In this study, cinnamic acid (CA) conjugated hydroxypropyl chitosan (HPCS) derivatives (HPCS-CA) with different degrees of substitution (DS) were successfully synthesized. The reaction was divided into two steps: the first step was to modify chitosan (CS) to HPCS, and the second step was to graft CA onto HPCS. Structural characterization and properties were carried out employing elemental analysis, Fourier transform infrared (FT-IR) spectroscopy, ultraviolet-visible (UV-vis) spectroscopy, nuclear magnetic resonance (NMR) spectra, X-ray diffraction (XRD), and thermogravimetric analysis (TGA). The solubility test revealed the better water solubility of derivatives than CS. In addition, in vitro antibacterial and antibiofilm tests were performed. As expected, HPCS-CA derivatives exhibited good antibacterial activity against Staphylococcus aureus (S. aureus) and Escherichia coli (E. coli). The MIC and MBC of HPCS-CA derivatives could reach 256 μg/mL and 512 μg/mL, respectively. Confocal laser scanning microscopy (CLSM) analysis proved the inhibitory effect of HPCS-CA derivatives on S. aureus and E. coli biofilms by disrupting the formation of biofilms, reducing the thickness of biofilms, and the number of live bacteria. These results suggest the potential applicability of HPCS-CA derivatives in the treatment of biofilm-associated infections and provide a practical strategy for the design of novel CS-based antibacterial materials.