Assessing small-sized D-fructose-functionalized micelles for active targeting of triple-negative breast cancer

Triple-negative breast cancer (TNBC) is one of the most aggressive and clinically challenging subtypes of breast cancer. Its high heterogeneity and lack of actionable molecular targets limit therapeutic options, leading to poor patient outcomes. Given the reported overexpression of the fructose transporter GLUT5 (Glucose Transporter type 5) in breast cancer cells, D-fructose (FRUC) has been proposed as a ligand for active targeting; however, its effectiveness remains uncertain. Here, we investigate passive and fructose-mediated active targeting of TNBC using small-sized supramolecular micelles in vitro and in vivo . We designed two micelle formulations, one incorporating a D-fructose headgroup, using amphiphiles composed of pentacosadiynoic acid as the hydrophobic core and polyethylene glycol as a spacer. These micelles (∼10 nm) were characterized by nuclear magnetic resonance, electron microscopy, and dynamic light scattering, and their stability was confirmed under various conditions. Cellular internalization studies, flow cytometry, and confocal microscopy revealed that PEGylated micelles exhibited superior intracellular accumulation compared to fructose-functionalized micelles, suggesting that D-fructose may hinder endocytic uptake rather than enhance it. In vivo biodistribution analysis in tumor-bearing mice showed similar tumor accumulation for both formulations, with FRUC-Mic exhibiting a transiently higher presence at 1-h post-administration, though this difference was not statistically significant. Although this study does not validate D-fructose as an effective ligand for active targeting, both micelle formulations showed key advantages, including Immunoglobulin G-sized particles, high stability, and facile synthesis, enabling scalable production and clinical translation. Moreover, their ability to solubilize, stabilize, and deliver hydrophobic dyes and cytotoxic agents underscores their potential as theranostic nanoplatforms for TNBC treatment. PDA-based micellar nanocarriers, functionalized with or without D-fructose ligand, were engineered to facilitate a comparative evaluation of active versus passive targeting strategies in triple-negative breast cancer (TNBC), exhibiting promising drug delivery performance in preliminary in vitro and in vivo studies.