and nNOS in the myocardium. We concluded that nitric oxide and endothelin pathways are implicated in the postresuscitative cardioprotective effects of TTM. The third study compared survival and hemodynamic effects of low-dose amio-darone and vasopressin to vasopressin in a porcine hypovolemic CA model. The study was designed to evaluate whether resuscitation with amiodarone and vasopressin compared to vasopressin alone would have an impact on resuscitation success, survival, and hemodynamic parameters after hemorrhagic CA. We found that combined resuscitation with amiodarone and vasopressin after hemorrhagic circulatory arrest resulted in greater 3-hour survival, better preserved hemodynamic parameters and smaller myocardial injury compared to resuscitation with vasopressin only. In our fourth study we planned to compare hemodynamic parameters between the treatment group (milrinone, esmolol and vasopressin; MEV) and control group (vasopressin only) during resuscitation from prolonged cardiac arrest in piglets. The study was designed to demonstrate if MEV treatment improved hemodynamics or cardiac damage compared to controls. We demonstrated that MEV treatment reduced cardiac injury compared with vasopressin alone.

Includes bibliographical references (leaves 135-150) / It is increasingly becoming apparent that elections, as component parts of democratic processes, do not always guarantee a smooth transfer of power and stable democratic cultures in many contemporary polities. Whenever election outcomes are disputed, Governments of National Unity (GNU) are presented as strategic conflict prevention and resolution models. Yet, the GNU’s ability to nurture democracy in situations where it remains weak or non-existent is extremely limited. This dissertation therefore uses Zimbabwe’s case study to examine how its recent internal political turmoil had overwhelming implications for its legitimacy and democracy. Through qualitative research, the study reveals that despite partially reconciling Zimbabwe’s political protagonists, the GNU neither permanently resolved the crisis nor addressed significant related issues; thus, creating a paradox of legitimacy for the country. The dissertation concludes with recommendations for further research on matters of legitimacy in power-sharing scenarios such as Zimbabwe’s. / Political Sciences / M.A. (International politics)

La détection précoce de l’athérosclérose, avant le développement de ses séquellespathologiques, comme l’infarctus du myocarde, l’angine ou l’accident cérébrauxvasculaire(ACV), représente un important défi au niveau de la médecine diagnostiqueactuelle. Malgré les récentes avances technologiques, les maladies cardiovasculairesdemeurent la principale cause de décès dans les pays occidentaux et la détection à unstage plus précoce s’avère nécessaire pour permettre une intervention thérapeutiqueadéquate. Notre étude se concentre sur la détection de l’athérosclérose, plusspécifiquement la vulnérabilité de la plaque, grâce à l’imagerie moléculaire combinée àl’observation pathologique. Afin de prédire la rupture de la plaque, l’imageriemoléculaire a émergé comme outil diagnostique puissant suite au développementcroissant de sondes ayant de l’affinité pour les molécules cibles du processusd’athérosclérose. Comme résultantes, ces molécules sélectives possédant une forteaffinité pour des cibles surexprimées durant le processus de formation de la plaque,comme l’αvβ3 par exemple, devrait représentées des sondes prometteuses pour ladétection de l’athérosclérose.Objectif L’objectif global de notre étude était d’évaluer et de prédire lavulnérabilité de la plaque d’athérome à l’aide de différents marqueurs moléculaires. Leprincipal objectif de notre recherche était d’évaluer la possibilité de détecter précocementla plaque en utilisant une ITOP (integrin targeted optical probe). Cette sonde synthétiquenouvellement développée et ciblant l’intégrine αvβ3 avait déjà démontré une affinité etspécificité particulièrement élevée pour le récepteur de l’αvβ3 dans le cancer. Nousavons également exploré la relation entre cette sonde et l’observation pathologique desplaques d’athéromes sur le modèle animale WHHL et sur des plaques humainesprovenant de différents patients.Procédure et Résultats Les expériences ont été réalisées sur un total de 12 lapinsWatanabe hyperlipidémiques de souche WHHL (Watanabe heritable hyperlipidemic) et 1lapin contrôle NZW (New Zealand White). Premièrement, notre ITOP, marquée avec lafluorescéine isothiocyanate (FITC), a été utilisée pour détecter in vitro et ex vivo laprésence du récepteur de l’αvβ3. La microscopie à fluorescence a révélé un importantmarquage de la plaque d’athérome, lequel était absent dans les tissus provenant des lapinscontrôles NZW. Le marquage a été détecté au niveau de segments de plaques provenantde deux régions distinctes de