Macromolecular microparticles and methods of production and use

- Epic Therapeutics, Inc.

Microparticles formed by mixing a macromolecule with a polymer at a pH near the isoelectric point of the macromolecule and incubating the mixture in the presence of an energy source for a predetermined length of time. The microparticles are composed of homogeneously distributed, intertwined macromolecule and polymer. Each microparticle allows aqueous fluids to enter and allows solubilized macromolecule and polymer to exit the microparticle and may be formulated to provide a sustained release of macromolecule and polymer from the interior of the microparticle when placed in an appropriate aqueous medium, such as under physiological conditions. Methods of production and methods of use for research, diagnostics and therapeutics are provided.

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Claims

1. A microparticle comprising a macromolecule and a polymer in an aqueous solution, wherein the concentration of the macromolecule in the microparticle is at least 40% and less than 100% by weight, and wherein the microparticle does not contain any oil.

2. The microparticle of claim 1 wherein the concentration of the polymer in the microparticle is greater than 0% and less than or equal to 30% by weight.

3. The microparticle of claim 1 wherein the macromolecule is a molecule having or capable of having a tertiary and quaternary structure.

4. The microparticle of claim 3 wherein the macromolecule is selected from the group consisting of a protein, peptide, polypeptide, carbohydrate, polysaccharide, protein conjugate, nucleic acid, virus, virus particle, and mixtures thereof.

5. The microparticle of claim 1 further comprising a pharmaceutical compound.

6. The microparticle of claim 1 wherein the polymer is water soluble or soluble in a water miscible solvent.

7. The microparticle of claim 1 wherein the polymer is selected from the group consisting of carbohydrate-based polymers, polyaliphatic alcohols, poly(vinyl) polymers, polyacrylic acids, polyorganic acids, polyamino acids, co-polymers, block co-polymers, tert-polymers, polyethers, naturally occurring polymers, polyimids, surfactants, polyesters, branched polymers, cyclo-polymers, polyaldehydes and mixtures thereof.

8. The microparticle of claim 1 further comprising a biomolecule bound inside or on the surface of the microparticle.

9. The microparticle of claim 1 wherein the macromolecule is released from the microparticle under physiological conditions.

10. The microparticle of claim 9 wherein the released macromolecule is biologically active.

11. The microparticle of claim 1 further comprising luteinizing hormone releasing hormone (LHRH) or an analog thereof, wherein the analog has LHRH agonist or antagonist activity.

12. The microparticle of claim 11 wherein the luteinizing hormone releasing hormone analog is leuprolide.

13. The microparticle of claim 1 wherein the polymer is selected from the group consisting of dextrans, polydextroses, chitins, starches and hetastarch.

14. The microparticle of claim 1 further comprising a small molecule or compound bound inside or on the surface of the microparticle by non-covalent interactions.

15. The microparticle of claim 1 further comprising dextran sulfate.

16. A method of making microparticles comprising combining a macromolecule and a polymer in an aqueous solution at a pH near the isoelectric point of the macromolecule and exposing the solution to an energy source for a sufficient amount of time to form microparticles.

17. The method of claim 16 wherein the energy source is heat.

18. The method of claim 16 wherein the energy source is heat and the macromolecule and polymer are heated for between approximately 5 minutes and 24 hours at a temperature between approximately 37.degree. C. and 90.degree. C.

19. The method of claim 16 wherein the pH is within 2 pH units of the pI of the macromolecule.

20. The method of claim 16 wherein the macromolecule is a molecule having or capable of having a tertiary and quaternary structure.

21. The method of claim 20 wherein the macromolecule is selected from the group consisting of a protein, peptide, polypeptide, carbohydrate, polysaccharide, protein conjugate, nucleic acid, virus, virus particle, and mixtures thereof.

22. The method of claim 16 further comprising adding a pharmaceutical compound.

23. The method of claim 16 wherein the polymer is water soluble or soluble in a water miscible solvent and dehydrates the macromolecule.

