Materials and methods for nerve repair

• US 20030072749A1
• Filed: 08/13/2002
• Published: 04/17/2003
• Est. Priority Date: 08/13/2001
• Status: Active Grant

• Current Assignee: University of Florida Research Foundation, Incorporated (State University System of Florida)
• Original Assignee: University of Florida Research Foundation, Incorporated (State University System of Florida)
• Inventors: Muir, David F.
• Granted Patent: US 7,851,447 B2
• Time in Patent Office: Days
• US Class Current: 424/94.62
• CPC Class Codes: A61K 2300/00 Mixtures or combinations of active ingredients, wherein at least one active ingredient is fully defined in groups A61K31/00 - A61K41/00
• CPC Class Codes: A61K 35/30 Nerves; Brain; Eyes; Corneal cells; Cerebrospinal fluid; Neuronal stem cells; Neuronal precursor cells; Glial cells; Oligodendrocytes; Schwann cells; Astroglia; Astrocytes; Choroid plexus; Spinal cord tissue
• CPC Class Codes: A61K 38/51 Lyases (4)
• CPC Class Codes: A61L 2430/32 for nerve reconstruction
• CPC Class Codes: A61L 27/227 Other specific proteins or polypeptides not covered by A61L27/222, A61L27/225 or A61L27/24
• CPC Class Codes: A61L 27/3604 characterised by the human or animal origin of the biological material, e.g. hair, fascia, fish scales, silk, shellac, pericardium, pleura, renal tissue, amniotic membrane, parenchymal tissue, fetal tissue, muscle tissue, fat tissue, enamel
• CPC Class Codes: A61L 27/3675 Nerve tissue, e.g. brain, spinal cord, nerves, dura mater
• CPC Class Codes: A61L 27/3683 subjected to a specific treatment prior to implantation, e.g. decellularising, demineralising, grinding, cellular disruption/non-collagenous protein removal, anti-calcification, crosslinking, supercritical fluid extraction, enzyme treatment
• CPC Class Codes: A61L 27/3687 characterised by the use of chemical agents in the treatment, e.g. specific enzymes, detergents, capping agents, crosslinkers, anticalcification agents
• CPC Class Codes: A61L 27/383 Nerve cells, e.g. dendritic cells, Schwann cells
• CPC Class Codes: A61L 27/3878 Nerve tissue, brain, spinal cord, nerves, dura mater
• CPC Class Codes: A61L 31/047 Other specific proteins or polypeptides not covered by A61L31/044 - A61L31/046
• CPC Class Codes: A61P 25/00 Drugs for disorders of the nervous system
• CPC Class Codes: A61P 25/02 for peripheral neuropathies
• CPC Class Codes: A61P 43/00 Drugs for specific purposes, not provided for in groups A61P1/00-A61P41/00

Abstract

The subject invention pertains to compositions and methods for promoting repair of damaged nerve tissue. The compositions and methods of the subject invention can be employed to restore the continuity of nerve interrupted by disease, traumatic events or surgical procedures. Compositions of the subject invention comprise one or more chondroitin sulfate proteoglycan (CSPG)-degrading enzymes that promote axonal penetration into damaged nerve tissue. The invention also concerns methods for promoting repair of damaged nerve tissue using the present compositions and nerve tissue treated according to such methods. The invention also pertains to kits for nerve repair.

Claims

1. A method for promoting repair of a damaged nerve, wherein said method comprises applying at least one chondroitin sulfate proteoglycan-degrading enzyme to the damaged nerve.

2. The method according to claim 1, wherein the chondroitin sulfate proteoglycan-degrading enzyme is selected from the group consisting of chondroitinase, hyalurondiase, and matrix metalloproteinase, or a combination thereof.

3. The method according to claim 1, wherein the chondroitin sulfate proteoglycan-degrading enzyme is selected from the group consisting of chondroitinase ABC, chondroitinase A, chondroitinase C, chondroitinase AC, hyaluronidase, matrix metalloproteinase-2, and matrix metalloproteinase-9, or a combination thereof.

