Quality by design enabled development & in-vitro assessment of a Nanoemulgel formulation for Nose-to-Brain delivery of Nintedanib for glioblastoma multiforme treatment.

Glioblastoma multiforme (GBM) is a deadly malignant brain tumor that spreads uncontrollably and invades the surrounding brain parenchyma. GBM treatment remains challenging due to the rigid blood-brain barrier, limiting therapeutic entry into the brain. Therefore, the current study focused on formulating a Nintedanib (Nint) loaded in-situ Nanoemulgel (Nint-Nanoemulgel) and exploring its permeation and therapeutic potential under in-vitro models to address these limitations. Nint-Nanoemulgel was optimized through the QbD-enabled Box-Behnken design. Optimized Nint-Nanoemulgel revealed significant globule size (27.4 ± 0.8  nm), PDI (0.17 ± 0.01), % encapsulation efficiency (93.5 ± 3.5 %), zeta potential (-4.7 ± 0.6 mV), %T (98.2 ± 0.2 %), pH (6.0 ± 0.2), and viscosity (2.59 ± 0.24 cP) at 25 °C. A cumulativein-vitro release study revealed 87.4 ± 1.9 % Nint release through Nanoemulgel after 12 h while 90.1 ± 2.1 % release after 24 h.The cytotoxicity potential of developed Nint-Nanoemulgel was screened in GBM cell lines, demonstrating a > 2-fold reduction in IC50 than plain Nint. However, after treatment with 100 µM of Nint-Nanoemulgel, % growth inhibition was found to be 91.0 ± 1.0 %, 92.1 ± 1.3 %, and 82.7 ± 1.0 % in LN229, U87, and U138 cell lines, respectively. Further,in-vitro cellular uptake exhibited significant coumarin cellular internalization through nanoformulations against coumarin solution. Moreover, clonogenic and scratch assay studies demonstrated the ability of Nint-NE to inhibit cell proliferation and colony formation. However, the outcomes of the live-dead assay demonstrated more cell death in Nint-nanoformulation-treated spheroids than in Nint-treated spheroids. Nint-Nanoemulgel improved intracellular permeation and demonstrated a 2-fold increase in Nint transport across the RPMI-2650 epithelial monolayer. Finally, favorable outcomes of intranasal Nint-Nanoemulgel could provide a novel avenue for the safe and effective delivery of Nint in GBM patients.