The carboxymethyl dextran-y-cyclodextrin (CMD-yCD) conjugate was prepared as the carrier for the delivery of the poorly water-soluble anticancer drug, doxorubicin (DOX). The conjugate could form self-assembled nanoparticles (315 nm in diameter) in an aqueous solution, which might be due to the hydrogen bonding among yCD molecules in the conjugate. DOX was effectively encapsulated into CMD-yCD nanoparticles (CMD-NPs) by the emulsion method. In particular, regardless of the feed amount of DOX, its loading efficiencies were always greater than 70%. CMD-NPs released DOX in a sustained manner, owing to the inclusion complex formation between DOX and yCD. When Cy5.5-labeled CMD-NPs were treated with SCC7 cancer cells, strong fluorescence signals were observed at the cytosol, indicating effective intracellular uptake. In addition, DOX-loaded CMD-NPs exhibited dose-dependent cytotoxicity to SCC7 cancer cells. However, the empty nanoparticles did not show toxicity to the cells, implying their high biocompatibility. Overall, these results suggest that the CMD-gammaCD conjugate could be a useful carrier for the delivery of DOX.