Chk1-Targeted Hesperidin Liposomes for Breast Cancer Therapy: An Integrated In Silico, <i>In Vitro</i> , and <i>In Vivo</i> Evaluation

Checkpoint 1 kinase (Chk1) regulates the cell cycle and facilitates DNA repair processes, which are essential for maintaining genomic integrity. In breast cancer, Chk1 is frequently overexpressed, enabling tumor cells to endure DNA damage. By blocking Chk1, a crucial regulator of the development and spread of malignancies, the current study investigates the potential of natural products to identify different treatment approaches for breast cancer. Based on the molecular docking, absorption, distribution, metabolism, excretion, and toxicity profiling and molecular dynamics simulations, hesperidin (NP621) interacted with important Chk1 residues, demonstrated good stability over 800 ns, and had a favorable binding energy of −9.6 kcal/mol. Liposomal encapsulation was employed to overcome hesperidin’s low solubility and poor bioavailability, thereby enhancing its stability and therapeutic potential in cancer treatment. The liposomal formulations were characterized by Fourier transform infrared, X-ray diffraction, and transmission electron microscopy (FTIR, XRD, particle size analysis, and TEM), confirming successful liposomal binding. Hesperidin encapsulated liposomes (HSP-LLPs) displayed remarkable cytotoxicity against the MCF-7 breast cancer cells (IC50 of 13.52 μg/mL) and significantly inhibited angiogenesis in the chorioallantoic membrane assay. HSP-LLPs reduced tumor burden, restored hematological and biochemical parameters, and improved tissue architecture in a 7,12-dimethylbenz[α]anthracene-induced rat model of breast cancer. These findings demonstrate that Chk1-targeted hesperidin liposomes represent a promising translational nanomedicine approach for breast cancer therapy.