Fabrication of in-silico optimized AS۱۴۱۱ aptamer functionalized cyclodextrin stabilized ß-lactoglobulin nanoparticles for targeted delivery and controlled release of Doxorubicin to cancer cells

Protein-based nanocarriers have attracted growing attention in drug delivery applications owing to their inherent biocompatibility, biodegradability, and functional adaptability. In the present study, a novel protein-based nanocarrier composed of ß-lactoglobulin (BLG) and B-cyclodextrin (CD) was designed and further functionalized with the AS۱۴۱۱ aptamer to enable targeted drug delivery. In addition, B-cyclodextrin nanosponges (CDNS) were explored as alternative carriers due to their superior drug encapsulation capacity compared with conventional B-cyclodextrin. This multifunctional platform combines the structural stability of ß-lactoglobulin nanoparticles (BNPs) with the high encapsulation efficiency of cyclodextrins and cyclodextrin nanosponges, along with the high affinity and selectivity provided by aptamer-mediated targeting, resulting in enhanced nanoparticle stability. Doxorubicin (DOX) was successfully loaded into BNP/CD/Apt and BNP/CDNS/Apt nanoparticles with high encapsulation efficiency. The developed formulations exhibited pH-responsive drug release behavior under mildly acidic conditions, mimicking the tumor microenvironment and enabling controlled and site-specific drug release. In vitro hemocompatibility assessments demonstrated excellent blood compatibility, as indicated by negligible hemagglutination, coagulation, and complement activation. Furthermore, the optimized nanocarriers showed significantly enhanced cytotoxic effects against MCF-۷ breast cancer cells compared with free DOX. This improvement was attributed to AS۱۴۱۱ aptamer-mediated targeting, which facilitated increased cellular uptake through specific binding to overexpressed nucleolin receptors on cancer cells, as well as the higher drug loading capacity of cyclodextrin nanosponges. Molecular docking and bioinformatic analyses supported the experimental findings, revealing stable interactions between the optimized AS۱۴۱۱ aptamer and the nucleolin receptor. Notably, the optimized ۲۵-nucleotide AS۱۴۱۱ aptamer exhibited strong binding affinity, reflected by an H-Dock score of -۳۱۹.۷۲. Overall, the DOX@BNP/CD/Apt and DOX@BNP/CDNS/Apt nanocarriers represent a promising approach for targeted cancer therapy, offering enhanced drug delivery efficiency, improved tumor specificity, and reduced systemic toxicity compared with conventional chemotherapeutic agents.

کلیدواژه ها:

B-lactogolubolin ، Doxorubicin ، Aptamer ، Cyclodextrin