Nanodrugs　composed　of　pure　pharmacologically　active　molecules　have　been　extensively　researched　as　emerging　nanomedicines　for　cancer　treatment.　However,　traditional　preparation　approaches　are　generally　limited　by　low　production　rates,　complicated　preparation　processes,　and　high　production　costs.　Here,　we　presented　a　simple　strategy　for　constructing　drug-based　nanoparticles　(2　＇　F-ASO/CPT　NPs)　consisting　of　pure　therapeutic　molecules　by　utilizing　2　＇-fluoro-substituted　antisense　oligonucleotide　(2　＇　F-ASO)-mediated　self-assembly　with　the　anticancer　agent　camptothecin　(CPT).　The　2　＇　F-substitution　could　enhance　the　intermolecular　interactions　between　DNA　and　CPT　through　electrostatic　and　dispersion　interactions.　The　resulting　2　＇　F-ASO/CPT　NPs　had　a　high　drug　loading　capacity　and　improved　drug　stability.　2　＇　F-ASO/CPT　NPs　could　specifically　respond　to　P-glycoprotein　(P-gp)　mRNA　and　inhibit　the　expression　of　P-gp,　protecting　CPT　from　P-gp-mediated　drug　efflux.　Therefore,　the　synergistic　anticancer　effect　caused　by　2　＇　F-ASO/CPT　NPs　could　be　successfully　achieved.　This　work　presented　a　new　2　＇　F-ASO-mediated　drug　self-assembly　strategy　for　carrier-free　chemotherapy-gene　therapy　with　the　potential　to　enhance　cancer　treatment　efficacy.