Formulations comprising antisense nucleotides to connexins
View Patent ↗A therapeutic and/or cosmetic formulation comprising at least one anti-sense polynucleotide to a connexin protein together with a pharmaceutically acceptable carrier or vehicle is useful in site specific down regulation of connexin protein expression, particularly in reduction of neuronal cells death, wound healing, reduction of inflammation, decrease of scar formation and skin rejuvenation and thickening.
1. A method of reducing migration of neutrophils to a wound on or within a human subject which comprises administering to said wound an effective amount of a connexin 43 antisense polynucleotide.
2. A method of decreasing neutrophils in a wound in a subject which comprises administering to a wound in or on within the subject an effective amount of a connexin 43 antisense polynucleotide.
3. A method according to claim 2 wherein the wound is on or within a tissue which is or has been subjected to physical trauma.
4. A method according to claim 1 or 2 wherein inflammation is decreased.
5. A method according to claim 1 or 2 wherein said polynucleotide is administered in an amount effective to decrease connexin 43 expression for at least about 4 to about 8 hours.
6. A method according to claim 1 wherein said polynucleotide is administered in an amount effective to reduce migration of neutrophils for at least about 4 to about 8 hours.
7. A method according to claim 2 wherein said polynucleotide is administered in an amount effective to reduce migration of neutrophils for at least about 4 to about 8 hours.
8. A method according to claim 2 wherein said polynucleotide is administered in an amount effective to decrease neutrophils for about 4 to about 8 hours.
9. A method according to claim 3 wherein said polynucleotide is administered in an amount effective to decrease neutrophils for about 4 to about 8 hours.
10. A method according to claim 3 wherein the trauma is a burn.
11. A method according to claim 3 wherein the tissue is skin.
12. A method according to claim 11 wherein said trauma is an incision or due to an incision.
13. A method according to claim 3 wherein the tissue is brain, spinal cord or optic nerve.
14. A method according to claim 6 or 8 wherein the wound is a burn.
15. A method according to claim 6 or 8 wherein the wound is a skin wound.
16. A method according to claim 3 wherein said wound on or within a tissue which is or has been subjected to physical trauma is an incision or due to an incision.
17. A method according to claim 3 wherein the tissue is brain, spinal cord or optic nerve.
18. A method according to claim 3 wherein the physical trauma is due to surgery.
19. A method according to claim 18 wherein said wound is an incision.
20. A method according to claim 3 wherein the wound on or within a tissue which is or has been subjected physical trauma is due to surgery.
21. A method according to claim 20 wherein said physical trauma is an incision.
22. A method according to claim 3 wherein said physical trauma is an incision or due to an incision.
23. A method according to claim 1 or 2 wherein said polynucleotide is an oligodeoxynucleotide.
24. A method according to claim 1 or 2 wherein said polynucleotide has from about 6 to about 40 nucleotides.
25. A method according to claim 24 wherein said polynucleotide has about 30 nucleotides.
26. A method according to claim 24 wherein said polynucleotide is selected from the group consisting of SEQ ID NOS: 1 to 3.
27. A method according to claim 24 wherein said polynucleotide is an unmodified phosphodiester oligomer.
28. A method according to claim 24 wherein said polynucleotide comprises SEQ ID NO:2.
29. A method according to claim 28 wherein said polynucleotide is an unmodified phosphodiester oligomer.
30. A method according to claim 28 wherein said polynucleotide is chemically modified.
31. A method according to claim 24 wherein said polynucleotide is a mixed backbone oligonucleotide.
32. A method according to claim 24 wherein said connexin antisense polynucleotide has at least about 70 percent homology to an antisense sequence of human connexin 43 mRNA.
33. A method according to claim 32 wherein said polynucleotide has at least about 80 percent homology.
34. A method according to claim 32 wherein said polynucleotide has at least about 90 percent homology.
35. A method according to claim 32 wherein said polynucleotide has at least about 95 percent homology.
36. A method according to claim 32 wherein said polynucleotide has at least about 97 percent homology.
37. A method according to claim 24 wherein said connexin antisense polynucleotide has at least about 70 percent homology to SEQ ID NO: 2.
