Hot Spot-Driven Discovery of β-Catenin/Tcf4 Interaction Inhibitors via FDA Drug Repurposing

Aberrant activation of the Wnt/β-catenin signaling pathway is a hallmark of colorectal cancer, primarily driven by the interaction between β-catenin and T-cell factor 4 (Tcf4). Due to the large and shallow interface (~4800 Ų) of this protein–protein interaction, it has historically been considered “undruggable.” However, emerging evidence highlights the presence of druggable hot spots within this interface. In this study, we employed a structure-based virtual screening approach to repurpose FDA-approved and investigational drugs targeting the β-catenin/Tcf4 interaction. Using physics-based molecular simulations and MM/GBSA binding energy calculations, we identified Leucovorin (Folinic acid), Carbenicillin, and Ceforanide as promising hit compounds with stable binding to critical hot spot residues (Asn430, Lys435, His470, Arg474, and Lys508) of β-catenin. Data includes MD simulations trajectories of these compounds.