Functionalized biomaterials for tissue regeneration

• US 9,273,101 B2
• Filed: 03/14/2012
• Issued: 03/01/2016
• 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
• Primary Examiner(s): Ha, Julie
• Application Number:
  US14/005,442
• Publication Number:
  US 20140066387A1
• Time in Patent Office: 1,448 Days
• Field of Search: 514/21.4, 530/324, 530/326, 530/350
• US Class Current: 1/1
• 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

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 are described.

Claims

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

(i) a heptapeptide domain comprising the amino acid sequence of

-KLPGWSG- SEQ ID NO;

10;

(ii) a spacer region comprising one to seven glycine amino acid units; and

(iii) a backbone region comprising at least one amino acid repetition unit selected from the group consisting of LDLK (SEQ ID NO;

     91), KFEF (SEQ ID NO;

     92), RADA (SEQ ID NO;

     93), LDLD (SEQ ID NO;

     94), and LKLK (SEQ ID NO;

     95).

2. The self-assembling peptide according to claim 1, wherein said backbone region comprises at least one LDLK (SEQ ID NO:

91) amino acid repetition unit.

3. The self-assembling peptide according to claim 2, wherein said backbone region comprises three LDLK (SEQ ID NO:

91) amino acid repetitions.

4. The self-assembling peptide according to claim 1, wherein said backbone region comprises at least two KFEF (SEQ ID NO:

92) amino acid repetition units.

5. The self-assembling peptide according to claim 4, wherein said backbone region comprises three KFEF (SEQ ID NO:

92) amino acid repetitions.

6. The self-assembling peptide according to claim 1, wherein said backbone region comprises at least one RADA (SEQ ID NO:

93) amino acid repetition unit.

7. The self-assembling peptide according to claim 6, wherein said backbone region comprises three RADA (SEQ ID NO:

93) amino acid repetitions.

8. The self-assembling peptide according to claim 1, wherein said backbone region comprises at least one LDLD (SEQ ID NO:

94) amino acid repetition unit.

9. The self-assembling peptide according to claim 8, wherein said backbone region comprises three LDLD (SEQ ID NO:

94) amino acid repetitions.

10. The self-assembling peptide according to claim 1, wherein said backbone region comprises at least one LKLK (SEQ ID NO:

95) amino acid repetition unit.

11. The self-assembling peptide according to claim 10, wherein said backbone region comprises three LKLK (SEQ ID NO:

95) amino acid repetitions.

12. The self-assembling peptide according to claim 1, wherein said spacer region comprises three glycine amino acid units.

13. The self-assembling peptide according to claim 1, wherein said self-assembling peptide comprises a modified N-terminus.

14. The self-assembling peptide according to claim 13, wherein said N-terminus modification is an acetylation.

15. The self-assembling peptide according to claim 1, wherein said self-assembling peptide comprises a modified C-terminus.

16. The self-assembling peptide according to claim 15, wherein said C-terminus has a CONH₂ group.

17. The self-assembling peptide according to claim 1, wherein said self-assembling peptide comprises the amino acid sequence of X-KLPGWSGGGGLDLKLDLKLDLK (SEQ ID NO:

85)-Y;

wherein X is NH₂ or Ac, and wherein Y is CONH₂ or COOH.

18. The self-assembling peptide according to claim 17, comprising:

NH₂-KLPGWSGGGGLDLKLDLKLDLK-CONH₂ SEQ ID NO;

34.

19. The self-assembling peptide according to claim 17, comprising:

Ac-KLPGWSGGGGLDLKLDLKLDLK-CONH₂ SEQ ID NO;

35.

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

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

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

23. A complex interwoven membrane comprising at least two tabular nanofibres according to claim 22.

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

25. 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.

26. The method according to claim 25, wherein said tissue is a tissue of the central nervous system.

27. 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.

28. 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.