Development of CIL-0X: A Platform to Enhance Delivery of Inhaled RNA Therapeutics to Lung Cells

PROJECT SUMMARY Chronic pulmonary disease, encompassing both rare and prevalent conditions such as primary ciliary dyskinesia, pulmonary fibrosis, cystic fibrosis, asthma, chronic obstructive pulmonary disease, non-cystic fibrosis bronchiectasis, and lung cancer, poses a global healthcare challenge necessitating novel therapeutics. Over 545 million suffer from these life-limiting diseases worldwide and more than 5.7 million succumb to disease progression each year. The field of ribonucleic acid (RNA), in delivery vehicles such as lipid nanoparticles (LNP), has shown great promise for correcting protein dysfunction and addressing gene defects, which is being harnessed for developing novel treatments for chronic pulmonary disease. Numerous clinical trials have been initiated in the last decade, investigating different therapeutic cargos and routes of delivery for multiple chronic pulmonary indications. To date, many have failed due to low transfection efficiency, which limits the therapeutic impact in lungs, the intended site of action. Particularly for inhaled RNA delivery, the airway mucus barrier presents an initial bottleneck that has not been sufficiently addressed by optimization of nanocarrier with hydrophilic PEG components. To address the mucus barrier and enhance delivery of inhaled RNA therapies, CILA Therapeutics proposes to employ an altogether different approach anchored on use of a novel transfection enhancing platform, CIL-0X, a first-of-its kind inhaled dry-powder microsphere mucolytic to temporarily disrupt the mucus gel matrix, enabling safe delivery of RNA-LNPs. Our innovation lies in the encapsulation chemistry and novel mode of action of our active ingredient. CILA’s preliminary investigations have demonstrated substantial increases (1 to 3-fold) in the transfection efficiency of mRNA-LNP (proprietary LNPs) in primary human bronchial epithelial cells grown at the Air-Liquid Interface. Furthermore, our agent is effective at reducing the viscoelasticity of sputum samples from patients with cystic fibrosis, primary ciliary dyskinesia and non‐cystic fibrosis bronchiectasis, lending weight to the broad applicability of our gateway opener platform. In this SBIR Phase I, we will translate our findings to in vivo models of healthy and mucus-producing mice, examining the transfection efficacy of standard luciferase mRNA-LNP. Firstly, in Aim 1, we will determine the maximum safe and tolerable dose (MTD) of inhaled CIL-0X in healthy A/J mice. In Aim 2 we will assess the efficacy of CIL-0X for improving mRNA-LNP transfection and establish tolerance of CIL-0X treatment in combination with mRNA-LNP in the healthy A/J mouse model. In Aim 3, we will quantify the impact of CIL-0X on transfection efficiency and delivery of inhaled mRNA-LNP in hyper-mucus-producing A/J mouse models. A successful outcome of this project will lead to establishing robust in vivo proof-of-concept, and lay the foundation for future research evaluating CIL-0X efficacy to improve delivery of PCD-specific RNA to lungs. We intend to commence research to combine CIL- 0X with mRNA-LNP as monotherapy and expand into treatment for pulmonary diseases.