Folic acid-targeted β-lactoglobulin nanocarriers for enhanced delivery of 5-fluorouracil and sodium butyrate in colorectal cancer treatment

Colorectal cancer (CRC) remains a significant public health concern, emphasizing the need for innovative therapeutic strategies to improve patient outcomes. This study aimed to develop a highly efficient nanocarrier for targeted drug delivery, enhancing drug efficacy while minimizing concentrations and limiting adverse effects. We synthesized protein-based beta-lactoglobulin (beta lg) nanoparticles (NPs), loaded with 5-fluorouracil (5-FU) and sodium butyrate (NaB), and further functionalized with folic acid (FA) for specific targeting of folate receptor- positive CRC cells. The beta lg-5-FU-NaB-FA nanoplatforms exhibited a well-defined size of 208 nm with a narrow size distribution (PDI approximate to 0.5). Zeta potential measurements showed a value of -11.4 mV, indicating stability and suitability for drug delivery. Scanning electron microscopy (SEM) and atomic force microscopy (AFM) confirmed the nanocarrier's spherical morphology and efficient distribution. Drug release profiles demonstrated that the NPs released more drugs at neutral to alkaline pH levels, attributed to pectin's ionization properties. The efficacy of the prepared beta lg-5-FU-NaB-FA nanoplatforms was investigated on HCT116 and Caco2 CRC cells, along with the normal cell line CRL-1831. The beta lg-5-FU-NaB-FA nanoplatforms exhibited remarkable cytotoxicity against both HCT116 and Caco2 CRC cells compared to free drugs, highlighting the efficacy of targeted delivery in folate receptor-positive cells. These NPs induce cell apoptosis and cell cycle arrest more effectively than free drugs, demonstrating their potential for targeted cancer therapy. Furthermore, a decrease in the expression of crucial genes involved in the Wnt signaling pathway was observed, which offers a valuable understanding of their underlying mechanism. Collectively, our results suggest that the FA-targeted beta lg nano- carriers represent a promising platform for the efficient and targeted delivery of 5-FU and NaB in folate receptor- positive CRC. This novel nanocarrier holds the potential to enhance therapeutic outcomes while minimizing side effects, providing a new avenue for the treatment of CRC.