Preparation of liposomes with peg-bound phospholipid on surface

• US 5,593,622 A
• Filed: 06/02/1995
• Issued: 01/14/1997
• Est. Priority Date: 08/11/1988
• Status: Expired due to Term

• Current Assignee: Terumo Kabushiki Kaisha (Terumo Corporation)
• Original Assignee: Terumo Kabushiki Kaisha (Terumo Corporation)
• Inventors: Yoshioka, Hiroshi, Goto, Hiroshi
• Primary Examiner(s): Lovering, Richard D.
• Application Number:
  US08/460,714
• Time in Patent Office: 592 Days
• Field of Search: 424/450, 428/402.2, 514/6, 514/832, 514/833, 264/4.3, 264/4.32
• US Class Current: 264/4.32
• CPC Class Codes: A61K 9/1271 Non-conventional liposomes, e.g. PEGylated liposomes, liposomes coated with polymers liposome as conjugate A61K47/6911
• CPC Class Codes: A61K 9/1272 with substantial amounts of non-phosphatidyl, i.e. non-acylglycerophosphate, surfactants as bilayer-forming substances, e.g. cationic lipids with cholesterol as the only non-phosphatidyl surfactant A61K9/127; lipids as modifying agent A61K47/543
• CPC Class Codes: A61P 7/08 Plasma substitutes; Perfusion solutions; Dialytics or haemodialytics; Drugs for electrolytic or acid-base disorders, e.g. hypovolemic shock artificial tears A61P27/04
• CPC Class Codes: Y10T 428/2984 Microcapsule with fluid core [includes liposome]

Abstract

Agents for inhibiting adsorption of proteins on the liposome surface and liposomes which are agglutination-free by binding said inhibiting agent on the surface are disclosed. The above-mentioned inhibiting agents comprise a hydrophobic moiety and a hydrophilic macromolecular chain moiety. Adsorption of plasma proteins on the liposomes is inhibited due to the hydrophilic moiety exposed on the liposome surface with a result that agglutination of the liposomes in plasma is prevented. Therefore, there is no danger of embolism in blood vessels inhibiting blood flow when the liposomes are introduced into the living body. Accordingly, the liposomes are especially highly useful as artificial erythrocytes for which a large dose of liposomes is needed for administration.

Moreover, when liposomes are introduced into the living body, antibody protein (immunoglobulin) to the liposome which is an antigen will be adsorbed on the liposome to produce foreign body recognition in the phagocytes (macro-phage) with a result that the liposome will be included in the macrophage and disappear within a short period of time. Thus, inhibition of the protein adsorption on liposome can delay disappearance of the liposome in plasma.

In addition, a method for preparing the above-described liposomes is disclosed.

Claims

1. A process for the preparation of a liposome comprising PEG-bound phospholipid on the surface thereof, wherein said PEG-bound phospholipid extends outwardly, and does not substantially extend inwardly from the surface of said liposome, said process comprising preparing a liposome and adding PEG-bound phospholipid to said liposome thereby fixing the PEG-bound phospholipid on the surface of said liposome.

2. The process according to claim 1, wherein the amount of said PEG-bound phospholipid is 0.1 to 50 mol % based on the molar ratio of the phospholipid of said liposome.

3. The process according to claim 2, wherein the amount of said PEG-bound phospholipid is 0.5 to 20 mol % based on the molar ratio of the phospholipid of said liposome.

4. The process according to claim 3, wherein the amount of said PEG-bound phospholipid is 1 to 5 mol % based on the molar ratio of the phospholipid of said liposome.

5. The process according to claim 1, wherein said liposome has an average particle size of 0.1 to 1 micron.

6. The process according to claim 1, wherein the PEG chain length of said PEG-bound phospholipid is in the range of 5 to 1000 moles in terms of the average degree of polymerization.

7. The process according to claim 6, wherein the PEG chain length of said PEG-bound phospholipid is in the range of 40 to 200 moles in terms of the average degree of polymerization.

8. The process according to claim 1, wherein the phospholipid of said PEG-bound phospholipid is phosphatidylcholine, sphingomyelin, phosphatidylethanolamine or phosphatidylserine.

9. The process according to claim 8, wherein the phospholipid of said PEG-bound phospholipid is phosphatidylethanolamine.

10. A process for the preparation of a liposome comprising PEG-bound phospholipid on the surface thereof, wherein said PEG-bound phospholipid extends outwardly and does not substantially extend inwardly from the surface of said liposome, said process comprising preparing a liposome containing a phospholipid with a reaction-active functional group and adding PEG which is activated at one end to said liposome thereby allowing the PEG to couple with the phospholipid of said liposome.

11. The process according to claim 10, wherein the amount of said PEG-bound phospholipid is 0.1 to 50 mol % based on the molar ratio of the phospholipid of said liposome.

12. The process according to claim 11, wherein the amount of said PEG-bound phospholipid is 0.5 to 20 mol % based on the molar ratio of the phospholipid of said liposome.

13. The process according to claim 12, wherein the amount of said PEG-bound phospholipid is 1 to 5 mol % based on the molar ratio of the phospholipid of said liposome.

14. The process according to claim 10, wherein said liposome has an average particle size of 0.1 to 1 micron.

15. The process according to claim 10, wherein the PEG chain length of said PEG-bound phospholipid is in the range of 5 to 1000 moles in terms of the average degree of polymerization.

16. The process according to claim 15, wherein the PEG chain length of said PEG-bound phospholipid is in the range of 40 to 200 moles in terms of the average degree of polymerization.

17. The process according to claim 10, wherein the phospholipid of said PEG-bound phospholipid is phosphatidylcholine, sphingomyelin, phosphatidylethanolamine or phosphatidylserine.

18. The process according to claim 17, wherein the phospholipid of said PEG-bound phospholipid is phosphatidylethanolamine.