NEDD4-IN-1 (Compound 32) is a potent inhibitor of NEDD4, with an IC50 of 0.12 μM. It demonstrates significant antiproliferative activity in the H292 cell line and exhibits high plasma protein binding and moderate to high in vitro clearance in both human and mouse models. The compound shows exceptional stability in mouse plasma, alongside excellent efficacy, selectivity, and favorable in vitro ADME properties. In mouse models, its systemic plasma clearance surpasses hepatic blood flow. NEDD4-IN-1 is applicable for anticancer research.