The　development　of　a　highly　effective　and　low　toxic　drug　is　very　beneficial　for　precise　cancer　therapy.　Herein,　taking　advantage　of　the　hypoxic　nature　of　tumors,　we　carefully　designed　and　synthesized　the　first　hypoxia-activated　single-molecule　prodrug　BDP-CPT-NO2　for　fluorescence　imaging-guided　chemo-photothermal　dual-mode　therapy.　The　prodrug　contains　the　chemotherapeutic　drug　camptothecin　(CPT)　and　a　boron　dipyrromethene　(BDP)-based　photothermal　agent,　which　are　connected　through　a　nitroreductase　(NTR)-responsive　linker　(a　nitro　unit),　and　shows　no　fluorescence　and　extremely　low　cytotoxicity　under　normoxia.　However,　the　nitro　in　the　prodrug　could　be　reduced　to　amino　by　exogenous　NTR　with　nicotinamide　adenine　dinucleotide　(NADH)　in　PBS　buffer　solution　and　endogenous　NTR　in　hypoxic　cancer　cells,　which　is　followed　by　a　self-immolative　reaction　that　leads　to　the　release　of　CPT　and　BDP-COOH.　This　process　is　accompanied　by　significantly　enhanced　BDP　and　CPT　fluorescence,　which　are　due　to　NTR-induced　ester-to-carboxylate　conversion　and　fluorescence　resonance　energy　transfer　(FRET)　interruption,　respectively.　Concomitantly,　BDP-CPT-NO2　displays　powerful　combined　chemo-PTT　therapeutic　efficacy,　as　evidenced　in　human　cervical　cancer　HeLa　and　human　liver　cancer　HepG2　cells,　as　well　as　in　mice　bearing　H22　tumors.　This　study　provides　a　promising　strategy　for　effective　and　selective　cancer　treatment　by　hypoxia-activated　prodrugs　caged　within　organic　photothermal　agents.　©　2024　Elsevier　B.V.