38. A method according to claim 37 wherein said polynucleotide has at least about 80 percent homology.
39. A method according to claim 37 wherein said polynucleotide has at least about 90 percent homology.
40. A method according to claim 37 wherein said polynucleotide has at least about 95 percent homology.
41. A method according to claim 37 wherein said polynucleotide has at least about 97 percent homology.
42. A method according to claim 24 wherein said connexin 43 antisense polynucleotide is sufficiently complementary to connexin 43 mRNA to form a duplex having a melting point of greater than 20° C. under physiological conditions.
43. A method according to claim 42 wherein said duplex has a melting point of greater than 30° C.
44. A method according to claim 42 wherein said duplex has a melting point of greater than 40° C.
45. A method according to claim 24 wherein said connexin 43 antisense polynucleotide hybridizes to connexin 43 mRNA under conditions of the medium to high stringency.
46. A method according to claim 24 wherein said connexin 43 antisense polynucleotide hybridizes to connexin mRNA under conditions of medium to high stringency which comprise 0.03 M sodium chloride and 0.03 M sodium citrate at from about 50° C. to about 60° C.
47. A method according to claim 28 wherein the wound is a burn.
48. A method according to claim 28 wherein the wound is a skin wound.
49. A method according to claim 48 wherein said wound is an incision or due to an incision.
50. A method according to claim 28 wherein the wound is due to surgery.
51. A method according to claim 50 wherein said wound is an incision.
52. A method according to claim 28 wherein said wound is an incision or due to an incision.
53. A method according to claim 28 wherein the wound is on or within brain.
54. A method according to claim 28 wherein the wound is or within spinal cord.
55. A method according to claim 28 wherein the wound is or within optic nerve.
56. A method according to claim 39 wherein the wound is a burn.
57. A method according to claim 39 wherein the wound is a skin wound.
58. A method according to claim 39 wherein said wound is an incision or due to an incision.
59. A method according to claim 37 wherein the wound is due to surgery.
60. A method according to claim 59 wherein said wound is an incision.
61. A method according to claim 39 wherein said wound is an incision or due to an incision.
62. A method according to claim 39 wherein the wound is on or within brain.
63. A method according to claim 39 wherein the wound is on or within spinal cord.
64. A method according to claim 34 wherein the wound is on or within optic nerve.
65. A method according to claim 1 or 2 wherein said polynucleotide is an unmodified phosphodiester oligomer which has about 30 nucleotides and which comprises SEQ ID NO: 2.
66. A method according to claim 65 wherein the wound site is on or within a tissue which is or has been subjected to physical trauma.
67. A method according to claim 66 wherein the trauma is a burn.
68. A method according to claim 66 wherein the physical trauma is to skin.
69. A method according to claim 68 wherein said physical trauma is an incision or due to an incision.
70. A method according to claim 66 wherein the physical trauma is due to surgery.
71. A method according to claim 70 wherein said wound is an incision.
72. A method according to claim 66 wherein said wound is an incision or due to an incision.
73. A method according to claim 66 wherein the tissue is brain.
74. A method according to claim 66 wherein the tissue is spinal cord.
75. A method according to claim 66 wherein the tissue is optic nerve.
76. A method according to claim 1 or 2 wherein the wound is a brain wound.
77. A method according to claim 1 or 2 wherein the wound is a spinal cord wound.
78. A method according to claim 1 or 2 wherein the wound is an optic nerve wound.
79. A method according to claim 1 or 2 wherein the wound is a burn.
80. A method according to claim 1 or 2 wherein the wound is a skin wound.
81. A method according to claim 80 wherein said wound is an incision or due to an incision.
82. A method according to claim 1 or 2 wherein the wound is due to surgery.
83. A method according to claim 82 wherein said wound is an incision.