Tailoring lipid nanoparticle dimensions through manufacturing processes.

Area of Science:

• Biotechnology
• Nanomedicine
• Drug Delivery

Background:

• Lipid nanoparticles (LNPs) are crucial delivery vehicles for nucleic acid therapies, including mRNA and siRNA.
• Physicochemical properties like size, encapsulation, and charge significantly influence LNP efficacy.
• Current LNP formulations often utilize sizes under 100 nm.

Purpose of the Study:

• To investigate the impact of aqueous-to-lipid phase ratios on LNP size and critical quality attributes.
• To evaluate how varying LNP sizes affect mRNA expression both in vitro and in vivo.
• To determine the optimal LNP size range for enhanced therapeutic efficacy.

Main Methods:

• Formulation of LNPs using ALC-0315 with controlled aqueous-to-organic lipid phase ratios.
• Characterization of LNP size and other critical quality attributes.
• In vitro expression studies in HEK293 and THP-1 cells.
• In vivo expression studies in BALB/c mice.

Main Results:

• Precise control over LNP size was achieved by adjusting phase ratios.
• In HEK293 cells, a linear correlation was observed between LNP size and mRNA expression.
• In THP-1 cells, maximal expression was seen with LNPs up to 120 nm.
• In vivo studies in mice indicated reduced expression for LNPs >120 nm, with robust expression between 60-120 nm.

Conclusions:

• LNP size can be precisely tuned via phase ratio adjustments.
• Expression of mRNA cargo within LNPs demonstrates a size-dependent effect, with robustness up to 120 nm.
• These findings suggest that larger LNPs (>100 nm) may offer improved therapeutic potential.