Granted Patent B2

US 8,617,613 · App. 10/592,769 · Granted Dec 31, 2013

Abstract

Claims (38)

1. A method of preparing a composition for the sustained release of a polypeptide comprising the steps of:

a) forming a mixture by combining an aqueous phase comprising a sugar and a water soluble polypeptide with an oil phase comprising a biocompatible polymer and a solvent for the polymer;

b) forming a water-in-oil emulsion of the mixture from step a), wherein the inner emulsion droplet size is less than about 0.5 microns;

c) adding a coacervation agent to the mixture to form embryonic microparticles, wherein said coacervation agent is silicone oil added in an amount sufficient to achieve a silicone oil to polymer solvent ratio of from about 0.75:1 to about 2:1;

d) transferring the embryonic microparticles to a quench solvent to harden the microparticles;

e)collecting the hardened microparticles; and

f) drying the hardened microparticles.

2. The method of claim 1 , wherein the ratio of silicone oil to polymer solvent is about 1.5:1.

3. The method of claim 1 , wherein the polymer is present in the oil phase at about 10% w/v or less.

4. The method of claim 1 , wherein the 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 and blends and copolymers thereof.

5. The method of claim 1 , wherein the polypeptide is selected from the group consisting of: glucagon, glucagon-like peptides, exendins, agonists of glucagon like peptides, vasoactive intestinal peptide, immunoglobulins, antibodies, cytokines, interleukins, macrophage activating factors, interferons, erythropoietin, tumor necrosis factor, colony stimulating factors, insulin, enzymes, tumor suppressors, blood proteins, follicle stimulating hormone, growth hormone, adrenocorticotropic hormone, and luteinizing hormone releasing hormone, NGF, EGF, gastrin, GRH, defensin, enkephalins, and muteins, analogs, deletion and substitution variants and pharmaceutically acceptable salts thereof.

6. A method of preparing a composition for the sustained release of exendin-4 comprising the steps of:

a) forming a mixture by combining an aqueous phase comprising water soluble exendin-4 polypeptide and sucrose with an oil phase comprising a biocompatible 50:50 PLGA 4A polymer in methylene chloride;

b) forming a water-in-oil emulsion of the mixture from step a), herein the inner emulsion droplet size is less than about 0.5 microns;

c) adding a coacervation agent to the mixture to form embryonic microparticles, wherein said coacervation agent is silicone oil added in an amount sufficient to achieve a silicone oil to polymer solvent ratio of from about 0.75:1 to about 2:1, and wherein the silicone oil is added to the water-in-oil emulsion in from about 3 minutes to about 5 minutes and the coacervation mixture is held for less than or about 1 minute;

d) transferring the embryonic microparticles to a quench solvent to harden the microparticles, wherein the quench solvent is a heptane/ethanol mixture and the transfer time is less than or about 3 minutes;

e) collecting the hardened microparticles; and

f) drying the hardened microparticles.

7. The method of claim 6 , wherein the water-in-oil emulsion of step b) is formed by homogenization.

8. A method of preparing a composition for the sustained release of a polypeptide comprising the steps of:

a) forming a mixture by combining an aqueous phase comprising a sugar and a water soluble polypeptide with an oil phase comprising a biocompatible polymer and a solvent for the polymer, wherein the polypeptide has a concentration of about 30 g to about 45 g per 600 g water;

b) homogenizing the mixture of step a) to form a water-in-oil emulsion of the mixture, wherein an inner emulsion is formed having a droplet size of about 0.4 microns;

c) adding a coacervation agent to the mixture to form embryonic microparticles, wherein said coacervation agent is silicone oil added in an amount sufficient to achieve a silicone oil to polymer solvent ratio of from about 0.75:1 to about 2:1;

d) transferring the embryonic microparticles to a quench solvent to harden the microparticles;

e) collecting the hardened microparticles; and

f) drying the hardened microparticles.

9. The method of claim 1 , wherein the inner emulsion droplet size is less than or equal to about 0.4 microns,

10. The method of claim 9 , wherein the mixture of step a) is homogenized to form the water-in-oil emulsion of the mixture.

11. The method of claim 1 , wherein in step c), the silicone oil is added in an amount sufficient to achieve a silicone oil to polymer solvent ratio of from about 1:1 to about 1.5:1.

12. The method of claim 1 , wherein the quench solvent is a heptane/ethanol solvent mixture.

13. The method of claim 1 , wherein the total load of the sugar and the polypeptide in the composition is about 10% (w/w) or less.

14. The method of claim 1 , further comprising the step of g) sieving the hardened microparticles through a sieve.

15. The method of claim 6 , wherein the inner emulsion droplet size is about 0.2 to 0.4 microns.

16. The method of claim 6 , wherein in step c), the silicone oil is added in an amount sufficient to achieve a silicone oil to polymer solvent ratio of from about 1:1 to about 1.5:1.

17. The method of claim 8 , wherein in step c), the silicone oil is added in an amount sufficient to achieve a silicone oil to polymer solvent ratio of from about 1:1 to about 1.5:1.

18. The method of claim 1 , wherein the polypeptide is selected from a glucoregulatory peptide and pharmaceutically acceptable salts thereof.

19. The method of claim 18 , wherein the glucoregulatory peptide is selected from glucagon, glucagon-like peptides, exendins, agonists of glucagon-like peptides, vasoactive intestinal peptide, insulin, and growth hormone.

20. The method of claim 19 , wherein the glucoregulatory peptide is selected from GLP-1, GLP-2, exendin-3, exendin-4, and combinations thereof.

Assignments (9)