© 2022 Informa UK Limited, trading as Taylor & Francis Group.This work aimed to create an extract of Melissa officinalis L. with strong antiradical efficacy, characterize it, and enhance its long-term efficacy by developing an ethosomal formulation. DPPH and ABTS assays were used to test the antiradical activity of extracts with different ethanol ratios obtained from the aerial part. Phytochemical characterization of the extract with the highest activity, ethyl acetate fraction of 60% ethanol extract, was analyzed by HPLC. The active ethyl acetate fraction was loaded into ethosomes, and characterization and release studies of the formulation were performed. The released extract from the formulation exhibited substantial antiradical action as well as inhibition of collagenase (71.5%) and elastase (75.5%) enzymes. The toxicity of the active extract and the formulation was determined in the mouse fibroblast cell line. This study successfully developed a long-term antioxidant and enzyme inhibitor formulation containing M. officinalis, which stands out for its medicinal properties.