4. The method according to claim 1, wherein the chondroitin sulfate proteoglycan-degrading enzyme is selected from the group consisting of chondroitinase ABC, chondroitinase A, chondroitinase C, and chondroitinase AC, or a combination thereof.

5. The method according to claim 1, wherein the chondroitin sulfate proteoglycan-degrading enzyme is a chondroitinase.

6. The method according to claim 1, wherein said method further comprises co-applying a tissue adhesive to the damaged nerve.

7. The method according to claim 6, wherein the tissue adhesive is a biological glue.

8. The method according to claim 6, wherein the tissue adhesive is selected from the group consisting of fibrin glue and platelet gel.

9. The method according to claim 1, wherein said method further comprises applying a biologically active agent to the damaged nerve.

10. The method according to claim 9, wherein the biologically active agent is a growth factor.

11. The method according to claim 10, wherein the growth factor is selected from the group consisting of nerve growth factor, fibroblast growth factor-1, fibroblast growth factor-2, epidermal growth factor, ciliary neurotrophic factor, brain-derived neurotrophic factor, neurotrophin-3, neurotrophin-4, neurotrophin-5, insulin-like growth factor-1, insulin-like growth factor-2, transforming growth factor, glial growth factor-2, vascular endothelial growth factor, granulocyte-macrophage colony stimulating factor, and lymphocyte infiltrating factor/cholinergic differentiating factor.

12. The method according to claim 1, wherein said method further comprises applying cells to the damaged nerve.

13. The method according to claim 12, wherein the cells are stem cells.

14. The method according to claim 12, wherein the cells are Schwann cells.

15. The method according to claim 1, wherein the chondroitin sulfate proteoglycan-degrading enzyme is applied to the damaged nerve at the site of damage or adjacent to the site of damage.

16. The method according to claim 1, wherein the nerve damage comprises a disruption in the continuity of at least one of the nerve sheaths of the damaged nerve selected from the group consisting of basal lamina, perineurium, and epineurium.

17. The method according to claim 1, wherein the nerve damage is a neurotmesis.

18. The method according to claim 1, wherein the nerve damage is a neurotmesis, wherein said method further comprises coapting the damaged nerve, and wherein the chondroitin sulfate proteoglycan-degrading enzyme is applied to the site of coaptation.

19. The method according to claim 1, wherein the damaged nerve is partially or completely severed, having a proximal end and a distal end, and wherein the chondroitin sulfate proteoglycan-degrading enzyme is applied to the proximal end of the damaged nerve, or to the distal end of the damaged nerve, or to both the proximal end and distal end of the damaged nerve.

20. The method according to claim 19, wherein said method further comprises applying a tissue adhesive to the proximal end of the damaged nerve, or to the distal end of the damaged nerve, or to both the proximal end and distal end of the damaged nerve.

21. The method according to claim 19, wherein said method further comprises suturing together the proximal end and distal end of the damaged nerve.

22. The method according to claim 1, wherein the chondroitin sulfate proteoglycan-degrading enzyme causes lysis of chondroitin-4-sulfate, chondroitin-6-sulfate, or both chondroitin-4-sulfate, and chondroitin-6-sulfate within the damaged nerve.

23. The method according to claim 1, wherein the damaged nerve is partially or completely severed, having a proximal end and a distal end, and wherein the chondroitin sulfate proteoglycan-degrading enzyme decreases the latency of axonal ingress into the distal end of the nerve.

24. The method according to claim 1, wherein the damaged nerve is partially or completely severed, having a proximal end and a distal end, and wherein the chondroitin sulfate proteoglycan-degrading enzyme increases the rate of axonal ingress into the distal end of the damaged nerve.

25. The method according to claim 1, wherein the chondroitin sulfate proteoglycan-degrading enzyme is applied topically to the damaged nerve.

26. The method according to claim 1, wherein the nerve damage is a neurotmesis, and wherein the chondroitin sulfate proteoglycan-degrading enzyme is applied topically to either side of the neurotmesis.

27. The method according to claim 1, wherein the chondroitin sulfate proteoglycan-degrading enzyme is injected into the damaged nerve.

