Gankyrin is a seven ankyrin repeat containing oncoprotein overexpressed in numerous cancer types such as lung, breast, prostate, colorectal, and many more. Gankyrin binds to S6ATPase to form the 19S regulatory cap of the 26S proteasome that is responsible for the degradation of polyubiquitylated proteins. Gankyrin facilitates a wide range of essential protein-protein interactions using its featureless concave surface, many of which have been identified in the onset of cancer development. Interestingly, gankyrin acts as a chaperone for the RING E3 ubiquitin ligase MDM2 to enhance ubiquitylation of the tumor suppressor protein p53. The overexpression of gankyrin has also been shown to increase p53 degradation leading to enhanced cancer cell migration and tumorigenesis. It is hypothesized that small molecule inhibitors of the gankyrin~MDM2 interaction could disrupt tumor formation and growth, but the precise mechanism for how these molecules disrupt gankyrin’s structure and/or fold remain unclear. Here, we describe the biophysical and biochemical impacts of various small molecule drugs on gankyrin using Isothermal Titration Calorimetry (ITC), Circular Dichroism (CD), Site Directed Mutagenesis (SDM), Nano Differential Scanning Calorimetry (DSC), and Nuclear Magnetic Resonance (NMR) spectroscopy to pinpoint where these small molecules bind to and affect the 3D structure of gankyrin. The specificity of these designed drugs for gankyrin was also assessed using similar biochemical analysis on other ankyrin repeat containing proteins such as RFXANK, ANKRA2, Trabid, and DARPin. The overarching goal of this interdisciplinary collaborative project between the Spratt Lab at Clark University and the Muth Lab at St. John’s University aims to produce and characterize potent small molecule inhibitors of gankyrin that will induce apoptosis in cancer cells.