IMPROVED WOUND HEALING COMPOSITIONS AND TREATMENTS
This invention concerns improved methods, uses, and kits for treating chronic wounds through the administration of anti-connexin agents, particularly anti-connexin 43 antisense polynucleotides. The methods, uses, and kits of the invention are based on the surprising and unexpected discovery that chronic wounds that do not increase or decrease in size by more than a pre-determined amount during a pre-treatment phase are more amenable to successful treatment than wounds whose size varies outside the target range during the pre-treatment phase.
1 - 47 . (canceled)
48 . A method of treating a refractory wound, comprising:
a. measuring the size a skin wound upon initial presentation for treatment to obtain a first size measurement;
b. administering standard of care such as compression bandaging and/or off-loading to the wound;
c. measuring the size of the wound after 2-4 weeks of administering the standard of care to obtain a second size measurement;
d. determining that the second size indicator of the wound is within a predetermined range from about −30 to about +35% of the first size measurement, thereby identifying a refractory wound; and
e. administering to the refractory wound a pharmaceutical composition comprising a pharmaceutical composition comprising a pharmaceutical carrier suitable for topical administration of an anti-connexin polynucleotide to a connexin selected from connexin 26, connexin 30 and connexin 43.
49 . A method of treating a refractory venous leg ulcer, comprising administering to the ulcer a pharmaceutical carrier suitable for topical administration and about 3-30 mg/mL of an anti-connexin polynucleotide to a connexin selected from connexin 26, connexin 30 and connexin 43.
50 . An article of manufacture for use in treating a refractory wound, comprising a receptacle containing a composition comprising a pharmaceutical carrier suitable for topical administration having about 20-23% of a nonionic polyoxyethylene-polyoxypropylene and about 3-30 mg/mL of an anti-connexin oligodeoxynucleotide to a connexin selected from connexin 26, connexin 30 and connexin 43, and, and instructions for the treatment of the refractory wound, by topically administering the composition to or in the proximity of the wound.
51 . A method of detecting a refractory wound with an increased likelihood of complete closure following topical administration to the recalcitrant wound of a composition comprising a pharmaceutical carrier suitable for topical administration of an anti-connexin oligodeoxynucleotide to a connexin selected from connexin 26, connexin 30 and connexin 43, the method comprising:
a. measuring the size of a skin wound upon initial presentation for treatment, to obtain a first size measurement;
b. administering standard of care, such as compression bandaging and/or off-loading to the wound;
c. measuring the size of the skin wound after about 2-4 weeks of administering the standard of care, to obtain a second size measurement;
d. detecting that the second size measurement is within −30% to +35% of the first size measurement, thereby detecting a refractory wound having an increased likelihood of complete closure following topical administration to the recalcitrant wound of a pharmaceutical composition comprising a pharmaceutical carrier suitable for topical administration having about 20-23% of a nonionic polyoxyethylene-polyoxypropylene and about 3-30 mg/mL of an anti-connexin oligodeoxynucleotide to a connexin selected from connexin 26, connexin 30 and connexin 43; and
e. administering the pharmaceutical composition to the wound.
52 . The method of claim 48 , wherein measuring the first or second size indicator further comprises using planimetry, digitizing techniques, stereophotogrammetry, a ruler, wound tracing, a handheld laser scanner or a camera.
53 . A method according to claim 52 , wherein the refractory wound is selected from the group consisting of a venous ulcers, venous stasis ulcers, arterial ulcers, pressure ulcers, diabetic ulcers, diabetic foot ulcers, vasculitic ulcers, decubitus ulcers, burn ulcers, trauma-induced ulcers, infectious ulcers, mixed ulcers, and pyoderma gangrenosum.
54 . A method according to claim 48 , wherein the chronic wound is a venous leg ulcer.
55 . A method according to claim 48 , comprising a plurality of administrations of the pharmaceutical composition.
