Development and Optimization of Osimertinib-loaded Biodegradable Polymeric Nanoparticles Enhance In-vitro Cytotoxicity in Mutant EGFR NSCLC Cell Models and In-vivo Tumor Reduction in H1975 Xenograft Mice Models.

Abstract

Osimertinib (OMB), a third-generation EGFR inhibitor, specifically and irreversibly inhibits EGFRT790M mutant form. Nevertheless, its clinical use is limited due to poor solubility, low absorption, and oral bioavailability. To overcome the low therapeutic capabilities of the free drug, we developed OMB-loaded PCL or CHS nanoparticles and characterized them. Among fifteen devised nanoparticle formulations (Npfs), OMB-PCL-f3, f9, and OMB-CHS-f3 showed great characteristics such as particle size (ranges from 101.3 ± 8.2 to 119.7 ± 10.4 nm), zeta potential (-36.4 ± 3.2 to -31.7 ± 3.9 mV), and polydispersity index (0.227 ± 0.037 to 0.261 ± 0.025). The % entrapment (91.25 ± 5.84 to 95.25 ± 5.88) and drug loading (29.64 ± 2.38 to 33.59 ± 2.36) indicated the formulation optimization. OMB-CHS-f3 demonstrated long-term in-vitro release, with a % cumulative OMB release of 99.99 ± 2.67 within 24 h, and the cytotoxicity of OMB-CHS-f3 showed 2.6- and 2.4-fold superior activity in mutant EGFR harboring H1975 and PC-9 cells, respectively, compared to plain OMB. Quantitative assessment of OMB cellular uptake from OMB-CHS-f3 showed superior drug accumulation of 81.59 ± 5.8% and 77.31 ± 4.6% in H1975 and PC-9 cells which was more than OMB-CHS-f9 and plain OMB. Flow cytometric cell cycle analysis revealed that OMB-CHS-f3 triggered G2/M phase arrest greater than OMB-PCL-f9 and plain OMB. In vivo, OMB-CHS-f3 Npf treatment reduced tumor size and body weight gain compared to Tagrisso treatment (p < 0.05). These findings showed that chitosan-coated OMB Npfs might improve outcomes by overcoming complications, including resistance and disease recurrence in NSCLC patients.