24. The method of claim 16 wherein the polymer is selected from the group consisting of carbohydrate-based polymers, polyaliphatic alcohols, poly(vinyl) polymers, polyacrylic acids, polyorganic acids, polyamino acids, co-polymers, block co-polymers, tert-polymers, polyethers, naturally occurring polymers, polyimids, surfactants, polyesters, branched polymers, cyclo-polymers, polyaldehydes, and mixtures thereof.

25. The method of claim 16 further comprising binding a biomolecule inside or on the surface of the microparticle.

26. The method of claim 16 wherein the macromolecule is released from the microparticle under physiological conditions.

27. The method of claim 26 wherein the released macromolecule is biologically active.

28. The method of claim 16 wherein the aqueous solution further comprises luteinizing hormone releasing hormone (LHRH) or an analog thereof, wherein the analog has LHRH agonist or antagonist activity.

29. The method of claim 28 wherein the luteinizing hormone releasing hormone analog is leuprolide.

30. The method of claim 16 wherein the polymer is selected from the group consisting of dextrans, polydextroses, chitins, starches and hetastarch.

31. The method of claim 16 wherein the microparticle further comprises a small molecule or compound bound inside or on the surface of the microparticle by non-covalent interactions.

32. The method of claim 16 wherein the aqueous solution further comprises dextran sulfate.

33. A microparticle prepared by a method comprising combining a macromolecule with a polymer in an aqueous solution at a pH near the isoelectric point of the macromolecule and exposing the solution to an energy source for a sufficient amount of time to form the microparticle.

34. The microparticle of claim 33 wherein the energy source is heat.

35. The microparticle of claim 33 wherein the energy source is heat and the macromolecule and polymer are heated for between approximately 5 minutes and 24 hours at a temperature between approximately 37.degree. C. and 90.degree. C.

36. The microparticle of claim 33 wherein the pH is within 2 pH units of the pI of the macromolecule.

37. The microparticle of claim 33 wherein the concentration of the macromolecule in the microparticle is at least 40% and less than 100% by weight.

38. The microparticle of claim 33 wherein the concentration of the polymer in the microparticle is greater than 0% and less than or equal to 30% by weight.

39. The microparticle of claim 33 wherein the macromolecule is a molecule having or capable of having a tertiary and quaternary structure.

40. The microparticle of claim 33 wherein the macromolecule is selected from the group consisting of a protein, peptide, polypeptide, carbohydrate, polysaccharide, protein conjugate, nucleic acid, virus, virus particle, and mixtures thereof.

41. The microparticle of claim 33 wherein the method further comprises adding a pharmaceutical compound.

42. The microparticle of claim 33 wherein the polymer is water soluble or soluble in a water miscible solvent and dehydrates the macromolecule.

43. The microparticle of claim 33 wherein the polymer is selected from the group consisting of carbohydrate-based polymers, polyaliphatic alcohols, poly(vinyl) polymers, polyacrylic acids, polyorganic acids, polyamino acids, co-polymers, block co-polymers, tert-polymers, polyethers, naturally occurring polymers, polyimids, surfactants, polyesters, branched polymers, cyclo-polymers, polyaldehydes, and mixtures thereof.

44. The microparticle of claim 33 wherein the method further comprises binding a biomolecule inside or on the surface of the microparticle.

45. The microparticle of claim 33 wherein the macromolecule is released from the microparticle under physiological conditions.

46. The microparticle of claim 45 wherein the released macromolecule is biologically active.

47. The microparticle of claim 33 wherein the aqueous solution further comprises luteinizing hormone releasing hormone (LHRH) or an analog thereof, wherein the analog has LHRH agonist or antagonist activity.

48. The microparticle of claim 47 wherein the luteinizing hormone releasing hormone analog is leuprolide.

49. The microparticle of claim 33 wherein the polymer is selected from the group consisting of dextrans, polydextroses, chitins, starches and hetastarch.

50. The microparticle of claim 33 further comprising a small molecule or compound bound inside or on the surface of the microparticle by non-covalent interactions.

