Osmotically stabilized microbubble preparations

• US 6,258,339 B1
• Filed: 05/05/1997
• Issued: 07/10/2001
• Est. Priority Date: 07/30/1993
• Status: Expired due to Term

• Current Assignee: Targeson, Inc.
• Original Assignee: Alliance Pharmaceutical Corporation
• Inventors: Schutt, Ernest G., Weers, Jeffry G., Anderson, Charles David, Kinner, Rene Alta, Evitts, David P.
• Primary Examiner(s): Hartley, Michael G.
• Application Number:
  US08/841,847
• Time in Patent Office: 1,527 Days
• Field of Search: 424/9.52, 424/9.51, 600/431, 600/458
• US Class Current: 424/9.51
• CPC Class Codes: A61K 49/223 Microbubbles, hollow microspheres, free gas bubbles, gas microspheres
• CPC Class Codes: A61K 49/227 Liposomes, lipoprotein vesicles, e.g. LDL or HDL lipoproteins, micelles, e.g. phospholipidic or polymeric
• CPC Class Codes: G02B 6/0006 Coupling light into the fibre in general G02B6/4298

Abstract

A microbubble preparation formed of a plurality of microbubbles comprising a first gas and a second gas surrounded by a membrane such as a surfactant, wherein the first gas and the second gas are present in a molar ratio of from about 1:100 to about 1000:1, and wherein the first gas has a vapor pressure of at least about (760−x) mm Hg at 37° C., where x is the vapor pressure of the second gas at 37° C., and wherein the vapor pressure of each of the first and second gases is greater than about 75 mm Hg at 37° C.; also disclosed are methods for preparing microbubble compositions, including compositions that rapidly shrink from a first average diameter to a second average diameter less than about 75% of the first average diameter and are stabilized at the second average diameter; kits for preparing microbubbles; and methods for using such microbubbles as ultrasound contrast agents.

Claims

1. A microbubble preparation comprising an aqueous medium having dispersed therein a plurality of osmotically stabilized microbubbles, said microbubbles comprising:

a generally spherical microbubble membrane containing at least one nonfluorocarbon modifier gas and at least one gas osmotic agent, wherein said modifier gas and said oas osmotic aoent are present in a molar ratio from about 1;

100 to about 1,000;

1 wherein said ratio is effective to stabilize said microbubble preparation, with the proviso that said modifier gas is not water vapor, and wherein said gas osmotic agent is selected from the group consisting of perfluoropentane, perfluorocyclopentane, perfluoromethylcyclopentane, perfluorodimethylcyclobutane, perfluorohexane, perfluorocyclohexane, perfluoroheptane, perfluorocycloheptane, perfluoromethylcyclohexane, perfluorodimethylcyclopentane, perfluorotrimethylcyclobutane, perfluorotriethylamine and combinations thereof.

2. The microbubble preparation of claim 1 wherein said nonfluorocarbon modifier gas is selected from the group consisting of nitrogen, oxygen, carbon dioxide and mixtures thereof.

3. The microbubble preparation of claim 1 wherein said nonfluorocarbon modifier gas is air.

4. The microbubble preparation of claim 1, wherein said gas osmotic agent has a water solubility of not more than about 0.5 mM at 25°

C. and one atmosphere.

5. The microbubble preparation of claim 1 wherein the microbubble membrane comprises a surfactant.

6. The microbubble preparation of claim 5 wherein said surfactant is selected from the group consisting of nonionic surfactants, neutral surfactants, anionic surfactants, neutral fluorinated surfactants, anionic fluorinated surfactants and combinations thereof.

7. The microbubble preparation of claim 5 wherein said surfactant is a non-Newtonian surfactant.

8. The microbubble preparation of claim 5 wherein said surfactant is selected from the group consisting of polyoxypropylene polyoxyethylene copolymers, sugar esters, fatty alcohols, aliphatic amine oxides, hyaluronic acid aliphatic esters, hyaluronic acid aliphatic ester salts, dodecyl poly(ethyleneoxy)ethanol, nonylphenoxy poly(ethyleneoxy)ethanol, hydroxy ethyl starch, hydroxy ethyl starch fatty acid esters, dextrans, dextran fatty acid esters, sorbitol, sorbitol fatty acid esters, gelatin, serum albumins, phospholipids, polyoxyethylene fatty acids esters, polyoxyethylene fatty alcohol ethers, polyoxyethylated sorbitan fatty acid esters, glycerol polyethylene glycol oxystearate, glycerol polyethylene glycol ricinoleate, ethoxylated soybean sterols, ethoxylated castor oil, cholesterol, oleic acid, sodium oleate and combinations thereof.

