Drug Development Geared to Discover, Validate, and Potentiate Cancer Cell Vulnerabilities

Cancers continue to have a large burden on individual health and our society at large. To achieve the constant growth, they have a unique set of metabolic needs. While many redundancies exist in biosynthetic pathways in cells, understanding how a cell uses one pathway over another is important to identifying weaknesses unique to that cancer. Furthermore, cutting off alternate pathways can lead to synthetic lethal combinations, which can differentially kill the cancer cells while leaving the non-cancerous cells alive. Here I will present work highlighting four different areas of drug development against cancer targets. First, I will present my work identifying novel small molecule compounds to inhibit ENTPD5. This work was done with recombinant protein produced with a variety of different methods and screening this enzyme against two libraries of compounds. The first consisted of over 20,000 molecules and used to identify novel compounds that inhibit this enzyme, and the second was a library based off the initial hits. These initial hits were further followed up in cell culture to show that they slow down cancer cell growth, and western blots confirm that the compound effects the downstream products of the enzyme. In a second project, I studied the interactions made between the enzyme DHODH and small molecule inhibitors. These inhibitors were discovered by our collaborators at UCLA using a phenotypic, high throughput screen. I purified recombinant DHODH enzyme, optimized around published crystallography conditions, and was able to obtain high resolution data showing one inhibitor specifically interacting with the protein. I later identified a novel DHODH crystallization condition which allowed for the co-crystallization of DHODH with the second inhibitor identified in the screen. Finally, I will explore an enzyme being developed as a novel biologic treatment of cancer. In the first of these chapters, I will highlight the identification of an in vitro mechanism of resistance as well as use novel NMR based metabolite tracing developed by a collaborator to highlight differences in cell metabolism when undergoing treatment. In the final chapter, I will highlight the synergistic effects of combining the novel biologic treatment with a known anti-cancer treatment.