58P Novel nanoACPA Delivery System Presents Antitumor Effect on Ectopic Non-Small Cell Lung Cancer Xenograft Model

Abstract

Non-small cell lung cancer (NSCLC) constitutes 87% of all lung cancer cases as a late diagnosed tumor with highest mortality rate. We previously reported cannabinoid 1 (CB1) receptor-mediated apoptotic effect of CB1 agonist Arachidonoylcyclopropylamide (ACPA) and generated a novel polycaprolactone-based ACPA nanoparticular system (NanoACPA) acting on NSCLC through PI3K/Akt and JNK pathways in vitro (PCT/TR2020/050618). Here, we hypothesized that NanoACPA technology platform selectively targeting CB1 receptor might have a chemotherapeutic potential inhibiting caspase-mediated PI3K/Akt pathway in vivo on a subcutaneous NSCLC nude mice model. Optimized NanoACPA formulation was freshly prepared by nanoprecipitation. Subcutaneously injected luciferase-expressing A549 cells induced NSCLC nodules on 6–8-week-old male BALB/c nude mice (Ethics Committee Approval #2021/15). Bioluminescence imaging assessed tumour size alterations and metastasis on days 7-21 in free ACPA, NanoACPA, solvent and saline (control)-treated mice besides 2D calliper measurements. Tumour nodules and potential metastatic loci were excised for histomorphometry and immunohistochemistry to analyse proliferation (Ki67) and apoptotic indices (TUNEL) and Western blot for caspases, Akt and p-Akt. NanoACPA more prominently decreased tumor size from days 7-21 by bioluminescence imaging and calliper measurement compared to control (p<0.05). No metastatic foci were noted in distant organs of chemotherapeutic-applied and control mice on days 7-21. ACPA and NanoACPA reduced proliferative and induced apoptotic indices in tumor foci comparing to controls by Ki-67 and TUNEL (p<0.05). NanoACPA induced caspase 9, cleaved caspase 3 and inhibited p-Akt. This study reveals the first in vivo validation of the novel NanoACPA platform acting on caspase-mediated PI3K/Akt pathway for NSCLC treatment on an ectopic xenograft model. The innovative synthetic non-psycoactive cannabinoid-based candidate might present a strong chemotherapeutic potency at a low dose for NSCLC, therefore may deserve scale up at GMP level for translational phase studies to assess further safety on a high technological infrastructure.