Charged peptide-amphiphile solutions and self-assembled peptide nanofiber networks formed therefrom

• US 7,534,761 B1
• Filed: 08/21/2003
• Issued: 05/19/2009
• Est. Priority Date: 08/21/2002
• Status: Expired due to Fees

• Current Assignee: Northwestern University
• Original Assignee: Northwestern University
• Inventors: Anthony, Shawn G., Spoerke, Erik D., Niece, Krista L., Stupp, Samuel I.
• Primary Examiner(s): Tsang; Cecilia J
• Assistant Examiner(s): Cordero Garcia; Marcela M
• Application Number:
  US10/645,304
• Time in Patent Office: 2,098 Days
• Field of Search: None
• US Class Current: 514/1.1
• 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: B82Y 30/00 Nanotechnology for materials or surface science, e.g. nanocomposites
• 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 2319/00 Fusion polypeptide
• CPC Class Codes: C07K 7/08 having 12 to 20 amino acids gastrins C07K14/595; somatostatins C07K14/655; melanotropins C07K14/68
• CPC Class Codes: Y10S 977/795 Composed of biological material

Abstract

The present invention provides a system of self-assembling peptide amphiphiles with an absolute net charge of 3 or greater whose design and function may be patterned after proteins involved in vertebrate mineralization or other tissue forming processes. This molecular system preferably consists of a hydrophobic hydrocarbon tail attached to a relatively hydrophilic peptide sequence. Self-assembly of this peptide amphiphile may be induced through pH variation, divalent ion addition, or dehydration. Variations of structural peptide sequences in the peptide amphiphile may enable the assembled nanofibers to be reversibly cross-linked for more or less structural stability, or may allow for control of the rate of self-assembly.

Claims

1. A peptide amphiphile composition comprising:

a hydrophobic component having a single alkyl group greater than six carbons in length; and

a hydrophilic component selected from the group consisting of CCCCGGGSS*DS*D (SEQ ID NO;

1), AAAAGGGSS*DS*D (SEQ ID NO;

2), SLSLGGGSS*DS*D (SEQ ID NO;

3), CCCCGGGS*S*DS*D (SEQ ID NO;

4), AAAAGGGS*S*DS*D (SEQ ID NO;

5), SLSLGGGS*S*DS*D (SEQ ID NO;

6), CCCCGGGDSS*DS* (SEQ ID NO;

7), AAAAGGGDSS*DS* (SEQ ID NO;

8), SLSLGGGDSS*DS* (SEQ ID NO;

9), CCCCGGGDS*S*DS* (SEQ ID NO;

10), AAAAGGGDS*S*DS* (SEQ ID NO;

11), SLSLGGGDS*S*DS* (SEQ ID NO;

12), CCCCGGGSDS*DS* (SEQ ID NO;

13), AAAAGGGSDS*DS* (SEQ ID NO;

14), SLSLGGGSDS*DS* (SEQ ID NO;

15), CCCCGGGS*DS*DS* (SEQ ID NO;

16), AAAAGGGS*DS*DS* (SEQ ID NO;

17), SLSLGGGS*DS*DS* (SEQ ID NO;

18), CCCCGGGDS*DS*D (SEQ ID NO;

19), AAAAGGGDS*DS*D (SEQ ID NO;

20), and SLSLGGGDS*DS*D (SEQ ID NO;

21), wherein S* is phosphorylated serine,

wherein said hydrophilic component is covalently bonded to said hydrophobic component in said peptide amphiphile, said hydrophilic component having a net charge at physiological pH, and wherein said peptide amphiphile self-assembles to form a cylindrical micelle.

2. The peptide-amphiphile compositions of claim 1, wherein the net charge on the peptide amphiphile is positive.

3. The peptide-amphiphile compositions of claim 1, wherein the net charge on the peptide amphiphile is negative.

4. The composition of claim 3, wherein the negative net charge on the peptide amphiphile is from −

4 to −

7.

5. The composition of claim 3, wherein the negative net charge on the peptide amphiphile is −

7 or more negative.

6. The composition of claim 1, wherein the peptide component of said peptide-amphiphile includes residue with a functional moiety capable of intermolecular covalent bond formation.

7. The composition of claim 6, wherein said residue is cysteine.

19. A composition comprising

one or more nanofibers formed from charged self-assembled peptide amphiphiles of claim 1, said peptide amphiphiles having a hydrophobic segment covalently bonded to a hydrophilic segment, said peptide amphiphile having a net absolute charge greater than 3 at substantially physiological pH.

20. The composition of claim 19 further including a substrate, said nanofibers covering at least a portion of said substrate.

21. The composition of claim 19 further including osteoblastic cells on said nanofibers.

22. The composition of claim 19 further including a crystalline material having a crystal axis preferentially oriented with respect to the length of said nanofiber.

23. The composition of claim 19 further including osteoblastic cells and a mineral on said nanofibers.

24. The composition of claim 19 wherein said nanofibers are preferentially oriented on at least a portion of the substrate.

25. A method of stimulating mineralization in a patient with tissue engineered material comprised of:

administering a peptide amphiphile composition of claim 1 to a site on said patient in need thereof,

said peptide amphiphile capable of stimulating mineralization of said site, said peptide amphiphile compositions having a net charge at physiological pH.

