We developed bis-guanidinium-based ionophores and ONBlinked proanionophore. Mechanistic studies revealed that compound 2 followed carrier-mediated ion transport properties via the phosphate/Cl -antiport pathway. The 1 H NMR titrations and lucigenin-based fluorescence assay established that the cleavage of photosensitive ONB-moiety leads to efficient activation of transmembrane anion transport. The photoinduced generation of the active anionophore from its proanionophore leads to an enhancement of cancer cell death. Therefore, lipophilic anionophores capable of transporting phosphate/Cl -ions can provide a future platform for photodynamic therapy for chemotherapeutic studies.