Küçük Hücreli Dışı Akciğer Kanseri için Kannabinoid Yüklü İlaç Taşıma Sistemlerinin Geliştirilmesi ve Validasyonu

Abstract

Lung cancer has a high mortality rate worldwide with its advanced invasive behavior and metastatic capacity. FDA-approved or currently tested targeted agents inhibit PI3K/Akt, Ras/MEK/Erk and JNK pathways but with low success due to rapid progression of the disease and systemic side-effects complicating the treatment process. Cannabinoid system components emerge as drug candidates for non-small cell lung cancer (NSCLC) treatment due to anti-tumoral capabilities, however, their stability, bioavailability and therapeutic efficacy should be evaluated. Our group previously presented the antiproliferative and apoptotic effective low dose window of synthetic specific CB1 receptor agonist Arachidonylcyclopropylamide (ACPA) and generated a novel polycaprolactone (PCL)-based nanoparticle system in conjunction with its metabolic pathway in NSCLC in vitro. If a novel high technology-controlled release platform increasing the bioavailability of ACPA at a low IC50 on NSCLC could be provided by in vivo animal model-based safety and tolerability tests with low/no systemic side effect, it might be offered for further clinical trials as a prospective therapeutic. In the thesis, firstly, the pharmacokinetic profile and immune response of ACPA and ACPA-loaded PCL-based nanomedicine by LC-MS/MS and complete blood count, respectively, were assessed in wild-type Balb/c mice. NSCLC induction was then accomplished by subcutaneous luciferase-expressing A549 injection to athymic nude mice. Tumor growth assessment following i.p. nanomedicine applications was performed by bioluminescence imaging (BLI) and caliper measurements. Tumor nodules excised following 7-21 days of application were subjected to histomorphometry, immunohistochemistry, TUNEL and Western blot. Injectable NanoACPA increased systemic exposure to tissues 5.5 times and maximum plasma concentration 6 times higher compared to free ACPA by significantly decreasing volume of distribution and clearance substantially improving bioavailability with a slight change in CBC parameters. Tumor nodules reached a homogenous size by caliper measurement and BLI in all groups on day 35. Weekly-applied NanoACPA provided 67% of tumor growth reduction compared to control on day 7, whereas, every 2-day-applied NanoACPA exhibited 50% and 96% decrease in tumor growth comparing to control, respectively, on days 14-21. NanoACPA presented maximum efficiency on reducing the tumor area and inflammation on day 21 compared to control and experimental groups with no fibrotic alterations or necrotic areas in other organs by histomorphometry. A potent effect of NanoACPA lasted for at least two days on ectopic NSCLC nude mice through Akt/PI3K, Ras/MEK/Erk and JNK pathways that diminished the Ki-67 proliferative and promoted TUNEL apoptotic cell scores on days 7-21. The output reveals that the NanoACPA platform could be an efficacious chemotherapeutic for NSCLC in the clinical practice, due to its in vivo antitumoral efficacy and improved systemic bioavailability at a safe low dose window. The Technology Readiness Level of the novel platform increased from 4 to 6 by simulating the real biological environment and is ready for forther GLP/GMP-based scale-up and phase trials to make the national participation possible to the international chemotherapeutics market.