Stabilizing formulations

• US 20060115472A1
• Filed: 08/12/2005
• Published: 06/01/2006
• Est. Priority Date: 08/13/2004
• Status: Active Grant

• Current Assignee: Wyeth LLC (Pfizer Inc.)
• Original Assignee: Wyeth (Pfizer Inc.)
• Inventors: MacMillan, Shannon B., Kantor, Angela, Li, Li
• Granted Patent: US 8,871,201 B2
• Time in Patent Office: Days
• US Class Current: 424/133.100
• CPC Class Codes: A61K 9/2886 having two or more different drug-free coatings; Tablets of the type inert core-drug layer-inactive layer of the type active core-drug layer-inactive layer A61K9/209
• CPC Class Codes: A61P 35/00 Antineoplastic agents
• CPC Class Codes: C07K 16/065 Purification, fragmentation
• CPC Class Codes: C07K 16/2803 against the immunoglobulin superfamily
• CPC Class Codes: C07K 16/2896 against molecules with a "CD"-designation, not provided for elsewhere
• CPC Class Codes: C07K 16/30 from tumour cells
• CPC Class Codes: C07K 2317/24 containing regions, domains or residues from different species, e.g. chimeric, humanized or veneered

Abstract

Formulations are described that are suitable for storage of proteins, such as antibodies, over a relatively broad range of protein concentrations, pH, and buffer types. Also described are methods of storing a protein and methods of identifying a suitable formulation for storage of a specific protein. In general, a formulation contains low to no surfactant, no to relatively low salt concentrations, and requires a relatively low buffer concentration.

Claims

1. A formulation comprising

(a) an isolated protein;

and

(b) an aqueous solution having pH 4.0 to pH 8.0,

wherein the formulation does not contain a cryoprotectant or surfactant, and the protein is stable for at least 3 weeks at −

80°

C. to 8°

C.

2. The formulation of claim 1, wherein the formulation comprises less that 150 mM salt.

3. The formulation of claim 1, wherein the protein is an antibody.

4. The formulation of claim 1, wherein the protein is a polyclonal antibody.

5. The formulation of claim 1, wherein the protein is a monoclonal antibody.

6. The formulation of claim 1, wherein the protein is an anti-CD22 antibody, an anti-Lewis Y antibody or an anti-5T4 antibody.

7. The formulation of claim 1, wherein the protein is a recombinant protein.

8. The formulation of claim 1, wherein the protein is a therapeutic protein.

9. The formulation of claim 1, wherein the aqueous solution is sterile water.

10. The formulation of claim 1, wherein the aqueous solution further comprises a buffer.

11. The formulation of claim 10, wherein the buffer is an acetate buffer, succinate buffer, phosphate buffer, or citrate buffer.

12. The formulation of claim 10, wherein the buffer concentration is about 0 mM to 150 mM.

13. The formulation of claim 10, wherein the buffer concentration is about 50 mM to 100 mM.

14. The formulation of claim 10, wherein the buffer concentration is about 1 mM to 50 mM.

15. The formulation of claim 1, wherein the protein is stable for at least 1 year at 5°

C. or the protein is stable for at least 3 years at 5°

C.

16. The formulation of claim 1, wherein the protein is present at a concentration of from about 1 mg/ml to about 200 mg/ml, about 1 mg/ml to about 100 mg/ml, about 20 mg/ml to about 200 mg/ml, or about 20 mg/ml to about 100 mg/ml.

17. The formulation of claim 1, wherein the protein is present at a concentration of from about 10 mg/ml to about 30 mg/ml, 25 mg/ml to about 30 mg/ml, about 10 mg/ml to about 50 mg/ml, or about 20 mg/ml to about 50 mg/ml.

18. The formulation of claim 1, wherein the protein has a pI of at least 6.0, 7.0, or 8.0.

19. The formulation of claim 1, wherein the formulation is stored at about 0°

C. to about 8°

C. or from about 0°

C. to about 5°

C.

20. The formulation of claim 1, wherein the pH of the formulation is about pH 5.0 to about pH 6.0.

21. The formulation of claim 1, wherein the pH of the formulation is about pH 5.0 to about pH 6.0 and the formulation is stored at from about −

80°

C. to 5°

C.

22. The formulation of claim 1, wherein the protein is a humanized anti-CD22 antibody and aqueous solution is about 20 mM succinate, pH 6.0.