l’aorte ascendante et descendante dans chaque lapin. Lesignal a été détecté principalement au niveau de l’adventitia et de l’intima proximale desvaisseaux aortiques, correspondant directement à l’expression de l’intégrine αvβ3,déterminée par essai immunochimique avec un anticorps contre l’αvβ3. De plus, uneforte association s’est révélée entre le niveau de marquage de la sonde ciblant l’αvβ3 etl’épaisseur de l’adventitia. Deuxièmement, nous avons évalué notre sonde sur deséchantillons humains affectés par l’athérosclérose et comparé les résultats avec uneévaluation morphologique. Nous avons remarqué la même tendance que chez le lapin, soiun marquage plus important lorsque l’adventitia s’épaissi. Finalement, nous avons testé lasonde sur des artères coronaires provenant d’une autopsie d’un patient affecté par le ADHIESet comparé les résultats avec l’évaluation morphologique de leurs artèrescoronaires. Nous avons trouvé un lien entre la morphologie de la plaque et la prévalenced’anévrysmes coronaires chez ces patients.Conclusion L’expression de l’αvβ3 est reliée à la foi aux processus inflammatoires età la sténose. Notre ITOP à marqué efficacement in vitro le premier type de plaqued’athérome classé comme avancé (type IV) et pouvant produire des manifestationscliniques. En combinaison avec l’imagerie noninvasive détectant la sténose, il pourraits’avéré utile dans la détection de la plaque vulnérable. / Purpose The detection of early atherosclerosis, before the development of its later sequelae of myocardial infarction, angina or stroke, constitutes an important challenge in current diagnostic medicine. Despite all the recent technological advances, cardiovascular disease remains the leading cause of death in the Western World and needs to be detected at an earlier stage to allow for more timely therapeutic intervention. This study is focusing on the detection of atherosclerosis or more specifically plaque vulnerability with the help of molecular imaging and pathological observation. Effectively, to predict plaque rupture, molecular imaging has emerged as a powerful diagnostic tool, consequent to the development of a growing number of new probes with affinity for key molecular targets. As a result, such selective molecule with high affinity for overexpressed target in plaque formation, as αvβ3 integrin, should have promise as a probe for imaging atherosclerosis. With the help of molecular imaging combined with pathological observations, we can better comprehend, predict, and detect plaque vulnerability and rupture. Objectives The overall objective of this study is to evaluate different molecular tools to predict the vulnerability of the atheromatous plaque. The major objective of the research was to investigate the possibility of detecting atherosclerotic plaque by using a newly developed synthetic αvβ3 integrin targeted optical probe (ITOP) showing particularly high affinity and specificity for the αvβ3 receptor. We also investigate the relation between this probe and pathological observation of atherosclerotic plaques from WHHL animal model and different human samples. Procedures and Results For this study, experiments were performed on 12 Watanabe heritable hyperlipidemic (WHHL) rabbits and 1 New Zealand White (NZW) rabbits for control. First, our ITOP labeled with fluorescein isothiocyanate was used for detecting the presence of αvβ3 receptors in vitro and ex vivo on a Watanabe rabbit model. Fluorescence microscopy demonstrated a strong labeling of atherosclerotic plaques, which was absent in tissue from normal NZW rabbits. Segments of plaque accumulation from two distinct regions of ascending and descending aortas were labeled in each rabbit. The signal was found principally in the adventitia and proximal intima of the aortic vessel, corresponding directly to the expression of integrin αvβ3 as determined by antibody assay. Moreover, there was a close association between the level of labeling with the αvβ3 targeted probe and the thickness of the adventitia. Secondly, the ITOP was evaluated on human atherosclerotic samples, and was found to efficiently labeled atherosclerotic plaques. Moreover, we observed the same tendency as in the Watanabe rabbit: the ITOP intensity correlated with the degree of adventitial thickening. Finally, we tested the ITOP on Job's Syndrome coronary arteries, and have been able to detect a plaque corresponding to the first type of advanced atherosclerosis (type IV). We also found a relationship between plaque morphology and predisposition to aneurysms in Job's syndrome. Conclusions αvβ3 expression is related to inflammatory and stenotic processes. Our ITOP can efficiently label in vitro the first type of advanced atherosclerotic plaque. In combination with noninvasive imaging techniques that evaluate stenosis, it has great potential for the detection of vulnerable plaque.