56 . A method according to claim 55 , wherein the pharmaceutical composition is applied repeatedly until wound closure is achieved.
57 . A method according to claim 56 , wherein the administrations are periodic.
58 . A method according to claim 57 , wherein the periodic administrations occur at regularly scheduled intervals, optionally, daily, every other day, twice weekly, weekly, twice monthly, and monthly.
59 . A method according to claim 48 , wherein the anti-connexin 43 agent is an anti-connexin 43 polynucleotide.
60 . A method according to claim 48 , wherein the anti-connexin 43 polynucleotide, optionally an oligodeoxynucleotide or modified oligodeoxynucleotide comprises from about 18 to about 32 nucleotides.
61 . A method according to claim 48 , wherein the subject is a mammal, optionally a human.
62 . The method of claim 48 , wherein the pharmaceutical composition comprises a pharmaceutically acceptable carrier and an anti-connexin 43 polynucleotide present at a concentration selected from the following: from about 10-200 μM, 200-300 μM, 300-400 μM, 400-500 μM, 500-600 μM, 600-700 μM, 700-800 μM, 800-900 μM, 900-1000 or 1000-1500 μM, or 1500 μM-2000 μM, 2000 μM-3000 μM, 3000 μM-4000 μM, 4000 μM-5000 μM, 5000 μM-6000 μM, 6000 μM-7000 μM, 7000 μM-8000 μM, 8000 μM-9000 μM, 9000 μM-10,000 μM, 10,000 μM-11,000 μM, 11,000 μM-12,000 μM, 12,000 μM-13,000 μM, 13,000 μM-14,000 μM, 14,000 μM-15,000 μM, 15,000 μM-20,000 μM, 20,000 μM-30,000 μM, or from about 30,000 μM-50,000 μM.
63 . The method according to claim 59 , wherein the anti-connexin 43 polynucleotide is selected from: an oligodeoxynucleotide, a modified oligodeoxynucleotide, an unmodified oligodeoxynucleotide, an antisense polynucleotide, an unmodified antisense polynucleotide, and a modified antisense polynucleotide.
64 . The method of claim 59 , wherein the anti-connexin 43 polynucleotide is a sequence selected from SEQ ID NOS: 1-3, or a sequence having up to about 100 nucleotides of a sequence complementary to SEQ.ID.NO. 134 or SEQ.ID.NO. 135.
65 . The method of claim 59 , wherein the anti-connexin 43 polynucleotide is a sequence selected from SEQ ID NOS: 1 and 2.
66 . The method of claim 59 , wherein the anti-connexin 43 polynucleotide is an antisense polynucleotide having at least about 70 percent homology with SEQ ID NOS: 1 and 2.
67 . The method of claim 59 , wherein the anti-connexin 43 polynucleotide is an antisense polynucleotide that hybridizes to connexin 43 mRNA under conditions of medium to high stringency.
68 . The method according to claim 67 , wherein said antisense polynucleotide has a sequence selected from SEQ.ID.NO:1-3, or a sequence having up to about 40 nucleotides of a sequence complementary to SEQ.ID.NO. 134 or SEQ.ID.NO. 135.
69 . The method according to claim 67 , wherein said antisense polynucleotide is selected from:
1.
(SEQ ID NO: 1)
GTA ATT GCG GCA AGA AGA ATT GTT TCT GTC;
and
2.
(SEQ ID NO: 2)
GTA ATT GCG GCA GGA GGA ATT GTT TCT GTC
3.
(SEQ ID NO: 3)
GGC AAG AGA CAC CAA AGA CAC TAC CAG CAT.
70 . The method according to claim 67 , wherein said antisense polynucleotide has from about 15 to about 35 nucleotides and is sufficiently complementary to connexin 43 mRNA to form a duplex having a melting point greater than 20° C. under physiological conditions.
71 . The method according to claim 67 , wherein the antisense polynucleotide has from about 15 to about 35 nucleotides and has at least about 70 percent homology to an antisense sequence of connexin 43 mRNA.