28. The method according to claim 1, wherein the nerve damage is a neurotmesis, and wherein the chondroitin sulfate proteoglycan-degrading enzyme is injected into either side of the neurotmesis.

29. The method according to claim 1, wherein the chondroitin sulfate proteoglycan-degrading enzyme is applied with a pharmaceutically acceptable carrier.

30. The method according to claim 29, wherein the pharmaceutically acceptable carrier is selected from the group consisting of liquids, gels, foams, solids, and polymers.

31. The method according to claim 29, wherein the pharmaceutically acceptable carrier is selected from the group consisting of membranes, sponges, fibrous structures, powders, fleeces, and particles.

32. The method according to claim 29, wherein the pharmaceutically acceptable carrier is a controlled release system.

33. The method according to claim 29, wherein the pharmaceutically acceptable carrier is a surgical cuff, and wherein the surgical cuff at least partially encircles the damaged nerve when applied.

34. The method according to claim 1, wherein the chondroitin sulfate proteoglycan-degrading enzyme is a chondroitinase, and wherein the chondroitinase is applied to the damaged nerve within a concentration range from about 10 units/mL to about 1000 units/mL.

35. The method according to claim 34, wherein the chondroitin sulfate proteoglycan-degrading enzyme is a chondroitinase, and wherein the chondroitinase is applied to the damaged nerve within a concentration range from about 100 units/mL to about 500 units/mL.

36. The method according to claim 1, wherein the chondroitin sulfate proteoglycan-degrading enzyme is a matrix metalloprotinase, and wherein the matrix metalloproteinase is applied to the damaged nerve within a concentration range from about 0.1 μ

g/mL to about 100 μ

g/mL.

37. The method according to claim 36, wherein the chondroitin sulfate proteoglycan-degrading enzyme is a matrix metalloprotinase, and wherein the matrix metalloproteinase is applied to the damaged nerve within a concentration range from about 10 μ

g/mL to about 50 μ

g/mL.

38. The method according to claim 1, wherein the damaged nerve is a damaged peripheral nerve.

39. The method according to claim 1, wherein the chondroitin sulfate proteoglycan-degrading enzyme is a bacterial enzyme.

40. The method according to claim 1, wherein the chondroitin sulfate proteoglycan-degrading enzyme is a mammalian enzyme.

41. The method according to claim 1, wherein the chondroitin sulfate proteoglycan-degrading enzyme is a recombinant enzyme.

42. The method according to claim 1, wherein said application of chondroitin sulfate proteoglycan-degrading enzyme comprises administering a genetically modified host to the damaged nerve, wherein the host has been genetically modified with a polynucleotide encoding the chondroitin sulfate proteoglycan-degrading enzyme.

43. The method according to claim 42, wherein the genetically modified host is a cell.

44. The method according to claim 43, wherein the cell is autologous with the damaged nerve.

45. The method according to claim 43, wherein the cell is allogeneic with the damaged nerve.

46. The method according to claim 43, wherein the cell is xenogeneic with the damaged nerve.

47. The method according to claim 1, wherein the nerve damage was caused by disease.

48. The method according to claim 1, wherein the nerve damage was caused by trauma.

49. The method according to claim 1, wherein the nerve damage was caused by a surgical procedure.

50. The method according to claim 1, wherein the damaged nerve is mammalian nerve.

51. The method according to 50, wherein the damaged nerve is human.

52. A method for promoting repair of a damaged nerve that has been partially or completely severed, wherein the nerve has a proximal end and a distal end, said method comprising applying chondroitinase to the proximal end of the nerve, or to the distal end of the nerve, or to both the proximal and distal ends of the nerve.

53. The method according to claim 52, wherein said method further comprises applying a tissue adhesive to the proximal end of the nerve, or to the distal end of the nerve, or to both the proximal and distal ends of the nerve.

54. The method according to claim 52, wherein the chondroitinase and the tissue adhesive are applied within the same formulation.

55. The method according to claim 52, wherein the tissue adhesive is fibrin glue or platelet gel.

56. The method according to claim 52, wherein the chondroitinase is selected from the group consisting of chondroitinase ABC, chondroitinase A, chondroitinase C, and chondroitinase AC, or a combination thereof.

