Liposomal apparatus and manufacturing methods
View Patent ↗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.
1. A process for producing a lipid vesicle encapsulating one or more therapeutic agents within the lipid vesicle, said process comprising:
providing an aqueous solution in a first reservoir;
providing an organic lipid solution in a second reservoir, wherein the lipids present in said organic lipid solution are solubilized in a lower alkanol at a concentration of about 75% v/v to 100% v/v, wherein said aqueous solution and/or said organic lipid solution comprises one or more therapeutic agents, and wherein said one or more therapeutic agents is selected from the group consisting of a nucleic acid, a protein, an antigen, paclitaxel, protax III, taxol, and mixtures thereof; and
mixing said organic lipid solution with said aqueous solution by introducing said organic lipid solution and said aqueous solution into a mixing environment at about equal flow rates;
wherein said mixing instantaneously produces a lipid vesicle encapsulating said one or more therapeutic agents within the lipid vesicle by diluting said lower alkanol to a concentration of between 45% v/v to about 60% v/v; and
wherein the mixing environment includes a mixing chamber, wherein said aqueous solution and said organic lipid solution are introduced into the mixing chamber at an angle of between about 27° and about 180° relative to each other and mixed within the mixing chamber.
2. The process of claim 1 , further comprising diluting said lipid vesicle with a buffer solution wherein said lipid vesicle undergoes a continuous stepwise dilution to further stabilize the lipid vesicle.
3. The process of claim 1 , wherein said nucleic acid is selected from the group consisting of a plasmid, an antisense polynucleotide, a ribozyme, mRNA, tRNA, snRNA, siRNA, pre-condensed DNA, and mixtures thereof.
4. The process of claim 1 , wherein said aqueous solution comprises said nucleic acid, protein, antigen, or mixtures thereof.
5. The process of claim 1 , wherein said organic lipid solution comprises said paclitaxel, protax III, taxol, or mixtures thereof.
6. The process of claim 1 , wherein said angle is between about 27° and about 90° or between about 90° and about 180°.
7. The process of claim 1 , wherein said lower alkanol is selected from the group consisting of methanol, ethanol, propanol, butanol, pentanol, isomers thereof, and mixtures thereof.
8. The process of claim 1 , wherein the lipids present in said organic lipid solution are solubilized in said lower alkanol at a concentration of between about 80% v/v to about 90% v/v or 100% v/v.
9. The process of claim 1 , wherein the lipids present in said organic lipid solution comprise a phospholipid, cholesterol, a PEG-lipid, and a cationic lipid.
10. The process of claim 2 , further comprising concentrating said lipid vesicle by tangential flow ultrafiltration.
11. An apparatus for producing a lipid vesicle encapsulating one or more therapeutic agents within the lipid vesicle, said apparatus comprising:
a first reservoir for holding an aqueous solution;
a second reservoir for holding an organic lipid solution, wherein the lipids present in said organic lipid solution are solubilized in a lower alkanol at a concentration of about 75% v/v to 100% v/v, wherein said aqueous solution and/or said organic lipid solution comprises one or more therapeutic agents, and wherein said one or more therapeutic agents is selected from the group consisting of a nucleic acid, a protein, an antigen, paclitaxel, protax III, taxol, and mixtures thereof; and
a pump mechanism configured to pump said aqueous solution and said organic lipid solution into a mixing environment at about equal flow rates;
wherein said organic lipid solution mixes with said aqueous solution in the mixing environment to instantaneously produce a lipid vesicle encapsulating said one or more therapeutic agents within the lipid vesicle by diluting said lower alkanol to a concentration of between 45% v/v to about 60% v/v; and
wherein the mixing environment includes a mixing chamber, wherein said aqueous solution and said organic lipid solution are introduced into the mixing chamber at an angle of between about 27° and about 180° relative to each other and mixed within the mixing chamber.
12. The apparatus of claim 11 , wherein said nucleic acid is selected from the group consisting of a plasmid, an antisense polynucleotide, a ribozyme, mRNA, tRNA, snRNA, siRNA, pre-condensed DNA, and mixtures thereof.
13. The apparatus of claim 11 , wherein said aqueous solution comprises said nucleic acid, protein, antigen, or mixtures thereof.
14. The apparatus of claim 11 , wherein said organic lipid solution comprises said paclitaxel, protax III, taxol, or mixtures thereof.
15. The apparatus of claim 11 , wherein said angle is between about 27° and about 90° or between about 90° and about 180°.
16. The apparatus of claim 11 , wherein said lower alkanol is selected from the group consisting of methanol, ethanol, propanol, butanol, pentanol, isomers thereof, and mixtures thereof.
17. The apparatus of claim 11 , wherein the lipids present in said organic lipid solution are solubilized in said lower alkanol at a concentration of between about 80% v/v to about 90% v/v or 100% v/v.
18. The apparatus of claim 11 , wherein the lipids present in said organic lipid solution comprise a phospholipid, cholesterol, a PEG-lipid, and a cationic lipid.
19. The apparatus of claim 11 , wherein the apparatus further comprises a tangential flow ultrafiltration system.
20. The apparatus of claim 11 , wherein the pump mechanism comprises a peristaltic pump.