Composition for sustained release of non-aggregated erythropoietin

• US 5,674,534 A
• Filed: 06/07/1995
• Issued: 10/07/1997
• Est. Priority Date: 06/11/1992
• Status: Expired due to Fees

• Current Assignee: Alkermes Controlled Therapeutics Inc. II (Alkermes plc)
• Original Assignee: Alkermes Pharma Ireland Limited (Alkermes plc)
• Inventors: Bernstein, Howard, Zale, Stephen E., Burke, Paul A., Brickner, Avram
• Primary Examiner(s): Azpuru, Carlos A.
• Application Number:
  US08/483,318
• Time in Patent Office: 853 Days
• Field of Search: 264/4.6, 428/402.2, 428/402.21, 424/491, 424/497, 424/501, 424/502
• US Class Current: 424/501
• CPC Class Codes: A61K 38/1816 Erythropoietin [EPO]
• CPC Class Codes: A61K 47/02 Inorganic compounds
• CPC Class Codes: A61K 47/26 Carbohydrates, e.g. sugar alcohols, amino sugars, nucleic acids, mono-, di- or oligo-saccharides; Derivatives thereof, e.g. polysorbates, sorbitan fatty acid esters or glycyrrhizin
• CPC Class Codes: A61K 9/0019 Injectable compositions; Intramuscular, intravenous, arterial, subcutaneous administration; Compositions to be administered through the skin in an invasive manner non-active ingredients are additionally classified in A61K47/00
• CPC Class Codes: A61K 9/1611 Inorganic compounds
• CPC Class Codes: A61K 9/1647 Polyesters, e.g. poly(lactide-co-glycolide)
• CPC Class Codes: A61K 9/1658 Proteins, e.g. albumin, gelatin
• 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 9/5052 Proteins, e.g. albumin
• CPC Class Codes: A61K 9/5153 Polyesters, e.g. poly(lactide-co-glycolide)
• CPC Class Codes: A61K 9/5169 Proteins, e.g. albumin, gelatin
• CPC Class Codes: Y10T 428/2984 Microcapsule with fluid core [includes liposome]
• CPC Class Codes: Y10T 428/2985 Solid-walled microcapsule from synthetic polymer

Abstract

A composition, and methods of forming and using said composition, for the sustained release of non-aggregated, biologically active, erythropoietin (EPO). The sustained release composition of this invention comprises a polymeric matrix of a biocompatible polymer and particles of biologically active, aggregation-stabilized EPO, wherein said particles are dispersed within the biocompatible polymer. The method of the invention for producing a composition for the sustained release of biologically active EPO, includes dissolving a biocompatible polymer in a polymer solvent to form a polymer solution, dispersing particles of biologically active, aggregation-stabilized EPO in the polymer solution, and then solidifying the polymer to form a polymeric matrix containing a dispersion of said EPO particles. The method for using a composition of the invention is a method for providing a therapeutically effective blood level of biologically active, non-aggregated erythropoietin in a subject for a sustained period. In this method, a subject is administered an effective dose of the sustained release composition of the present invention.

Claims

1. A composition for the sustained release of biologically active, non-aggregated erythropoietin from a polymeric matrix, comprising:

a) a biodegradable polymer; and

b) particles of biologically active, aggregation-stabilized erythropoietin, wherein said particles include erythropoietin in contact with a salting-out salt, and wherein said erythropoietin particles are dispersed within the polymeric matrix.

2. A sustained release composition of claim 1 wherein the salting-out salt comprises a salt containing a cation selected from the group consisting of Mg⁺², Li⁺, Na⁺, K⁺, NH₄⁺ and combinations thereof.

3. A sustained release composition of claim 1 wherein the salting-out salt comprises a salt containing an anion selected from the group consisting of SO₄^-2, HPO₄^-2, acetate, titrate, tartrate, Cl^-, NO₃^-, ClO₃^-, I^-, ClO₄^-, SCN^- and combinations thereof.

4. A sustained release composition of claim 1 wherein the salting-out salt is ammonium sulfate.

5. A sustained release composition of claim 1 wherein the biodegradable polymer is selected from the group consisting of poly(lactides), poly(glycolides), poly(lactide-co-glycolides), poly(lactic acid)s, poly(glycolic acid)s;

poly(lactic acid-co-glycolic acid)s, polycaprolactone, polycarbonates, polyesteramides, polyanhydrides, poly(amino acids), polyorthoesters, polycyanoacrylates, poly(p-dioxanone), poly(alkylene oxalate)s, biodegradable polyurethanes, blends and copolymers thereof.

6. A sustained release composition of claim 5 wherein said polymer comprises poly(lactide-co-glycolide).

7. A sustained releage composition of claim 1 further comprising a metal cation component, wherein the metal cation component is not contained in said erythropoietin particles, and wherein the metal cation component is dispersed within the biodegradable polymer to modulate the release of erythropoietin from the polymeric matrix.

8. A sustained release composition of claim 7 wherein the metal cation component is selected from the group consisting of magnesium hydroxide, magnesium carbonate, calcium carbonate, zinc carbonate, magnesium acetate, zinc acetate, magnesium sulfate, zinc sulfate, magnesium chloride, zinc chloride, zinc citrate, magnesium citrate and combinations thereof.