9. The microbubble preparation of claim 1 wherein the microbubble membrane comprises a liposome.

10. The microbubble preparation of claim 1 wherein the microbubble membrane comprises a proteinaceous material.

11. The microbubble preparation of claim 10 wherein said proteinaceous material is albumin.

12. The microbubble preparation of claim 1 wherein said gas osmotic agent is perfluoropentane.

13. The microbubble preparation of claim 12 wherein said nonfluorocarbon modifier gas is selected from the group consisting of nitrogen, oxygen, carbon dioxide and mixtures thereof.

14. The microbubble preparation of claim 12 wherein said nonfluorocarbon modifier gas is air.

15. The microbubble preparation of claim 12 wherein the microbubble membrane comprises a surfactant.

16. The microbubble preparation of claim 15 wherein said surfactant is selected from the group consisting of nonionic surfactants, neutral surfactants, anionic surfactants, neutral fluorinated surfactants, anionic fluorinated surfactants and combinations thereof.

17. The microbubble preparation of claim 15 wherein said surfactant is a fluorinated surfactant.

18. The microbubble preparation of claim 12 wherein the microbubble membrane comprises a proteinaceous material.

19. The microbubble preparation of claim 1 wherein said gas osmotic agent is perfluoropentane and said nonfluorocarbon modifier gas is nitrogen.

20. The microbubble preparation of claim 19 wherein the microbubble membrane comprises a surfactant.

21. The microbubble preparation of claim 19 wherein said surfactant is selected from the group consisting of nonionic surfactants, neutral surfactants, anionic surfactants, neutral fluorinated surfactants, anionic fluorinated surfactants and combinations thereof.

22. The microbubble preparation of claim 19 wherein said surfactant is a non-Newtonian surfactant.

23. The microbubble preparation of claim 19 wherein said surfactant is a fluorinated surfactant.

24. The microbubble preparation of claim 19 wherein said surfactant is selected from the group consisting of polyoxypropylene polyoxyethylene copolymers, sugar esters, fatty alcohols, aliphatic amine oxides, hyaluronic acid aliphatic esters, hyaluronic acid aliphatic ester salts, dodecyl poly(ethyleneoxy)ethanol, nonylphenoxy poly(ethyleneoxy)ethanol, hydroxy ethyl starch, hydroxy ethyl starch fatty acid esters, dextrans, dextran fatty acid esters, sorbitol, sorbitol fatty acid esters, gelatin, serum albumins, phospholipids, polyoxyethylene fatty acids esters, polyoxyethylene fatty alcohol ethers, polyoxyethylated sorbitan fatty acid esters, glycerol polyethylene glycol oxystearate, glycerol polyethylene glycol ricinoleate, ethoxylated soybean sterols, ethoxylated castor oil, cholesterol oleic acid, sodium oleate and combinations thereof.

25. The microbubble preparation of claim 1 wherein said gas osmotic agent is perfluorohexane.

26. The microbubble preparation of claim 25 wherein said nonfluorocarbon modifier gas is selected from the group consisting of nitrogen, oxygen, carbon dioxide and mixtures thereof.

27. The microbubble preparation of claim 25 wherein said nonfluorocarbon modifier gas is air.

28. The microbubble preparation of claim 25 wherein the microbubble membrane comprises a surfactant.

29. The microbubble preparation of claim 28 wherein said surfactant is selected from the group consisting of nonionic surfactants, neutral surfactants, anionic surfactants, neutral fluorinated surfactants, anionic fluorinated surfactants and combinations thereof.

30. The microbubble preparation of claim 28 wherein said surfactant is selected from the group consisting of polyoxypropylene polyoxyethylene copolymers, sugar esters, fatty alcohols, aliphatic amine oxides, hyaluronic acid aliphatic esters, hyaluronic acid aliphatic ester salts, dodecyl poly(ethyleneoxy)ethanol, nonylphenoxy poly(ethyleneoxy)ethanol, hydroxy ethyl starch, hydroxy ethyl starch fatty acid esters, dextrans, dextran fatty acid esters, sorbitol, sorbitol fatty acid esters, gelatin, serum albumins, phospholipids, polyoxyethylene fatty acids esters, polyoxyethylene fatty alcohol ethers, polyoxyethylated sorbitan fatty acid esters, glycerol polyethylene glycol oxystearate, glycerol polyethylene glycol ricinoleate, ethoxylated soybean sterols, ethoxylated castor oil, cholesterol oleic acid, sodium oleate and combinations thereof.

