METHOD OF TREATING AN INFLAMMATORY DISEASE BY DOUBLE STRANDED RIBONUCLEIC ACID
What is disclosed is the use of a formulation comprising double stranded ribonucleic acid (dsRNA) in the manufacture of a medicament for treating an inflammatory disease in a mammal and inhibiting production of tumor necrosis factor-α (TNF-α) in the mammal.
1 .- 18 . (canceled)
19 . A double-stranded ribonucleic acid (dsRNA) molecule, comprising a sense strand and an antisense strand that form a double stranded region of about 25 base pairs to about 30 base pairs, wherein the antisense strand is complementary to a tumor necrosis factor-α (TNF-α) mRNA comprising a nucleic acid sequence of UAGGGUCGGAACCCAAGCUUA (SEQ ID NO: 135).
20 . The dsRNA molecule of claim 19 wherein the TNF-α mRNA comprises a sequence selected from the group consisting of GCGUGGAGCUGAGAGAUAA (SEQ ID NO: 109); GCCUGUAGCCCAUGUUGUA (SEQ ID NO: 110); GGUAUGAGCCCAUCUAUCU (SEQ ID NO: 111); CCAGGGACCUCUCUCUAAU (SEQ ID NO: 112); GCCCGACUAUCUCGACUUU (SEQ ID NO: 113); UGACAAGCCUGUAGCCCAU (SEQ ID NO: 114); GGUCUACUUUGGGAUCAUU (SEQ ID NO: 115); CCCAGGGACCUCUCUCUAA (SEQ ID NO: 116); AAUCGGCCCGACUAUCUCGACUU (SEQ ID NO: 117); AAUGGCGUGGAGCUGAGAGAU (SEQ ID NO: 118); AACCUCCUCUCUGCCAUCAAG (SEQ ID NO: 119); AACUGAAAGCAUGAUCCGGGA (SEQ ID NO: 120); AAUCUCGACUUUGCCGAGUCU (SEQ ID NO: 121); AAGGGUGACCGACUCAGCGCU (SEQ ID NO: 122); AAUCAGCCGCAUCGCCGUCUC (SEQ ID NO: 123); AACCCAUGUGCUCCUCACCCA (SEQ ID NO: 124); AAGCUCCAGUGGCUGAACCGC (SEQ ID NO: 125); AAGUCAGAUCAUCUUCUCGAA (SEQ ID NO: 126); AAGGGACCUCUCUCUAAUCAG (SEQ ID NO: 127); CCUCAGCCUCUUCUCCUUCCUGA (SEQ ID NO: 128); AAUCCUCAGCCUCUUCUCCUU (SEQ ID NO: 129); AACCAAUGCCCUCCUGGCCAA (SEQ ID NO: 130); CUGAUUAAGUUGUCUAAACAA (SEQ ID NO: 131); CCGACUCAGCGCUGAGAUCAA (SEQ ID NO: 132); CUUGUGAUUAUUUAUUAUUUA (SEQ ID NO: 133); AAGCCUGUAGCCCAUGUUGUA (SEQ ID NO: 134); CUGAAAGCAUGAUCCGGGA (SEQ ID NO: 136); AGGCGGUGCUUGUUCCUCA (SEQ ID NO: 137); CCACCACGCUCUUCUGCCU (SEQ ID NO: 138); AGGGACCUCUCUCUAAUCA (SEQ ID NO: 139); UGACAAGCCUGUAGCCCAU (SEQ ID NO: 140); GCCUGUAGCCCAUGUUGUA (SEQ ID NO: 141); UAGCCCAUGUUGUAGCAAA (SEQ ID NO: 142); CCAAUGCCCUCCUGGCCAA (SEQ ID NO: 143); CCAAUGGCGUGGAGCUGAG (SEQ ID NO: 144); GGCGUGGAGCUGAGAGAUA (SEQ ID NO: 145); GCGUGGAGCUGAGAGAUAA (SEQ ID NO: 146); GCCUGUACCUCAUCUACUC (SEQ ID NO: 147); CCUCCUCUCUGCCAUCAAG (SEQ ID NO: 148); GGUAUGAGCCCAUCUAUCU (SEQ ID NO: 149); GCUGGAGAAGGGUGACCGA (SEQ ID NO: 150); GAGAAGGGUGACCGACUCA (SEQ ID NO: 151); GCCCGACUAUCUCGACUUU (SEQ ID NO: 152); GCAGGUCUACUUUGGGAUC (SEQ ID NO: 153); GGUCUACUUUGGGAUCAUU (SEQ ID NO: 154); UGGGAUCAUUGCCCUGUGA (SEQ ID NO: 155); GGUCGGAACCCAAGCUUAG (SEQ ID NO: 156); CCAGAAUGCUGCAGGACUU (SEQ ID NO: 157); GAGAAGACCUCACCUAGAA (SEQ ID NO: 158); GAAGACCUCACCUAGAAAU (SEQ ID NO: 159); CCAGAUGUUUCCAGACUUC (SEQ ID NO: 160); CUAUUUAUGUUUGCACUUG (SEQ ID NO: 161); UCUAAACAAUGCUGAUUUG (SEQ ID NO: 162); and GACCAACUGUCACUCAUU (SEQ ID NO: 163).
