Self-Assembly of Peptide-Amphiphile Nanofibers under Physiological Conditions

• US 20080177033A1
• Filed: 09/28/2007
• Published: 07/24/2008
• Est. Priority Date: 02/15/2002
• Status: Active Grant

• Current Assignee: Elia Beniash, Jeffrey D. Hartgerink, Samuel I. Stupp
• Original Assignee: Elia Beniash, Jeffrey D. Hartgerink, Samuel I. Stupp
• Inventors: Stupp, Samuel I., Hartgerink, Jeffrey D., Beniash, Elia
• Granted Patent: US 7,745,708 B2
• Time in Patent Office: Days
• US Class Current: 530/328
• 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

1-33. -33. (canceled)

34. A method of making a cylindrical micellar nanofiber composed of beta-sheets comprising the steps of:

(a) providing a peptide amphiphile molecule in an aqueous solution comprising a hydrophobic or hydrocarbon segment covalently linked to a peptide segment; and

(b) subjecting said peptide amphiphile molecule to conditions that induce formation of a cylindrical micellar nanofiber composed of beta-sheets.

35. The method of claim 34, wherein said conditions comprise placing the aqueous solution on a surface and removing the aqueous component from said solution.

36. The method of claim 35, wherein said removal results from passive drying.

37. The method of claim 34, wherein said conditions comprise physiological conditions.

38. The method of claim 34, wherein said conditions comprise contacting the aqueous solution with a reagent that induces gelation of the peptide amphiphile molecule.

39. The method of claim 38, wherein said reagent comprises a bodily fluid.

40. The method of claim 38, wherein said reagent comprises a metal ion selected from the group consisting of Ca, Mg, Cd, Fe, and Zn divalent ions, Al, Fe, and Gd trivalent ions, and combination of said ions.

41. The method of claim 38, wherein said conditions further comprise the step of introducing an oxidizing agent that forms a disulfide bond between peptide amphiphile molecules.

42. The method of claim 41, wherein said conditions further comprise the step of introducing a reducing agent that cleaves said disulfide bond.

43. The method of claim 42, wherein said reducing agent comprises a hydroxide ion.

44. The method of claim 34, wherein said peptide amphiphile molecule has a net charge at substantially physiological pH.

45. The method of claim 34, wherein said peptide segment further comprises at least one phosphorylated residue.

46. The method of claim 45, wherein said phosphorylated residue is phosphoserine.

47. The method of claim 34, wherein said peptide segment further comprises a residue with a functional moiety capable of intermolecular covalent bond formation.

48. The method of claim 47, wherein said residue is cysteine.

49. The method of claim 48, wherein said peptide segment further comprises at least one phosphoserine residue.

50. The method of claim 49, wherein said peptide segment further comprises at least one glycine residue between said cysteine and phosphoserine residues.

51. The method of claim 34, wherein said peptide segment comprises a bioactive epitope sequence selected from RGD, KGE, IKVAV (SEQ ID NO:

1) and YIGSR (SEQ ID NO;

     20).

52. The method of claim 34, 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)

53. The method of claim 34, wherein said hydrophobic or hydrocarbon segment comprises an alkyl moiety composed of at least six carbon units.

54. The method of claim 34, wherein said hydrophobic or hydrocarbon segment comprises a C₆ to C₂₂ linear hydrocarbon.

55. The method of claim 34, wherein said hydrophobic or hydrocarbon segment is linked to the N-terminus of said peptide segment.

56. The method of claim 34, wherein said cylindrical micellar nanofiber has dimensional and functional characteristics mimetic of collagen fibrils.

57. The method of claim 34, wherein said cylindrical micellar nanofiber has a conical shape comprised of a narrow hydrophobic or hydrocarbon tail at one end and a bulkier peptide region at the other.

58. The method of claim 34, wherein said cylindrical micellar nanofiber displays a secondary structure that includes both beta-sheets and alpha-helices.