31. The microbubble preparation of claim 28 wherein said surfactant comprises a mixture of a phospholipid and a polyoxyethylene-polyoxypropylene copolymer.

32. The microbubble preparation of claim 25 wherein the microbubble membrane comprises a proteinaceous material.

33. The microbubble preparation of claim 32 wherein said proteinaceous material is albumin.

34. The microbubble preparation of claim 1 wherein said gas osmotic agent is perfluorohexane and said nonfluorocarbon modifier gas is nitrogen.

35. The microbubble preparation of claim 34 wherein the microbubble membrane comprises a surfactant.

36. The microbubble preparation of claim 35 wherein said surfactant is selected from the group consisting of nonionic surfactants, neutral surfactants, anionic surfactants, neutral fluorinated surfactants, anionic fluorinated surfactants and combinations thereof.

37. The microbubble preparation of claim 35 wherein said surfactant is a non-Newtonian surfactant.

38. The microbubble preparation of claim 35 wherein said surfactant is selected from the group consisting of polyoxypropylene polyoxyethylene copolymers, sugar esters, fatty alcohols, aliphatic amine oxides, hyaluronic acid aliphatic esters, hyaluronic acid aliphatic ester salts, dodecyl poly(ethyleneoxy)ethanol, nonylphenoxy poly(ethyleneoxy)ethanol, hydroxy ethyl starch, hydroxy ethyl starch fatty acid esters, dextrans, dextran fatty acid esters, sorbitol, sorbitol fatty acid esters, gelatin, serum albumins, phospholipids, polyoxyethylene fatty acids esters, polyoxyethylene fatty alcohol ethers, polyoxyethylated sorbitan fatty acid esters, glycerol polyethylene glycol oxystearate, glycerol polyethylene glycol ricinoleate, ethoxylated soybean sterols, ethoxylated castor oil, cholesterol, oleic acid, sodium oleate and combinations thereof.

39. The microbubble preparation of claim 35 wherein said surfactant comprises a mixture of a phospholipid and a polyoxyethylene-polyoxypropylene copolymer.

40. The microbubble preparation of claim 39 wherein said microbubble membrane further comprises a hydroxy ethel starch.

84. The microbubble preparation of claim 1, wherein the molar ratio of said modifier gas and said gas osmotic agent is between about 1:

100 and 1;

1.

85. The microbubble preparation of claim 1, wherein the molar ratio of said modifier gas and said gas osmotic agent is between about 1:

10 and 1;

1.

86. The microbubble of claim 1, in which the molar ratio of said modifier gas and said gas osmotic agent is greater than 1:

1.

87. The microbubble of claim 86, wherein the modifier gas is a non-fluorocarbon and the gas osmotic agent is a fluorocarbon.

88. The microbubble preparation of claim 1, wherein said plurality of osmotically stabilized microbubbles have a diameter from about 1 to 10 μ

m.

89. The microbubble preparation of claim 88, wherein the diameter of said plurality of osmotically stabilized microbubbles is about 6 μ

m.

41. A microbubble preparation comprising an aqueous medium having dispersed therein a plurality of osmotically stabilized microbubbles, said microbubbles comprising:

a generally spherical microbubble membrane containing at least one modifier gas and at least one gas osmotic agent, wherein said modifier gas osmotic agent are present in a molar ratio from about 1;

100 to about 1.000;

1, wherein said ratio is effective to stabilize said mircrobubble preparation, with the proviso that said modifier gas is not water vapor, and wherein said gas osmotic agent comprises the vapor of a compound which is a liquid at 37°

C. and 760 Torr.

42. The microbubble preparation of claim 41 wherein said gas osmotic agent is selected from the group consisting of perfluorohexane, perfluorocyclohexane, perfluoroheptane, perfluorocycloheptane, perfluoromethylcyclohexane, perfluorodimethylcyclopentane, perfluorotrimethylcyclobutane, perfluorotriethylamine and combinations thereof.

43. The microbubble preparation of claim 41 wherein said modifier gas is a fluorocarbon gas.

44. The microbubble preparation of claim 41 wherein said fluorocarbon modifier gas is selected from the group comprising perfluoropropane, perfluorobutane, perfluorocyclobutane, perfluoromethylcyclobutane, perfluoropentane and perfluorocyclopentane.

