Preparation and uses of multi-phase microspheres

• US 5,288,502 A
• Filed: 10/16/1991
• Issued: 02/22/1994
• Est. Priority Date: 10/16/1991
• Status: Expired due to Fees

• Current Assignee: Board of Regents of the University of Texas System (University of Texas System)
• Original Assignee: University of Texas System
• Inventors: Iwata, Motokazu, McGinity, James W.
• Primary Examiner(s): Michl, Paul R.
• Assistant Examiner(s): Kulkosky, Peter F.
• Application Number:
  US07/779,173
• Time in Patent Office: 860 Days
• Field of Search: 424/489, 424/490, 424/491, 424/488, 424/484, 428/402.2, 428/402.21, 428/402.22
• US Class Current: 424/484
• CPC Class Codes: A61K 9/1647 Polyesters, e.g. poly(lactide-co-glycolide)
• CPC Class Codes: A61K 9/1694 resulting in granules or microspheres of the matrix type containing more than 5% of excipient
• CPC Class Codes: A61K 9/5031 obtained otherwise than by reactions only involving carbon-to-carbon unsaturated bonds, e.g. polyethylene glycol, poly(lactide-co-glycolide)
• CPC Class Codes: A61K 9/5089 Processes
• CPC Class Codes: Y10T 428/2984 Microcapsule with fluid core [includes liposome]
• CPC Class Codes: Y10T 428/2985 Solid-walled microcapsule from synthetic polymer
• CPC Class Codes: Y10T 428/2987 Addition polymer from unsaturated monomers only

Abstract

Multi-phase polymeric microspheres containing a molecular compound dispersed in a polymeric matrix are described. Methods for preparing the multi-phase microspheres are also described, which includes a multiple emulsion solvent evaporation technique. Drug loading efficiencies between 80 to 100% were achieved using the described methods. Particular ratios of the W/O emulsion to polymer, and concentration of surfactant and dispersion media (mineral oil) provide highly efficient multi-phase microspheres. In particular embodiments, the multi-phase microspheres feature a high loading efficiency of water-soluble drugs, and also eliminates partitioning of the water soluble agent into the polymer acetonitrile (solvent) phase, thus preventing low encapsulation efficiency. The described multi-phase microspheres possess efficient drug loading, release properties, and drug stability, and also provide a vehicle for long term therapeutic release of a biologically active molecule for therapeutically effective periods of time. Molecular compounds which may be incorporated within the multi-phase microspheres include both water-soluble and water-insoluble drugs, proteins (e.g. TNF), peptides, and chemicals. The molecular compound is protected within an oily droplet, and contact with polymer, surfactant, organic solvents, and other potentially denaturing agents is prevented.

Claims

1. A delivery system for a protein, peptide, or drug with biodegradable multi-phase microspheres, said microspheres comprising a protein, peptide, or drug contained within a fixed oil and an essentially water insoluble, biodegradable polymeric matrix comprised of polylactic acid or polylactic glycolic acid wherein the polymeric matrix surrounds the fixed oil of the microsphere and wherein the fixed oil contains the protein, peptide, or drug.

5. The delivery system of claim 1 wherein the molecular compound is a protein, a peptide, a chemical or a dug.

6. The delivery system of claim 1, 2, 3 or 4 wherein the multi-phase microsphere is about 150μ

in size.

7. The delivery system of claim 1, 2, 3 or 4 wherein the multi-phase microsphere is about between 50μ

to 100μ

in size.

8. The delivery system of claim 1 wherein the molecular compound is water soluble and pharmacologically active.

9. The delivery system of claim 8 wherein the molecular compound is prepared as an aqueous solution together in a fixed oil to provide a microemulsion.

10. The delivery system of claim 1 wherein the molecular compound is an enzyme.

11. The delivery system of claim 1 wherein the molecular compound is selected from the group consisting of interferon, macrophage activation factor, interleukin, colony stimulating factor, tumor degenerating factor, epidermal growth factor, erythropoietin, tissue plasminogen activator, insulin, urokinase, luteinizing hormone releasing hormone, superoxide dismutase, or an antibody or an enzyme.

12. The delivery system of claim 1 wherein the molecular compound is chlorpheniramine maleate, diphenhydramine hydrochloride, procainamide hydrochloride, promazine hydrochloride, or brilliant blue.

13. The delivery system of claim 1, 2, 3 or 4 wherein the multi-phase microspheres are comprised of a biodegradable polymer of lactic acid or glycolic acid.

14. The delivery system of claim 1, 2, 3 or 4 wherein the microspheres are comprised of a polymer poly (d,l)-lactic, poly (l)-lactic acid, or copolymers of lactic and glycolic acid.

15. The delivery system of claim 1, 2, 3 or 4 wherein the fixed oil is safflower, soybean, cottonseed, peanut, sesame or cod liver oil.

