Plant-mediated nanoparticles gaining importance due to their broad spectrum of pharmacological applications in comparison to the synthetic drugs. The nanoparticles amalgamated using plant extracts have been receiving much attention due to the robust phytochemicals and therapeutic potential. Thus, we herein report the biosynthesis and pharmacological evaluation (anti-oxidant, anti-inflammatory, anti-platelet, and anti-cancer properties) of Decalepis hamiltonii leaves extract copper ferrite nanoparticles (DHLE-CuFe2O4 NPs). The characterization of DHLE-CuFe2O4 NPs was carried out using Powder X-ray diffraction (PXRD), Fourier transform infrared spectroscopy (FTIR), Scanning electron microscopy (SEM), Energy-dispersive X-ray analysis (EDAX), High-resolution transmission electron microscopy (HR-TEM), and Vibrating sample magnetometer (VSM). DHLE-CuFe2O4 NPs exhibited anti-oxidant activity by scavenging 1,1-diphynyl-2-picrylhydrazyl (DPPH) radicals (66.46%) and reduced ferric to ferrous ions at the concentration of 150 μg/mL. Furthermore, DHLE-CuFe2O4 NPs restored the NaNO2-induced oxidative stress markers such as lipid peroxidation, protein carbonyl content, total thiol, and anti-oxidant enzyme (catalase and super oxide dismutase) activities in RBCs. DHLE-CuFe2O4 NPs were non-toxic as it did not cause RBCs lysis, while it exhibited anti-inflammatory activity by inhibiting heat-induced hemolysis, egg albumin, and bovine serum albumin denaturation. In addition, DHLE-CuFe2O4 NPs inhibited ADP and epinephrine-induced platelet aggregation. Most importantly, DHLE-CuFe2O4 NPs showed anti-cancer potential by eliciting cytotoxic effect to MCF-7 cells in a dose-dependent manner. Further, DHLE-CuFe2O4 NPs caused apoptosis to MCF-7 cells which was confirmed by annexin V/PI staining test and flow cytometer. In conclusion, DHLE-CuFe2O4 NPs regulate oxidative stress-induced red blood cell damage, thrombosis, inflammation, and MCF-7 cells growth.