2024 AIChE Annual Meeting
(414f) Two Nitrogen-Based Ionizable Lipid System for mRNA Delivery to T Lymphocytes
Authors
To address these concerns, we designed a two-nitrogen library of lipids. It was hypothesized that, at the same lipid-tertiary-amine to mRNA-phosphate (N/P) ratio, the two nitrogen atoms in the lipid would be utilized to encapsulate mRNA at a relatively low molecular weight, which in turn could reduce the total lipid dosage compared to a one-nitrogen lipid. This class of lipids could potentially mitigate the side effects posed by single-nitrogen lipid while maintaining the same performance level. We screened the library of lipids against hard-to-transfect Jurkat human T cells in vitro. We found that three lead LNPs, distinct in structure, performed well. The LNPs were optimized by evaluating the effect of cholesterol and its analogues. Furthermore, the cell uptake and endolytic pathways were determined for the lead LNPs. The three lead LNPs were then tested in vivo using Balb/c mice. Lipid 1 LNPs showed translation efficiency in the liver while Lipid 2 LNPs showed higher protein translation in spleen relative to SM-102-based LNP. Overall, the library of lipids showed promising results for mRNA delivery with reduced total lipid dosage.