10139483Therapeutic liposome composition and method of preparation

• US 6,936,272 B2
• Filed: 12/10/2001
• Issued: 08/30/2005
• Est. Priority Date: 10/11/1996
• Status: Expired due to Term

• Current Assignee: ALZA Corporation (Johnson & Johnson)
• Original Assignee: ALZA Corporation (Johnson & Johnson)
• Inventors: Martin, Francis J., Zalipsky, Samuel
• Primary Examiner(s): Kishore, Gollamudi S.
• Application Number:
  US10/016,324
• Publication Number:
  US 20020172711A1
• Time in Patent Office: 1,359 Days
• Field of Search: 424/450, 424 43- 46, 424/1.21, 424/9.321, 424/9.51, 424/417, 424/94.3, 436/829, 935/54
• US Class Current: 424/450
• CPC Class Codes: A61K 47/544 Phospholipids
• CPC Class Codes: A61K 47/61 the organic macromolecular compound being a polysaccharide or a derivative thereof
• CPC Class Codes: A61K 47/62 the modifying agent being a protein, peptide or polyamino acid
• CPC Class Codes: A61K 47/6849 the antibody targeting a receptor, a cell surface antigen or a cell surface determinant
• CPC Class Codes: A61K 47/6911 the form being a liposome
• CPC Class Codes: A61K 47/6913 the liposome being modified on its surface by an antibody
• CPC Class Codes: A61K 51/1234 Liposomes
• CPC Class Codes: A61K 9/127 Liposomes
• 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: C07K 16/2854 against selectins, e.g. CD62
• CPC Class Codes: C07K 2317/55 Fab or Fab'
• CPC Class Codes: Y10S 424/812 Liposome comprising an antibody, antibody fragment, antigen, or other specific or nonspecific immunoeffector
• CPC Class Codes: Y10S 436/829 Liposomes, e.g. encapsulation

Abstract

A fusogenic liposome composition for delivering a liposome-entrapped compound into the cytoplasm of a target cell is described. The liposomes have an outer surface coating of chemically releasable hydrophilic polymer chains which shield hydrophobic polymers on the liposome outer surface. Release of the hydrophilic polymer chains exposes the hydrophobic polymers for interaction with outer cell membranes of the target cells to promote fusion of the liposome with the target cells. Also disclosed is a method for using the composition to deliver a compound to target cells, and a method for selecting suitable hydrophobic polymers for use in the composition.

Claims

1. A method of administering a therapeutic agent, comprising,

administering via inhalation liposomes formed of vesicle-forming lipids and having a coating of hydrophilic polymer chains on the piopsome outer surface, said liposomes having an entrapped therapeutic agent.

2. The method of claim 1, wherein the vesicle-forming lipid is selected from the group consisting of hydrogenated soy phosphatidyicholine, distearoylphosphatidylcholine, sphingomyelin, diacyl glycerol, phosphatidyl ethanolamine, phosphatidylglycerol, distearylphosphatidylcholine, and distearyl phosphatidylethanolamine.

3. The method of claim 1, wherein said liposomes further contain a cationic lipid shielded by said coating of hydrophilic polymer chains, said cationic lipid being effective to impart a positive liposome-surface charge.

4. The method of claim 3, wherein the cationic lipid is selected from the group consisting of 1,2-dioleyloxy-3-(trimethylamino) propane, N-[1-(2,3,-ditetradecyloxy)propyl[-N,N-dimethyl-N-hydroxyethylammonium bromide, N-[1-(2,3,-dioleyloxy) propyl-N,N-dimethyl-N-hydroxy ethylammonium bromide, N-[1-(2,3-dioleyloxy) propyl]-N,N,N-trimethylammonium chloride;

3β

[N-(N′

,N′

-dimethylaminoethane) carbamoly]cholesterol; and

dimethyldioctadecylammonium.

5. The method of claim 3, wherein the cationic lipid is a neutral lipid derivatized with a cationic lipid.

6. The method of claim 1, wherein said hydrophilic polymer coating is composed of hydrophilic polymers selected from the group consisting of polyvinylpyrrolidone, polyvinylmethylether, polymethyloxazoline, polyethyloxazoline, polyhydroxypropyloxazoline, polyhydroxypropylmethacrylamide, polymethacrylamide, polydimethylacrylamide, polyhydroxypropylmethacrylate, polyhydroxyethylacrylate, hydroxymethylcellulose, hydroxyethylcellulose, polyethyleneglycol, and polyaspartamide.

