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:
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.
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