21 . The dsRNA molecule of claim 19 wherein the dsRNA molecule is formulated with a polynucleotide delivery-enhancing peptide.
22 . The dsRNA molecule of claim 21 wherein the peptide consists of an amino acid sequence of KGSKKAVTKAQKKDGKKRKRSRKESYSVYVYKVLKQ (SEQ ID NO: 59).
23 . The dsRNA molecule of claim 21 wherein the peptide comprises an amino acid sequence is selected from the group consisting of KGSKKAVTKAQKKDGKKRKRSRKESYSVYVYKVLKQ (SEQ ID NO: 59); KKAVTKAQKKDGKKRKRSRKESYSVYVYKVLKQ (SEQ ID NO: 165); VTKAQKKDGKKRKRSRKESYSVYVYKVLKQ (SEQ ID NO: 166); AQKKDGKKRKRSRKESYSVYVYKVLKQ (SEQ ID NO: 167); KDGKKRKRSRKESYSVYVYKVLKQ (SEQ ID NO: 168); KKRKRSRKESYSVYVYKVLKQ (SEQ ID NO: 169); KRSRKESYSVYVYKVLKQ (SEQ ID NO: 170); RKESYSVYVYKVLKQ (SEQ ID NO: 171); SYSVYVYKVLKQ (SEQ ID NO: 172); VYVYKVLKQ (SEQ ID NO: 173) and YKVLKQ (SEQ ID NO: 174).
24 . The dsRNA molecule of claim 21 wherein the peptide is complexed or conjugated to the dsRNA.
25 . The dsRNA molecule of claim 21 wherein the formulation further comprises a cationic lipid.
26 . The dsRNA molecule of claim 25 wherein the cationic lipid is optionally combined with a non-cationic lipid.
27 . The dsRNA molecule of claim 26 wherein the non-cationic lipid is a neutral lipid.
28 . The dsRNA molecule of claim 27 wherein the neutral lipid is dioleoylphosphatidylethanolamine (DOPE), diphytanoylphosphatidylethanolamine (DPhPE), or cholesterol.
29 . The dsRNA molecule of claim 19 wherein the dsRNA molecule is used in conjunction with a further therapeutic agent, wherein the further therapeutic agent is selected from the group consisting of a non-steroidal anti-inflammatory drug (NSAID), methotrexate, gold compound, D-penicillamine, antimalarial, sulfasalazine, glucocorticoid, infliximab, and entracept.
30 . A pharmaceutical composition, comprising a peptide conjugated to a double stranded ribonucleic acid (dsRNA) molecule, wherein the dsRNA molecule comprises a sense strand and an antisense strand that form a double stranded region of about 25 base pairs to about 30 base pairs, wherein the antisense strand is complementary to a TNF-α mRNA comprising a nucleic acid sequence of UAGGGUCGGAACCCAAGCUUA (SEQ ID NO: 135).
31 . The composition of claim 30 wherein the TNF-α mRNA comprises a sequence selected from the group consisting of GCGUGGAGCUGAGAGAUAA (SEQ ID NO: 109); GCCUGUAGCCCAUGUUGUA (SEQ ID NO: 110); GGUAUGAGCCCAUCUAUCU (SEQ ID NO: 111); CCAGGGACCUCUCUCUAAU (SEQ ID NO: 112); GCCCGACUAUCUCGACUUU (SEQ ID NO: 113); UGACAAGCCUGUAGCCCAU (SEQ ID NO: 114); GGUCUACUUUGGGAUCAUU (SEQ ID NO: 115); and CCCAGGGACCUCUCUCUAA (SEQ ID NO: 116).
