Dual Ionizable Lipids (ALC0315 and SM102) in a Novel Lipid Nanoparticle Formulation Enhance In Vivo Delivery and Time‐Dependent Expression of Messenger RNA and Self‐Amplifying RNA

Jun 9, 2026ChemMedChem

Two special lipids in a new nanoparticle improve delivery and timing of messenger and self-amplifying RNA in living organisms

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mRNA Technology on OpenScience ↗PubMed ↗DOI ↗

Abstract

LNP-5 achieved high encapsulation efficiencies (>80%) for both mRNA and saRNA.

LNP-5 is a five-component lipid nanoparticle formulation designed for effective mRNA and saRNA delivery.
Physicochemical analysis indicated a controlled size of 80-120 nm and low polydispersity (<0.2) for LNP-5.
In vivo studies showed that LNP-5 had superior mRNA delivery efficiency and enhanced target protein expression compared to LNP-4.
LNP-mRNA delivery resulted in rapid and intense but transient protein expression, while LNP-saRNA led to a gradual increase in protein production peaking at day 7 post-injection.
Quantitative RT-PCR demonstrated that saRNA exhibited a 4.3-fold higher copy number relative to mRNA by day 7, indicating active intracellular replication.

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This study introduces a novel five-component LNP formulation incorporating ionizable lipid combination ALC-0315 and SM-102 with cholesterol, DSPC, and DMG-PEG-2000. LNP-5 was designed for efficient delivery of mRNA and saRNA. We employed a Design of Experiments approach and microfluidics to optimize the formulation parameters, achieving high encapsulation efficiencies (>80%) for both RNA types. Physicochemical characterization by dynamic light scattering confirmed a controlled size (80-120 nm) and low polydispersity (<0.2). The morphology of investigated LNP-5 was studied by STEM in comparison with LNP-4 (with SM-102 or ALC-0315). Notably, LNP-5 demonstrated superior in vivo mRNA delivery efficiency and enhanced target protein expression compared to conventional LNP-4. The study compares the in vivo performance of mRNA and saRNA platforms formulated in novel LNP-5 lipid nanoparticles containing dual ionizable lipids. While LNP-mRNA delivery provides rapid and intense but transient reporter expression, LNP-saRNA mediates a gradual increase in protein production, peaking at day 7 post-injection. Quantitative RT-PCR analysis reveals superior saRNA persistence and a 4.3-fold higher copy number relative to mRNA by day 7, confirming active intracellular replication of the saRNA platform. These findings demonstrate that LNP-5 efficiently delivers both platforms, with saRNA offering a more sustained therapeutic window due to its self-amplifying properties.