57. The method according to claim 52, wherein the nerve damage was caused by disease.

58. The method according to claim 52, wherein the nerve damage was caused by trauma.

59. The method according to claim 52, wherein the nerve damage was caused by a surgical procedure.

60. A pharmaceutical composition comprising at least one chondroitin sulfate proteoglycan-degrading enzyme and a pharmaceutically acceptable carrier.

61. The pharmaceutical composition of claim 60, wherein said chondroitin sulfate proteoglycan-degrading enzyme is selected from the group consisting of chondroitinase, hyalurondiase, and matrix metalloproteinase, or a combination thereof.

62. The pharmaceutical composition of claim 60, wherein said chondroitin sulfate proteoglycan-degrading enzyme is selected from the group consisting of chondroitinase ABC, chondroitinase A, chondroitinase C, chondroitinase AC, hyaluronidase, matrix metalloproteinase-2, and matrix metalloproteinase-9, or a combination thereof.

63. The pharmaceutical composition of claim 60, wherein said chondroitin sulfate proteoglycan-degrading enzyme is selected from the group consisting of chondroitinase ABC, chondroitinase A, chondroitinase C, and chondroitinase AC, or a combination thereof.

64. The pharmaceutical composition of claim 60, wherein said chondroitin sulfate proteoglycan-degrading enzyme is a chondroitinase.

65. The pharmaceutical composition of claim 60, wherein said chondroitin sulfate proteoglycan-degrading enzyme is a chondroitinase within a concentration range from about 10 units/mL to about 1000 units/mL.

66. The pharmaceutical composition of claim 60, wherein said chondroitin sulfate proteoglycan-degrading enzyme is a chondroitinase within a concentration range from about 100 units/mL to about 500 units/mL.

67. The pharmaceutical composition of claim 60, wherein said chondroitin sulfate proteoglycan-degrading enzyme is a matrix metalloproteinase within a concentration range from about 0.1 μ

g/mL to about 100 μ

g/mL.

68. The pharmaceutical composition of claim 60, wherein said chondroitin sulfate proteoglycan-degrading enzyme is a matrix metalloproteinase within a concentration range from about 10 μ

g/mL to about 50 μ

g/mL.

69. The pharmaceutical composition of claim 60, wherein said pharmaceutical composition further comprises a tissue adhesive.

70. The pharmaceutical composition of claim 69, wherein said tissue adhesive is a biological glue.

71. The pharmaceutical composition of claim 69, wherein said tissue adhesive is selected from the group consisting of fibrin glue and platelet gel.

72. The pharmaceutical composition of claim 60, wherein said pharmaceutically acceptable carrier is selected from the group consisting of solids, liquids, gels, creams, salves, and foams.

73. The pharmaceutical composition of claim 60, wherein said pharmaceutically acceptable carrier is a saline solution.

74. The pharmaceutical composition of claim 60, wherein said pharmaceutically acceptable carrier is a polymer.

75. The pharmaceutical composition of claim 60, wherein said pharmaceutically acceptable carrier is a controlled release system.

76. The pharmaceutical composition of claim 60, wherein said pharmaceutically acceptable carrier is a surgical cuff.

77. A pharmaceutical composition comprising at least one chondroitin sulfate proteoglycan-degrading enzyme and a tissue adhesive.

78. The pharmaceutical composition of claim 77, wherein said chondroitin sulfate proteoglycan-degrading enzyme is a chondroitinase.

79. The pharmaceutical composition of claim 77, wherein said tissue adhesive comprises fibrin glue.

80. The pharmaceutical composition of claim 77, wherein said tissue adhesive comprises platelet gel.

81. A surgical cuff to promote repair of a damaged nerve, wherein said cuff comprises at least one chondroitin sulfate proteoglycan-degrading enzyme and a sleeve to be applied to a nerve graft or damaged nerve.

82. The surgical cuff of claim 81, wherein said sleeve comprises a tube, and wherein said tube defines a lumen for insertion of a nerve graft or damaged nerve.

