Development and in silico, in vitro and in vivo biological evaluation of flutamide-loaded potato starch-chitosan-lactoferrin nanoparticles as promising candidates for targeted colorectal cancer therapy

Flutamide-loaded potato starch-chitosan-lactoferrin nanoparticles (FLU-PS-CS-LF-NPs) were explored for targeted delivery against colorectal cancer therapy. The best nanoparticles, with a diameter of 186.8 ± 63.12 nm and a zeta potential of +5.77 ± 1.56 mV, showed passive tumor targeting via the EPR effect and enhanced mucoadhesion. Drug entrapment and physicochemical integrity were ascertained by spectroscopic investigations (FTIR, XRD, Raman, and NMR). Thermal and microscopic analysis established morphological homogeneity and a change in crystalline form, supporting enhanced solubility. MTT assay showed enhanced cytotoxicity against HCT-116 cells (IC50 = 6.77 μg/mL), and flow cytometry confirmed increased apoptosis and G2/M phase cell cycle arrest. Other results showed decreased ROS and mitochondrial damage, effective cellular uptake, and strong anti-metastatic activity. Antibacterial and fecal culture tests showed inhibition of CRC-associated microbiota, while biocompatibility tests established safety and nontoxicity. In vivo colon-targeting and pH-dependent delivery were achieved, and the formulation remained stable for 6 months. Results indicate that FLU-PS-CS-LF-NPs are a potential nanocarrier system for effective, safe, and targeted therapy of colorectal cancer.