RNA interference mediated inhibition of catenin (cadherin-associated protein), beta 1 (CTNNB1) gene expression using short interfering nucleic acid (SINA)
View Patent ↗The present invention relates to compounds, compositions, and methods for the study, diagnosis, and treatment of traits, diseases and conditions that respond to the modulation of CTNNB1 gene expression and/or activity, and/or modulate a beta-catenin gene expression pathway. Specifically, the invention relates to double-stranded nucleic acid molecules including small nucleic acid molecules, such as short interfering nucleic acid (siNA), short interfering RNA (siRNA), double-stranded RNA (dsRNA), micro-RNA (miRNA), and short hairpin RNA (shRNA) molecules that are capable of mediating or that mediate RNA interference (RNAi) against CTNNB1 gene expression.
1. A double-stranded short interfering nucleic acid (siNA) molecule that inhibits the expression of cadherin-associated protein, beta 1 (CTNNB1), wherein
(a) the siNA comprises a sense strand and an antisense strand;
(b) each strand is independently 19-24 nucleotides in length; and
(c) at least one strand comprises a nucleotide sequence comprising at least 17 contiguous nucleotides of SEQ NO: 194.
2. A double-stranded short interfering nucleic acid (siNA) molecule that inhibits the expression of cadherin-associated protein, beta 1 (CTNNB1), wherein
(a) the siNA comprises a sense strand and an antisense strand;
(b) each strand is independently 19-24 nucleotides in length; and
(c) the antisense strand comprises a nucleotide sequence comprising at least 17 contiguous nucleotides 5′-ACGACUAGUUCAGUUGCUU-3′ (SEQ ID NO: 194).
3. A double-stranded short interfering nucleic acid (siNA) molecule that inhibits the expression of cadherin-associated protein, beta 1 (CTNNB1), wherein
(a) the siNA comprises a sense strand and an antisense strand;
(b) each strand is independently 19-24 nucleotides in length; and
(c) the antisense strand comprises a nucleotide sequence comprising at least 17 contiguous nucleotides of 5′-AAGCAACUGAACUAGUCGU-3′ (SEQ ID NO: 5107).
4. The double-stranded short interfering nucleic acid (siNA) molecule of claim 3 , wherein the sense strand comprises a nucleotide sequence comprising at least 17 contiguous nucleotides of 5′-ACGACUAGUUCAGUUGCUU-3′ (SEQ ID NO: 194); and wherein one of more of the nucleotides are chemically modified.
5. The double-stranded short interfering nucleic acid (siNA) molecule according to any one of claims 1 - 3 , wherein at least one nucleotide is a chemically modified nucleotide.
6. The double-stranded short interfering nucleic acid (siNA) molecule according to any one of claims 1 - 4 , further comprising at least one non-nucleotide.
7. The double-stranded short interfering nucleic acid (siNA) molecule according to any one of claims 1 - 4 , wherein at least one nucleotide comprises a universal base.
8. The double-stranded short interfering nucleic acid (siNA) molecule according to any one of claims 1 - 4 , comprising at least one phosphorothioate internucleotide linkage.
9. The double-stranded short interfering nucleic acid (siNA) molecule according to any one of claims 1 - 4 , comprising a cap on the 3′-end, 5′-end or both 3′ and 5′ ends of at least one strand.
10. The double-stranded short interfering nucleic acid (siNA) molecule according to any one of claims 1 - 4 , comprising one or more 3′-overhang nucleotides on one or both strands.
11. The double-stranded short interfering nucleic acid (siNA) molecule according to any one of claims 1 - 4 , wherein the 5′ end of the antisense strand is phosphorylated.
12. The double-stranded short interfering nucleic acid (siNA) molecule of claim 10 , wherein the 3′-overhang nucleotides on at least one strand are 2′-O-methyl nucleotides.
13. The double-stranded short interfering nucleic acid (siNA) molecule of claim 12 , wherein the 2′-O-methyl nucleotides are linked with a phosphorothioate internucleotide linkage.
14. The double-stranded short interfering nucleic acid (siNA) molecule of claim 5 , wherein the chemically modified nucleotide is a 2′-deoxy-2′-fluoro nucleotide.
15. The double-stranded short interfering nucleic acid (siNA) molecule of claim 5 , wherein the chemically modified nucleotide is a 2′-deoxy nucleotide.
16. The double-stranded short interfering nucleic acid (siNA) molecule of claim 5 , wherein the chemically modified nucleotide is a 2′-O-alkyl nucleotide.
17. The double-stranded short interfering nucleic acid (siNA) molecule according to any one of claims 1 - 4 , wherein five or more pyrimidine nucleotides in one or both strands are 2′-deoxy-2′-fluoro pyrimidine nucleotides.
18. The double-stranded short interfering nucleic acid (siNA) molecule according to any one of claims 1 - 4 , wherein five or more pyrimidine nucleotides in one or both strands are 2′-O-methyl pyrimidine nucleotides.
