Identifying Key Factors Affecting mRNA-Lipid Nanoparticles Drug Product Formulation Stability

Area of Science:

• Biotechnology
• Pharmaceutical Sciences
• Nanomedicine

Background:

• Messenger RNA-lipid nanoparticles (mRNA-LNPs) are critical for mRNA vaccines and gene therapies.
• Their long-term stability depends on physicochemical properties and formulation components.
• Optimizing formulation is key to preserving mRNA-LNP integrity and effectiveness.

Purpose of the Study:

• To systematically evaluate the long-term stability of mRNA-LNP formulations under various storage conditions.
• To investigate the impact of different ionizable lipids and polyethylene glycol (PEG)-lipids on LNP stability.
• To identify primary failure modes and degradation pathways affecting mRNA-LNP integrity.

Main Methods:

• Systematic evaluation of LNP formulations with varied ionizable lipids (Dlin-MC3-DMA, ALC-0315) and PEG-lipids (DMG-PEG2k, ALC-0159).
• Storage of formulations at -80 °C, -20 °C, 5 °C, and 25 °C in Tris buffer (pH 7.4) for 12 months.
• Analysis of sixteen quality attributes, including particle size, mRNA encapsulation, lipid oxidation, and transfection efficiency.

Main Results:

• LNPs stored at -80 °C and -20 °C showed acceptable stability.
• Storage at 5 °C led to aggregation, reduced in vivo expression, and mRNA degradation.
• Storage at 25 °C resulted in complete loss of transfection within six months.
• Primary failure modes included lipid degradation (DSPC, MC3 N-oxidation) and subvisible particulates.
• Higher Tris buffer concentration accelerated 5'-cap hydrolysis.

Conclusions:

• Deep-cold storage (≤-20 °C) is essential for preserving mRNA-LNP integrity.
• Optimized formulation components are critical for enhancing LNP shelf-life.
• Low-ionic-strength buffers are recommended to prevent mRNA degradation.
• Findings provide insights for designing next-generation LNPs with improved stability.