45. The microbubble preparation of claim 41 wherein said modifier gas is a nonfluorocarbon gas.

46. The microbubble preparation of claim 45 wherein said nonfluorocarbon modifier gas is selected from the group consisting of nitrogen, oxygen, carbon dioxide and mixtures thereof.

47. The microbubble preparation of claim 45 wherein said nonfluorocarbon modifier gas is air.

48. The microbubble preparation of claim 41 wherein the microbubble membrane comprises a surfactant.

49. The microbubble preparation of claim 48 wherein said surfactant is selected from the group consisting of nonionic surfactants, neutral surfactants, anionic surfactants, neutral fluorinated surfactants, anionic fluorinated surfactants and combinations thereof.

50. The microbubble preparation of claim 48 wherein said surfactant is a non-Newtonian surfactant.

51. The microbubble preparation of claim 48 wherein said surfactant is selected from the group consisting of polyoxypropylene polyoxyethylene copolymers, sugar esters, fatty alcohols, aliphatic amine oxides, hyaluronic acid aliphatic esters, hyaluronic acid aliphatic ester salts, dodecyl poly(ethyleneoxy)ethanol, nonylphenoxy poly(ethyleneoxy)ethanol, hydroxy ethyl starch, hydroxy ethyl starch fatty acid esters, dextrans, dextran fatty acid esters, sorbitol, sorbitol fatty acid esters, gelatin, serum albumins, phospholipids, polyoxyethylene fatty acids esters, polyoxyethylene fatty alcohol ethers, polyoxyethylated sorbitan fatty acid esters, glycerol polyethylene glycol oxystearate, glycerol polyethylene glycol ricinoleate, ethoxylated soybean sterols, ethoxylated castor oil, cholesterol, oleic acid, sodium oleate and combinations thereof.

52. The microbubble preparation of claim 41 wherein the microbubble membrane comprises a liposome.

53. The microbubble preparation of claim 41 wherein the microbubble membrane comprises a proteinaceous material.

54. The microbubble preparation of claim 53 wherein said proteinaceous material is albumin.

90. The microbubble preparation of claim 41, wherein the molar ratio of said modifier gas and said gas osmotic agent is between about 1:

100 and 1;

1.

91. The microbubble preparation of claim 41, wherein the molar ratio of said modifier gas and said gas osmotic agent is between about 1:

10 and 1;

1.

92. The microbubble preparation of claim 41, in which the molar ratio of said modifier gas and said gas osmotic agent is greater than 1:

1.

93. The microbubble preparation of claim 92, wherein the modifier gas is a non-fluorocarbon and the gas osmotic agent is a fluorocarbon.

94. The microbubble preparation of claim 41, wherein said plurality of osmotically stabilized microbubbles have a diameter from about 1 to 10 μ

m.

95. The microbubble preparation of claim 94, wherein the diameter of said plurality of osmotically stabilized microbubbles is about 6 μ

m.

55. A microbubble preparation comprising an aqueous medium having dispersed therein a plurality of osmotically stabilized microbubbles, said microbubbles comprising:

a generally spherical microbubble membrane comprising proteinaceous material containing at least one nonfluorocarbon modifier gas and at least one gas osmotic agent having from about three to about eight carbon atoms, wherein said modifier gas and said gas osmotic agent are present in a molar ratio from about 1;

100 to about 1,000;

1, wherein said ratio is effective to stabilize said microbubble preparation, with the proviso that said modifier gas is not water vapor.

56. The microbubble preparation of claim 55 wherein said gas osmotic agent has a water solubility of not more than about 0.5 mM at 25°

C. and one atmosphere.

57. The microbubble preparation of claim 55 wherein said gas osmotic agent comprises a fluorocarbon.

58. The microbubble preparation of claim 57 wherein said fluorocarbon gas osmotic agent is selected from the group consisting of perfluoropropane, perfluorobutane, perfluorocyclobutane, perfluoromethylcyclobutane, perfluoropentane, perfluorocyclopentane, perfluoromethylcyclopentane, perfluorodimethylcyclobutanes, perfluorohexane, perfluorocyclohexane, perfluoroheptane, perfluorocycloheptane, perfluoromethylcyclohexane, perfluorodimethylcyclopentane, perfluorotrimethylcyclobutane, perfluorotriethylamine and combinations thereof.

59. The microbubble preparation of claim 55 wherein said nonfluorocarbon modifier gas is selected from the group consisting of nitrogen, oxygen, carbon dioxide and mixtures thereof.