51. The microparticle of claim 33 wherein the aqueous solution further comprises dextran sulfate.

52. A method for delivering a therapeutic agent to a patient comprising administering to the patient a therapeutically effective amount of a microparticle comprising a macromolecule and a polymer in an aqueous solution, wherein the concentration of the macromolecule in the microparticle is at least 40% and less than 100% by weight, wherein the macromolecule comprises a therapeutic agent.

53. The delivery method of claim 52 wherein the macromolecule is selected from the group consisting of enzymes, recombinant proteins, immunoglobulins and mixtures thereof.

54. The delivery method of claim 52 wherein a biomolecule specific for a target site is bound inside or on the surface of the microparticle.

55. The delivery method of claim 52 wherein the macromolecule is coupled to a pharmaceutical compound.

56. The delivery method of claim 52 wherein the method of administration is by injection.

57. The delivery method of claim 52 wherein the method of administration is by inhalation.

58. The delivery method of claim 37 wherein the microparticle further comprises luteinizing hormone releasing hormone (LHRH) or an analog thereof, wherein the analog has LHRH agonist or antagonist activity.

59. The delivery method of claim 58 wherein the luteinizing hormone releasing hormone analog is leuprolide.

60. The delivery method of claim 37 wherein the polymer is selected from the group consisting of dextrans, chitins, starches and hetastarch.

61. The delivery method of claim 37 wherein the microparticle further comprises a small molecule or compound bound inside or on the surface of the microparticle by non-covalent interactions.

62. The delivery method of claim 37 wherein the microparticle further comprises dextran sulfate.

63. A method for forming a microparticle comprising:

(1) forming an aqueous mixture containing a protein and a carbohydrate-based polymer; and
(2) exposing the aqueous mixture to an energy source for a sufficient amount of time to form the microparticle;
wherein the protein is present in the aqueous mixture in an amount sufficient to form a microparticle that contains at least 40% and less than 100% by weight protein.

64. The method of claim 63, wherein the energy comprises heat.

65. The method of claim 63, further comprising the step of:

(3) contacting the microparticle with a solution of luteinizing hormone releasing hormone (LHRH) or an analog thereof, wherein the analog has LHRH agonist or antagonist activity, to incorporate the luteinizing hormone releasing hormone or analog thereof into the microparticle.

66. The method of claim 65, wherein the step of contacting the microparticle with the solution of luteinizing hormone releasing hormone or an analog thereof results in a yield of incorporation of from 83% to 90%.

67. The microparticle produced by the process of claim 63.

68. The microparticle produced by the process of claim 65.

69. A microparticle comprising:

(1) a protein; and
(2) a carbohydrate-based polymer,

70. The microparticle of claim 69, wherein the protein comprises albumin.

71. The microparticle of claim 69, wherein the carbohydrate-based polymer represents greater than 0% and less than or equal to 30% by weight of the microparticle.

72. The microparticle of claim 69, wherein the carbohydrate-based polymer comprises hetastarch.

73. The microparticle of claim 69, wherein the protein comprises albumin and the carbohydrate-based polymer comprises hetastarch.

74. The microparticle of claim 69, further comprising luteinizing hormone releasing hormone (LHRH) or an analog thereof, wherein the analog has LHRH agonist or antagonist activity.

75. A microparticle comprising:

(1) a protein;
(2) a carbohydrate-based polymer, and
(3) a luteinizing hormone releasing hormone (LHRH) or an analog thereof;

76. The microparticle of claim 75, wherein the carbohydrate-based polymer comprises dextran sulfate.

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Patent History
Patent number: 5849884
Type: Grant
Filed: Aug 19, 1996
Date of Patent: Dec 15, 1998
Assignee: Epic Therapeutics, Inc. (Norwood, MA)
Inventors: James E. Woiszwillo (Milford, MA), Larry R. Brown (Newton, MA), Terrence L. Scott (Winchester, MA), Jie Di (Norwood, MA), Judith Sudhalter (Newton, MA), Charles D. Blizzard (West Roxbury, MA)
Primary Examiner: Nancy Degen
Law Firm: Jones' Askew, LLP
Application Number: 8/699,586