26. The method of claim 25, wherein said net charge on the peptide amphiphile is positive.

27. The method of claim 25, wherein said net charge on the peptide amphiphile is negative.

28. The method of claim 27, wherein the negative net charge on the peptide amphiphile is −

4 or more negative.

29. The method of claim 27, further comprising the step of adding an agent to induce self assembly of said peptide amphiphiles at said site.

30. The method of claim 25, wherein peptide-amphiphile includes and amino acid selected from the group consisting of serine, phosphorylated serine, and aspartic acid.

31. The method of claim 25, wherein the peptide-amphiphile includes a residue with a functional moiety capable of intermolecular covalent bond formation.

32. The method of claim 31, wherein the functional moiety is cysteine.

8. A peptide-amphiphile compound comprising:

an alkyl tail greater than six carbons in length covalently bonded to a hydrophilic component selected from the group consisting of CCCCGGGSS*DS*D (SEQ ID NO;

1), AAAAGGGSS*DS*D (SEQ ID NO;

2), SLSLGGGSS*DS*D (SEQ ID NO;

3), CCCCGGGS*S*DS*D (SEQ ID NO;

4), AAAAGGGS*S*DS*D (SEQ ID NO;

5), SLSLGGGS*S*DS*D (SEQ ID NO;

6), CCCCGGGDSS*DS* (SEQ ID NO;

7), AAAAGGGDSS*DS* (SEQ ID NO;

8), SLSLGGGDSS*DS* (SEQ ID NO;

9), CCCCGGGDS*S*DS* (SEQ ID NO;

10), AAAAGGGDS*S*DS* (SEQ ID NO;

11), SLSLGGGDS*S*DS* (SEQ ID NO;

12), CCCCGGGSDS*DS* (SEQ ID NO;

13), AAAAGGGSDS*DS* (SEQ ID NO;

14), SLSLGGGSDS*DS* (SEQ ID NO;

15), CCCCGGGS*DS*DS* (SEQ ID NO;

16), AAAAGGGS*DS*DS* (SEQ ID NO;

17), SLSLGGGS*DS*DS* (SEQ ID NO;

18), CCCCGGGDS*DS*D (SEQ ID NO;

19), AAAAGGGDS*DS*D (SEQ ID NO;

20), and SLSLGGGDS*DS*D (SEQ ID NO;

21), wherein S* is phosphorylated serine,

wherein the peptide-amphiphile compound self-assembles to form a cylindrical micelle.

9. The peptide-amphiphile compound of claim 8, wherein said peptide amphiphile has a positive net charge.

10. The peptide-amphiphile compound of claim 8, wherein said peptide amphiphile has a negative net charge.

11. The compound of claim 10, wherein the negative net charge on the peptide amphiphile is from −

4 to −

7.

12. The compound of claim 10, wherein the negative net charge on the peptide amphiphile is more negative than −

7.

13. The compound of claim 10, wherein the structural peptide includes a residue with a functional moiety capable of intermolecular covalent bond formation.

14. A composition comprising:

an aqueous solution of at least one charged peptide amphiphile, said charged peptide amphiphile having a hydrophobic segment consisting of having a single alkyl group greater than six carbons in length and a hydrophilic component selected from the group consisting of CCCCGGGSS*DS*D (SEQ ID NO;

1), AAAAGGGSS*DS*D (SEQ ID NO;

2), SLSLGGGSS*DS*D (SEQ ID NO;

3), CCCCGGGS*S*DS*D (SEQ ID NO;

4), AAAAGGGS*S*DS*D (SEQ ID NO;

5), SLSLGGGS*S*DS*D (SEQ ID NO;

6), CCCCGGGDSS*DS* (SEQ ID NO;

7), AAAAGGGDSS*DS* (SEQ ID NO;

8), SLSLGGGDSS*DS* (SEQ ID NO;

9), CCCCGGGDS*S*DS* (SEQ ID NO;

10), AAAAGGGDS*S*DS* (SEQ ID NO;

11) SLSLGGGDS*S*DS* (SEQ ID NO;

12), CCCCGGGSDS*DS* (SEQ ID NO;

13), AAAAGGGSDS*DS* (SEQ ID NO;

14), SLSLGGGSDS*DS* (SEQ ID NO;

15), CCCCGGGS*DS*DS* (SEQ ID NO;

16), AAAAGGGS*DS*DS* (SEQ ID NO;

17), SLSLGGGS*DS*DS* (SEQ ID NO;

18), CCCCGGGDS*DS*D (SEQ ID NO;

19), AAAAGGGDS*DS*D (SEQ ID NO;

20), and SLSLGGGDS*DS*D (SEQ ID NO;

21), wherein S* is phosphorylated serine, wherein said hydrophilic component is covalently bonded to said hydrophobic component in said peptide amphiphile, said peptide amphiphile having a net charge at substantially physiological pH; and

an agent for inducing said charged peptide amphiphiles to self assemble into a cylindrical micelle.

15. The composition of claim 14, wherein the net charge of said peptide amphiphile is positive.

16. The composition of claim 14, wherein the net charge of said peptide amphiphile is negative.

17. The composition of claim 14 wherein the agent includes solvent removal from the peptide amphiphile solution.

18. The composition of claim 16, wherein the agent inducing self assembly is chosen from the group consisting of oppositely charged peptide amphiphiles, cations, and anions.