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Nanoparticle green synthesis is an emerging scientific field that focuses on the production of nanoparticles by living cells. In this work, the potential of extracts of alfalfa leaves (Medicago sativa) as affordable and fabricated zirconium nanoparticles (ZrNPs) was evaluated for the first time. alfalfa alcoholic extract and zirconium precursor were used to produce ZrNPs, the production and characteristics of nanoparticles were measured using the Tyndall effect, UV‐vis, FT‐IR, EDX, DLS, and SEM tests, antioxidant and antibacterial properties, and toxicity Measurement of nanoparticles on breast cancer (MDA‐MB‐468) and human healthy cells (HFF) was investigated. The optimization of nanoparticle production was evaluated with ANN and GN algorithms. The analysis showed that nanoparticles have spherical‐like ZrNPs shapes with an average of 50 nm size. in addition, ZrNPs have significant cytotoxic effects against human breast cancer (MDA‐MB‐468) cells, and Human foreskin fibroblast (HFF) cells exhibited the highest resistance to ZrNPs. There was a significant agreement between the results of the designed ANN‐GA model and the experimental data. In this research, ZrNP was successfully created using alfalfa extract as a renewable source of stabilizing and reducing agents with an environmentally friendly approach, and they showed significant antioxidant, antibacterial, and anticancer properties. [ABSTRACT FROM AUTHOR]