Ligand-targeted emulsions carrying bioactive agents

• US 6,676,963 B1
• Filed: 10/27/2000
• Issued: 01/13/2004
• Est. Priority Date: 10/27/2000
• Status: Expired due to Fees

• Current Assignee: Barnes-Jewish Hospital (BJC Health System)
• Original Assignee: Barnes-Jewish Hospital (BJC Health System)
• Inventors: Lanza, Gregory M., Wickline, Samuel A.
• Primary Examiner(s): KISHORE, GOLLAMUDI S
• Application Number:
  US09/697,796
• Time in Patent Office: 1,173 Days
• Field of Search: 424/450, 424/9.5, 424/1.21, 424/1.37, 424/9.321, 424/9.51, 424/417, 424/94.3, 436/829, 514/937-943
• US Class Current: 424/450
• CPC Class Codes: A61K 47/6913 the liposome being modified on its surface by an antibody
• CPC Class Codes: A61K 47/6925 the form being a microcapsule, nanocapsule, microbubble or nanobubble
• CPC Class Codes: A61K 9/1075 Microemulsions or submicron emulsions; Preconcentrates or solids thereof; Micelles, e.g. made of phospholipids or block copolymers A61K9/0026 takes precedence
• CPC Class Codes: B82Y 5/00 Nanobiotechnology or nanomedicine, e.g. protein engineering or drug delivery

Abstract

A composition for use in delivering a bioactive agent to targeted tissues or cells comprises: (a) a site-specific targeting ligand; (b) a lipid encapsulated oil in water emulsion; and (c) a bioactive agent in or on the surface of the outer monolayer of the emulsion; the ligand being conjugated directly or indirectly to the emulsion and the composition providing facilitated delivery of the bioactive agent through prolonged association and increased contact of the ligand-bound lipid encapsulated emulsion particles with the lipid bilayer of the target tissues or cells. The composition may also comprise a lipid encapsulated oil in water emulsion and a combination site-specific targeting ligand/bioactive agent. Methods for improved delivery of a bioactive agent to targeted tissues or cells are also disclosed.

Claims

1. A composition for use in delivering a bioactive agent to targeted tissues or cells comprising:

(a) a lipid-encapsulated oil in water emulsion of fluorochemical particles, wherein the particles have an outer monolayer of lipid and wherein the lipid outer monolayer further comprises a primer material;

(b) a bioactive agent in or on the surface of the lipid outer monolayer of the particles; and

(c) a site-specific targeting ligand, wherein the ligand is covalently coupled to the primer material;

wherein the primer material is selected from the group consisting of 1-ethyl-3-(3-N-N-dimethylaminopropyl)carbodiimide hydrochloride, 1-cyclohexyl-3-(2-morpholinoethyl)carbodiimide methyl-p-toluenesulfonate, and 1,2-diacyl-sn-glycerol-3-phosphoethanolamine-N-(4-p-maleimidephenyl)-butyramide.

2. A composition as set forth in claim 1 wherein said site-specific targeting ligand is selected from the group consisting of antibodies, antibody fragments, peptides, asialoglycoproteins, polysaccharides, aptamers, nucleic acids, peptidomimetics, mimetics and drugs.

3. A composition as set forth in claim 2 wherein said site-specific targeting ligand is an antibody.

4. A composition as set forth in claim 1 wherein said fluorochemical is a fluorocarbon.

5. A composition as set forth in claim 4 wherein said fluorocarbon is perfluorooctylbromide.

6. A composition as set forth in claim 1 wherein said fluorochemical is a liquid with a boiling point above approximately 30°

C.

7. A composition as set forth in claim 6 wherein said fluorochemical liquid has a boiling point above approximately 90°

C.

8. A composition as set forth in claim 1 wherein said bioactive agent is selected from the group consisting of chemotherapeutic agents, drugs, genetic materials, nucleic acid-based therapy, protein or peptide therapy, radioactive isotopes or combinations thereof.