32 . The composition of claim 30 wherein the TNF-α mRNA comprises a sequence selected from the group consisting of AAUCGGCCCGACUAUCUCGACUU (SEQ ID NO: 117); AAUGGCGUGGAGCUGAGAGAU (SEQ ID NO: 118); AACCUCCUCUCUGCCAUCAAG (SEQ ID NO: 119); AACUGAAAGCAUGAUCCGGGA (SEQ ID NO: 120); AAUCUCGACUUUGCCGAGUCU (SEQ ID NO: 121); AAGGGUGACCGACUCAGCGCU (SEQ ID NO: 122); AAUCAGCCGCAUCGCCGUCUC (SEQ ID NO: 123); AACCCAUGUGCUCCUCACCCA (SEQ ID NO: 124); AAGCUCCAGUGGCUGAACCGC (SEQ ID NO: 125); AAGUCAGAUCAUCUUCUCGAA (SEQ ID NO: 126); AAGGGACCUCUCUCUAAUCAG (SEQ ID NO: 127); CCUCAGCCUCUUCUCCUUCCUGA (SEQ ID NO: 128); AAUCCUCAGCCUCUUCUCCUU (SEQ ID NO: 129); AACCAAUGCCCUCCUGGCCAA (SEQ ID NO: 130); CUGAUUAAGUUGUCUAAACAA (SEQ ID NO: 131); CCGACUCAGCGCUGAGAUCAA (SEQ ID NO: 132); CUUGUGAUUAUUUAUUAUUUA (SEQ ID NO: 133); AAGCCUGUAGCCCAUGUUGUA (SEQ ID NO: 134); and UAGGGUCGGAACCCAAGCUUA (SEQ ID NO: 135).
33 . The composition of claim 30 wherein the TNF-α mRNA comprises a sequence selected from the group consisting of CUGAAAGCAUGAUCCGGGA (SEQ ID NO: 136); AGGCGGUGCUUGUUCCUCA (SEQ ID NO: 137); CCACCACGCUCUUCUGCCU (SEQ ID NO: 138); AGGGACCUCUCUCUAAUCA (SEQ ID NO: 139); UGACAAGCCUGUAGCCCAU (SEQ ID NO: 140); GCCUGUAGCCCAUGUUGUA (SEQ ID NO: 141); UAGCCCAUGUUGUAGCAAA (SEQ ID NO: 142); CCAAUGCCCUCCUGGCCAA (SEQ ID NO: 143); CCAAUGGCGUGGAGCUGAG (SEQ ID NO: 144); GGCGUGGAGCUGAGAGAUA (SEQ ID NO: 145); GCGUGGAGCUGAGAGAUAA (SEQ ID NO: 146); GCCUGUACCUCAUCUACUC (SEQ ID NO: 147); CCUCCUCUCUGCCAUCAAG (SEQ ID NO: 148); GGUAUGAGCCCAUCUAUCU (SEQ ID NO: 149); GCUGGAGAAGGGUGACCGA (SEQ ID NO: 150); GAGAAGGGUGACCGACUCA (SEQ ID NO: 151); GCCCGACUAUCUCGACUUU (SEQ ID NO: 152); GCAGGUCUACUUUGGGAUC (SEQ ID NO: 153); GGUCUACUUUGGGAUCAUU (SEQ ID NO: 154); UGGGAUCAUUGCCCUGUGA (SEQ ID NO: 155); GGUCGGAACCCAAGCUUAG (SEQ ID NO: 156); CCAGAAUGCUGCAGGACUU (SEQ ID NO: 157); GAGAAGACCUCACCUAGAA (SEQ ID NO: 158); GAAGACCUCACCUAGAAAU (SEQ ID NO: 159); CCAGAUGUUUCCAGACUUC (SEQ ID NO: 160); CUAUUUAUGUUUGCACUUG (SEQ ID NO: 161); UCUAAACAAUGCUGAUUUG (SEQ ID NO: 162); and GACCAACUGUCACUCAUU (SEQ ID NO: 163).