24. A method for providing a therapeutically effective blood level of biologically active, non-aggregated erythropoietin in a subject for a sustained period, comprising administering to the subject a dose of the sustained release composition of claim 1.

25. A method of claim 24 wherein the salting-out salt comprises a salt containing a cation selected from the group consisting of Mg⁺², Li⁺, Na⁺, K⁺, NH₄⁺ and combinations thereof.

26. A method of claim 24 wherein the salting-out salt comprises a salt containing an anion selected from the group consisting of SO₄^-2, HPO₄^-2, acetate, citrate, tartrate, Cl^-, NO₃^-, ClO₁^-, I^-, ClO₄^-, SCN^- and combinations thereof.

27. A method of claim 24 wherein the salting-out salt is ammonium sulfate.

28. A method of claim 24 wherein the biologically active, aggregation-stabilized erythropoietin particles are formed by lyophilizing a solution containing:

a) biologically active erythropoietin;

b) a salting-out salt; and

c) a buffer with a pH between about 4 and about 8.

29. A method of claim 28 wherein the salting-out salt is ammonium sulfate.

9. A composition for the sustained release of biologically active, non-aggregated erythropoietin from a polymeric matrix, comprising:

a) unblocked poly(lactide-co-glycolide) with a molecular weight of at least about 10,000 Daltons;

b) particles of biologically active, aggregation-stabilized erythropoietin, wherein said particles include erythropoietin in contact with a salting-out salt, wherein the percentage of weight of the composition attributable to said erythropoietin particles is about 5%, and wherein said erythropoietin particles are dispersed within the polymeric matrix; and

c) magnesium carbonate particles dispersed in the polymeric matrix, in an effective amount.

10. A composition of claim 9 wherein the salting-out salt is present at a concentration of at least about 10% of the total solids in the erythropoietin particles.

11. A method for forming a composition for the sustained release of biologically active erythropoietin from a polymeric matrix, comprising the steps of:

a) dissolving a biodegradable polymer in a polymer solvent to form a polymer solution;

b) dispersing particles of biologically active, aggregation-stabilized erythropoietin in the polymer solution, wherein said particles include erythropoietin in contact with a salting-out salt; and

c) solidifying the polymer to form a polymeric matrix containing a dispersion of said erythropoietin particles.

12. A method of claim 11 wherein the salting-out salt comprises a salt containing an anion selected from the group consisting of SO₄^-2, HPO₄^-2, acetate citrate, tartrate, Cl^-, NO₃^-, ClO₃^-, I^-, ClO₄^-, SCN^- and combinations thereof.

13. A method of claim 11 wherein the salting-out salt comprises a salt containing a cation selected from the group consisting Mg⁺², Li⁺, Na⁺, K⁺, NH₄⁺ and combinations thereof.

14. A method of claim 11 wherein the salting-out salt is ammonium sulfate.

15. The method of claim 11 further comprising the step of dispersing a metal cation component within the polymer solution, wherein the metal cation component is not contained in the erythropoietin particles.

16. A method of claim 15 wherein the metal cation component is multivalent.

17. A method of claim 16 wherein the metal cation component is selected from the group consisting of magnesium hydroxide, magnesium carbonate, calcium carbonate, zinc carbonate, magnesium acetate, zinc acetate. magnesium sulfate, zinc sulfate, magnesium chloride, zinc chloride. zinc citrate, magnesium citrate and combinations thercof.

18. A method for forming microparticles for the sustained release of biologically active, non-aggregated erythropoietin, comprising the steps of:

a) mixing biologically active erythropoietin with a salting-out salt and with a buffer, wherein said buffer has a pH between about 4 and about 8, to form an aggregation-stabilizing mixture; and

b) lyophilizing said mixture to form biologically active, aggregation-stabilized erythropoietin;

c) dispersing particles of biologically active, aggregation-stabilized erythropoietin in a polymer solution to form a dispersion;

d) freezing droplets of the dispersion to form microparticles; and

e) contacting the microparticles with a liquid non-solvent, which is miscible with the polymer solvent, whereby the polymer solvent is extracted from the microparticles, thereby forming microparticles for the sustained release of biologically active, non-aggregated erythropoietin.

19. The method of claim 18 wherein the salting-out salt is ammonium sulfate.

20. Biologically active, aggregation-stabilized erythropoietin, comprising a lyophilizate of a solution containing:

a) biologically active erythropoietin;

b) a salting-out salt; and

c) a buffer with a pH between about 4 and about 8.

21. A biologically active, aggregation-stabilized erythropoietin of claim 20 wherein the salting-out salt comprises a salt containing an anion selected from the group consisting of:

SO₄^-2, HPO₄^-2, acetate, citrate, tartrate, Cl^-, NO₃^-, ClO₃, I^-, ClO₄^-, SCN^- and combination thereof.

22. A biologically active, aggregation-stabilized erythropoietin of claim 20 wherein the salting-out salt comprises a salt containing a cation selected from the group consisting of Mg⁺², Li⁺, Na⁺, K⁺, NH₄⁺ and combinations thereof.

23. A biologically active, aggregation-stabilized erythropoietin of claim 20 wherein the salting-out salt is ammonium sulfate.