Aminoglycoside Drugs as Adjuvants to Enhance siRNA/mRNA Delivery by Lipid Nanoparticles

Nucleic acid drugs (such as siRNA and mRNA) hold broad prospects in disease treatment, but their clinical application is limited by low delivery efficiency, particularly due to insufficient lysosomal escape. This study aims to explore aminoglycoside small-molecule drugs as adjuvants to enhance the lysosomal escape efficiency of lipid nanoparticles (LNPs) for nucleic acid drug delivery, thereby improving therapeutic effects and providing new strategies for gene therapy. In this study, five FDA-approved aminoglycoside small molecules were selected as adjuvants to enhance the lysosomal escape of LNPs/siRNA and improve the siRNA delivery effect. <i>In vitro</i> results confirmed that aminoglycoside drugs significantly enhanced the delivery efficiency of LNPs/siRNA, reducing luciferase expression to as low as 22.15%. Mechanistic studies revealed that kanamycin sulfate markedly reduced siRNA colocalization with lysosomes and destabilized lysosomal membranes under acidic conditions (pH 6.2), increasing hemolytic effects by 3.6-fold. Additionally, kanamycin sulfate significantly improved the silencing efficacy of LNPs/siEGFR on EGFR mRNA and protein expression and boosted LNPs/mSARS-CoV-2-mediated spike protein expression. Therefore, aminoglycoside small molecules (particularly kanamycin) can effectively enhance nucleic acid delivery efficiency by disrupting endosomal membrane structures, providing a novel adjuvant strategy for the clinical application of siRNA and mRNA drugs.