# US 11,298,320 B2 · Liposomal apparatus and manufacturing methods > Agents: this data is available as JSON from the patentgrep API and MCP server — see https://patentgrep.com/llms.txt - Number: US 11,298,320 B2 - Document ID: US11298320B2 - Status: Granted - Kind: B2 - Application number: 17/330,209 - Filed: 2021-05-25 - Published: 2021-09-09 - Granted: 2022-04-12 - Earliest claimed: 2002-06-28 - Assignees: Arbutus Biopharma Corporation - Applicants: Arbutus Biopharma Corporation - Inventors: Ian MacLachlan; Lloyd B. Jeffs; Lorne R. Palmer; Cory Giesbrecht - Web page: https://patentgrep.com/patent/US11298320B2 ## CPC - A61K 9/1271: Non-conventional liposomes, e.g. PEGylated liposomes or liposomes coated or grafted with polymers (liposomes as conjugates {A61K47/6911}) - A61K 9/127: Synthetic bilayered vehicles, e.g. liposomes or liposomes with cholesterol as the only non-phosphatidyl surfactant - A61K 9/1277: Preparation processes; Proliposomes - A61K 31/7084: Compounds having two nucleosides or nucleotides, e.g. nicotinamide-adenine dinucleotide, flavine-adenine dinucleotide - A61K 31/7088: Compounds having three or more nucleosides or nucleotides - A61K 47/10: Alcohols; Phenols; Salts thereof, e.g. glycerol; Polyethylene glycols [PEG]; Poloxamers; PEG/POE alkyl ethers - A61K 47/24: containing atoms other than carbon, hydrogen, oxygen, halogen, nitrogen or sulfur, e.g. cyclomethicone or phospholipids - A61K 47/44: Oils, fats or waxes according to two or more groups of A61K47/02-A61K47/42; Natural or modified natural oils, fats or waxes, e.g. castor oil, polyethoxylated castor oil, montan wax, lignite, shellac, rosin, beeswax or lanolin (synthetic glycerides, e.g. medium-chain triglycerides, A61K47/14) - A61P 31/12: Antivirals - A61P 35/00: Antineoplastic agents - A61P 43/00: Drugs for specific purposes, not provided for in groups A61P1/00-A61P41/00 - A61K: PREPARATIONS FOR MEDICAL, DENTAL OR TOILETRY PURPOSES (devices or methods specially adapted for bringing pharmaceutical products into particular physical or administering forms A61J3/00; chemical aspects of, or use of materials for deodorisation of air, for disinfection or sterilisation, or for bandages, dressings, absorbent pads or surgical articles A61L; soap compositions C11D) - A61P: SPECIFIC THERAPEUTIC ACTIVITY OF CHEMICAL COMPOUNDS OR MEDICINAL PREPARATIONS ## Abstract The present invention provides apparatus and processes for producing liposomes. By providing a buffer solution in a first reservoir, and a lipid solution in a second reservoir, continuously diluting the lipid solution with the buffer solution in a mixing chamber produces a liposome. The lipid solution preferably comprises an organic solvent, such as a lower alkanol. ## Claims ### Claim 1 An apparatus for producing a lipid vesicle encapsulating a nucleic acid within the lipid vesicle, the apparatus comprising: a first reservoir containing an aqueous solution including a nucleic acid; a second reservoir containing an organic lipid solution, wherein the lipids present in the organic lipid solution are solubilized in a lower alkanol at a concentration of about 75% v/v to 100% v/v; and a pump mechanism configured to pump the aqueous solution and the organic lipid solution into a mixing chamber at different flow rates relative to each other; wherein the mixing chamber is configured such that the aqueous solution and the organic lipid solution are introduced into the mixing chamber as opposing flows at about 180° relative to each other and mixed within the mixing chamber to instantaneously produce a lipid vesicle encapsulating the nucleic acid within the lipid vesicle by diluting the concentration of the lower alkanol in the organic lipid solution. ### Claim 2 (depends on 1) The apparatus of claim 1, wherein the lipids present in the organic lipid solution comprise a phospholipid, cholesterol, a PEG-lipid, and a cationic lipid. ### Claim 3 (depends on 2) The apparatus of claim 2, wherein a plurality of lipid vesicles are produced and said plurality of lipid vesicles have a mean particle diameter of less than 150 nm. ### Claim 4 (depends on 2) The apparatus of claim 2, wherein the aqueous solution includes an acidic buffer. ### Claim 5 (depends on 2) The apparatus of claim 2, wherein the aqueous solution includes a buffer having a pH lower than the pKa of the cationic lipid component. ### Claim 6 (depends on 2) The apparatus of claim 2, wherein the aqueous solution and the organic lipid solution are mixed within the mixing chamber to produce a plurality of lipid vesicles at a pH range of about 3.5 to about 6. ### Claim 7 (depends on 2) The apparatus of claim 2, wherein the aqueous solution and the organic lipid solution are mixed within the mixing chamber to produce a plurality of lipid vesicles at a pH range of about 3.5 to about 8. ### Claim 8 (depends on 1) The apparatus of claim 1, wherein the nucleic acid comprises an RNA. ### Claim 9 (depends on 8) The apparatus of claim 8, wherein the lower alkanol comprises ethanol and wherein the lipids present in the organic lipid solution comprise a phospholipid, cholesterol, a PEG-lipid, and a cationic lipid. ### Claim 10 (depends on 9) The apparatus of claim 9, wherein the aqueous solution includes an acidic buffer. ### Claim 11 (depends on 9) The apparatus of claim 9, wherein the aqueous solution includes a buffer having a pH lower than the pKa of the cationic lipid component. ### Claim 12 (depends on 9) The apparatus of claim 9, wherein the aqueous solution and the organic lipid solution are mixed within the mixing chamber to produce a plurality of lipid vesicles at a pH range of about 3.5 to about 6 and said plurality of lipid vesicles have a mean particle diameter of less than 150 nm. ### Claim 13 (depends on 8) The apparatus of claim 8, wherein the RNA comprises an mRNA. ### Claim 14 (depends on 13) The apparatus of claim 13, wherein the lower alkanol comprises ethanol and wherein the lipids present in the organic lipid solution comprise a phospholipid, cholesterol, a PEG-lipid, and a cationic lipid. ### Claim 15 (depends on 14) The apparatus of claim 14, wherein the aqueous solution includes an acidic buffer. ### Claim 16 (depends on 14) The apparatus of claim 14, wherein the aqueous solution and the organic lipid solution are mixed within the mixing chamber to produce a plurality of lipid vesicles at a pH range of about 3.5 to about 6 and said plurality of lipid vesicles have a mean particle diameter of less than 150 nm. ### Claim 17 (depends on 14) The apparatus of claim 14, wherein the aqueous solution and the organic lipid solution are mixed within the mixing chamber to produce a plurality of lipid vesicles at a pH range of about 3.5 to about 8. ### Claim 18 An apparatus for producing a lipid vesicle encapsulating a nucleic add within the lipid vesicle, the apparatus comprising: a first reservoir containing an aqueous solution including a nucleic acid; a second reservoir containing an organic lipid solution, wherein the lipids present in the organic lipid solution are solubilized in a lower alkanol at a concentration of about 75% v/v to 100% v/v; and a pump mechanism configured to pump the aqueous solution and the organic lipid solution into a mixing chamber at different flow rates relative to each other; wherein the mixing chamber is configured such that the aqueous solution and the organic lipid solution are introduced into the mixing chamber at an angle of between 90° and 180° relative to each other and mixed within the mixing chamber to instantaneously produce a lipid vesicle encapsulating the nucleic acid within the lipid vesicle by diluting the concentration of the lower alkanol in the organic lipid solution. ### Claim 19 (depends on 18) The apparatus of claim 18, wherein the lower alkanol comprises ethanol and wherein the lipids present in the organic lipid solution comprise a phospholipid, cholesterol, a PEG-lipid, and a cationic lipid. ### Claim 20 (depends on 19) The apparatus of claim 19, wherein the aqueous solution includes an acidic buffer having a pH lower than the pKa of the cationic lipid component. ### Claim 21 (depends on 20) The apparatus of claim 20, wherein the aqueous solution and the organic lipid solution are mixed within the mixing chamber to produce a plurality of lipid vesicles at a pH range of about 3.5 to about 6 and said plurality of lipid vesicles have a mean particle diameter of less than 150 nm. ### Claim 22 (depends on 20) The apparatus of claim 20, wherein the aqueous solution and the organic lipid solution are mixed within the mixing chamber to produce a plurality of lipid vesicles at a pH range of about 3.5 to about 8. ### Claim 23 (depends on 18) The apparatus of claim 18, wherein the nucleic acid comprises an RNA. ### Claim 24 (depends on 23) The apparatus of claim 23, wherein the lipids present in the organic lipid solution comprise a phospholipid, cholesterol, a PEG-lipid, and a cationic lipid. ### Claim 25 (depends on 24) The apparatus of claim 24, wherein the aqueous solution includes an acidic buffer. ### Claim 26 (depends on 24) The apparatus of claim 24, wherein the aqueous solution and the organic lipid solution are mixed within the mixing chamber to produce a plurality of lipid vesicles at a pH range of about 3.5 to about 8. ### Claim 27 (depends on 23) The apparatus of claim 23, wherein the RNA comprises an mRNA. ### Claim 28 (depends on 27) The apparatus of claim 27, wherein the lipids present in the organic lipid solution comprise a phospholipid, cholesterol, a PEG-lipid, and a cationic lipid. ### Claim 29 (depends on 28) The apparatus of claim 28, wherein the aqueous solution includes a buffer having a pH lower than the pKa of the cationic lipid component. ### Claim 30 (depends on 28) The apparatus of claim 28, wherein the aqueous solution and the organic lipid solution are mixed within the mixing chamber to produce a plurality of lipid vesicles at a pH range of about 3.5 to about 6 and said plurality of lipid vesicles have a mean particle diameter of less than 150 nm.