9. A composition as set forth in claim 8 wherein said bioactive agent is a chemotherapeutic agent.

10. A composition as set forth in claim 1 wherein said lipid outer monolayer is composed of a material selected from the group consisting of a natural or synthetic phospholipid, a fatty acid, cholesterol, lysolipid, sphingomyelin, tocopherol, glucolipid, stearylamine, cardiolipin, a lipid with ether or ester linked fatty acids and a polymerized lipid.

11. A composition as set forth in claim 1 wherein said lipid outer monolayer also contains an additional surfactant incorporated therein for stabilizing said emulsion.

12. A composition as set forth in claim 11 wherein said additional surfactant is selected from the group consisting of nonionic and amphoteric surfactants.

13. A composition as set forth in claim 11 wherein said surfactant contains a cationic lipid to facilitate adhesion of said bioactive agent to said emulsion particles.

14. A composition as set forth in claim 13 wherein said cationic lipid is selected from the group consisting of N-[1-(2,3-dioleoyloxy)propyl]-N,N,N-trimethylammonium chloride, 1,2-dioleoyloxy-3-(trimethylammonio)propane, 1,2-dioleoyl-3-(4′

-trimethylammonio)butanoyl-sn-glycerol, 1,2-diacyl-3-trimethylammonium-propane, 1,2-diacyl-3-dimethylammonium-propane, and 3β

-[N′

,N′

-dimethylaminoethane)-carbamol]cholesterol-HCl.

15. A composition as set forth in claim 1 wherein said emulsion contains an emulsifying and/or solubilizing agent.

16. A composition as set forth in claim 1 wherein said emulsion particles have a diameter in the range of approximately 0.01 to 10 microns.

17. A composition as set forth in claim 16 wherein said emulsion particles have a diameter in the range of approximately 0.1 to 0.5 microns.

18. A composition for use in delivering a bioactive agent to targeted tissues or cells comprising:

(a) a lipid-encapsulated oil in water emulsion of fluorochemical particles, wherein the particles have an outer monolayer of lipid and wherein the lipid outer monolayer further comprises a primer material; and

(b) a combination site-specific targeting ligand/bioactive agent in or on the surface of the lipid outer monolayer of the particles;

wherein said combination ligand/bioactive agent is covalently coupled to the primer material;

wherein the primer material is selected from the group consisting of 1-ethyl-3-(3-N-N-dimethylaminopropyl)carbodiimide hydrochloride, 1-cyclohexyl-3-(2-morpholinoethyl)carbodiimide methyl-p-toluenesulfonate, and 1,2-diacyl-sn-glycerol-3-phosphoethanolamine-N-(4-p-maleimidephenyl)-butyramide.

19. A composition as set forth in claim 18 wherein said combination ligand/bioactive agent is selected from the group consisting of antibodies, antibody fragments, peptides and mimetics.

20. A composition as set forth in claim 19 wherein said combination ligand/bioactive agent is an antibody.

21. A composition as set forth in claim 18 wherein said fluorochemical is a fluorocarbon.

22. A composition as set forth in claim 18 wherein said fluorochemical is a liquid with a boiling point above approximately 30°

C.

23. A composition as set forth in claim 22 wherein said fluorochemical liquid has a boiling point above approximately 90°

C.

24. A composition as set forth in claim 18 wherein said lipid outer monolayer is composed of a material selected form the group consisting of a natural or synthetic phospholipid, a fatty acid, cholesterol, lysolipid, sphingomyelin, tocopherol, glucolipid, stearylamine, cardiolipin, a lipid with ether or ester linked fatty acids and a polymerized lipid.

25. A composition as set forth in claim 18 wherein said lipid outer monolayer also contains an additional surfactant incorporated therein for stabilizing said emulsion.

