In this study, nanogels based on the conjugation of heparin and poloxamer P407 (HR-P407) with different grafting ratios were successfully synthesised. The structural properties of products were evaluated by Fourier-transform infrared spectroscopy (FT-IR) and proton nuclear magnetic resonance (1H-NMR). The morphology and size of nanoparticles were assessed by dynamic light scattering (DLS) method, which showed that particle size distribution was from 144 nm to 214 nm. The HR-P407 nanogel systems were biocompatible and non-toxic to healthy cells. While free PTX displayed rapid and potent cytotoxic effects, PTX released from HR-P407 (1:10)/PTX demonstrated a gradual and sustained cytotoxicity. This controlled release mechanism allowed prolonged drug exposure, reducing acute toxicity and enhancing cytotoxicity over time. The results suggest that the synthesised nanogels can be used as an effective hydrophobic anticancer drug carrier system.