BIOMATERIALS HAVING NANOSCALE LAYERS AND COATINGS

• US 20090214616A1
• Filed: 03/02/2007
• Published: 08/27/2009
• Est. Priority Date: 03/03/2006
• Status: Active Grant

• Current Assignee: Washington University In St Louis
• Original Assignee: Washington University In St Louis
• Inventors: Elbert, Donald, Kaneda, Megan, Wacker, Brad, Scott, Evan, Alford, Shannon
• Granted Patent: US 8,790,678 B2
• Time in Patent Office: Days
• US Class Current: 424/423
• CPC Class Codes: A61L 2300/254 Enzymes, proenzymes
• CPC Class Codes: A61L 2300/608 having two or more layers
• CPC Class Codes: A61L 2400/12 Nanosized materials, e.g. nanofibres, nanoparticles, nanowires, nanotubes; Nanostructured surfaces
• CPC Class Codes: A61L 2420/06 Coatings containing a mixture of two or more compounds
• CPC Class Codes: A61L 27/18 obtained otherwise than by reactions only involving carbon-to-carbon unsaturated bonds
• CPC Class Codes: A61L 27/34 Macromolecular materials
• CPC Class Codes: A61L 27/50 Materials characterised by their function or physical properties , e.g. injectable or lubricating compositions, shape-memory materials, surface modified materials
• CPC Class Codes: A61L 27/52 Hydrogels or hydrocolloids
• CPC Class Codes: A61L 27/54 Biologically active materials, e.g. therapeutic substances A61L27/227 takes precedence
• CPC Class Codes: A61L 31/10 Macromolecular materials
• CPC Class Codes: A61L 31/14 Materials characterised by their function or physical properties , e.g. injectable or lubricating compositions, shape-memory materials, surface modified materials
• CPC Class Codes: C08L 71/02 Polyalkylene oxides
• CPC Class Codes: C12Y 207/01091 Sphinganine kinase (2.7.1.91)

Abstract

The invention generally relates to substrates and surfaces having substrates. Generally speaking, the substrates may be thinly layered substrates, and the surfaces may comprise thinly layered substrates. Additionally, the substrates may comprise a multifunctional water soluble polymer and a lipoprotein and the surfaces may comprise a multifunctional water soluble polymer and a lipoprotein.

Claims

1. A substrate, the substrate having a plurality of cross-linked, distinctly-formed layers, the distinctly-formed layers comprising multifunctional, water soluble polymer layers and protein layers, the protein having at least one lipid binding site, each protein layer being disposed between two layers of the water soluble polymer, whereby each protein layer cross-links the water soluble polymer layers together to form the plurality of distinctly-formed layers.

2. The substrate of claim 1, wherein the substrate has a porous matrix with an average pore diameter from about 1 nm to 100 μ

m.

3. The substrate of claim 2, wherein the substrate is a hydrogel.

4. The substrate of claim 1, wherein the water soluble polymer is selected from the group consisting of a hydrophilic polymer, cellulose derivative, polysaccharide, and poly(amino acid).

5. The substrate of claim 1, wherein the protein is albumin or a high density lipoprotein particle comprising Apo A-1 and lipids.

6. The substrate of claim 1, wherein the protein has from about 2 to about 7 lipid binding sites.

7. The substrate of claim 1, wherein the protein is a lipoprotein selected from the group consisting of chylomicrons, very low density lipoproteins, intermediate density lipoproteins, low density lipoproteins, and high density lipoproteins.

8. The substrate of claim 4, wherein the water soluble polymer is polyethylene glycol.

9. The substrate of claim 1, further comprising a therapeutic molecule non covalently conjugated to the protein.

10. The substrate of claim 1, wherein the substrate further comprises a therapeutic molecule that converts an endogenous precursor form of a molecule to the active form of the endogenous molecule.

11. The substrate of claim 10, wherein the therapeutic molecule is an enzyme.

12. The substrate of claim 11, wherein the enzyme is a recombinant enzyme having an added GST tag.

13. The substrate of claim 11, wherein the enzyme is sphingosine kinase.

14. The substrate of claim 13, wherein the sphingosine kinase has an amino acid sequence of SEQ ID NO. 1 or SEQ ID NO:

2.