7. The method of claim 6, wherein said hydrophilic polymer coating is composed of polyethylene glycol chains having a molecular weight of between about 500 Daltons and about 10,000 Daltons.

8. The method of claim 1 wherein between about 1 mole percent and about 20 mole percent of the vesicle-forming lipids are derivatized with said hydrophilic polymer chains.

9. The method of claim 1, wherein at least a portion of the hydrophilic polymer chains are joined by a chemically releasable bond.

10. The method of claim 9, wherein said releasable bond is a disulfide bond.

11. The method of claim 9, wherein said releasable bond is a pH sensitive chemical linkage.

12. The method of claim 1, wherein the liposomes are composed of between about 70-90 mole percent hydrogenated soy phosphatidylcholine, about 1-20 mole percent distearylphosphatidylcholine derivatized with polyethyleneglycol and about 1-50 mole percent cholesterol.

13. The method of claim 1, wherein the liposome is about 0.1 to about 10 microns.

14. The method of claim 1, wherein the agent entrapped in the lipid vesicles is a polynucleotide capable of expressing a selected protein, when taken up by a target cell.

15. The method of claim 1, wherein the agent entrapped in the liposomes is an oligonucleotide or oligonucleotide analog effective for sequence-specific binding to cellular RNA or DNA.

16. The method of claim 1, wherein the agent entrapped in the liposomes is selected from the group consisting of DNA, proteins, and peptides.

17. The method of claim 1, wherein the agent entrapped in the liposomes is selected from the group consisting of antibiotics, antivirals, and antitumor drugs.

18. The method of claim 1, wherein said liposomes further contain a ligand attached to the distal end of at least a portion of said hydrophilic polymer chains.

19. The method of claim 1, wherein the liposomes further include a ligand attached the polar head group of at least a portion of the vesicle-forming lipids of the liposome.

20. The method of claim 18 or 19, wherein the ligand is an antibody or an antibody fragment.

21. The method of claim 20, wherein the ligand is a Fab′

fragment of an antibody.

22. The method of claim 20, wherein the ligand is a single chain Fv antibody.

23. The method of claim 18 or 19, wherein the ligand specifically binds to an extracellular domain of a growth factor receptor.

24. The method of claim 23, wherein the receptor is selected from the group consisting of epidermal growth factor receptor, basic fibroblast growth factor receptor and vascular endothelial growth factor receptor.

25. The method of claim 18 or 19, wherein the ligand binds a receptor selected from the group consisting of E-selectin receptor, L-selectin receptor, P-selectin receptor, folate receptor, CD4 receptor, α

β

integrin receptors and chemokine receptors.

26. The method of claim 18 or 19, wherein the ligand is selected from the group consisting of folic acid, pyridoxal phosphate, sialyl Lewis*, transferrin, epidermal growth factor, basic fibroblast growth factor, vascular endothelial growth factor, VCAM-1, ICAM-1, PECAN-1, and RGD peptides.

27. The method of claim 18 or 19, wherein the ligand is selected from the group consisting of water soluble vitamins, apolipoproteins, insulin, galactose, Mac-1, PECAM-1/CD31, fibronectin, osteopontin, RGD sequences of matrix proteins, HIV GP 120/41 domain peptomers, GP120C4 domain peptomers, T cell tropic isolates, SDF-1 chemokines, Macrophage tropic isolates, anti-cell surface receptor antibodies or fragments thereof, pyridoxyl ligands, RGD peptide mimetics, and anti-E-selectin Fab.

28. The method of claim 27, wherein the anti-cell surface receptor antibodies or fragments thereof is selected from the group consisting of anti-selectin and anti-VEGF pyridoxyl.

29. The method of claim 27, wherein the pyridoxyl ligand is selected from the group consisting of pyridoxal, pyridoxine, pyridoxamine, pyridoxal 5′

-phosphate and N-(4′

-pyridoxyl)amines.

30. The method of claim 1, wherein said liposomes are further comprised of a lipid derivatized by a diblock copolymer composed of a hydrophobic polymer chain covalently bound to the lipid and a hydrophilic polymer chain, the hydrophobic and hydrophilic chains being joined by a bond effective to release the hydrophilic polymer chains in response to an existing or an induced physiologic condition, thereby exposing the hydrophobic polymer chains.

31. The method of claim 30, wherein said hydrophobic polymer is selected from the group consisting of polypropylene oxide, polyethylene, polypropylene, polycarbonate, polystyrene, polysulfone, polyphenylene oxide and polytetramethylene ether.