Molecular engineering to elevate reactive oxygen species generation for synergetic damage on lipid droplets and mitochondria

Photodynamic therapy (PDT), characterized by high treatment efficiency, absence of drug resistance, minimal trauma , and few side effects , has gradually emerged as a novel and alternative clinical approach compared to traditional surgical resection, chemotherapy and radiation. Whereas, considering the limited diffusion distance and short lifespan of reactive oxygen species (ROS), as well as the hypoxic tumor microenvironment , it is crucial to design photosensitizers (PSs) with suborganelle specific targeting ability and low-oxygen dependence for accurate and highly efficient photodynamic therapy. In this study, we have meticulously designed three PSs, namely CIH , CIBr , and CIPh , based on molecular engineering. Theoretical calculation demonstrate that the three compounds possess good molecular planarity with calculated S 1 -T 1 energy gaps (Δ E S1-T1 ) of 1.04 eV for CIH , 0.92 eV for CIBr , and 0.84 eV for CIPh respectively. Notably, CIPh showcases remarkable dual subcellular targeting capability towards lipid droplets (LDs) and mitochondria owing to the synergistic effect of lipophilicity derived from coumarin's inherent properties combined with electropositivity conferred by indole salt cations. Furthermore , CIPh demonstrates exclusive release of singlet oxygen ( 1 O 2 )and highly efficient superoxide anion free radicals(O 2 ⦁− ) upon light irradiation supported by its smallest S1-T1 energy gap (Δ E S1-T1  = 0.84 eV). This leads to compromised integrity of LDs along with mitochondrial membrane potential , resulting in profound apoptosis induction in HepG2 cells. This successful example of molecular engineering guided by density functional theory (DFT) provides valuable experience for the development of more effective PSs with superior dual targeting specificity. It also provides a new idea for the development of advanced PSs with efficient and accurate ROS generation ability towards fluorescence imaging-guided hypoxic tumor therapy.