	US 12,064,515 B2
	Process of preparing mRNA-loaded lipid nanoparticles
Shrirang Karve, Lexington, MA (US); Zarna Patel, Lexington, MA (US); Yi Zhang, Lexington, MA (US); Ashish Sarode, Lexington, MA (US); Rebecca L. Goldman, Lexington, MA (US); Frank DeRosa, Lexington, MA (US); and Michael Heartlein, Lexington, MA (US)
Assigned to TRANSLATE BIO, INC., Waltham, MA (US)
Filed by Translate Bio, Inc., Lexington, MA (US)
Filed on May 5, 2022, as Appl. No. 17/737,712.
Application 17/737,712 is a continuation of application No. 16/553,747, filed on Aug. 28, 2019, granted, now 11,357,726.
Claims priority of provisional application 62/725,765, filed on Aug. 31, 2018.
Claims priority of provisional application 62/724,582, filed on Aug. 29, 2018.
Prior Publication US 2022/0395459 A1, Dec. 15, 2022
Int. Cl. A61K 9/51 (2006.01); A61K 9/127 (2006.01); A61K 9/14 (2006.01); A61K 31/7105 (2006.01)

CPC A61K 9/1277 (2013.01) [A61K 9/141 (2013.01); A61K 9/51 (2013.01); A61K 31/7105 (2013.01)]

19 Claims

1. A process of encapsulating messenger RNA (mRNA) in lipid nanoparticles comprising:

mixing a solution comprising pre-formed lipid nanoparticles and mRNA such that lipid nanoparticles encapsulating mRNA are formed,

wherein the pre-formed lipid nanoparticles and the mRNA are present in the solution at a concentration of about 0.1 mg/ml,

wherein the pre-formed lipid nanoparticles are provided in a 10% wt/volume trehalose solution buffer,

wherein said pre-formed lipid nanoparticles comprise one or more cationic lipids; 1,2-dioleyl-sn-glycero-3-phosphoethanolamine (DOPE) or 1,2-distearoyl-sn-glycero-3-phosphocholine (DSPC); cholesterol; and a PEG-modified lipid that is DMG-PEG, and

wherein said PEG-modified lipid is in an amount no greater than 3% of total lipids in the lipid nanoparticles.