A novel 5-fluorouracil (5-FU)-based anticancer prodrugs consisting of pyrene and oligonucleotide trimer having 5-halouracils were designed. The UVA-induced electron injection from photoexcited pyrene followed by the radical-induced C1' hydrogen abstraction of 5-fluoro-2'-deoxyuridine is expected to cause the elimination of 5-FU. Since the prodrug clearly suppressed the proliferation of HeLa cells in an UVA-responsive manner, we have successfully developed a novel 5-FU prodrug.