23. The formulation of claim 1, wherein the protein is an anti-Lewis Y antibody and the aqueous solution is about 10 mM Na citrate, pH 5.5 and about 75 mM NaCl.

24. The formulation of claim 1, wherein the protein is an anti-5T4 antibody and the aqueous solution is about 10 mM Na acetate, pH 5.5.

25. The formulation of claim 1, wherein stability is determined by assaying at least one of the percentage of high molecular weight species, the percentage of low molecular weight species, or the percentage of acidic species compared to a control.

26. The formulation of claim 1, wherein the protein is purified at least 95%.

27. The formulation of claim 1, wherein the formulation is sterile.

28. A method of storing a protein, the method comprising

(a) providing a formulation of claim 1;

(b) placing the formulation at a selected temperature; and

(c) maintaining the formulation at the selected temperature, wherein the protein in the formulation is stable for at least one week.

29. The method of claim 28, wherein the protein is stable for at least one month, three months, one year, five years, seven years, or ten years.

30. The method of claim 29, wherein the protein is stable at a temperature of about −

80°

C. or about 2°

C. to about 8°

C.

31. The method of claim 28, wherein the protein is stored at a temperature of about −

80°

C. and the protein is stable for at least five years, seven years, or ten years.

32. The method of claim 28, wherein the formulation is frozen at the selected temperature and after step (c), the protein is rapidly thawed.

33. The method of claim 28, wherein the formulation is frozen in (b), and the freezing is fast freezing.

34. The method of claim 28, wherein the protein is an antibody.

35. The method of claim 28, wherein the protein is a monoclonal antibody.

36. The method of claim 28, wherein the protein is an antibody an anti-CD22 antibody, an anti-Lewis Y antibody or an anti-5T4 antibody.

37. The method of claim 34, wherein the antibody is an intermediate substance.

38. The method of claim 28, wherein the protein concentration is at least 15 mg/ml, at least 20 mg/ml, or at least 30 mg/ml.

39. The method of claim 28, wherein upon recovery from storage, the protein has at least 70% of the activity of a reference.

40. The method of claim 28, wherein upon recovery from storage, the formulation comprises less than or equal to 5% high molecular weight products compared to a reference.

41. The method of claim 28, wherein upon recovery from storage, the formulation comprises less than or equal to 10% high molecular weight products and aggregates compared to a reference.

49. A polypeptide produced by a method comprising storage in a formulation of claim 1.

50. The method of claim 49, wherein the polypeptide is a protein.

51. The method of claim 49, wherein the protein is an antibody.

52. A polypeptide for use in manufacturing a modified protein, wherein the polypeptide is stored in a formulation of claim 1.

53. The polypeptide of claim 52, wherein the polypeptide is an antibody.

54. The polypeptide of claim 52, wherein the polypeptide is used for manufacturing a conjugated protein.

55. The polypeptide of claim 54, wherein the protein is conjugated to a toxin.

42. A method of determining a favorable formulation for storing a selected isolated protein, the method comprising

(a) providing a selected isolated protein;

(b) storing the selected isolated protein in a series of formulations comprising an aqueous solution between about pH 5.0 and about pH 8.0 and comprising between about 0 mM to 150 mM buffer;

(c) determining at least one parameter of stability; and

(d) identifying a formulation in which the protein is stable, wherein a formulation in which the protein is stable is a favorable formulation for storing the isolated selected protein.

43. The method of claim 42, wherein a decrease in stability is indicated by an increase in the amount of high molecular weight protein in the sample compared to a control.

44. The method of claim 43, wherein a condition is selected in which not more than 0.5%, 0.2%, or 0.1% of the protein is an high molecular weight species.

45. The method of claim 42, wherein the parameter of stability is activity assayed by enzyme-linked immunoassay and the activity is at least 50% of a control or is at least 80% of a control in a stable formulation.

46. The method of claim 42, wherein the parameter of stability is the presence of high molecular weight species and a stable formulation comprises not more than 5% high molecular weight species compared to a control.

47. The method of claim 42, wherein the parameter of stability is the presence of high molecular weight species and protein aggregates, and a stable formulation comprises not more than 10% high molecular weight species and protein aggregates or not more than 15% high molecular weight species and protein aggregates combined compared to a control.

48. The method of claim 42, wherein the parameter of stability is the ratio of acidic and basic proteins, and the ratio in a stable formulation is not more than 15% different from a control.