15. The substrate of claim 1, wherein the substrate further comprises a polypeptide that promotes adhesion of cells.

16. The substrate of claim 15, wherein the polypeptide comprises the amino acid sequence RGD.

17. The substrate of claim 1, wherein the protein is a recombinant protein having the amino acid sequence of a native protein, but with the addition of amino acid residues that react with the water soluble polymer to form cross-links between the protein layers and the water soluble polymer layers.

20. The substrate of claim 1, wherein the substrate is from about 90 percent to about 99 percent protein rejecting.

21. The substrate of claim 1, where the substrate has from about 4 to about 50 distinctly-formed layers.

22. The substrate of claim 9, wherein the substrate is contacted in vivo with a subject'"'"'s blood or tissue.

23. The substrate of claim 22, wherein less than about 90 percent of the therapeutic molecule is released from the substrate in about 24 hours after the substrate has been contacted in vivo with a subject'"'"'s blood or tissue.

24. The substrate of claim 22, wherein the therapeutic molecule is released from the substrate in a controlled release formulation such that at least some of the therapeutic molecule is released over a period of about thirty days after the substrate has been contacted in vivo with a subject'"'"'s blood or tissue.

25. The substrate of claim 1, wherein the substrate is used on a medical device selected from the group consisting of a cardiovascular device, an artificial blood vessel, an artificial bone joint, a scaffold that supports tissue growth, a biosensor, and a percutaneous device.

26. The substrate of claim 1, wherein the polymer layer comprising the polymer is reacted with an agent to form a reactive group disposed on the polymer selected from the group consisting of sulfones, sulfoxides, sulfonates, sulfonamides, phosphonates, and phosphonamides

18. The substrate of claim 18, wherein the added amino acid residue is cysteine.

19. The substrate of claim 18, wherein the recombinant protein has an amino acid sequence comprising SEQ ID NO. 3 or SEQ ID NO:

4.

27. A surface, the surface being coated with a plurality of cross-linked, distinctly-formed layers, the distinctly-formed layers comprising multifunctional, water soluble polymer layers and protein layers, the protein having at least one lipid binding site, each protein layer being disposed between two layers of the water soluble polymer, whereby each protein layer cross-links the water soluble polymer layers together to form the plurality of distinctly-formed layers.

28. The surface of claim 27, wherein the surface is a material selected from the group consisting of silver, gold, stainless steel, titanium, glass, silicon containing material, cadmium, palladium, platinum, iron, nickel, cobalt, iron oxide, titanium oxide, silicon oxide, and copper.

29. The surface of claim 27, wherein the coating on the surface is from about 1 nm to 100 μ

m in thickness.

30. The surface of claim 27, wherein the water soluble polymer is selected from the group consisting of a hydrophilic polymer, cellulose derivative, polysaccharide, and poly(amino acid).

31. The surface of claim 27, wherein the protein is albumin or a high density lipoprotein particle comprising Apo A-1 and lipids.

32. The surface of claim 27, wherein the protein has from about 2 to about 7 lipid binding sites.

33. The surface of claim 27, wherein the protein is a lipoprotein selected from the group consisting of chylomicrons, very low density lipoproteins, intermediate density lipoproteins, low density lipoproteins, and high density lipoproteins.

34. The surface of claim 27, wherein the water soluble polymer is polyethylene glycol.

35. The surface of claim 27, further comprising a therapeutic molecule conjugated to the protein.

36. The surface of claim 27, wherein the surface further comprises a therapeutic molecule that converts an endogenous precursor form of a molecule to the active form of the endogenous molecule.

37. The surface of claim 36, wherein the therapeutic molecule is an enzyme.

38. The surface of claim 37, wherein the enzyme is a recombinant enzyme having an added GST tag.

39. The surface of claim 36, wherein the enzyme is sphingosine kinase.

40. The surface of claim 36, wherein the sphingosine kinase has an amino acid sequence of SEQ ID NO:

1 or SEQ ID NO;

2.

