Mitotropic triphenylphosphonium doxorubicin-loaded core-shell nanoparticles for cellular and mitochondrial sequential targeting of breast cancer.

[Display omitted] • New chitosan-hyaluronic acid (CS-HA) core–shell nanoparticles (NPs) were developed. • The CS-HA NPs are the first sequential targeting delivery system of doxorubicin. • Mitotropic triphenylphosphonium-doxorubicin conjugate was synthesized. • The drug conjugate-loaded NPs were tested for sequential targeting of breast cancer. • In-vivo studies confirmed efficiency of CS-HA core–shell NPs for dual-stage delivery. Targeted therapy exploits cancerous niches' properties including acidic extracellular environment, hypoxic tumor core, and over expression of tumor-specific surface antigens. The present study aims to develop and evaluate a sequential targeted core–shell nanoparticulate (NPs) system for treatment of breast cancer. Sequential (double-stage) targeting was achieved at the cellular-level through employing the selective CD44- receptor binding hyaluronic acid (HA), followed by subcellular mitochondrial drug-delivery using the mitotropic triphenylphosphonium-conjugated doxorubicin (DOX-TPP+). NPs were prepared through incorporation of the electrostatic-complexes of DOX.HCl/DOX-TPP+ with tripolyphosphate (STPP-) into chitosan (CS) forming the core that was further coated with HA shell. Physicochemical characterization techniques namely; FTIR, DSC, DLS, morphological evaluation and spectroscopic assessments were implemented. Moreover, the drug entrapment efficiency (EE%), loading capacity (LC%), drug release profile and kinetics were investigated. Lastly, to validate the biological efficiency of the developed NPs, cytotoxic activity was evaluated as well as flow cytometric analyses to assess apoptosis induction and cell-cycle arrest were studied. Results showed that, the obtained core–shell NPs possessed a spherical shape with a mean size of 220–280 nm and attained high EE% and LC%. In-vitro cytotoxicity evaluations demonstrated successful apoptosis induction and cell-cycle abrogation. Moreover, in-vivo studies on Solid Ehrlich carcinoma (SEC)-bearing mice confirmed the efficient anticancer activity of the mitotropic DOX-TPP+-loaded NPs. Conclusively, the developed core–shell NPs proved efficient in sequential targeting of DOX to breast cancer. [ABSTRACT FROM AUTHOR]