19. The double-stranded short interfering nucleic acid (siNA) molecule according to any one of claims 1 - 4 , wherein five or more purine nucleotides in one or both strands are 2′-deoxy-2′-fluoro purine nucleotides.
20. The double-stranded short interfering nucleic acid (siNA) molecule according to any one of claims 1 - 4 , wherein five or more purine nucleotides in one or both strands are 2′-O-methyl purine nucleotides.
21. The double-stranded short interfering nucleic acid (siNA) molecule of claim 17 , wherein five or more purine nucleotides in one or both strands are 2′-O-methyl purine nucleotides.
22. The double-stranded short interfering nucleic acid (siNA) molecule of claim 18 , wherein five or more purine nucleotides in one or both strands are 2′-deoxy-2′-fluoro nucleotides.
23. A double-stranded short interfering nucleic acid (siNA) molecule that inhibits the expression of cadherin-associated protein, beta 1 (CTNNB1), comprising SEQ ID NOS: 6372 and 6374.
24. A double-stranded short interfering nucleic acid (siNA) molecule that inhibits the expression of cadherin-associated protein, beta 1 (CTNNB1), comprising SEQ ID NOS: 6372 and 6373.
25. A composition comprising:
(a) a double-stranded short interfering nucleic acid (siNA) molecule having SEQ ID NOS:6372 and 6374;
(b) (13Z,16Z)-N,N-dimethyl-3-nonyldocosa-13,16-dien-1-amine;
(c) cholesterol;
(d) DSPC; and
(e) PEG-DMG.
26. A composition comprising:
(a) a double-stranded short interfering nucleic acid (siNA) molecule having SEQ ID NOS: 6372 and 6373;
(b) (13Z,16Z)-N,N-dimethyl-3-nonyldocosa-13,16-dien-1-amine;
(c) cholesterol;
(d) DSPC; and
(e) PEG-DMG.
27. A double-stranded short interfering nucleic acid (siNA) molecule that inhibits the expression of cadherin-associated protein, beta 1 (CTNNB 1), comprising SEQ ID NOS: 6372 and 6371.
28. A double-stranded short interfering nucleic acid (siNA) molecule that inhibits the expression of cadherin-associated protein, beta 1 (CTNNB 1), comprising SEQ ID NOS: 1912 and 1913.
29. A double-stranded short interfering nucleic acid (siNA) molecule that inhibits the expression of cadherin-associated protein, beta 1 (CTNNB 1), comprising SEQ ID NOS: 1840 and 1841.
30. A composition comprising:
(a) a double-stranded short interfering nucleic acid (siNA) molecule having SEQ ID NOS: 6372 and 6371;
(b) (13Z,16Z)-N,N-dimethyl-3-nonyldocosa-13,16-dien-1-amine;
(c) cholesterol;
(d) DSPC; and
(e) PEG-DMG.
31. A composition comprising:
(a) a double-stranded short interfering nucleic acid (siNA) molecule having SEQ ID NOS: 1912 and 1913;
(b) (13Z,16Z)-N,N-dimethyl-3-nonyldocosa-13,16-dien-1-amine;
(c) cholesterol;
(d) DSPC; and
(e) PEG-DMG.
32. A composition comprising:
(a) a double-stranded short interfering nucleic acid (siNA) molecule having SEQ ID NOS: 1840 and 1841;
(b) (13Z,16Z)-N,N-dimethyl-3-nonyldocosa-13,16-dien-1-amine;
(c) cholesterol;
(d) DSPC; and
(e) PEG-DMG.
33. The double-stranded short interfering nucleic acid (siNA) molecule according to any one of claims 23 , 24 , and 27 - 29 , comprising one or more phosphorothioate internucleotide linkages.
34. A composition comprising the double-stranded short interfering nucleic acid (siNA) molecule according to any one of claims 1 - 4 , 23 , 24 , and 27 - 29 in a pharmaceutically acceptable carrier or diluent.
35. The composition according to any one of claims 25 , 26 , and 30 - 32 , wherein the (13Z,16Z)-N,N-dimethyl-3-nonyldocosa -13,16-dien-1-amine, cholesterol, DSPC, and PEG-DMG have a molar ratio of 50:30: 10:2 respectively.
36. The composition according to any one of claims 25 , 26 , and 30 - 32 , further comprising sucrose, trehalose, or any combination thereof.
37. A method of treating a human subject suffering from a condition which is mediated by the action, or by loss of action, of cadherin-associated protein, beta 1 (CTNNB 1), comprising administering to said subject an effective amount of the double-stranded short interfering nucleic acid (siNA) molecule of any one of claims 1 - 4 , 23 , 24 , and 27 - 29 .
38. The method according to claim 37 , wherein the condition is cancer.
39. The method of claim 37 , wherein the subject is a human.