83. The surgical cuff of claim 82, wherein said tube has a slit extending longitudinally along the length of said tube.

84. The surgical cuff of claim 83, wherein said slit has first and second abutting edges.

85. The surgical cuff of claim 84, wherein said first abutting edge is in separable contact with said second abutting edge, and wherein said first and second abutting edges can be separated to expose said lumen.

86. The surgical cuff of claim 81, wherein said surgical cuff further comprises means for delivering said chondroitin sulfate proteoglycan-degrading enzyme to the damaged nerve.

87. The surgical cuff of claim 86, wherein said chondroitin sulfate proteoglycan-degrading enzyme delivering means comprises a polymer.

88. The surgical cuff of claim 87, wherein said chondroitin sulfate proteoglycan-degrading enzyme delivering means comprises an expression system for producing said chondroitin sulfate proteoglycan-degrading enzyme.

89. The surgical cuff of claim 88, wherein said expression system comprises a genetically modified cell, wherein said genetically modified cell comprises a polynucleotide encoding said chondroitin sulfate proteoglycan-degrading enzyme.

90. The surgical cuff of claim 81, wherein said sleeve is biodegradable.

91. The surgical cuff of claim 81, wherein said cuff further comprises living cells seeded upon said sleeve.

92. The surgical cuff of claim 91, wherein said living cells comprise Schwann cells.

93. The surgical cuff of claim 91, wherein said living cells are genetically modified with a polynucleotide encoding said chondroitin sulfate proteoglycan-degrading enzyme.

94. The surgical cuff of claim 81, wherein said sleeve is flexible.

95. The surgical cuff of claim 81, wherein said sleeve has a facing surface for contacting said nerve graft or said damaged nerve, and wherein said cuff further comprises a tissue adhesive on at least a portion of said facing surface.

96. The surgical cuff of claim 95, wherein said tissue adhesive comprises a biological glue.

97. The surgical cuff of claim 95, wherein said tissue adhesive comprises fibrin glue.

98. The surgical cuff of claim 95, wherein said tissue adhesive comprises platelet gel.

99. The surgical cuff of claim 81, wherein said chondroitin sulfate proteoglycan-degrading enzyme is selected from the group consisting of chondroitinase, hyalurondiase, and matrix metalloproteinase, or a combination thereof.

100. The surgical cuff of claim 81, wherein said chondroitin sulfate proteoglycan-degrading enzyme is selected from the group consisting of chondroitinase ABC, chondroitinase A, chondroitinase C, chondroitinase AC, hyaluronidase, matrix metalloproteinase-2, and matrix metalloproteinase-9, or a combination thereof.

101. The surgical cuff of claim 81, wherein said chondroitin sulfate proteoglycan-degrading enzyme is selected from the group consisting of chondroitinase ABC, chondroitinase A, chondroitinase C, and chondroitinase AC, or a combination thereof.

102. The surgical cuff of claim 81, wherein said chondroitin sulfate proteoglycan-degrading enzyme is a chondroitinase.

103. The surgical cuff of claim 81, wherein said sleeve comprises a sheet to be contacted with said nerve graft or said damaged nerve.

104. The surgical cuff of claim 103, wherein said sheet is a flat sheet.

105. The surgical cuff of claim 103, wherein said sheet is a self-curling sheet, and wherein said sheet automatically encircles said nerve graft or damaged nerve when contacted with said nerve graft or damaged nerve.

106. A kit promoting repair of damage nerve tissue, said kit comprising a first compartment comprising at least one chondroitin sulfate proteoglycan-degrading enzyme, and a second compartment comprising a tissue adhesive.

107. The kit of claim 106, wherein said chondroitin sulfate proteoglycan-degrading enzyme is selected from the group consisting of chondroitinase, hyalurondiase, and matrix metalloproteinase, or a combination thereof.

108. The kit of claim 106, wherein said chondroitin sulfate proteoglycan-degrading enzyme is a chondroitinase.

109. The kit of claim 106, wherein said tissue adhesive is selected from the group consisting of fibrin glue and platelet gel.

110. The kit of claim 106, wherein said kit further comprises a third compartment for mixing said chondroitin sulfate proteoglycan-degrading enzyme and said tissue adhesive.