VWK147 is a second-generation HSP90 C-terminal domain (CTD) inhibitor. It acts on the CTD dimerization interface, preventing the dimerization of HSP90CTD, disrupting the binding of the co-chaperone protein PPID with HSP90CTD, and inhibiting dimerization-dependent functions of HSP90. VWK147 reduces the protein levels of HSP90 client proteins ULK1, RIPK1, and CDK4 without inducing a heat shock response. It induces cell death, including apoptosis (apoptosis), in both cisplatin-sensitive and resistant urothelial carcinoma cells. VWK147 causes LC3-II accumulation, inhibits autophagosome-lysosome fusion to block classical autophagy (autophagy), and triggers non-classical LC3 lipidation independent of the ULK1 and PIK3C3 complex. VWK147 is applicable to research on urothelial carcinoma.