16. The delivery system of claim 1 wherein the molecular compound is tumor necrosis factor.

2. A delivery system for a water-soluble protein, peptide, or drug with biodegradable multi-phase microspheres, said microspheres comprising a microemulsion of a water soluble protein, peptide, or drug, a fixed oil and a biodegradable, essentially water insoluble polymeric matrix of polylactic acid or polylactic glycolic acid, said polymeric matrix surrounding the microemulsion of fixed oil and the protein, peptide, or molecular compound.

42. The method of claim 2 wherein the ratio of microemulsion to polymer is between about 0.25 to 1.0 grams to 1 by weight.

3. A delivery system for cytokines with biodegradable multi-phase microspheres, said microspheres comprising a cytokine contained within a fixed oil, and an essentially water insoluble, biodegradable polymeric matrix of polylactic acid or polylactic glycolic acid, wherein the polymeric matrix surrounds the fixed oil and wherein the fixed oil contains the cytokine.

4. A delivery system for tumor necrosis factor with biodegradable multi-phase microspheres said microspheres comprising a lyophilized microemulsion of an aqueous solution of tumor necrosis factor in a fixed oil, and an essentially water insoluble, biodegradable polymeric matrix of polylactic acid or polylactic glycolic acid, wherein the polymeric matrix surrounds said microemulsion of tumor necrosis factor and fixed oil.

17. The delivery system of claim 4 or 6 wherein the microemulsion consists of lyophilized tumor necrosis factor in a fixed oil and the polymeric matrix is poly-lactic acid (PLGA).

18. The delivery system of claim 17 wherein the fixed oil is soybean oil and includes aluminum monostearate.

19. A method for providing sustained release of a protein, peptide, or drug in an animal comprising:

preparing a formulation of microspheres, said microspheres containing the protein, peptide, or drug in a fixed oil, and a biodegradable essentially water insoluble polymeric matrix of polylactic acid or polylactic glycolic acid; and

administering an amount of the formulation effective to provide sustained release of the protein, peptide, or drug in the animal for a prescribed period of time, wherein the polymeric matrix surrounds the fixed oil and the fixed oil contains the protein, peptide, or drug.

20. The method of claim 19 wherein the water-soluble molecular compound is a protein, a peptide, a chemical, or a drug.

21. The method of claim 19 wherein the molecular compound is a therapeutic agent.

22. The method of claim 19 wherein the molecular compound is CPM, DPH, PMZ, TNF or PRC.

23. The method of claim 19 wherein the formulation is administered parenterally.

24. The method of claim 19 wherein the formulation is administered intramuscularly.

25. The method of claim 19 wherein the protein, peptide, or drug is water-soluble and is prepared as an aqueous solution together in a fixed oil to provide a microemulsion and wherein the polymeric matrix surrounds the microemulsion.

26. The method of claim 19 wherein sustained release of the molecular compound is provided for up to 1 year.

27. The method of claim 19 wherein sustained release of the molecular compound is provided for about 3 months.

28. The method of claim 19 wherein the molecular compound is water-soluble.

29. A method for preparing multi-phase microspheres containing a protein, peptide, or dug comprising:

mixing an aqueous solution of the protein, peptide, or drug with a fixed oil to form a W/O microemulsion;

mixing polylactic acid or polylactic glycolic acid and a polymer solvent together to form a polymer solution;

dispersing the microemulsion into the polymer solution to form a W/O/"O" emulsion;

mixing the W/O/"O" emulsion together in a dispersion oil which is incompatible with the polymer solvent to form a multiple emulsion;

agitating and removing the solvent from the multiple emulsion to form hardened microspheres; and

washing and drying the hardened microspheres to form multi-phase microspheres containing the water-soluble protein, peptide, or drug.

30. The method of claim 29 wherein the fixed oil of the microemulsion is soybean, cottonseed, peanut, sesame cod liver oil or safflower oil.

31. The method of claim 29 wherein the polymer solvent is acetonitrile or methylene chloride.

32. The method of claim 29 wherein the polymer is PLA or PLGA.

33. The method of claim 29 wherein the polymer solution comprises PLA and acetonitrile.

34. The method of claim 29 wherein the molecular compound is a protein, a peptide, a chemical or a drug.

35. The method of claim 29 wherein the microspheres are about 150μ

in size.

36. The method of claim 29 wherein the microspheres are between about 50μ

and about 100μ

in size.

37. The method of claim 29 wherein the molecular compound is water soluble and is selected from the group consisting of CPM, DPH, brilliant blue, PMZ, and PRC.

38. The method of claim 29 wherein the microemulsion comprises about 1% w/w Tween 80 of the microemulsion.

39. The method of claim 29 wherein the microemulsion comprises about 2% aluminum monostearate and about 5% W/W Span 80 of the microemulsion.

40. The method of claim 29 wherein the multiple emulsion comprises about 0.25% to 2% w/w Span 80.

41. The method of claim 29 wherein the molecular compound is:

insulin;

LHRH;

a steroid;

atriopeptin III;

monoclonal antibodies;

TNF;

interferon;

macrophage activating factor;

interleukins;

colony stimulating factor;

tumor degenerating factor;

epidermal growth factor;

erythroprotein;

tissue plasminogen activator;

urokinase;

superoxide dismutase or

P-450.

