Pressure ulcers are commonly associated with microbial infections on the wounds which need an effective wound dressing. However, the silver dressings have shown promising result but they have toxicity and argyria. Hence, this study aimed to develop and characterize chitosan-polyethylene glycol (PEG) nanocomposite hydrogel loaded with phomopsidione as an antimicrobial dressing. The hydrogel being synthesized was analyzed with transmission and scanning electron microscopes. Drug release and mechanical properties were studied having confirmed the functional groups with Fourier transform infrared (FTIR) spectroscopy. Finally, antimicrobial activities were evaluated against the clinical wound pathogens. The developed hydrogel was soft, flexible and elastic, having nanospheres of chitosan-PEG but no sign of aggregation under the electron microscopes. Releasing of phomopsidione from the nanocomposite hydrogel was slow and gradual following the first order of kinetic. On average, 34 μg/mL phomopsidione released per hour and 67.9% active ingredients delivered into the surrounding medium over the study period. Although, the bioactivity activity of the hydrogel was narrow-spectrum, it showed significant results against all Gram-negative bacteria and Candida utilis with 99.99% reduction of microbial growth. The findings reveal that the phomopsidione loaded hydrogel has a great promise to act as an antimicrobial dressing for chronic wounds.