Method of making liposomes, liposome compositions made by the methods, and methods of using the same

1. A method of making liposomes with an entrapped agent, comprising:

solubilizing at least about 10 w/w % of vesicle-forming lipids in about 1-12 w/w % of a solvent miscible with water at room temperature to form a lipid solution;

separately solubilizing about 2.5-25 w/w % of the agent in about 25-65 w/w % of an aqueous solvent and 0.005-0.01 w/w % of EDTA at room temperature to form an agent containing solution;

filtering the agent containing solution;

injecting a stream of the lipid solution into the agent containing solution while mixing; and

allowing the resulting lipid and agent containing solution to hydrate for at least one hour with frequent mixing;

wherein the w/w % is based on the weight of the liposome formulation with the entrapped agent,

wherein the resulting liposomes retain at least about 65-100% of the entrapped agent after at least one month storage.

2. The method of claim 1 , wherein the solvent miscible with water comprises an alcohol.

3. The method of claim 1 , further comprising adding more lipid or a thickener to form a gel after the hydration step.

4. The method of claim 1 , wherein the vesicle-forming lipids comprises at least about 45-50% phosphatidylcholine.

5. The method of claim 4 , wherein the phosphatidylcholine is derived from egg or soy.

6. The method of claims 1 - 4 , wherein the agent is ascorbic acid or a salt thereof.

7. The method of claim 1 , wherein the agent is glutathione.

8. The method of claim 1 , wherein the agent is alpha lipoic acid.