Compositions and methods for inhibiting expression of IKK-B gene
View Patent ↗The invention relates to a doable-stranded ribonucleic acid (dsRNA) for inhibiting the expression of the IKK-B gene, comprising an antisense strand having a nucleotide sequence which is less that 30 nucleotides in length, generally 19-25 nucleotides in length, and which is substantially complementary to at least a part of the IKK-B gene. The invention also relates to a pharmaceutical composition comprising the dsRNA together with a pharmaceutically acceptable carrier; methods for treating diseases, caused by the expression or activation of the IKK-B gene using the pharmaceutical composition; and methods for inhibiting the expression of the IKK-B gene in a cell.
1. A double-stranded ribonucleic acid (dsRNA) for inhibiting the expression of a human IKK-B gene, wherein said dsRNA comprises a sense strand and an antisense strand and a duplex structure between 15 and 30 base pairs in length, the antisense strand comprising at least 17 contiguous nucleotides of the nucleotide sequence of SEQ ID NO:24.
2. The dsRNA of claim 1 , wherein said dsRNA comprises at least one modified nucleotide.
3. The dsRNA of claim 2 , wherein said modified nucleotide is chosen from the group of a 2′-O-methyl modified nucleotide, a nucleotide comprising a 3′-phosphorothioate group, and a terminal nucleotide linked to a cholesteryl derivative or dodecanoic acid bisdecylamide group.
4. The dsRNA of claim 2 , wherein said modified nucleotide is chosen from the group of a 2′-deoxy-2′-fluoro modified nucleotide, a 2′-deoxy-modified nucleotide, a locked nucleotide, an abasic nucleotide, 2′-amino-modified nucleotide, 2′-alkyl-modified nucleotide, morpholino nucleotide, a phosphoramidate, and a non-natural base comprising nucleotide.
5. A cell comprising the dsRNA of claim 1 .
6. A pharmaceutical composition for inhibiting the expression of the IKK-B gene, comprising the dsRNA of claim 1 and a pharmaceutically acceptable carrier.
7. A method for inhibiting the expression of the IKK-B gene in a cell, the method comprising:
(a) introducing into the cell the dsRNA of claim 1 ; and
(b) maintaining the cell produced in step (a) for a time sufficient to obtain degradation of the mRNA transcript of the IKK-B gene, thereby inhibiting expression of the IKK-B gene in the cell.
8. A method of treating inflammation comprising administering to a patient in need thereof a therapeutically effective amount of the dsRNA of claim 1 .
9. A vector for inhibiting the expression of the IKK-B gene, said vector comprising a regulatory sequence operably linked to a nucleotide sequence that encodes at least one strand of the dsRNA of claim 1 .
10. A cell comprising the vector of claim 9 .
11. The dsRNA of claim 1 , wherein the sense strand comprises at least 15 contiguous nucleotides of the nucleotide sequence of SEQ ID NO: 23.
12. The dsRNA of claim 11 , wherein the antisense strand comprises the nucleotide sequence of SEQ ID NO:24 and the sense strand comprises the nucleotide sequence of SEQ ID NO:23.
13. The dsRNA of claim 11 , wherein said sense strand consists of SEQ ID NO:23 and said antisense strand consists of SEQ ID NO:24.
14. The dsRNA of claim 1 , wherein the antisense strand comprises at least 17 contiguous nucleotides of the nucleotide sequence CCCUGAUCCmAGCUCCUUGGTT (SEQ ID NO: 30) wherein the trailing lower case “m” denotes a nucleotide containing a 2′O-Methyl substitution and the last “T” has a phosphorothioate modification.
15. The dsRNA of claim 1 , wherein the antisense strand comprises the nucleotide sequence CCCUGAUCCmAGCUCCUUGGTT (SEQ ID NO: 30) wherein the trailing lower case “m” denotes a nucleotide containing a 2′O-Methyl substitution and the last “T” has a phosphorothioate modification.
16. The dsRNA of claim 1 , wherein the antisense strand consistsof the nucleotide sequence CCCUGAUCCmAGCUCCUUGGTT (SEQ ID NO: 30) wherein the trailing lower case “m” denotes a nucleotide containing a 2′O-Methyl substitution and the last “T” has a phosphorothioate modification.
17. The dsRNA of claim 1 , wherein the sense strand comprises at least 15 contiguous nucleotides of the nucleotide sequence CmCmAAGGAGCmUmGGAUmCmAGGGTT (SEQ ID NO: 29) wherein a trailing lower case “m” denotes a nucleotide containing a 2′O-Methyl substitution and the last “T” has a phosphorothioate modification.
18. The dsRNA of claim 1 , wherein the sense strand comprises the nucleotide sequence CmCmAAGGAGCmUmGGAUmCmAGGGTT (SEQ ID NO: 29) wherein a trailing lower case “m” denotes a nucleotide containing a 2′O-Methyl substitution and the last “T” has a phosphorothioate modification.
19. The dsRNA of claim 1 , wherein the sense strand consists of the nucleotide sequence CmCmAAGGAGCmUmGGAUmCmAGGGTT (SEQ ID NO: 29) wherein a trailing lower case “m” denotes a nucleotide containing a 2′O-Methyl substitution and the last “T” has a phosphorothioate modification.
20. The dsRNA of claim 1 , wherein the antisense strand comprises the nucleotide sequence CCCUGAUCCmAGCUCCUUGGTT (SEQ ID NO: 30) and the sense strand comprises the nucleotide sequence CmCmAAGGAGCmUmGGAUmCmAGGGTT (SEQ ID NO: 29) wherein in both the sense and antisense sequences, a trailing lower case “m” denotes a nucleotide containing a 2′O-Methyl substitution and the last “T” has a phosphorothioate modification.
21. The dsRNA of claim 1 , wherein the antisense strand consists of the nucleotide sequence CCCUGAUCCmAGCUCCUUGGTT (SEQ ID NO: 30) and the sense strand consists of the nucleotide sequence CmCmAAGGAGCmUmGGAUmCmAGGGTT (SEQ ID NO: 29) wherein in both the sense and antisense sequences, a trailing lower case “m” denotes a nucleotide containing a 2′O-Methyl substitution and the last “T” has a phosphorothioate modification.
22. The dsRNA of claim 1 , wherein said dsRNA, upon contact with a cell expressing said IKK-B, inhibits expression of said IKK-B gene by at least 25%.
23. The dsRNA of claim 1 , wherein said dsRNA, upon contact with a cell expressing said IKK-B, inhibits expression of said IKK-B gene by at least 40%.
24. The dsRNA of claim 1 , comprising at least one 2′-O-methyl modified nucleotide.
25. The dsRNA of claim 1 , comprising at least one single-stranded nucleotide overhang of 1 to 4 nucleotides.
26. The dsRNA of claim 1 conjugated to at least one non-ligand group.
27. The pharmaceutical composition of claim 6 , wherein the carrier is a lipid carrier.
28. The dsRNA of claim 1 , wherein the antisense strand comprises at least 18 contiguous nucleotides of the nucleotide sequence of SEQ ID NO:24.
29. The dsRNA of claim 1 , wherein the antisense strand comprises at least 19 contiguous nucleotides of the nucleotide sequence of SEQ ID NO:24.
30. The dsRNA of claim 1 , wherein the antisense strand comprises at least 20 contiguous nucleotides of the nucleotide sequence of SEQ ID NO:24.