34 . The composition of claim 30 wherein the peptide comprises an amino acid sequence selected from the group consisting of KGSKKAVTKAQKKDGKKRKRSRKESYSVYVYKVLKQ (SEQ ID NO: 59); KKAVTKAQKKDGKKRKRSRKESYSVYVYKVLKQ (SEQ ID NO: 165); VTKAQKKDGKKRKRSRKESYSVYVYKVLKQ (SEQ ID NO: 166); AQKKDGKKRKRSRKESYSVYVYKVLKQ (SEQ ID NO: 167); KDGKKRKRSRKESYSVYVYKVLKQ (SEQ ID NO: 168); KKRKRSRKESYSVYVYKVLKQ (SEQ ID NO: 169); KRSRKESYSVYVYKVLKQ (SEQ ID NO: 170); RKESYSVYVYKVLKQ (SEQ ID NO: 171); SYSVYVYKVLKQ (SEQ ID NO: 172); VYVYKVLKQ (SEQ ID NO: 173); and YKVLKQ (SEQ ID NO: 174).
35 . The composition of claim 30 wherein the peptide consists of an amino acid sequence of KGSKKAVTKAQKKDGKKRKRSRKESYSVYVYKVLKQ (SEQ ID NO: 59).
36 . The composition of claim 30 wherein the formulation further comprises a cationic lipid.
37 . The composition of claim 36 wherein the cationic lipid is optionally combined with a non-cationic lipid.
38 . The composition of claim 37 wherein the non-cationic lipid is a neutral lipid.
39 . The composition of claim 38 wherein the neutral lipid is dioleoylphosphatidylethanolamine (DOPE), diphytanoylphosphatidylethanolamine (DPhPE), or cholesterol.
40 . A method of treating or preventing an inflammatory disease, comprising administering a dsRNA molecule according to claim 1 in an amount sufficient to treat or prevent the inflammatory disease.
41 . The method of claim 40 wherein the dsRNA molecule is formulated with a polynucleotide delivery-enhancing peptide.
42 . The method of claim 41 wherein the peptide consists of an amino acid sequence of KGSKKAVTKAQKKDGKKRKRSRKESYSVYVYKVLKQ (SEQ ID NO: 59).
43 . The method of claim 41 wherein the peptide comprises an amino acid sequence is selected from the group consisting of KGSKKAVTKAQKKDGKKRKRSRKESYSVYVYKVLKQ (SEQ ID NO: 59); KKAVTKAQKKDGKKRKRSRKESYSVYVYKVLKQ (SEQ ID NO: 165); VTKAQKKDGKKRKRSRKESYSVYVYKVLKQ (SEQ ID NO: 166); AQKKDGKKRKRSRKESYSVYVYKVLKQ (SEQ ID NO: 167); KDGKKRKRSRKESYSVYVYKVLKQ (SEQ ID NO: 168); KKRKRSRKESYSVYVYKVLKQ (SEQ ID NO: 169); KRSRKESYSVYVYKVLKQ (SEQ ID NO: 170); RKESYSVYVYKVLKQ (SEQ ID NO: 171); SYSVYVYKVLKQ (SEQ ID NO: 172); VYVYKVLKQ (SEQ ID NO: 173) and YKVLKQ (SEQ ID NO: 174).
44 . The method of claim 41 wherein the peptide is complexed or conjugated to the dsRNA.
45 . The method of claim 41 wherein the formulation further comprises a cationic lipid.
46 . The method of claim 45 wherein the cationic lipid is optionally combined with a non-cationic lipid.
47 . The method of claim 46 wherein the non-cationic lipid is a neutral lipid.
48 . The method of claim 47 wherein the neutral lipid is dioleoylphosphatidylethanolamine (DOPE), diphytanoylphosphatidylethanolamine (DPhPE), or cholesterol.
49 . The method of claim 40 wherein the subject is a human.
50 . The method of claim 40 wherein the dsRNA molecule is administered to the circulation of the subject.
51 . The method of claim 50 , wherein the dsRNA molecule is administered intravenously.
52 . The method of claim 40 wherein the dsRNA molecule is administered to blood leucocytes.
53 . The method of claim 52 wherein the leucocytes are monocytes.
54 . The method of claim 40 wherein the dsRNA molecule is administered in conjunction with a further therapeutic agent, wherein the further therapeutic agent is selected from the group consisting of a non-steroidal anti-inflammatory drug (NSAID), methotrexate, gold compound, D-penicillamine, antimalarial, sulfasalazine, glucocorticoid, infliximab, and entracept.
55 . The method of claim 40 wherein the inflammatory disease is a systemic disease.
56 . The method of claim 40 wherein the inflammatory disease is rheumatoid arthritis.
57 . The method of claim 40 wherein the inflammatory disease is psoriasis.
58 . The method of claim 40 wherein the inflammatory disease is psoriatic arthritis.