26. A composition as set forth in claim 25 wherein said surfactant contains a cationic lipid to facilitate adhesion of said ligand/bioactive agent to said emulsion particles.

27. A composition as set forth in claim 18 wherein said emulsion contains an emulsifying agent and/or solubilizing agent.

28. A composition as set forth in claim 18 wherein said emulsion particles have a diameter in the range of approximately 0.10 to 10 microns.

29. A composition as set forth in claim 28 wherein said emulsion particles have a diameter in the range of approximately 0.1 to 0.5 microns.

58. A composition as set forth in claim 26 wherein said cationic lipid is selected from the group consisting of N-[1-(2,3-dioleoyloxy)propyl]-N,N,N-trimethylammonium chloride, 1,2-dioleoyloxy-3-(trimethylammonio)propane, 1,2-dioleoyl-3-(4′

-trimethylammonio)butanoyl-sn-glycerol, 1,2-diacyl-3-trimethylammonium-propane, 1,2-diacyl-3-dimethylammonium-propane, and 3β

-[N′

,N′

-dimethylaminoethane-carbamol]cholesterol-HCl.

30. A method for improved delivery of a bioactive agent to targeted tissues or cells comprising administering to said tissues or cells a composition comprising:

(a) a lipid-encapsulated oil in water emulsion of fluorochemical particles, wherein the particles have an outer monolayer of lipid and wherein the lipid outer monolayer further comprises a primer material;

(b) a bioactive agent in or on the surface of the lipid outer monolayer of the particles; and

(c) a site-specific targeting ligand, wherein the ligand is covalently coupled to the primer material;

wherein the primer material is selected from the group consisting of 1-ethyl-3-(3-N-N-dimethylaminopropyl)carbodiimide hydrochloride, 1-cyclohexyl-3-(2-morpholinoethyl)carbodiimide methyl-p-toluenesulfonate, and 1,2-diacyl-sn-glycerol-3-phosphoethanolamine-N-(4-p-maleimidephenyl)-butyramide.

31. A method as set forth in claim 30 wherein said site-specific targeting ligand is selected from the group consisting of antibodies, antibody fragments, peptides, asialoglycoproteins, polysaccharides, aptamers, nucleic acids, peptidomimetics, mimetics and drugs.

32. A method as set forth in claim 31 wherein said site-specific targeting ligand is an antibody.

33. A method as set forth in claim 30 wherein said fluorochemical is a fluorocarbon.

34. A method as set forth in claim 33 wherein said fluorocarbon is perfluorooctylbromide.

35. A method as set forth in claim 30 wherein said fluorochemical is a liquid with a boiling point above approximately 30°

C.

36. A method as set forth in claim 35 wherein said fluorochemical liquid has a boiling point above approximately 90°

C.

37. A method as set forth in claim 30 wherein said bioactive agent is selected from the group consisting of chemotherapeutic agents, drugs, genetic materials, nucleic acid-based therapy, protein or peptide therapy, radioactive isotopes or combinations thereof.

38. A method as set forth in claim 37 wherein said bioactive agent is a chemotherapeutic agent.

39. A method as set forth in claim 30 wherein said lipid outer monolayer is composed of a material selected from the group consisting of a natural or synthetic phospholipid, a fatty acid, cholesterol, lysolipid, sphingomyelin, tocopherol, glucolipid, stearylamine, cardiolipin, a lipid with ether or ester linked fatty acids and a polymerized lipid.

40. A method as set forth in claim 30 wherein said lipid outer monolayer also contains an additional surfactant incorporated thereon for stabilizing said emulsion.

41. A method as set forth in claim 40 wherein said additional surfactant is selected from the group consisting of nonionic and amphoteric surfactants.

42. A method as set forth in claim 40 wherein said surfactant contains a cationic lipid to facilitate adhesion of said bioactive agent to said emulsion particles.

