Although countless work on the synthesis, encapsulation and toxicity of paclitaxel-loaded nanoparticles (NPs) has been reported, there have been few reports on the development of the nano-carrier materials of the paclitaxel (PTX) delivery system with therapeutic effects. In this study, we synthesized PTX-loaded tocopheryl succinate-conjugated chitosan oligosaccharide (CSO–TOS) NPs for synergistic chemotherapy. The resultant PTX-loaded CSO–TOS NPs were about 116 nm in diameter with spherical shape and high encapsulation efficiency. Fluorescence microscopy and flow cytometry studies showed that PTX-loaded CSO–TOS NPs had excellent cellular uptake ability by human glioma U87 cells. In vitro cytotoxicity studies revealed that the PTX-loaded CSO–TOS NPs were more potent than free PTX. The therapeutic effects of carrier materials were also studied and a synergistic effect between TOS and PTX was achieved. Cytoarchitecture studies demonstrated a similar cytoskeleton pattern before and after PTX-NPs were loaded into bone marrow derived macrophages. In vivo pharmacokinetic results indicated that the PTX-loaded CSO–TOS NPs had a longer systemic circulation time and slower plasma elimination rate than those of Taxol^®. These results suggested that CSO–TOS could be a potential vehicle for delivering hydrophobic chemotherapeutic drugs to tumors.