Method for producing sustained-release formulations

Claim 1

A method of making a microparticle comprising: (a) providing a low salt-containing aqueous polypeptide composition at a pH about 5.5 or greater comprising biologically active human insulin-like growth factor-1 (IGF-1) or biologically active analog thereof, wherein said IGF-1 analog has at least 80% amino acid sequence identity to the amino acid sequence of human IGF-1, and wherein said human IGF-1 or analog thereof is present in a concentration of 250 mg/ml to 500 mg/ml; (b) adding said low salt-containing polypeptide composition to a solution of a polymer comprising a poly(α-hydroxy acid), wherein said polymer is present at a concentration of about 1%-30% in an organic solvent and further wherein said IGF-1 or analog thereof is present at 0.1% to about 40% (w/w) in the polymer/polypeptide solution; (c) emusifying the polymer/polypeptide solution to form an emulsion; (d) adding an emulsion stabilizer to the emulsion to form microparticles; (e) removing organic solvent from the stabilized emulsion; and (f) recovering the microparticles.

Claim 2

A method of making a microparticle comprising: (a) providing a low salt-containing aqueous polypeptide composition at a pH about 5.5 or greater comprising biologically active human insulin-like growth factor-1 (IGF-1) or biologically active analog thereof, wherein said IGF-1 analog has at least 80% amino acid sequence identity to the amino acid sequence of human IGF-1, and wherein said human IGF-1 or analog thereof is present in a concentration of 250 mg/ml to 500 mg/ml; (b) adding said low salt-containing polypeptide composition to a solution of a polymer comprising a poly(α-hydroxy acid), wherein said polymer is present at a concentration of about 1%-30% in an organic solvent and further wherein said IGF-1 or analog thereof is present at 0.1% to about 40% (w/w) in the polymer/polypeptide solution; (c) emusifying the polymer/polypeptide solution to form an emulsion; (d) adding an emulsion stabilizer to the emulsion to form microparticles; (e) removing organic solvent from the stabilized emulsion; and (f) recovering the microparticles.

Claim 10

A method of making a microparticle comprising: (a) preparing a low salt-containing aqueous polypeptide composition at a pH about 5.5 or greater comprising biologically active human insulin-like growth factor-1 (IGF-1) or biologically active analog thereof, wherein said IGF-1 analog has at least 95% amino acid sequence identity to the amino acid sequence of human IGF-1, and wherein said human IGF-1 or analog thereof is present in a concentration of 250 mg/ml to 500 mg/ml; (b) cooling said low salt-containing polypeptide composition to a temperature of about 2° C. to about 8° C.; (c) adding said cooled polypeptide composition to a solution of a poly(α-hydroxy acid) polymer selected from the group consisting of poly(L-lactide), poly(D, L-lactide) and poly(D,L-lactide-co-glycolide), wherein said polymer is present at a concentration of about 5%-20% in methylene chloride and further wherein said IGF-1 or analog thereof is present at about 3% to about 20% (w/w) in the polymer/polypeptide solution; (d) emusifying the polymer/polypeptide solution to form an emulsion; (e) adding polyvinyl alcohol as an emulsion stabilizer to the emulsion to form microparticles; (f) removing methylene chloride from the stabilized emulsion; and (g) recovering the microparticles.

Claim 11

A method of making a microparticle comprising: (a) preparing a low salt-containing aqueous polypeptide composition at a pH about 5.5 or greater comprising biologically active human insulin-like growth factor-1 (IGF-1) or biologically active analog thereof, wherein said IGF-1 analog has at least 95% amino acid sequence identity to the amino acid sequence of human IGF-1, and wherein said human IGF-1 or analog thereof is present in a concentration of 250 mg/ml to 500 mg/ml; (b) cooling said low salt-containing polypeptide composition to a temperature of about 2° C. to about 8° C.; (c) adding said cooled polypeptide composition to a solution of a poly(α-hydroxy acid) polymer selected from the group consisting of poly(L-lactide), poly(D, L-lactide) and poly(D,L-lactide-co-glycolide), wherein said polymer is present at a concentration of about 5%-20% in methylene chloride and further wherein said IGF-1 or analog thereof is present at about 3% to about 20% (w/w) in the polymer/polypeptide solution; (d) emusifying the polymer/polypeptide solution to form an emulsion; (e) adding polyvinyl alcohol as an emulsion stabilizer to the emulsion to form microparticles; (f) removing methylene chloride from the stabilized emulsion; and (g) recovering the microparticles.

Claim 18

The method of any one of claims 9 , 14 or 15 , wherein said low salt-containing polypeptide composition in step (a) is present at a pH from about 5.5 to about 9.0.