60. The microbubble preparation of claim 55 wherein the microbubble membrane further comprises a surfactant.

61. The microbubble preparation of claim 60 wherein said surfactant is selected from the group consisting of nonionic surfactants, neutral surfactants, anionic surfactants, neutral fluorinated surfactants, anionic fluorinated surfactants and combinations thereof.

62. The microbubble preparation of claim 60 wherein said surfactant is a non-Newtonian surfactant.

63. The microbubble preparation of claim 60 wherein said surfactant is a fluorinated surfactant.

64. The microbubble preparation of claim 60 wherein said surfactant is selected from the group consisting of polyoxypropylene polyoxyethylene copolymers, sugar esters, fatty alcohols, aliphatic amine oxides, hyaluronic acid aliphatic esters, hyaluronic acid aliphatic ester salts, dodecyl poly(ethyleneoxy)ethanol, nonylphenoxy poly(ethyleneoxy)ethanol, hydroxy ethyl starch, hydroxy ethyl starch fatty acid esters, dextrans, dextran fatty acid esters, sorbitol, sorbitol fatty acid esters, gelatin, phospholipids, polyoxyethylene fatty acids esters, polyoxyethylene fatty alcohol ethers, polyoxyethylated sorbitan fatty acid esters, glycerol polyethylene glycol oxystearate, glycerol polyethylene glycol ricinoleate, ethoxylated soybean sterols, ethoxylated castor oil, cholesterol, oleic acid, sodium oleate and combinations thereof.

65. The microbubble preparation of claim 55 wherein said proteinaceous material is albumin.

66. The microbubble preparation of claim 65 wherein said gas osmotic agent comprises perfluoropropane.

67. The microbubble preparation of claim 66 wherein said modifier gas comprises nitrogen.

68. The microbubble preparation of claim 66 wherein said modifier gas comprises oxygen.

69. The microbubble preparation of claim 66 wherein said modifier gas comprises air.

70. The microbubble preparation of claim 66 wherein said modifier gas comprises carbon dioxide.

71. The microbubble preparation of claim 65 wherein said gas osmotic agent comprises perfluoropropane and said modifier gas comprises nitrogen.

72. The microbubble preparation of claim 65 wherein said gas osmotic agent comprises perfluoropropane and said modifier gas comprises oxygen.

73. The microbubble preparation of claim 65 wherein said gas osmotic agent comprises perfluoropropane and said modifier gas comprises a mixture of oxygen and nitrogen.

74. The microbubble preparation of claim 65 wherein said gas osmotic agent comprises perfluoropropane and said modifier gas comprises air.

75. The microbubble preparation of claim 65 wherein said gas osmotic agent comprises perfluorohexane.

76. The microbubble preparation of claim 75 wherein said modifier gas comprises nitrogen.

77. The microbubble preparation of claim 75 wherein said modifier gas comprises oxygen.

78. The microbubble preparation of claim 75 wherein said modifier gas comprises air.

79. The microbubble preparation of claim 75 wherein said modifier gas comprises carbon dioxide.

80. The microbubble preparation of claim 65 wherein said gas osmotic agent comprises perfluorohexane and said modifier gas comprises nitrogen.

81. The microbubble preparation of claim 65 wherein said gas osmotic agent comprises perfluorohexane and said modifier gas comprises oxygen.

82. The microbubble preparation of claim 65 wherein said gas osmotic agent comprises perfluorohexane and said modifier gas comprises a mixture of oxygen and nitrogen.

83. The microbubble preparation of claim 65 wherein said gas osmotic agent comprises perfluorohexane and said modifier gas comprises air.

96. The microbubble preparation of claim 55, wherein the molar ratio of said modifier gas and said gas osmotic agent is between about 1:

100 and 1;

1.

97. The microbubble preparation of claim 55, wherein the molar ratio of said modifier gas and said gas osmotic agent is between about 1:

10 and 1;

1.

98. The microbubble preparation of claim 55, in which the molar ratio of said modifier gas and said gas osmotic agent is greater than 1:

1.

99. The microbubble preparation of claim 98, wherein the modifier gas is a non-fluorocarbon and the gas osmotic agent is a fluorocarbon.

100. The microbubble preparation of claim 55, wherein said plurality of osmotically stabilized microbubbles have a diameter from about 1 to 10 μ

m.

101. The microbubble preparation of claim 100, wherein the diameter of said plurality of osmotically stabilized microbubbles is about 6 μ

m.