Norcantharidin-Loaded Liposomes Potentiate Radiotherapy by Reprogramming Tumor Vasculature toward Normalization

Anti-angiogenic drugs are theorized to suppress tumor growth by blocking neovascularization. Notably, low-dose regimens may transiently normalize tumor vasculature. However, their rapid systemic clearance limits sustained vascular modulation. This study developed norcantharidin-loaded liposomes (NCTD-LPs) to prolong tumor vascular normalization in 4T1 tumor-bearing BALB/c mice, thereby alleviating hypoxia and enhancing radiotherapy efficacy. NCTD-LPs were synthesized via thin-film dispersion and characterized for particle size (140.94 ± 13.23 nm), zeta potential (29.54 ± 2.60 mV), drug-loading capacity (41.81 ± 5.75%), and encapsulation efficiency (15.89% ± 1.28%). In the BALB/c model, dynamic changes in vascular markers (CD31, α-SMA) and hypoxia marker (HIF-1α) were quantified to identify the optimal vascular normalization window. Radiotherapy was administered during this window and compared with conventional regimens to evaluate therapeutic synergy. The coordinated changes in CD31, α-SMA and HIF-1α identified day 10 post-treatment as the optimal period for vascular normalization. The combined NCTD-LPs+radiation group achieved a tumor inhibition rate of 43.60%, significantly surpassing radiation group (18.13%) or NCTD-LPs group (17.24%). Mechanistically, NCTD-LPs stabilized tumor neovasculature, curbed aberrant angiogenesis, and mitigated vascular leakage, collectively improving oxygen perfusion and radiation sensitivity. This study pioneers a liposome-enabled vascular normalization strategy to amplify radiotherapy outcomes, providing a translatable paradigm for combinatorial oncology therapies.