Chain architecture-engineered artesunate nanoassemblies target LONP1 to induce oxidative damage for enhanced anti-tumor therapy

Despite demonstrating significant anti-tumor potential as an artemisinin derivative, artesunate faces delivery efficiency challenges due to low water solubility and insufficient targeting specificity. To improve the delivery efficiency, we engineered three artesunate (ART) derivatives, AC 15 -L (linear), AC 15 -B (branched), and AC 15 -C (cyclic) with distinct aliphatic chain architectures. Unexpectedly, we observed that AC 15 -C exhibited superior cytotoxicity against 4T1 breast cancer cells, and had the highest binding affinity for Lon protease 1 (LONP1) (−72.6 kcal/mol). Subsequently, disulfide bond-containing lipid-PEG (DSPE-SS-PEG 2K ) modified chain architecture-engineered ART derivatives nanoassemblies (NAs) were developed to mitigate solubility-related limitations while enhancing targeting precision. Molecular docking and experimental validation demonstrated that ART derivatives inhibited LONP1 through hydrophobic interactions while preserved Fe 2+ -mediated Fenton-like reaction activity. In vitro and in vivo evaluations demonstrated that AC 15 -C NAs outperformed free ART and other NAs, suppressing 4T1 tumor growth via dual action: LONP1-directed mitochondrial proteostasis collapse and reactive oxygen species (ROS) amplification through Fe 2+ -ART interactions. This study elucidated a novel anti-tumor mechanism of ART through the rational design of derivatives with spatially configured aliphatic chains, and developed reduction-responsive NAs to provide an advanced delivery strategy. The targeted effect of the artesunate nanoassemblies (AC 15 -C NAs) designed through a chain structure on LONP1 was demonstrated, which was used for dual-action anti-tumor therapy: the Fenton-like reaction and the inhibition of LONP1, thereby achieving enhanced efficacy and tumor-specific delivery.