	US 11,938,227 B2
	Lipid nanoparticles encapsulation of large RNA
Yanjie Bao, San Diego, CA (US); Brenda Clemente, San Diego, CA (US); and Priya Prakash Karmali, San Diego, CA (US)
Assigned to Arcturus Therapeutics, Inc., San Diego, CA (US)
Filed by Arcturus Therapeutics, Inc., San Diego, CA (US)
Filed on Sep. 13, 2021, as Appl. No. 17/473,063.
Claims priority of provisional application 63/077,648, filed on Sep. 13, 2020.
Prior Publication US 2022/0168234 A1, Jun. 2, 2022
Int. Cl. A61K 9/00 (2006.01); A61K 9/51 (2006.01); A61K 31/00 (2006.01); A61K 31/7105 (2006.01)

CPC A61K 9/5192 (2013.01) [A61K 31/7105 (2013.01)]

20 Claims

1. A method of producing a lipid-encapsulated RNA nanoparticle, comprising the steps

a) flowing an aqueous solution comprising an RNA through a 1^sttube having an inner diameter (ID) of from about 0.01 inches to about 0.08 inches; wherein a pH of the aqueous solution is in a range from about 3.0 to about 4.5 with an optional NaCl concentration of up to about 300 mM;

wherein the RNA comprises from about 6,000 to about 13,000 nucleotides;

b) flowing an ethanol solution comprising lipids through a 2^ndtube having an ID of from about 0.01 inches to about 0.04 inches at a flow rate of about 0.2 to about 1 times a flow rate of the aqueous solution through the 1^sttube, wherein the lipids comprise a cationic lipid; and

c) mixing the ethanol solution with the aqueous solution;

wherein the mixing produces an output solution flowing in the 1^sttube comprising a turbulent flow of the RNA and the lipids in about 10% to 75% ethanol v/v; and

wherein the lipid-encapsulated RNA nanoparticles have a bilayer structure.