Novel piperazine‐based ionizable lipid nanoparticles allow the repeated dose of mRNA to fibrotic lungs with improved potency and safety

Abstract

AbstractmRNA‐based protein replacement therapy has received much attention as a novel intervention in clinical disease treatment. Lipid nanoparticles (LNPs) are widely used for their therapeutic potential to efficiently deliver mRNA. However, clinical translation has been hampered by the immunogenicity of LNPs that may aggravate underlying disease states. Here, we report a novel ionizable LNP with enhanced potency and safety. The piperazine‐based biodegradable ionizable lipid (244cis) was developed for LNP formulation and its level of protein expression and immunogenicity in the target tissue was evaluated. It was found that 244cis LNP enabled substantial expression of the target protein (human erythropoietin), while it minimally induced the secretion of monocyte chemoattractant protein 1 (MCP‐1) as compared to other conventional LNPs. Selective lung targeting of 244cis LNP was further investigated in tdTomato transgenic mice with bleomycin‐induced pulmonary fibrosis (PF). The repeated administration of 244cis LNP with Cre recombinase mRNA achieved complete transfection of lung endothelial cells (~80%) and over 40% transfection of Sca‐1‐positive fibroblasts. It was shown that 244cis LNP allows the repeated dose of mRNA without the loss of activity due to its low immunogenicity. Our results demonstrate that 244cis LNP has great potential for the treatment of chronic diseases in the lungs with improved potency and safety.

Development of Lipid Nanoparticle Formulation for the Repeated Administration of mRNA Therapeutics

“…The 244-cis ionizable lipid is a piperazine-based biodegradable ionizable lipid with a structure different from SM-102 (Fig. 4 A) [ 34 ]. 244-cis LNP is composed of DOPE (1,2-dioleoyl-sn-glycero-3-phosphoethanolamine), cholesterol, and C16 PEG ceramide (Fig.…”

“…To formulate LNPs, the ethanol phase containing lipid components and an aqueous phase containing RNA were mixed via a microfluidic mixing system (Ignite, PNI, Canada). mRNA-loaded 244-cis and SM-102 LNPs were formulated via a microfluidic device as previously reported [ 34 ]. Ionizable lipids (244cis, SM-102), cholesterol, helper lipids (DOPE, DSPC), and PEG lipid (C16-PEG 2000 ceramide, DMG-PEG 2000) were dissolved in ethanol.…”

Discovery of a Peptoid-Based Nanoparticle Platform for Therapeutic mRNA Delivery via Diverse Library Clustering and Structural Parametrization

“…High particle charge is known to direct LNP cargos to the lung, while moderate surface charges often localize in the liver. 46,47 The highest spleen expression was observed for cluster 3, which had significantly higher molecular weights and the largest number of total lipid monomers of any cluster. Interestingly, based on the hierarchy of clustering, cluster 3 splits from cluster 11 almost immediately, suggesting that they are the most chemically distinct from each other (Figure 2A) which is also reflected in their expression patterns.…”