Self-assembly of peptide-amphiphile nanofibers under physiological conditions

• US 7,745,708 B2
• Filed: 09/28/2007
• Issued: 06/29/2010
• Est. Priority Date: 02/15/2002
• Status: Expired due to Fees

• Current Assignee: Northwestern University
• Original Assignee: Northwestern University
• Inventors: Stupp, Samuel I., Hartgerink, Jeffrey D., Beniash, Elia
• Primary Examiner(s): Desai; Anand U
• Assistant Examiner(s): LIU, SAMUEL W
• Application Number:
  US11/863,975
• Publication Number:
  US 20080177033A1
• Time in Patent Office: 1,005 Days
• Field of Search: None
• US Class Current: 424/278.1
• 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: C07K 14/78 Connective tissue peptides, e.g. collagen, elastin, laminin, fibronectin, vitronectin or cold insoluble globulin [CIG]
• CPC Class Codes: Y10S 977/705 Protein or peptide
• CPC Class Codes: Y10T 428/12014 having metal particles
• CPC Class Codes: Y10T 428/24413 Metal or metal compound

Abstract

Peptide amphiphile compounds, compositions and methods for self-assembly or nanofibrous network formation under neutral or physiological conditions.

Claims

1. A method of making a cylindrical micellar nanofiber composed of beta-sheets of peptide segment of peptide amphiphile comprising the steps of:

(a) providing the peptide amphiphile molecule in an aqueous solution, wherein said peptide amphiphile molecule comprises a single C₆ to C₂₂ linear hydrocarbon segment covalently linked to a peptide segment comprising a bioactive epitope sequence selected from RGD, KGE, IKVAV (SEQ ID NO;

     1) and YIGSR (SEQ ID NO;

     20); and

(b) contacting said peptide amphiphile molecule with an effective amount of a metal ion sufficient to induce formation of a cylindrical micellar nanofiber composed of beta-sheets.

2. The method of claim 1, wherein said contacting step occurs under physiological conditions.

3. The method of claim 1, wherein said metal ion is present in a bodily fluid.

4. The method of claim 1, wherein the metal ion selected from the group consisting of Ca²⁺, Mg²⁺, Cd²⁺, Fe²⁺ and Zn²⁺ divalent ions, Al³⁺, Fe³⁺, and Gd³⁺ trivalent ions, and combination of said ions.

5. The method of claim 1, wherein said peptide amphiphile molecule has a net charge at substantially physiological pH.

6. The method of claim 1, wherein said peptide segment further comprises at least one phosphorylated residue.

7. The method of claim 6, wherein said phosphorylated residue is phosphoserine.

8. The method of claim 1, wherein said peptide segment further comprises a residue with a functional moiety capable of intermolecular covalent bond formation.

9. The method of claim 8, wherein said residue is cysteine.

10. The method of claim 9, wherein said peptide segment further comprises at least one phosphoserine residue.

11. The method of claim 10, wherein said peptide segment further comprises at least one glycine residue between said cysteine and phosphoserine residues.

12. The method of claim 1, wherein said peptide segment comprises a first peptide sequence selected from the group consisting of CCCCGGGS(p) (SEQ ID NO:

3), CCCCGGGS (SEQ ID NO;

     26), CCCCGGGE (SEQ ID NO;

     27), AAAAGGGS(p) (SEQ ID NO;

     28), AAAAGGGS (SEQ ID NO;

     29), GGGS(p) (SEQ ID NO;

     30), and GGGS (SEQ ID NO;

     31) and a second peptide sequence selected from RGD, KGE, IKVAV (SEQ ID NO;

     1) and YIGSR (SEQ ID NO;

     20), wherein S(p) stands for phosphorylated serine residue.

13. The method of claim 1, wherein said hydrocarbon segment is linked to the N-terminus of said peptide segment.

14. The method of claim 1, wherein said cylindrical micellar nanofiber has dimensional and functional characteristics mimetic of collagen fibrils.

15. The method of claim 1, wherein said cylindrical micellar nanofiber forms a conical shape comprised of a narrow hydrocarbon tail at one end and a bulkier peptide region at the other.

16. The method of claim 1, wherein said cylindrical micellar nanofiber displays a secondary structure that includes both beta-sheets and alpha-helices.