The aim of this research was to produce a film based on mucilage of the flower of Paneerak modified with licorice root and copper sulfate nanoparticles. The D-optimal statistical scheme was used to study the antioxidant, antimicrobial, X-ray diffraction (XRD), Fourier transform infrared (FTIR), scanning electron microscopy (SEM), and thermal decomposition (DSC) properties of the films. The results showed that with the increase of licorice and copper sulfate nanoparticles, the antioxidant activity of the film increases significantly (p<0.05). The results of the antimicrobial activity of the prepared films showed that the addition of licorice and copper sulfate nanoparticles caused the antimicrobial activity of the film against Escherichia coli and Staphylococcus aureus. X-ray diffraction analysis shows that copper sulfate nanoparticles are physically combined with the mucilage polymer of Paneerak flower and it reduces the crystal structure. The results of Fourier transform infrared (FTIR) confirm the physical presence of copper sulfate nanoparticles in the polymer matrix and have electrostatic interaction with the polymer network. The results of the scanning electron microscope (SEM) show that the surface of films containing licorice root copper sulfate nanoparticles are more heterogeneous than the mucilage of Paneerak flower. Licorice and copper sulfate nanoparticles were able to delay the thermal decomposition of the mucilage of Paneerak flower and increase the thermal stability of the mucilage film. Conclusion: Addition of nanoparticles of copper sulfate and licorice root to edible films based on mucilage of Paneerak flower improved antimicrobial, antioxidant, DSC and SEM.