<p>Hyaluronic acid receptor targeting is an innovative approach in cancer treatment. This work aims to characterize anticancer properties of Fe₃O₄ nanoparticles functionalized with glucose and co-conjugated with hyaluronic acid (HA) and Kaempferol (KAE) in triple negative breast cancer (TNBC) cells. The Fe₃O₄@Glu-HA-KAE NPs were characterized by FT-IR, XRD, EDS, SEM, TEM, DLS and zeta potential analyses. Cytotoxicity in MDA-MB-231 cells was evaluated using MTT assays. Apoptosis and cell cycle changes were analyzed by flow cytometry, Nuclear morphology was examined via AO/PI staining, and ROS production was measured in treated and control groups The FT-IR, XRD and EDS analyses confirmed the correct synthesis of Fe₃O₄@Glu-HA-KAE NPs. The NPs were spherical with a particle size of 10–60&#xa0;nm in their dried form and an average diameter of 276&#xa0;nm and a surface charge of -39.7 mV. Fe₃O₄@Glu-HA-KAE NPs exhibited dose- and time-dependent toxicity against TNBC cells and the 24-hour and 48-hour IC₅₀ of the NPs in the MDA-B-231 cells were 215 and 149&#xa0;µg/mL, respectively. In addition, the NPs caused cell cycle arrest at the sub-G1 phase, and increased cell apoptosis percentage to 65.1–68.1%. The synthesized NPs triggered significant nuclear alterations, enhanced ROS generation, and elevated cell death in TNBC cells.Furthermore, exposure to Fe₃O₄@Glu-HA-KAE NPs led to a 1.41-fold increase in <i>Caspase-8</i> expression, while <i>BCRT1</i> lncRNA transcript levels were markedly reduced to 0.73-fold, indicating that apoptosis-related mechanisms contribute to the observed cytotoxicity. This work demonstrates efficient anticancer properties of Fe₃O₄@Glu-HA-KAE NPs against TNBC cells, representing an innovative approach to combat TNBC.</p>