US 11,737,979 B2
Method of making lipid-encapsulated RNA nanoparticles
Priya Karmali, San Diego, CA (US); Padmanabh Chivukula, San Diego, CA (US); Joseph E. Payne, San Diego, CA (US); and Yanjie Bao, San Diego, CA (US)
Assigned to Arcturus Therapeutics, Inc., San Diego, CA (US)
Filed by Arcturus Therapeutics, Inc., San Diego, CA (US)
Filed on Mar. 18, 2020, as Appl. No. 16/823,212.
Claims priority of provisional application 62/820,496, filed on Mar. 19, 2019.
Prior Publication US 2020/0297634 A1, Sep. 24, 2020
Int. Cl. A61K 47/69 (2017.01); B82Y 5/00 (2011.01); A61K 9/127 (2006.01); A61K 9/14 (2006.01); A61K 47/60 (2017.01)
CPC A61K 9/1277 (2013.01) [A61K 9/145 (2013.01); A61K 47/60 (2017.08); A61K 47/69 (2017.08); B82Y 5/00 (2013.01)] 19 Claims
 
1. A method of producing a lipid-encapsulated RNA nanoparticle, comprising the steps
a) flowing an aqueous solution comprising an RNA through a 1st tube having an inner diameter (ID) of between about 0.1″ and 0.132″;
b) flowing an ethanol solution comprising lipids through a 2nd tube having an ID of between about 0.005″ and 0.02″ at one third the flow rate of the aqueous solution through the 1st tube, wherein the lipids comprise a cationic lipid; and
c) mixing the ethanol solution with the aqueous solution by flowing the ethanol solution and the aqueous solution into a mixing module consisting of the 2nd tube perpendicularly joined to the 1st tube;
wherein the mixing produces an output solution flowing in the 1st tube comprising a turbulent flow of the RNA and the lipids in between about 10% to 75% ethanol v/v;
wherein the lipid-encapsulated RNA nanoparticles have a bilayer structure;
wherein the RNA is encapsulated at greater than 98%;
wherein the polydispersity index is less than 0.09; and,
wherein the cationic lipid has a pKa of about 6 to about 7 and a structure of Formula I:

OG Complex Work Unit Chemistry
or a pharmaceutically acceptable salt or solvate thereof, wherein
R5 and R6 are each independently selected from the group consisting of a linear or branched C1-C31 alkyl, C2-C31 alkenyl or C2-C31 alkynyl and cholesteryl;
L5 and L6 are each independently selected from the group consisting of a linear C1-C20 alkyl and C2-C20 alkenyl;
X5 is —C(O)O— or —OC(O)—;
X6 is —C(O)O— or —OC(O)—;
X7 is S or O;
L7 is absent or lower alkyl;
R4 is a linear or branched C1-C6 alkyl; and
R7 and R8 are each independently selected from the group consisting of a hydrogen and a linear or branched C1-C6 alkyl.