43. A method as set forth in claim 30 wherein said emulsion contains an emulsifying and/or solubilizing agent.

44. A method as set forth in claim 30 wherein said emulsion particles have a diameter in the range of approximately 0.01 to 10 microns.

45. A method as set forth in claim 44 wherein said emulsion particles have a diameter in the range of approximately 0.1 to 0.5 microns.

59. A method as set forth in claim 42 wherein said cationic lipid is selected from the group consisting of N-[1-(2,3-dioleoyloxy)propyl]-N,N,N-trimethylammonium chloride, 1,2-dioleoyloxy-3-(trimethylammonio)propane, 1,2-dioleoyl-3-(4′

-trimethylammonio)butanoyl-sn-glycerol, 1,2-diacyl-3-trimethylammonium-propane, 1,2-diacyl-3-dimethylammonium-propane, and 3β

-[N′

,N′

-dimethylaminoethane-carbamol]cholesterol-HCl.

46. A method for improved delivery of a bioactive agent to targeted tissues or cells comprising administering to said tissues or cells a composition comprising:

(a) a lipid-encapsulated oil in water emulsion of fluorochemical particles, wherein the particles have an outer monolayer of lipid and wherein the lipid outer monolayer further comprises a primer material; and

(b) a combination site-specific targeting ligand/bioactive agent in or on the surface of the lipid outer monolayer of the particles;

wherein said combination ligand/bioactive agent is covalently coupled to the primer material;

wherein the primer material is selected from the group consisting of 1-ethyl-3-(3-N-N-dimethylaminopropyl)carbodiimide hydrochloride, 1-cyclohexyl-3-(2-morpholinoethyl)carbodiimide methyl-p-toluenesulfonate, and 1,2-diacyl-sn-glycerol-3-phosphoethanolamine-N-(4-p-maleimidephenyl)-butyramide.

47. A method as set forth in claim 46 wherein said combination ligand/bioactive agent is selected from the group consisting of antibodies, antibody fragments, peptides, and mimetics.

48. A method as set forth in claim 47 wherein said combination ligand/bioactive agent is an antibody.

49. A method as set forth in claim 46 wherein said fluorochemical is a fluorocarbon.

50. A method as set forth in claim 46 wherein said fluorochemical is a liquid with a boiling point above approximately 30°

C.

51. A method as set forth in claim 50 wherein said fluorochemical liquid has a boiling point above approximately 90°

C.

52. A method as set forth in claim 46 wherein said lipid outer monolayer is composed of a material selected from the group consisting of a natural or synthetic phospholipid, a fatty acid, cholesterol, lysolipid, sphingomyelin, tocopherol, glucolipid, stearylamine, cardiolipin, a lipid with ether or ester linked fatty acids and a polymerized lipid.

53. A method as set forth in claim 46 wherein said lipid outer monolayer also contains an additional surfactant incorporated therein for stabilizing said emulsion.

54. A method as set forth in claim 53 wherein said surfactant contains a cationic lipid to facilitate adhesion of said ligand/bioactive agent to said emulsion particles.

55. A method as set forth in claim 46 wherein said emulsion contains an emulsifying agent and/or solubilizing agent.

56. A method as set forth in claim 46 wherein said emulsion particles have a diameter in the range of approximately 0.01 to 10 microns.

57. A method as set forth in claim 56 wherein said emulsion particles have a diameter in the range of approximately 0.1 to 0.5 microns.

60. A method as set forth in claim 54 wherein said cationic lipid is selected from the group consisting of N-[1-(2,3-dioleoyloxy)propyl]-N,N,N-trimethylammonium chloride, 1,2-dioleoyloxy-3-(trimethylammonio)propane, 1,2-dioleoyl-3-(4′

-trimethylammonio)butanoyl-sn-glycerol, 1,2-diacyl-3-trimethylammonium-propane, 1,2-diacyl-3-dimethylammonium-propane, and 3β

-[N′

,N′

-dimethylaminoethane-carbamol]cholesterol-HCl.