Purpose: Recent studies have shown that flavonoids like Hesperetin can bind strongly to the ACE2 receptors abundant on lung cancer cells which induces apoptosis in cancer cells. Our prior studies in a murine lung tumor model shows immune treatment with AntiCD40 can induce anti-tumor effect to destroy treated as well as untreated metastatic tumors where radiation further induces this phenomenon by exposing tumor antigens to immune system. Radiation induces apoptosis, causing cancer cells to be fragmented and exposed to the immune system which is already activated by AntiCD40 antibody. In this study, we like to develop a treatment model for aerosol delivery of AntiCD40 along with Hesperetin nanoparticle (HNP) to lung cancer which will simplify this treatment system without any radiation or any injection method.

Methods: At first, we performed in-vitro survival assay analysis with different doses of Hesperetin (Sizma) (5 to 200 ug/ml) with or without radiation treatment (2 Gy) in human lung cancer cell line (A549). Next, we prepared Hesperetin nanoparticles (HNP) of different sizes 10-20 nm and 20-40 nm of diameters to confirm the in-vivo efficacy in a lung cancer murine model.

Results: Our in-vitro study shows treatment with 15 µg/ml concentration of Hesperetin can generate toxicity of 2 Gy radiation dose in A549 cell line. In further analysis with HNP shows 20-40 nm sized HNP demonstrates higher antitumor effect compared to 10-20 nm size HNPs. Also, we were able to successfully administer aerosol delivery of both HNP and Anti-CD40 in a murine orthotopic lung cancer model.