The application of metallic nanoparticles improves the yield and content of bioactive compounds in plants. The aim of the present study was to determine the effects of the foliar application of copper nanoparticles (CuO-NPs) in the yield and content of bioactive compounds in lettuce. Different concentrations of CuO-NPs (0, 0.5, 1, 2, 4, and 6 mg mL−1) were applied in lettuce. The yield, nutraceutical quality, and enzymatic activity were determined. Foliar spraying of CuO-NPs induced an increase in the biosynthesis of bioactive compounds. In addition to an increase in the activity of the enzymes superoxide dismutase (SOD) and catalase (CAT) in lettuce plants, there were no negative effects on yield. Therefore, with the application of CuO-NPs, better quality lettuces are produced for the human diet due to the higher production of bioactive compounds.