Waterborne polyurethane (WPU) prepolymer was synthesised by the reaction of poly(butylene itaconate) ester (PBI, Mn = 1109 g/mol), 1,6-hexanediol, dimethylol propionic acid (DMPA), 2,4-toluene diisocynate (TDI), hydroxyethyl acrylate (HEA), and absolute ethanol as blocking agent, triethylamine as neutralizer. Cross-linked WPU was synthesized by trimethylolpropane (TMP) as crosslinker. The influences of PBI, DMPA, and TMP content on WPU emulsions and films were investigated. The structure of WPU was determined by Fourier transform infrared (FTIR) spectra, thermal properties and glass transition temperature of WPU films were determined by thermogravimetric analysis (TGA) and differential scanning calorimetry (DSC), respectively, and morphology of the emulsion particles was observed by transmission electron microscopy (TEM). Through TGA, the heat resistance of the cross-linked WPU film was better than WPU film. By DSC analysis, glass transition temperature of cross-linked WPU film (21 °C) was higher than WPU film (10 °C).