No description available.

To address the challenge of accurate, low cost and robust biosensors for diabetes management and early detection of diabetes complications, we have developed novel, robust sensing chemistry (or receptors) for electrochemical POC biosensors. The biosensors have been developed for the bio-markers associated with diabetes management such as glycated haemoglobin (HbA1c), glycated albumin, glucose, biomarkers associated with diabetes complications such as microalbuminuria, urine creatinine and albumin-to-creatinine ratio (ACR) and biomarkers associated with anaemia and malnutrition conditions such as haemoglobin and serum albumin. For haemoglobin detection, a new POC bio sensing technique has been developed based on Aza-heterocyclic chemicals. The repeatability and accuracy of the biosensor have been tested on real pathology samples. The glycated form of haemoglobin, called glycated haemoglobin or HbA1c, is the gold standard test in diabetes management as it gives the 90-days average blood glucose value. We demonstrate a simple method for electrochemical detection of HbA1c by combining bosonic affinity principle along with aza-heterocyclic receptors. The technique has been verified on the real clinical patient samples. Albumin is the most abundant protein in the human blood. Human serum albumin (HSA) is either alone or an associative biomarker in several chronic diseases like necrosis, nephrosis, hepatitis, malnutrition, arthritis, immune disorders, cancer, diabetes and in some severe infections. In pathology laboratories, the serum albumin is usually tested on serum samples and not in whole blood samples. Since albumin is not a metalloproteinase, it is very difficult to develop electrochemical POC biosensor. We have developed a novel technique for the electrochemical detection of serum albumin in whole blood samples, by exploiting its binding property with redox active copper salts. The accuracy of technique has been verified on both real human blood plasma as well as whole blood samples. Glycated albumin, which is the glycated form of serum albumin, is emerging as a novel biomarker for diabetes management, as it gives the average blood glucose value of 15-20 days. It is also extremely useful in chronic kidney disease patients and patients with hemoglobinopathies where HbA1c can give the erroneous results. By combining the copper chemistry along with bosonic affinity principle, we present the first ever demonstration of glycated albumin sensing. Instant blood glucose monitoring is an integral part of diabetes management. Most of the glucometers available in the market are based on glucose oxidase enzyme. We have demonstrated a low cost non-enzymatic electrochemical technique for blood glucose detection using alkaline methylene blue chemistry. The accuracy of the technique has been verified on real human blood plasma samples. Glucometer is one of the most easily available POC biosensor and a useful tool for diabetes population. India has second largest diabetes population in the world. To analyse the accuracy of the POC glucometers which are available in Indian market, a comprehensive study was conducted. The results were compared with clinical accuracy guidelines using exhaustive statistical analysis techniques. The shortcomings of the commercial glucometers are elucidated, regarding different international standards. Diabetic nephropathy is one of the major diabetes complications and is the primary cause of chronic kidney disease (CKD). The presence of albumin in urine is a well-established biomarker for the early detection of diabetic nephropathy. We have developed a technique for electrochemical detection of microalbuminuria for point of care applications by exploring the binding property of human albumin with electrochemically active molecules like copper and hemin. Methylene blue mediated sensing technique has also been proposed. Urine Albumin-to creatinine ratio (ACR) is another variant of the microalbumuria test that can be done any time and does not suffer from the dilution factor of urine. Iron binding property of creatinine is exploited to develop creatinine biosensor, thus enabling POC ACR tests.