41. The surface of claim 27, wherein the surface further comprises a polypeptide that promotes adhesion of cells.

42. The surface of claim 41, wherein the polypeptide comprises the amino acid sequence RGD.

43. The surface of claim 27, wherein the protein is a recombinant protein having the amino acid sequence of a native protein, but with the addition of amino acid residues that react with the water soluble polymer to form cross-links between the protein layers and the water soluble polymer layers.

44. The surface of claim 43, wherein the added amino acid residue is cysteine.

45. The surface of claim 43, wherein the recombinant protein has an amino acid sequence comprising SEQ ID NO:

3 or SEQ ID NO;

4.

46. The surface of claim 27, wherein the surface is from about 90 percent to about 99 percent protein rejecting.

47. The surface of claim 27, where the surface has from about 4 to about 50 distinctly-formed layers.

48. The surface of claim 27, wherein the surface is contacted in vivo with a subject'"'"'s blood or tissue.

49. The surface of claim 27, wherein the surface is a medical device selected from the group consisting of a vascular graft a cardiovascular device, an artificial blood vessel, an artificial bone joint, a scaffold that support tissue growth, a biosensor, and a percutaneous device.

50. The surface of claim 27, wherein the polymer layer comprising the polymer is reacted with an agent to form a reactive group disposed on the polymer selected from the group consisting of sulfones, sulfoxides, sulfonates, sulfonamides, phosphonates, and phosphonamides.

51. A substrate, the substrate comprising a multifunctional water soluble polymer layer and a recombinant high density lipoprotein, whereby the recombinant high density lipoprotein cross-links the water soluble polymer to form the substrate.

52. The substrate of claim 51, further comprising a therapeutic molecule conjugated to the recombinant high density lipoprotein.

53. The substrate of claim 51, wherein the substrate further comprises a therapeutic molecule that converts an endogenous precursor form of a molecule to the active form of the endogenous molecule.

54. The substrate of claim 53, wherein the therapeutic molecule is an enzyme.

55. The substrate of claim 54, wherein the enzyme is sphingosine kinase.

56. The substrate of claim 54, wherein the sphingosine kinase has an amino acid sequence of SEQ ID NO:

1 or SEQ ID NO;

2.

57. The substrate of claim 51, wherein the substrate further comprises a polypeptide that promotes adhesion of cells.

58. The substrate of claim 57, wherein the polypeptide is comprised of the amino acid sequence RGD.

59. The substrate of claim 51, wherein the substrate is used on a medical device selected from the group consisting of a vascular graft a cardiovascular device, an artificial blood vessel, an artificial bone joint, a scaffold that supports tissue growth, a biosensor, and a percutaneous device.

60. A surface, the surface being coated with a substrate comprising a multifunctional water soluble polymer layer and a recombinant high density lipoprotein, whereby the recombinant high density lipoprotein cross-links the water soluble polymer.

61. The surface of claim 60, further comprising a therapeutic molecule conjugated to the recombinant high density lipoprotein.

62. The surface of claim 60, wherein the surface further comprises a therapeutic molecule that converts an endogenous precursor form of a molecule to the active form of the endogenous molecule.

63. The surface of claim 60, wherein the therapeutic molecule is an enzyme.

64. The surface of claim 63, wherein the enzyme is sphingosine kinase.

65. The surface of claim 63, wherein the sphingosine kinase has an amino acid sequence of SEQ ID NO:

1 or SEQ ID NO;

2.

66. The surface of claim 60, wherein the surface further comprises a polypeptide that promotes adhesion of cells.

67. The surface of claim 66, wherein the polypeptide is comprised of the amino acid sequence RGD.

68. The surface of claim 60, wherein the surface is a medical device selected from the group consisting of a vascular graft a cardiovascular device, an artificial blood vessel, an artificial bone joint, a scaffold that supports tissue growth, a biosensor, and a percutaneous device.