Use of proteins and peptides of the TGF-Beta superfamily for purification and therapeutic methods

• US 20080051340A1
• Filed: 06/01/2007
• Published: 02/28/2008
• Est. Priority Date: 06/02/2006
• Status: Active Grant

• Current Assignee: Wyeth (Pfizer Inc.)
• Original Assignee: Wyeth (Pfizer Inc.)
• Inventors: Brown, Christopher, Yaworsky, Paul, Zhang, Jimin, Lu, Zhijian, Liu, Wei, Shen, Emily Sheng-ming, Olland, Stephane
• Granted Patent: US 7,754,689 B2
• Time in Patent Office: Days
• US Class Current: 514/12
• CPC Class Codes: A61K 38/00 Medicinal preparations containing peptides peptides containing beta-lactam rings A61K31/00; cyclic dipeptides not having in their molecule any other peptide link than those which form their ring, e.g. piperazine-2,5-diones, A61K31/00; ergot alkaloids of the cyclic peptide type A61K31/48; containing macromolecular compounds having statistically distributed amino acid units A61K31/74; medicinal preparations containing antigens or antibodies A61K39/00; medicinal preparations characterised by the non-active ingredients, e.g. peptides as drug carriers, A61K47/00
• CPC Class Codes: A61P 17/02 for treating wounds, ulcers, burns, scars, keloids, or the like
• CPC Class Codes: C07K 1/22 Affinity chromatography or related techniques based upon selective absorption processes
• CPC Class Codes: C07K 14/475 Growth factors; Growth regulators
• CPC Class Codes: C07K 14/495 Transforming growth factor [TGF]
• CPC Class Codes: C07K 14/51 Bone morphogenetic factor; Osteogenins; Osteogenic factor; Bone-inducing factor

Abstract

Members of the TGF-β superfamily and peptide fragments based on member proteins are employed to purify solutions containing member proteins or as therapeutics.

Claims

1. Chromatography media, comprising:

a chromatography resin derivatized with a peptide that is at least 75% homologous to a portion of a protein that is a member of the TGF-β

superfamily.

2. The chromatography media of claim 1, wherein the member is a TGF-β

, a growth differentiation factor (GDF), a bone morphogenic protein, an activin, or an inhibin.

3. The chromatography media of claim 1, wherein the peptide is a substantially complete molecule of the protein.

4. The chromatography media of claim 1, wherein the peptide is at least 75% homologous to a portion of a finger-1 peptide from the member.

6. The chromatography media of claim 1, wherein the peptide is at least 85% homologous to a portion of a finger-1 peptide from the member.

8. The chromatography media of claim 1, wherein the peptide is at least 95% homologous to a portion of a finger-1 peptide from the member.

5. (canceled)

7. (canceled)

9. A method of purifying a sample including a growth factor that is a member of the TGF-β

superfamily, comprising;

providing a chromatography column containing a chromatography resin derivatized with a peptide that is at least 75% homologous to a portion of a protein that is a member of the TGF-β

superfamily;

loading the column with the sample under conditions in which the growth factor tends to aggregate; and

eluting the growth factor from the column under conditions in which the growth factor tends to solubilize.

10. The method of claim 9, wherein the peptide is not a portion of the growth factor.

11. The method of claim 9, wherein the member is a TGF-β

, a growth differentiation factor (GDF), a bone morphogenic protein, an activin, or an inhibin.

12. The method of claim 9, wherein the peptide is a substantially complete molecule of the protein.

13. The method of claim 9, wherein the peptide is at least 75% homologous to a portion of a finger-1 peptide from the member.

15. The method of claim 9, wherein the peptide is at least 85% homologous to a portion of a finger-1 peptide from the member.

17. The method of claim 9, wherein the peptide is at least 95% homologous to a portion of a finger-1 peptide from the member.

14. (canceled)

16. (canceled)

18-22. -22. (canceled)

23. A solution of bone morphogenic proteins in a solvent, comprising:

a predetermined concentration of a predetermined bone morphogenic protein; and

a predetermined concentration of a peptide that is at least 75% homologous to a portion of a finger-1 peptide of a member of the TGF-β

superfamily.

24. The solution of claim 23, wherein the member of the TGF-β

superfamily is not the predetermined bone morphogenic protein.

25. The solution of claim 23, wherein the member of the TGF-β

superfamily is the predetermined bone morphogenic protein.

26. The solution of claim 23, wherein the member is a TGF-β

, a growth differentiation factor (GDF), a bone morphogenic protein, an activin, or an inhibin.

28. The solution of claim 23, wherein the peptide is at least 85% homologous to the portion.

30. The solution of claim 23, wherein the peptide is at least 95% homologous to the portion.

27. (canceled)

29. (canceled)

31-38. -38. (canceled)

39. A composition, comprising:

a peptide that is at least 75% homologous to a portion of a finger-1 peptide of a member of the TGF-β

superfamily; and

a carrier.

40. The composition of claim 39, wherein the carrier is a solvent, a gel, a polymer, demineralized bone, a suture, a surgical mesh, a ceramic, a micelle, or any combination of the above.

41. The composition of claim 39, wherein the carrier is a buffer solution.

42. The composition of claim 39, wherein the carrier comprises collagen.

43. The composition of claim 39, wherein the carrier is a collagen sponge.

44. The composition of claim 39, wherein the carrier is a cellulose-based material.

45. The composition of claim 39, wherein the carrier comprises one or more of alkylcellulose (including hydroxyalkylcellulose), including methylcellulose, ethylcellulose, hydroxyethylcellulose, hydroxypropylcellulose, hydroxypropyl-methylcellulose, and carboxymethylcellulose.

46. The composition of claim 39, wherein the carrier comprises polyglyconate, hyaluronic acid, polylactic acid, poly(ethylene glycol), sodium alginate, poly(ethylene glycol), polyoxyethylene oxide, carboxyvinyl polymer, and poly(vinyl alcohol).

47. The composition of claim 39, wherein the peptide is stabilized by one or more of a conjugated poly(propylene glycol), a conjugated sialyl group, a conjugated polysialyl chain, incorporation of modified amino acids, incorporation of non-natural amino acids, interchain covalent links, intrachain covalent links, interchain non-covalent links, interchain non-covalent links, and a presentation enhancer.

48. The composition of claim 39, wherein the member is a TGF-β

, a growth differentiation factor (GDF), a bone morphogenic protein, an activin, or an inhibin.

49. The composition of claim 39, wherein the member is BMP-2, BMP-12, BMP-9, GDF-19, BMP-3, BMP-15, GDF-9, GDF-3, BMP-4, a TGF-β

, or GDF-8.

51. The composition of claim 39, wherein the peptide is at least 85% homologous to the portion.

53. The composition of claim 39, wherein the peptide is at least 95% homologous to the portion.

50. (canceled)

52. (canceled)

54-89. -89. (canceled)