Use of metal based anticancer medication began with the clinical approval of cisplatin in 1978. Research led to the development of six platinum based drug candidates which are in use around the world. However there is a great need to develop better treatment strategies. The present work entitled “Cytotoxicity of Metal Based Anticancer Active Complexes and Their Targeted Delivery Using Nanoparticles” is an effort to prepare cytotoxic metal complexes based on platinum(IV) and copper(II) and deliver them selectively to cancer cells using a targeting ligand, biotin, with two different delivery vehicles, viz. PEGylated polyamidoamine dendrimer (PAMAM) and gold nanoparticles (AuNPs). Chapter 1 provides a brief introduction to cancer and its characteristic features, followed by a short description about different treatment modalities in clinical practice. An account of the development of anticancer drugs starting from purely organic drugs to the field of metal based anticancer drugs is discussed. An overview of the available targeting strategies are discussed with specific examples. The section ends with the scope of the present work. Platinum based anticancer drugs currently in use contain platinum in the +2 oxidation state. These drugs showed side effects and are often ineffective against resistant cells, especially in the latter stages of treatment. A recent focus of metal based anticancer drug research is the development of platinum(IV) systems which shows promise to have greater activity in cancer cells in a reducing environment. Reported platinum(IV) dual drugs contain the components of “cisplatin” or an analogue along with an active organic drug. But there are no known dual drugs based on platinum(IV) that would generate a cytotoxic metal complex along with cisplatin. In Chapter 2, a bimetallic dual drug (M4) (Figure 1), the first of its kind, with components of cisplatin and copper bis(thiosemicarbazone) has been prepared (Figure 1). The components and the bimetallic complex were characterized using several spectroscopic techniques. The dual drug M4 was found to be highly cytotoxic (IC50 1.3 M) against HeLa cells and was better than cisplatin (IC50 6.8 M). The bimetallic complex turned out to be better than the mixture (IC50 7.2 M) of individual drugs which indicated possible synergism of the released cisplatin and the copper bis(thiosemicarbazone) from the dual drug. Figure 1: Structure of the platinum(IV) and copper bis(thiosemicarbazone) complexes. A novel approach towards conjugation of platinum(IV) drugs to a carrier has been developed using a malonate moiety (Figure 2). The bis(butyric acid) complex, Pt(NH3)2(OCOC3H7)2Cl2 (M1), was taken as model complex to demonstrate the conjugation strategy. The complex M4 was also conjugated to the partially PEGylated 5th generation PAMAM dendrimers. Figure 2: Schematic representation of the platinum(IV) drug conjugated PAMAM dendrimer. The cytotoxicity of M4 was reduced to a small extent on conjugation to the dendrimer. In the presence of 5 mM sodium ascorbate as a reducing agent, sustained release (40 %) of the drug was shown to occur over a period of 48 h by the drug release study. The reduction in cytotoxicity of the dendrimer conjugates could be due to incomplete release of the active drug. Unfortunately, no enhanced activity was observed with the additional targeting ligand, biotin. The drug uptake study revealed that the dendrimer conjugates were successful in entering cancer cells. There was no preferential uptake with biotin conjugated dendrimers which explained the similar cytotoxicity of dendrimer conjugates with and without biotin. Different delivery vehicles showed varied efficiency in delivering the pay load (drugs) to the cancer site. In this connection, PEGylated gold nanoparticles have shown good promise as a drug delivery vehicle. In Chapter 3, M1 and M4 are both conjugated to malonate functionalized PEGylated gold nanoparticles (30 nm). Biotin was also attached to the AuNPs for targeting HeLa cells. Figure 3: Schematic representation of the platinum(IV) drug and biotin conjugated AuNPs. The AuNPs were highly stable in water without agglomeration. There was no shift in the Surface Plasmon Resonance (SPR) band after conjugation of the drug molecules and targeting ligands. TEM images and DLS measurements showed there was no change in particle size. Drug conjugated AuNPs were also very stable in high salt concentrations as well as over a large range of pH. AuNPs with M1 were found to be less cytotoxic than the parent drug. Biotinylated AuNPs with M1 were more potent than non-biotinylated nanoparticles and increased cytotoxicity (35 %) was observed with biotin conjugation. Surprisingly, the enhanced activity of biotinylated AuNPs could not be correlated to the drug uptake study. The cytotoxicity of the bimetallic dual drug