US 12,458,604 B2
Methods of lipid nanoparticle manufacture and compositions derived therefrom
Michael Daro Buschmann, Fairfax, VA (US); Mikell Paige, Fairfax, VA (US); Suman Alishetty, Fairfax, VA (US); Manuel Carrasco, Fairfax, VA (US); Mohamad Gabriel Alameh, Fairfax, VA (US); and Drew Weissman, Fairfax, VA (US)
Assigned to The Trustees of the University of Pennsylvania, Philadelphia, PA (US); and George Mason Research Foundation, Inc., Fairfax, VA (US)
Filed by George Mason Research Foundation, Inc., Fairfax, VA (US); and The Trustees of the University of Pennsylvania, Philadelphia, PA (US)
Filed on Oct. 13, 2021, as Appl. No. 17/500,491.
Claims priority of provisional application 63/179,885, filed on Apr. 26, 2021.
Claims priority of provisional application 63/179,872, filed on Apr. 26, 2021.
Claims priority of provisional application 63/091,616, filed on Oct. 14, 2020.
Claims priority of provisional application 63/091,603, filed on Oct. 14, 2020.
Prior Publication US 2022/0235377 A1, Jul. 28, 2022
Int. Cl. A61K 9/51 (2006.01); A61K 9/127 (2006.01); A61K 31/7088 (2006.01); A61K 31/7105 (2006.01); A61K 47/18 (2017.01); A61K 47/28 (2006.01); A61P 37/04 (2006.01); B82Y 5/00 (2011.01); C07C 229/16 (2006.01); C07D 207/09 (2006.01); C07D 211/26 (2006.01); C07D 295/13 (2006.01); C12N 15/88 (2006.01); A61K 48/00 (2006.01)
CPC A61K 9/5123 (2013.01) [A61K 9/127 (2013.01); A61K 31/7088 (2013.01); A61K 31/7105 (2013.01); A61K 47/18 (2013.01); A61K 47/28 (2013.01); A61P 37/04 (2018.01); B82Y 5/00 (2013.01); C07C 229/16 (2013.01); C07D 207/09 (2013.01); C07D 211/26 (2013.01); C07D 295/13 (2013.01); C12N 15/88 (2013.01); A61K 48/00 (2013.01)] 45 Claims
 
1. A method for making a lipid nanoparticle comprising a nucleic acid (“naLNP”) comprising:
a. Providing a nucleic acid solution comprising at least one nucleic acid at a nucleic acid concentration from about 1 to about 3 mg/ml;
b. Providing a lipid solution comprising at least one lipid at a lipid concentration from about 50 mM to about 100 mM; and
c. Combining a portion of the nucleic acid solution and a portion of the lipid solution to create a mixing solution comprising a mixing nitrogen-phosphate ratio and a lipid:nucleic acid ratio; and
d. Optionally adjusting the pH in the final mixed solution to physiological pH to obtain a pH-adjusted mixing solution; and
e. Obtaining the naLNPs from the mixed solution; and
wherein the naLNPs have a greater potency than a reference lipid nanoparticle (“refLNP”), wherein the refLNP comprises the at least one lipid and the at least one nucleic acid and is made by a reference LNP manufacturing method.