Abstract
<title>Abstract</title> <p>Protein nanoparticles offer several advantages as a drug delivery system, including increased stability and activity due to increased protection, biocompatibility, biodegradability, and the existence of multiple functional groups to carry large amounts of drugs and the possibility of attachment of targeting molecules. This study developed a novel, multifunctional nanocarrier for the targeted delivery of doxorubicin. Bovine β-lactoglobulin (BLG) nanoparticles were synthesized as a core, sequentially coated with poly-L-lysine (PLL) and hyaluronic acid (HA) to form BLGNP-DOX/PLL/HA. This design leverages HA's affinity for CD44 receptors, overexpressed on cancer cells, for active targeting. The optimized nanoparticles exhibited a spherical morphology with a hydrodynamic size of ~ 205 nm and a negative surface charge. The system demonstrated a high drug loading capacity and a pH- and hyaluronidase-responsive release profile, enabling controlled DOX release in the tumor microenvironment. In vitro, the BLGNP-DOX/PLL/HA system showed enhanced cytotoxicity and significantly higher cellular uptake in MCF-7 breast cancer cells compared to free DOX and non-targeted nanoparticles (BLGNP-DOX), attributed to CD44 receptor-mediated endocytosis. The nanocarrier also exhibited excellent hemocompatibility, with minimal hemolysis and no significant activation of the coagulation or complement systems. In vivo studies in tumor-bearing mouse model revealed that BLGNP-DOX/PLL/HA effectively inhibited tumor growth and showed a favorable safety profile, with hematological and biochemical parameters largely within normal ranges. The results of this study indicated that the BLGNP/PLL/HA platform is a highly promising and biocompatible targeted drug delivery system for efficient cancer therapy.</p>