containing AuNPs were about 10-fold less and no increased activity was observed with the biotinylated conjugates. The reduced activity of AuNPs with the bimetallic drug was due to incomplete release from the AuNPs (20 % release after 48 h). But the release kinetics was very slow and sustained which might increase in vivo activity. The unexpected lower activity of biotinylated conjugates with copper bis(thiosemicarbazone) was suggestive of interference between bis(thiosemicarbazone) complex and the biotin receptor resulting in reduced drug uptake. Copper bis(thiosemicarbazone) complexes hold very good promise as a class of non-platinum anticancer drug candidates. However, they lack selectivity towards malignant cells. Recently, CuATSM has shown hypoxia selectivity and very good cytotoxicity resulting in 64CuATSM being used in advanced stages of clinical trials for imaging hypoxic cells. In Chapter 4, a copper bis(thiosemicarbazone) complex analogous to Cu(ATSM) with a redox active cleavable disulfide linker and a terminal carboxylic acid group (CuATSM-SS-COOH) was synthesised and characterised spectroscopically. The complex was highly cytotoxic and has an IC50 value (6.9 M) similar to that of cisplatin against HeLa cells. The complex was conjugated to PEGylated gold nanoparticles by amide coupling between the acid group from the drug molecule and the amine on the AuNPs (20 nm) for smart drug delivery. The gold nanoparticles were decorated with biotin for targeted delivery to the HeLa cells. Figure 4: Schematic representation of the CuATSM-SS-COOH and biotin decorated AuNPs. The CuATSM-SS-COOH was insoluble in water but conjugation to PEGylated gold nanoparticles made it water soluble. The drug molecules and biotin conjugated AuNPs were highly stable which was confirmed by TEM and DLS measurements. Similar to the study described in the previous chapter, these AuNPs were also stable in a wide range of pH and salt concentrations. In vitro glutathione (GSH) triggered release study demonstrated substantial release of the cytotoxic agent from the AuNPs (60 %) over a period of 48 h. In vitro cell viability study with HeLa cells showed reduced cytotoxicity (IC50 15 M) of AuNPs with and without biotin containing drug conjugates relative to the parent copper complex (IC50 6.9 M). The reduction of the cytotoxicity correlated well with the released amount of the active drug from the nanoconjugates over the same time period. In vivo studies demonstrated the effectiveness of these nanoparticle carriers as suitable vehicles as they exhibited nearly four-fold reduction of tumor volume without significant loss in body weight. Moreover, the biotin targeted nanoparticle showed significant (p < 0.5) reduction in tumor volume compared to the non-targeted gold nanoparticles. Thus, this smart linking strategy Can be extended to other cytotoxic complexes that suffer from non-specificity, low aqueous solubility and toxicity. Multinuclear anticancer active complexes do not act in the same way as that of their corresponding mononuclear analogues. In the case of multinuclear platinum complexes, the activity not only depends on the active moiety but also on the spacer length between the moieties. In Chapter 5, a series of multinuclear copper bis(thiosemicarbazone) complexes were prepared and characterised using different techniques. Figure 5: General structures of binuclear copper bis(thiosemicarbazone) complexes. All the complexes showed redox activity and have a very high negative reduction potential, i.e. these compounds would not be easily reduced in the biological medium and would remain as copper(II) species. As the concentration of the reducing agents are more within cancer cells, once these complexes are inside cells they would be reduced to Cu(I). These compounds were shown to be highly lipophilic from the large log P values. Unfortunately, these binuclear complexes were less active than similar mononuclear complexes. One possible reason for the reduced cytotoxicity of these complexes could be adherence of the complexes to the cell membrane due to the high lipophilicity of these complexes. Out of five different methylene spacers between two bis(thiosemicrarbazone) moieties, the complex with a three carbon spacer was shown to be the most active against HeLa cells. The complexes with five and six methylene spacers turn out to be noncytotoxic. Further experiments are necessary to reveal the mechanism of action in these complexes. In summary, bimetallic complexes can be very active and may be a way of overcoming drug resistance in platinum based therapy. A dual drug can be delivered using a malonate moiety and a disulfide linker. Gold nanoparticles are good delivery vehicles for these dual drugs and show great potential for improvement and translation to the next stage. (For figures pl refer the abstract pdf file)