Lipid 331 is a biodegradable cyclic ionizable cationic lipid (pKa = 7.53) used in the formulation of lipid nanoparticles (LNPs) for mRNA delivery both in vitro and in vivo. LNPs containing lipid 331 and encapsulating 5-methoxyuridine-modified mRNA encoding ovalbumin activate interferon regulatory factor (IRF) in a reporter assay using THP-1 monocytes. Additionally, intranasal administration of LNPs formulated with lipid 331, DOTAP, and mRNA encoding a fusion of the receptor-binding domain (RBD) of the SARS-CoV-2 spike glycoprotein with mouse complement C3d as an adjuvant significantly increases serum anti-SARS-CoV-2 spike glycoprotein RBD IgG titers in mice.
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