Combining the NanaPPI Toolbox and AI-Driven Virtual Inhibitor Screening for the p53-MDM2 Interaction

High-throughput screening for inhibitors of protein-protein interactions (PPIs) provides vital information for therapeutic intervention in diseases driven by aberrant PPIs. Traditionally, the discovery of PPI inhibitors involves sequential steps: in vitro screening with purified proteins and then cellular assessment of cell permeability and PPI disruption, which prolong the screening timeline and escalate costs. Here, we have advanced our NanaPPI technique to create a versatile toolkit compatible with at least 95% of commercial murine antibodies, facilitating the in situ quantification of cellular PPIs. By integrating AI-driven virtual screening with the NanaPPI toolbox, we successfully constructed the NaviScreen platform and rapidly identified a potent p53-MDM2 inhibitor, AB-460, from a library of 3 million compounds. AB-460 effectively inhibits the p53-MDM2 interaction, suppresses the proliferation of tumor cells, and reduces tumor volume in zebrafish by increasing p53 and p21 protein levels. This NaviScreen platform combines the high throughput of virtual screening with the in situ PPI quantification capabilities of NanaPPI, offering an efficient strategy for the discovery of cell-permeable and stable PPI inhibitors.