FUNCTIONALIZED BIOMATERIALS FOR TISSUE REGENERATION

• US 20140066387A1
• Filed: 03/14/2012
• Published: 03/06/2014
• Est. Priority Date: 03/15/2011
• Status: Active Grant

• Current Assignee: Nanomed3D S.R.L.
• Original Assignee: Nanomed3D S.R.L.
• Inventors: Gelain, Fabrizio, Vescovi, Angelo Luigi
• Granted Patent: US 9,273,101 B2
• Time in Patent Office: Days
• US Class Current: 514/21.4
• CPC Class Codes: C07K 14/001 by chemical synthesis
• CPC Class Codes: C07K 14/78 Connective tissue peptides, e.g. collagen, elastin, laminin, fibronectin, vitronectin or cold insoluble globulin [CIG]
• CPC Class Codes: C07K 7/06 having 5 to 11 amino acids
• CPC Class Codes: C07K 7/08 having 12 to 20 amino acids gastrins C07K14/595; somatostatins C07K14/655; melanotropins C07K14/68
• CPC Class Codes: C12N 5/0018 Culture media for cell or tissue culture media for specific animal cell type C12N5/06
• CPC Class Codes: G01N 33/5005 involving human or animal cells immunoassay G01N33/56966; immunoassays of protozoa G01N33/56905; protozoa in screening assays C12Q1/025
• CPC Class Codes: Y10T 442/30 Woven fabric [i.e., woven strand or strip material]

Abstract

The present invention concerns the field of functionalized self-assembling peptides suitable for obtaining hydrogels for use in a wide range of applications in the biomedical field, such as for the development of biomaterials for regenerative medicine and basic science research.

Claims

1. A self-assembling peptide (SAP) comprising:

a heptapeptide domain selected from the group consisting of;

2. The self-assembling peptide according to claim 1, wherein said backbone region comprises at least one amino acid repetition unit selected from the group consisting of LDLK, KFEF, RADA, LDLD, and LKLK.

3. The self-assembling peptide according to claim 2, wherein said backbone region comprises at least one LDLK amino acid repetition unit.

4. The self-assembling peptide according to claim 3, wherein said backbone region comprises three LDLK amino acid repetitions.

5. The self-assembling peptide according to claim 2, wherein said backbone region comprises at least two KFEF amino acid repetition units.

6. The self-assembling peptide according to claim 5, wherein said backbone region comprises three KFEF amino acid repetitions.

7. The self-assembling peptide according to claim 2, wherein said backbone region comprises at least one RADA amino acid repetition unit.

8. The self-assembling peptide according to claim 7, wherein said backbone region comprises three RADA amino acid repetitions.

9. The self-assembling peptide according to claim 2, wherein said backbone region comprises at least one LDLD amino acid repetition unit.

10. The self-assembling peptide according to claim 9, wherein said backbone region comprises three LDLD amino acid repetitions.

11. The self-assembling peptide according to claim 2, wherein said backbone region comprises at least one LKLK amino acid repetition unit.

12. The self-assembling peptide according to claim 11, wherein said backbone region comprises three LKLK amino acid repetitions.

13. The self-assembling peptide according to claim 1, wherein said spacer region comprises from one to seven glycine aminoacid units.

14. The self-assembling peptide according to claim 13, wherein said spacer region comprises three glycine aminoacid units.

15. The self-assembling peptide according to claim 1, wherein said heptapetide domain is selected from the group consisting of SEQ ID NO. 1, 2, 3 and 10.

16. The self-assembling peptide according to claim 1, wherein said heptapetide domain is SEQ ID NO. 1.

17. The self-assembling peptide according to claim 1 wherein the N-terminus is modified.

18. The self-assembling peptide according to claim 17, wherein said N-terminus modification is an acetylation.

19. The self-assembling peptide according to claim 1, wherein the C-terminus is modified.

20. The self-assembling peptide according to claim 19, wherein said C-terminus has a CONH₂ group.

21. The self-assembling peptide according to claim 1, selected from the group consisting of:

22. The self-assembling peptide according to claim 21, comprising:

23. The self-assembling peptide according to claim 21, comprising:

24. The self-assembling peptide according to claim 21, comprising:

25. The self-assembling peptide according to claim 21, comprising:

26. The self-assembling peptide according to claim 21, comprising:

27. The self-assembling peptide according to claim 21, comprising:

28. The self-assembling peptide according to claim 21, comprising:

29. The self-assembling peptide according to claim 21, comprising:

30. A hydrogel comprising the self-assembling peptides according to claim 1 and a hydrogelating ingredient.

31. A self-assembling peptide polymer comprising at least two self-assembling peptides according to claim 1.

32. A tabular nanofiber comprising at least two self-assembling peptides according to claim 1.

33. A complex interwoven membrane comprising at least two tabular nanofibres according to claim 32

34. A medicament comprising a self-assembling peptide polymer comprising at least two self-assembling peptides according to claim 1.

35. A method for using a self-assembling peptide polymer comprising at least two self-assembling peptides according to claim 1 as a scaffold in tissue regeneration comprising seeding the scaffold with cells and culturing the cells with the scaffold.

36. The method according to claim 35, wherein said tissue is a tissue of the central nervous system.

37. A method for using the self-assembling peptide according to claim 1, for use as a biomaterial and as a cell culture substrate comprising seeding the biomaterial with cells and culturing the cells with the biomaterial.

42. A method for using the heptapeptide according to claim 1, as a marker for neural stem cells comprising characterizing and isolating the neural stem cells.

38. A method for the selection and identification of a least one SAP sequence, or heptapeptide for biomaterial functionalization, comprising the steps of:

(a) contacting a bacteriophage library expressing a plurality of peptides with a cell;

(b) removing unbound bacteriophages;

(c) identifying the peptide presented by the bound bacteriophages;

(d) linking the peptide identified in step (c) to a spacer region and to a backbone region.

39. The method according to claim 38, wherein said cell of step (a) is selected from the group consisting of neural stem cells, endothelial cells, neurons, osteobalsts, hemotopoietic stem cells, adipocytes, condrocytes, keratinocytes, liver progentior cells, hepatocytes, embryonic and somatic stem cells, colon stem cells, myocytes and cardiomiocytes.

40. The method according to claim 38, wherein said cell of step (a) is a neural stem cell.

41. The method according to claim 38, wherein said SAP sequence is selected from the group consisting of: