Advantage and Core Benefit
- New concept to devise solvents for existing solvent dilution methods
- Solvent X were found to be uniformly effective in studies with several ionized lipids.
Background and Technology
The solvent dilution method is a standard method for preparing lipid nanoparticles used in mRNA vaccines. Specifically, mRNA-encapsulated lipid nanoparticles are generated by mixing a lipid solution containing PEG lipid, ionized lipids such as SM-102, phospholipids, and cholesterol dissolved in an organic solvent with an aqueous solution containing mRNA using the vortex method or microfluidic channels. The organic solvent used in the solvent dilution method must be (1) easy to solubilize lipid, (2) easily miscible with water, and (3) liquid at room temperature. Ethanol is commonly used as a conventional technology.
While conducting research on the placement of proteins and other molecules on the surface of lipid nanoparticles, the inventors noticed that the mRNA transfection efficiency was altered depending on the choice of solvent used in the preparation of lipid nanoparticles. They investigated the differences in mRNA transfection efficiency depending on the solvent and found that when Solvent X was used, the uniformity of mRNA-encapsulated lipid nanoparticles was improved, and the amount of protein expression by mRNA increased.
Data
 |
- Comparison of protein expression levels after addition of mRNA-encapsulated lipid nanoparticles of fluorescent protein to A549 and HepG2 cells showed that the use of Solvent X enhanced protein expression (A).
- When mRNA-encapsulated lipid nanoparticles consisting of various ionized lipids (SM-102, ALC-0315, Dlin-MC3-DMA) were prepared and added to A549 and HepG2 cells and the luciferase expression levels were measured, the protein expression was enhanced by the use of Solvent X in any of the ionized lipids.
- When mRNA lipid nanoparticles prepared with ethanol or Solvent X were administered intravenously or intramuscularly to mice and luciferase expression levels in each organ were measured 6 h later, Luc expression increased in many organs including liver (B, C).
Researcher
Dr. Koki Ogawa (Nagoya City University)
Expectations
We are interested in working with mRNA drug development companies, service providers, CROs, and CDMOs who are interested in licensing this invention and working on the practical application of Solvent X-based solvent dilution methods.
Project. WL-04808