Herpes simplex virus-1 (HSV-1) is the primary cause of infectious blindness. Despite impressive therapeutic outcomes of conventional treatments, HSV-1 drug resistance can be easily developed. Thus, more constructive strategies should be implemented. Led by this inspiration, this work describes the potential utility of a biodegradable silk fibroin/collagen (SF/Col) film combined with GI-20-loaded poly lactic-co-glycolic acid (PLGA) nanoparticle to provide efficient and sustained delivery platform for synthetic GI-20 peptide against HSV-1. A non-irritant film containing 90 % SF and 10 % Col incorporated with mentioned nanodrug showed some optimum physicochemical properties including loading efficiency (74.15 % ± 1.12), tensile strength (3.16 ± 0.67 MPa), water uptake ability (∼73 %), cytocompatibility (viable up to 35 µg/mL of GI-20), and sustained release paradigm (∼90 % within 14 days). Also, GI-20 peptide at concentration of 35 µg/mL could prophylactically attenuate viral titration by 5 log