Claim 19

A method of making a microparticle comprising: (a) preparing a low salt-containing aqueous polypeptide composition at a pH about 5.5 or greater comprising biologically active human insulin-like growth factor-1 (IGF-1) or biologically active analog thereof, wherein said IGF-1 analog has at least 80% amino acid sequence identity to the amino acid sequence of human IGF-1, and wherein said human IGF-1 or analog thereof is present in a concentration of 250 mg/ml to 500 mg/ml; (b) cooling said low salt-containing polypeptide composition to a temperature of about 2° C. to about 8° C.; (c) adding said cooled polypeptide composition to a solution of a poly(α-hydroxy acid) polymer selected from the group consisting of poly(L-lactide), poly(D, L-lactide) and poly(D,L-lactide-co-glycolide), wherein said polymer is present at a concentration of about 5%-20% in methylene chloride and further wherein said IGF-1 or analog thereof is present at about 3% to about 20% (w/w) in the polymer/polypeptide solution; (d) emusifying the polymer/polypeptide solution to form an emulsion; (e) adding polyvinyl alcohol as an emulsion stabilizer to the emulsion to form microparticles; (f) removing methylene chloride from the stabilized emulsion; and (g) recovering the microparticles.

Claim 20

A method of making a microparticle comprising: (a) preparing a low salt-containing aqueous polypeptide composition at a pH about 5.5 or greater comprising biologically active human insulin-like growth factor-1 (IGF-1) or biologically active analog thereof, wherein said IGF-1 analog has at least 80% amino acid sequence identity to the amino acid sequence of human IGF-1, and wherein said human IGF-1 or analog thereof is present in a concentration of 250 mg/ml to 500 mg/ml; (b) cooling said low salt-containing polypeptide composition to a temperature of about 2° C. to about 8° C.; (c) adding said cooled polypeptide composition to a solution of a poly(α-hydroxy acid) polymer selected from the group consisting of poly(L-lactide), poly(D, L-lactide) and poly(D,L-lactide-co-glycolide), wherein said polymer is present at a concentration of about 5%-20% in methylene chloride and further wherein said IGF-1 or analog thereof is present at about 3% to about 20% (w/w) in the polymer/polypeptide solution; (d) emusifying the polymer/polypeptide solution to form an emulsion; (e) adding polyvinyl alcohol as an emulsion stabilizer to the emulsion to form microparticles; (f) removing methylene chloride from the stabilized emulsion; and (g) recovering the microparticles.

Claim 27

The method of any one of claims 17 , 22 or 23 , wherein said low salt-containing polypeptide composition in step (a) is present at a pH from about 5.5 to about 9.0.

Claim 28

A method of making a microparticle comprising: (a) preparing a low salt-containing aqueous polypeptide composition at a pH about 5.5 or greater comprising biologically active human insulin-like growth factor-1 (IGF-1) or biologically active analog thereof, wherein said IGF-1 analog has at least 90% amino acid sequence identity to the amino acid sequence of human IGF-1, and wherein said human IGF-1 or analog thereof is present in a concentration of 250 mg/ml to 500 mg/ml; (b) cooling said low salt-containing polypeptide composition to a temperature of about 2° C. to about 8° C.; (c) adding said cooled polypeptide composition to a solution of a poly(α-hydroxy acid) polymer selected from the group consisting of poly(L-lactide), poly(D, L-lactide) and poly(D,L-lactide-co-glycolide), wherein said polymer is present at a concentration of about 5%-20% in methylene chloride and further wherein said IGF-1 or analog thereof is present at about 3% to about 20% (w/w) in the polymer/polypeptide solution; (d) emusifying the polymer/polypeptide solution to form an emulsion; (e) adding polyvinyl alcohol as an emulsion stabilizer to the emulsion to form microparticles; (f) removing methylene chloride from the stabilized emulsion; and (g) recovering the microparticles.

Claim 29

A method of making a microparticle comprising: (a) preparing a low salt-containing aqueous polypeptide composition at a pH about 5.5 or greater comprising biologically active human insulin-like growth factor-1 (IGF-1) or biologically active analog thereof, wherein said IGF-1 analog has at least 90% amino acid sequence identity to the amino acid sequence of human IGF-1, and wherein said human IGF-1 or analog thereof is present in a concentration of 250 mg/ml to 500 mg/ml; (b) cooling said low salt-containing polypeptide composition to a temperature of about 2° C. to about 8° C.; (c) adding said cooled polypeptide composition to a solution of a poly(α-hydroxy acid) polymer selected from the group consisting of poly(L-lactide), poly(D, L-lactide) and poly(D,L-lactide-co-glycolide), wherein said polymer is present at a concentration of about 5%-20% in methylene chloride and further wherein said IGF-1 or analog thereof is present at about 3% to about 20% (w/w) in the polymer/polypeptide solution; (d) emusifying the polymer/polypeptide solution to form an emulsion; (e) adding polyvinyl alcohol as an emulsion stabilizer to the emulsion to form microparticles; (f) removing methylene chloride from the stabilized emulsion; and (g) recovering the microparticles.