43. The method of claim 29 wherein the molecular compound is unstable in water, and wherein the microemulsion is lyophilized prior to dispersing the microemulsion into the polymer solution to form a W/O/"W" emulsion.

44. The method of claim 43 wherein the molecular compound is TNF.

45. A method for preparing microspheres containing a water-soluble drug comprising:

preparing a first mixture of a water-soluble drug in water, gelatin and Tween 80 to form an aqueous phase;

preparing a second mixture of an amount of aluminum stearate and a volume of a fixed oil to produce a 2% w/w aluminum stearate and about 5% w/w Span 80 oil phase;

combining the first mixture with the second mixture to form a coarse W/O emulsion;

processing the coarse W/O emulsion into a fine W/O microemulsion;

preparing a third mixture of polylactic acid or polylactic glycolic acid and a solvent;

combining a quantity of the fine W/O microemulsion with the third mixture to form a W/O/"O" emulsion;

preparing a fourth mixture of an amount of Span 80 and an oil incompatible with the polymer solvent;

pouring the W/O/"O" emulsion into the fourth mixture to form a multiple emulsion;

agitating and evaporating the polymer solvent from the multiple emulsion to form hardened microspheres;

separating the hardened microspheres from the mixture; and

washing and drying the hardened microspheres to form multi-phase microspheres containing a water-soluble drug.

46. The method of claim 45 wherein the third mixture comprises less than 35% w/w of the polymer.

47. The method of claim 45 wherein the third mixture comprises about 33% w/w of a PLA biodegradable polymer in an acetonitrile solvent.

48. The method of claim 45 wherein the third mixture comprises about 31% w/w of a PLGA biodegradable polymer in an acetonitrile solvent.

49. The method of claim 45 wherein the water-soluble molecular compound is CPM, DPH, PMZ, or PRC.

50. The method of claim 45 wherein the quantity of the fine W/O emulsion is between about 0.25 to 1.0 g by weight of the W/O/"W" emulsion.

51. The method of claim 45 wherein the fourth mixture includes between about 0.25% to about 2% w/w Span 80.

52. The method of claim 45 wherein the fourth mixture includes about 0.254 to 0.5% w/w Span 80.

53. The method of claim 45 wherein the fourth mixture includes about 0.254 w/w Span 80.

54. A multi-phase microsphere for the delivery of a peptide, protein, or drug comprising a biodegradable essentially water insoluble polymeric matrix of polylactic acid or polylactic glycolic acid surrounding a protein, peptide, or drug contained within a fixed oil, said microsphere prepared by a process of:

preparing a first mixture of a water-soluble protein, peptide, or dug in water, gelatin and Tween 80 to form an aqueous base;

preparing a second mixture of an amount of aluminum stearate and a volume of a fixed oil to produce a 2% w/w aluminum stearate and about 5% w/w span 80 oil phase;

combining the first mixture with the second mixture to form a coarse W/O emulsion;

processing the coarse W/O emulsion into a fine W/O micro emulsion;

preparing a third mixture of a biodegradable essentially water-insoluble polymer polylactic acid of polylactic glycolic acid and a solvent;

combining a quantity of the fine W/O micro emulsion with the third mixture to form a W/O/"O" emulsion;

preparing a fourth mixture of an amount of span 80 and an oil incompatible with the polymer solvent;

pouring the W/O/"O" emulsion into the fourth mixture to form a multiple emulsion;

evaporating the solvent from the multiple emulsion to form hardened microspheres;

separating the hardened microspheres from the mixture; and

washing and drying the hardened microspheres to form multi-phase microspheres containing a water-soluble protein, peptide, or drug.

55. The microsphere of claim 54 wherein the fixed oil is safflower, soybean, cottonseed, peanut, sesame or cod liver oil.

56. The microsphere of claim 54 wherein the protein is tumor necrosis factor.

57. The microsphere of claim 54 wherein the biodegradable polymer is poly(d,l-lactic acid) or poly(d,l-lactic) co-glycolic lactic acid.

58. A biodegradable polymeric microsphere comprising an essentially water insoluble poly (d,l-lactic acid) or poly (d,l-lactic) co-glycolic acid polymeric matrix surrounding a protein-containing fixed oil droplet.

59. The biodegradable polymeric microsphere of claim 58 wherein the polymeric matrix is poly(d,l-lactic acid).

60. The biodegradable polymeric microsphere of claim 58 wherein the fixed oil is safflower, soybean, cottonseed, peanut, sesame or cod liver oil.

61. The biodegradable polymeric microsphere of claim 58 wherein the protein is tumor necrosis factor of interleukin-1.

62. The biodegradable polymeric microsphere of claim 58 wherein the microsphere has a diameter of between 50μ

and 200μ

.