Response surface methodology and artificial neural network-based modeling and optimization of phosphatidylserine targeted nanocarriers for effective treatment of cancer: In vitro and in silico studies

This study aimed to develop and evaluate phosphatidylserine (PS)-targeted nanoparticles for efficient delivery of methotrexate to tumor sites. Therefore, methotrexate-loaded formulations were designed and optimized by response surface methodology and validated by the genetic algorithm-artificial neural network. The optimized nanoparticles (MNP17) were further conjugated with annexin-v (ANX) using the carbodiimide coupling tech-nique to develop the ANX-conjugated nanoparticles (ANX-MNP). These formulations were evaluated for physi-cochemical properties such as drug entrapment, loading, mean particle size, surface charge, morphology, and in vitro drug release. The mean particle size of formulations MNP17 and ANX-MNP were found to be 121 and 146 nm, respectively. Drug entrapment and drug loading were found to be greater than 65% and 32%, respectively. The morphology of the nanoparticles was investigated by transmission electron microscopy and atomic force microscopy. In vitro drug release from formulation was found to be approximately 90% after 120 h. The release mechanism followed the Korsmeyer Peppas model and non-Fickian diffusion (0.5 < n < 0.85). In vitro cytotoxic studies showed significantly better effects of ANX-MNP than MNP17 and native drug in MDA-MB-231 cells. Qualitative and quantitative cellular uptake studies showed higher uptake of ANX-MNP than MNP17 by MDA-MB-231 cells. The formulations showed enhanced nuclear fragmentation, generation of reactive oxygen species, alterations in mitochondrial membrane potential, and decreased colony formation and Bcl-2 expressions in ANX-MNP as compared to MNP17 and native drug. These study findings demonstrated the potential of ANX-MNP as a carrier for improved drug delivery in cancer treatment. The utilization of physiological changes of cancer cells, i. e., PS on the outer leaflet of plasma membrane serves as a ligand to achieve the site-specific drug delivery via exploiting the affinity of ANX towards exposed PS is the novelty of present research.

Automated summary

This study developed PS-targeted nanoparticles for efficient delivery of methotrexate to tumor sites. The nanoparticles showed good drug entrapment and loading rates, and the ANX-MNP formulation exhibited better cellular uptake and cytotoxic effects compared to MNP17. The study demonstrates the potential of ANX-MNP as a carrier for improved drug delivery in cancer treatment.