In Drama Education mask work is undertaken and presented as both a methodology and knowledge base. There are numerous workshops and journal articles available for teachers that offer knowledge or implementation of mask work. However, empirical examination of the context or potential implementation of masks as a pedagogical tool remains undeveloped.

On a theoretical level, throughout both ancient and modern drama education and performance, masks have been seen as synonymous to the field of drama. The mask is an iconic theatrical symbol from the times of Socrates to Modern western theatres. Simply put, masks symbolise the adoption of the role and hold a central place in drama across time and culture.  Within Drama (as a field in itself), the use of mask have been used by influential drama theorists explicitly in specialist drama training. In schools, however, whilst referenced in official curricula internationally, there is no formal development of pedagogies for mask use in Drama, and little to no research in its potential impact upon the enacted curriculum.

This paper presents some methodologies of ‘how’ to apply masks offered through presenting theoretical, historical knowledge contexts. Two teacher workshops in mask application and pedagogical potentials is also further discussed.

Bioorthogonal chemistries are widely used to image and profile biomolecules in their native environments. While many bioorthogonal reactions have been reported in recent years, most cannot be used inside cells owing to reagent instabilities. Additionally, there are few reactions that can be used concurrently for multi-component labeling. Many of the most popular bioorthogonal reagents exhibit significant cross- reactivities. To address these limitations, I developed new classes of bioorthogonal reagents, including cyclopropenones, cyclopropenethiones, and functionalized phosphines.

In Chapter 1, I introduce the overall strategy involved in designing and optimizing new bioorthogonal reagents. I also highlight some of the current needs in bioorthogonal reactivity and recent advances that have been made in these areas. In Chapter 2, I describe the optimization of bioorthogonal cyclopropenones for use in intracellular environments. I synthesized a panel of disubstituted cyclopropenones and identified scaffolds that are inert to thiols and other biological nucleophiles. These substituted cyclopropenones react efficiently with a wide array of functionalized phosphines to form covalent adducts. I further developed cyclopropenone amino acids that can be used for recombinant protein production and labeling applications. Chapter 3 showcases the investigation of cyclopropenethiones as more rapidly reacting heteroanalogs of cyclopropenones. Cyclopropenethione-phosphine ligations proceed at markedly improved rates, and cyclopropenethiones are suitable for use in complex biological environments. Finally, in Chapter 4, I discuss the use of cyclopropenones as chemically activatable protein crosslinkers. Using a model split protein system, I demonstrated that cyclopropenones can serve as electrophilic traps to form covalent adducts between interacting biomolecules.

Overall, this thesis describes the optimization of cyclopropenones, cyclopropenethiones, and phosphines for use in biological environments. The stability and highly tunable nature of these reagents make them useful for a wide range of applications, including protein conjugation, cellular imaging, and biomolecule trapping. The unique reactivity of these reagents also presents new opportunities for multicomponent labeling. Based on their biocompatibility and versatility, I anticipate that the cyclopropenone and cyclopropenethione will be broadly adopted for applications in cells and living systems.

This dissertation details the chemistry work that has been done to support the Radu lab’s cancer research efforts. Although cancer research is typically performed via cell cultures, tissue cultures, mice studies, etc., there are many aspects that require specialized chemistry to accomplish. What is sometimes overlooked or quickly summarized is the tedious chemistry groundwork that may lead to the invention of a successful drug or tool compound, enabling interesting biology to be studied or observed. Even though publications generally focus on successful final outcomes, failed cases can provide important lessons going forward. For example, sometimes new compounds no longer target the desired protein, but can still treat cancer or elicit intriguing biological outcomes. Whether or not the projects detailed in this dissertation were successful is subjective and may not be determined in a short period of time. Nonetheless, the accurate documentation of this work hopefully may provide the opportunity for other scientists to learn from and build upon. This dissertation outlines the results of three independent projects all revolving around the development of novel cancer therapies. In Chapter One, new inhibitors of deoxycytidine kinase that were designed to improve upon drug-like properties such as solubility or ease of synthesis will be introduced. The new compounds will be compared to the clinically-viable lead, (R)-DI-87, and assessed in terms of potency, solubility, and ease of synthesis. At the end of Chapter One, how a dCK inhibitor possessing an amine handle, NMc-77, was used to construct potential deoxycytidine kinase degraders will also be described. In Chapter Two, HCT13, a thiosemicarbazone derivative, was made initially to improve upon Triapine’s ribonucleotide reductase-inhibiting activity. Surprisingly, small structural changes abolished the compound’s activity against ribonucleotide reductase, yet HCT13 remained a potent and effective drug against aggressive leukemia in mouse models. Lastly, Chapter Three will detail efforts to synthesize cysteine labeling compounds that can enable high-throughput studies of cysteine oxidative post-translational modifications. These tool compounds may help scientists gain more insight into various treatment efficacies and resistance mechanisms by elucidating changes in cysteine-mediated redox signaling.