Claim 36

The method of any one of claims 25 , 30 or 31 , wherein said low salt-containing polypeptide composition in step (a) is present at a pH from about 5.5 to about 9.0.

Claim 37

A method of making a microparticle comprising: (a) preparing a low salt-containing aqueous polypeptide composition at a pH about 5.5 or greater comprising biologically active human insulin-like growth factor-1 (IGF-1) wherein said human IGF-1 is present in a concentration of 250 mg/ml to 500 mg/ml; (b) cooling said low salt-containing polypeptide composition to a temperature of about 2° C. to about 8° C.; (c) adding said cooled polypeptide composition to a solution of a poly(α-hydroxy acid) polymer selected from the group consisting of poly(L-lactide), poly(D, L-lactide) and poly(D,L-lactide-co-glycolide), wherein said polymer is present at a concentration of about 5%-20% in methylene chloride and further wherein said IGF-1 is present at about 3% to about 20% (w/w) in the polymer/IGF-1 solution; (d) emusifying the polymer/IGF-1 solution to form an emulsion; (e) adding polyvinyl alcohol as an emulsion stabilizer to the emulsion under conditions that allow microparticles to form; (f) removing organic solvent from the stabilized emulsion; and (g) recovering the microparticles.

Claim 38

A method of making a microparticle comprising: (a) preparing a low salt-containing aqueous polypeptide composition at a pH about 5.5 or greater comprising biologically active human insulin-like growth factor-1 (IGF-1) wherein said human IGF-1 is present in a concentration of 250 mg/ml to 500 mg/ml; (b) cooling said low salt-containing polypeptide composition to a temperature of about 2° C. to about 8° C.; (c) adding said cooled polypeptide composition to a solution of a poly(α-hydroxy acid) polymer selected from the group consisting of poly(L-lactide), poly(D, L-lactide) and poly(D,L-lactide-co-glycolide), wherein said polymer is present at a concentration of about 5%-20% in methylene chloride and further wherein said IGF-1 is present at about 3% to about 20% (w/w) in the polymer/IGF-1 solution; (d) emusifying the polymer/IGF-1 solution to form an emulsion; (e) adding polyvinyl alcohol as an emulsion stabilizer to the emulsion under conditions that allow microparticles to form; (f) removing organic solvent from the stabilized emulsion; and (g) recovering the microparticles.

Claim 39

A method of making a microparticle comprising: (a) preparing a low salt-containing aqueous polypeptide composition at a pH about 5.5 or greater comprising biologically active human insulin-like growth factor-1 (IGF-1) wherein said human IGF-1 is present in a concentration of 250 mg/ml to 500 mg/ml; (b) cooling said low salt-containing polypeptide composition to a temperature of about 4° C.; (c) adding said cooled polypeptide composition to a solution of a poly(D,L-lactide-co-glycolide) polymer, wherein said polymer is present at a concentration of about 5%-20% in methylene chloride and further wherein said IGF-1 is present at about 3% to about 20% (w/w) in the polymer/IGF-1 solution; (d) emusifying the polymer/IGF-1 solution to form an emulsion; (e) adding polyvinyl alcohol as an emulsion stabilizer to the emulsion under conditions that allow microparticles to form; (f) removing organic solvent from the stabilized emulsion; and (g) recovering the microparticles.

Claim 40

A method of making a microparticle comprising: (a) preparing a low salt-containing aqueous polypeptide composition at a pH about 5.5 or greater comprising biologically active human insulin-like growth factor-1 (IGF-1) wherein said human IGF-1 is present in a concentration of 250 mg/ml to 500 mg/ml; (b) cooling said low salt-containing polypeptide composition to a temperature of about 4° C.; (c) adding said cooled polypeptide composition to a solution of a poly(D,L-lactide-co-glycolide) polymer, wherein said polymer is present at a concentration of about 5%-20% in methylene chloride and further wherein said IGF-1 is present at about 3% to about 20% (w/w) in the polymer/IGF-1 solution; (d) emusifying the polymer/IGF-1 solution to form an emulsion; (e) adding polyvinyl alcohol as an emulsion stabilizer to the emulsion under conditions that allow microparticles to form; (f) removing organic solvent from the stabilized emulsion; and (g) recovering the microparticles.

Claim 41

The method of any one of claims 33 or 34 , wherein said IGF-1 present in step (c) is present at about 3% to about 4% (w/w).

Claim 42

The method of any one of claims 33 or 34 , wherein said IGF-1 present in step (c) is present at about 18% to about 20% (w/w).

Claim 43

The method of any one of claims 33 or 34 , wherein said low salt-containing polypeptide composition in step (a) is present at a pH from about 5.5 to about 9.0.

Claim 44

The method of any one of claims 33 or 34 wherein said low salt-containing polypeptide composition in step (a) comprises arginine or an arginine analog.