Dual-targeted hyaluronic acid conjugates on lipid nanoparticles improve delivery and efficacy against glioblastoma: The linkage leads the fate

Glioblastoma (GB) presents a significant therapeutic challenge due to the barriers posed by the blood-brain barrier (BBB) and blood-tumor barrier (BTB), limiting effective drug delivery. This study explores modifying ultra-small nanostructured lipid carriers (usNLCs) co-loading atorvastatin (ATO) and curcumin (CUR), functionalized with hyaluronic acid (HA)-based conjugates to enhance targeted delivery to GB. To optimize targeting performance, dual-targeting peptides (c[RGDfK] and H₇K(R₂)₂) and folic acid (FA) were incorporated, using two distinct functionalization strategies: covalent bonding (NLCs^Conj) and electrostatic interaction (NLCs^Conj+). In vitro studies on U87 GB cells showed that NLCs^Conj exhibited superior cytotoxicity, achieving a 1.6-fold higher targeting efficiency than non-functionalized nanoparticles. A 2D-BBB model demonstrated higher GB cell internalization with NLCs^Conj but lower basolateral drug levels compared to usNLCs or NLCs^Conj+. In vivo biodistribution studies in RNU rats revealed that nanoparticles functionalized with hyaluronic acid-based conjugates exhibited approximately a 5-fold reduction in liver accumulation. Efficacy studies demonstrated that NLCs^Conj (2 i.d.) extended median survival by 71 days, achieving 8.5-fold greater efficacy than free drugs and 1.6-fold improvement over standard treatment with minimal side effects. These findings highlight the potential of HA-based surface modification (NLCs^Conj) to enhance the therapeutic index of NLCs, offering a promising strategy to overcome delivery barriers and improve outcomes in glioblastoma treatment.