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Chromolaena odorata (L.), locally named Siam weed, is a rich source of phyto-bioactives and has been traditionally used for wound healing. This study deals with the phytochemical investigation of the leaves of C. odorata (L.) for their antioxidant, anti-inflammatory, and antibacterial properties that contribute to the observed wound healing properties. The phytoconstituents were sequentially extracted from the powdered leaves of C. odorata (L.) and chemoprofiled using the LC-MS technique. Furthermore, copper oxide/copper nanoparticles (CuNPs) were synthesized by the use of water extracts from the leaves of C. odorata (L.), characterized and evaluated for their potential biological and wound healing impacts. In this relation, scutellarein, isosakuranetin, and rutin, along with 15 other phytoconstituents were identified. Higher antioxidant (79.2% at 200 µg/mL) and anti-inflammatory potentials (66.8%) were obtained for CuNPs (120 µg/mL), in the size range of 66.8 ± 24.8 nm. Antibacterial activity against methicillin-resistant Staphylococcus aureus and Pseudomonas sp. demonstrated mean zones of inhibition of 16.7 ± 4 mm and 17.73 ± 2 mm, respectively. Hence, the synthesized CuNPs would be desirable for inclusion in nanofibrous medical dressings. [ABSTRACT FROM AUTHOR]