Nanotechnology is swiftly becoming a revolutionary scientific discipline focused on the synthesis, characterization, and development of nanomaterials, which exhibit unique physicochemical properties, including a significantly enhanced surface area compared to their bulk counterparts (Dubchak et al., 2010, Mubarak et al., 2013, He et al., 2019). Nanomaterials have effective fundamental roles that contribute to improving industrial, agricultural, communications, and medical applications (Roco, 2011). Currently there are over 1000 commercial and scientific nanoproducts assessable in the global markets (Thakkar et al., 2010). Nanomaterials exhibit considerable variations in catalytic activity, thermal properties, melting point, conductivity, mechanical characteristics, and optical absorption (Schmid et al., 2017). The antimicrobial activity of metals positively influences metal oxides due to their ability to bind and target. The inherent antimicrobial properties of metal oxide nanoparticles, such as continuous mass, distinctive atomic structure, high reactivity, controlled pore size, and tailored particle morphology, have garnered significant research interest. Among these, CuO, ZnO, and Au2O3 nanoparticles are frequently employed due to their demonstrated antibacterial, anticancer, antiviral, and antioxidant activities (Rafique et al., 2017). CuO nanoparticles have sparked widespread interest owing to their optical, catalytic, mechanical, electrical, and biological capabilities. Moreover, biogenic synthesized CuO-NPs offer a wide range of applications, including antimicrobial, antifungal, and drug delivery. A variety of synthetic approaches have been enhanced for synthesizing copper and copper oxide nanoparticles, including electrochemical reduction, physical and other chemical techniques (Kareem et al., 2019), and electrochemical reduction and hybrid procedures (Yang et al., 2013). Furthermore, ecologically friendly, sustainable approaches using microorganisms and plant extracts as sustainable alternatives to limited chemical and physical methods (Sadeghi and Gholamhoseinpoor, 2015). Conventional synthesis routes frequently rely on harmful chemical agents that pose significant environmental risks, require significant energy, extended reaction periods, and external additions throughout the synthesis process. Implementing the sustainable bio-protocol is crucial since it provides cost-effectiveness, environmental friendliness, energy efficiency, and non-toxic chemicals (Qiu et al., 2018). The manufacturing of nanoparticles using plant extracts, “green synthesis,” is the future of medical applications. The potential to employ phytochemicals, which are essential sources of biocompatible and bioactive compounds, as natural reducing and stabilizing agents in the biosynthesis of nanomaterials presents a promising future for nanomaterial creation (Makarov et al., 2014). Lepidium sativum L. Medicinal-herbal plants have been considered as natural sources of active phenolic compounds such as polyphenols, flavonoids, phenolic acids, and other phytochemicals throughout the ages (Bhatia et al., 2021, Anand et al., 2019, Godlewska-Zyłkiewicz et al., 2020). The active phytochemicals have been shown to have antibacterial activity by reducing the formation of biofilms, preventing the attachment of bacteria to mucosal surfaces, as well as affecting the hydrophobicity of the cell surface, and decreasing the levels of glycolytic enzymes (Mani et al., 2020, Edis et al., 2019, Küünal et al., 2018). The plant extracts are characterized by garden cress seeds as well as sprouts being rich in compounds from these groups. In L. sativum seeds, phenolic compounds such as gallic acid, protocatechuic acid, coumaric acid, caffeic acid, coumaroylquinic acid, caffeoylquinic acid, quercetin, and derivatives of vanillic acid and kaempferol have been identified (Zia-Ul-Haq et al., 2012, Alqahtani et al., 2019). Isoflavonoids such as daidzin, genistein, ononin, daidzein, sissos-trin, genistein, formononetine, prunetin, and biochanin A are found in L. sativum sprouts. The most abundant are prunetin, formononetin, and biochanin A, followed by daidzein and ononin (Lapcik et al., 2006). In addition, avonole derivatives (kaempherol-sinapoyl-di-hexose-pen-tose, nonacylated kaempherol-dihexose-pentose, kaempherol-feruloyl-dihexoside-pentoside) and quercetin-sinapoyl-dihexose-pentoside were found (Oszmiański et al., 2013), demonstrating the presence of sinapic acid compounds and its derivatives: cis and trans forms of sinapic acid, as well as sinapine (sinapine acid and choline ester), 1-O-sinapoyl-β-glucose, and its isomer (Sadia Aroob et al.). Building on previous investigations of plant phytochemicals, our current research aimed to synthesize copper oxide nanoparticles from Lepidium sativum L. leaves for the first time, utilizing LS as a reducing and stabilizing agent to augment the inhibitory effects of CuO-NP (Akintelu et al.).