Halzen Nanoformulation Exhibits Selective Anticancer and Immunomodulatory Effects Through Nanoencapsulation of Phytochemicals

Phytochemical therapeutics are often limited by poor solubility and low bioavailability, restricting their clinical translation despite strong bioactivity. In this study, a multifunctional Halzen nanoformulation (HNF) was developed by encapsulating curcuminoids, withanolides, and thymol within a chitosan–whey protein nanoemulsion to enhance stability and biological performance. Physicochemical characterization confirmed nanoscale droplet size, uniform dispersion, and successful encapsulation. In vitro release studies demonstrated controlled, sustained, and pH-responsive release of encapsulated phytochemicals, while storage stability analysis confirmed preservation of composition and physicochemical integrity under long-term and accelerated conditions. Molecular dynamics simulations demonstrated that HNF forms a stable supramolecular architecture with favorable interactions at the plasma membrane interface, while molecular docking revealed strong binding affinities of key phytochemicals toward inflammation- and cancer-associated targets including IL-1β, CD40, HMOX1, STAT3, and MAPK8. HNF demonstrated strong antioxidant activity with significant reduction of intracellular ROS, selective cytotoxicity toward MDA-MB-231 breast cancer cells, and suppression of nitric oxide production in macrophages, while the blank formulation showed negligible effects under in vitro conditions. Overall, the results demonstrate that nanoencapsulation enhances the functional efficacy of phytochemicals and supports HNF as a promising platform for managing oxidative stress and inflammation-associated diseases, warranting further in vivo validation.