72 . The method according to claim 59 , wherein said anti-connexin agent is an RNAi or siRNA polynucleotide.
73 . The method according to claim 48 , which is formulated as a gel.
74 . The method according to claim 59 , wherein the pharmaceutical formulation is administered topically.
75 . The method according to claim 73 , wherein said gel is a polyoxyethylene-polyoxypropylene copolymer-based gel or a carboxymethylcellulose-based gel.
76 . The method according to claim 75 , wherein said gel is a pluronic gel.
77 . The method according to claim 76 , wherein said gel is a pluronic F-127.
78 . The method according to claim 48 , wherein the pharmaceutically acceptable carrier comprises an alginate.
79 . The method according to claim 59 , wherein the pharmaceutically acceptable carrier comprises a hydrogel.
80 . The method according to claim 79 , wherein the hydrogel comprises a hydrogel selected from the group consisting of hydrogels containing a cellulose derivative and hydrogels containing polyacrylic acid.
81 . The method according to claim 48 , wherein the pharmaceutically acceptable carrier is a cellulose-based carrier.
82 . The method according to claim 81 , wherein the pharmaceutically acceptable carrier comprises a cellulose-based carrier selected from the group consisting of hydroxyethyl cellulose, hydroxymethyl cellulose, carboxymethyl cellulose, hydroxypropylmethyl cellulose and mixtures thereof.
83 . The method according to claim 48 , wherein the composition is formulated for sustained release.
84 . The method according to claim 48 , wherein the composition is formulated for slow release, extended release, or controlled release.
85 . The method according to claim 48 , wherein the composition is a cream, ointment, emulsion, lotion, spray, salve, foam or paint.
86 . A kit comprising package material containing a composition for use in the method of claim 59 together with instructions for use in treating a refractory chronic wound.
87 . A kit according to claim 86 , wherein the wound is characterized at least in part by increased expression of connexin 43.
88 . A kit according to claim 86 , wherein the wound is characterized at least in part by inflammation.
89 . A kit according to claim 86 , wherein the wound is a dehiscent wound.
90 . A kit according to claim 86 , wherein the wound is a venous ulcer.
91 . A kit according to claim 86 , wherein the wound is a diabetic ulcer.
92 . A kit according to claim 86 , wherein the wound is a diabetic foot ulcer.
93 . A kit according to claim 86 , wherein the wound is a pressure ulcer.
94 . A kit according to claim 86 , wherein the wound is an arterial ulcer.
95 . A kit according to claim 86 , wherein the wound is a vasculitic ulcer.
96 . A kit according to claim 86 , wherein the wound is a skin ulcer resulting from trauma or a burn.
97 . A kit according to claim 86 , wherein the subject is diabetic.
98 . A kit according to claim 86 , wherein the subject has a venous valve malfunction and/or venous insufficiency.
99 . A kit according to claim 86 , wherein the subject has an arterial blockage.
100 . A kit according to claim 86 , wherein said composition is applied more than once.
101 . A kit according to claim 86 , wherein the instructions provide for administration of said composition about once per week.
102 . A kit according to claim 86 , wherein the instructions provide for bi-weekly administration of said composition.
103 . A kit according to claim 86 , wherein the instructions provide for administration of said composition every 3-7 days.
104 . A kit according to claim 86 , wherein the subject is a human.
105 . A kit according to claim 86 , wherein the subject is a non-human animal.
106 . A method of treating a subject having a wound not healing at an expected rate, comprising administering to the wound a composition comprising an anti-connexin 43 polynucleotide and a pharmaceutically acceptable carrier, wherein the amount of said anti-connexin 43 polynucleotide administered to said wound ranges from about 30 to about 500 μg per square centimeter of said wound, the improvement comprising determining whether the wound heals by more than about 30-35% during a 2-4 week run-in period during which the wound is treated by compression and, if not, applying said anti-connexin 43 composition to said wound.