Serpina1 siRNAs: compositions of matter and methods of treatment
The technology described herein relates to double-stranded ribonucleic acid (dsRNA) compositions targeting the Serpina 1 gene, and methods of using such dsRNA compositions to inhibit expression of Serpina 1.
1. A method of inhibiting Serpina 1 expression in a cell, the method comprising:
(a) introducing into the cell a Serpina-1 inhibiting double stranded ribonucleic acid (dsRNA) comprising:
i. a sense strand molecule which is 19-23 nucleotides in length comprising at least 19 contiguous nucleotides of SEQ ID NO:62, said sense strand comprising at least 10 nucleotides comprising a 2′-O-methyl modification;
ii. a separate antisense strand molecule which is 19-23 nucleotides in length comprising at least 19 contiguous nucleotides of SEQ ID NO: 106, said antisense strand comprising at least 3 nucleotides comprising a 2′O-methyl modification; and
iii. at least 19 of the nucleotides in each strand are modified; and
wherein the sense strand and antisense strand are two separate molecules; and
(b) maintaining the cell produced in step (a) for a time sufficient to obtain degradation of the mRNA transcript of a Serpina 1 gene, thereby inhibiting expression of the Serpina 1 gene in the cell.
2. The method of claim 1 , wherein the nucleotides in said antisense strand are all modified nucleotides.
3. The method of claim 1 , wherein said modified nucleotides comprise a modification selected from the group consisting of: 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.
4. The method of claim 1 , wherein said sense strand comprises at least one phosphorothioate linkage.
5. The method of claim 4 , wherein said sense strand comprises three phosphorothioate linkages.
6. The method of claim 1 , wherein said antisense strand comprises at least one phosphorothioate linkage.
7. The method of claim 6 , wherein said antisense strand comprises three phosphorothioate linkages.
8. The method of claim 1 , wherein said dsRNA comprises a ligand.
9. The method of claim 8 , wherein said ligand comprises at least one carbohydrate conjugate.
10. The method of claim 8 , wherein said ligand is attached to the 5′ end of said sense strand.
11. The method of claim 1 , wherein said sense strand comprises at least one inverted nucleotide linkage.
12. The method of claim 11 , wherein said sense strand comprises at least two inverted nucleotide linkages.
13. A method of inhibiting Serpina 1 expression in a cell, the method comprising:
(a) introducing into the cell a Serpina-1 inhibiting double stranded ribonucleic acid (dsRNA) comprising:
i. a sense strand molecule which is 19-23 nucleotides in length comprising:
A. at least 19 contiguous nucleotides of SEQ ID NO:62;
B. three phosphorothioate linkages;
C. a ligand attached to the 5′ end wherein said ligand comprises at least one carbohydrate conjugate;
D. at least one inverted base;
E. all modified nucleotides;
F. at least 10 nucleotides comprising a 2′-O-methyl modification; and
ii. a separate antisense strand molecule which is 19-23 nucleotides in length comprising:
A. at least 19 contiguous nucleotides of SEQ ID NO: 106;
B. three phosphorothioate linkages;
C. all modified nucleotides;
D. at least 3 nucleotides comprising a 2′-O-methyl modification; and
wherein the sense strand and antisense strand are two separate molecules; and
(b) maintaining the cell produced in step (a) for a time sufficient to obtain degradation of the mRNA transcript of a Serpina 1 gene, thereby inhibiting expression of the Serpina 1 gene in the cell.
14. A method of treating a disorder mediated by Serpina 1 expression comprising administering to a patient in need of such treatment a therapeutically effective amount of a Serpina-1 inhibiting double stranded ribonucleic acid (dsRNA) comprising:
i. a sense strand molecule which is 19-23 nucleotides in length comprising at least 19 contiguous nucleotides of SEQ ID NO:62, wherein at least 10 nucleotides comprise a 2′-O-methyl modification;
ii. a separate antisense strand molecule which is 19-23 nucleotides in length comprising at least 19 contiguous nucleotides of SEQ ID NO: 106, wherein at least 3 nucleotides comprise a 2′-O-methyl modification; and
iii. at least 19 of the nucleotides in each strand are modified wherein the sense strand and antisense strand are two separate molecule.
15. The method of claim 14 , wherein the disorder is Alpha 1 anti-trypsin deficiency liver disease.
16. The method of claim 14 , wherein the administration of the dsRNA to the patient causes a decrease in cirrhosis, fibrosis, and/or Serpina 1 protein accumulation in the liver.
17. The method of claim 14 , wherein the likelihood of hepatocellular carcinoma occurring in the patient is reduced.
18. The method of claim 14 , wherein the dsRNA is administered at a concentration of 0.01 mg/kg 5 mg/kg body-weight of the patient.
19. A method of treating a disorder mediated by Serpina 1 expression comprising administering to a patient in need of such treatment a therapeutically effective amount of a Serpina-1 inhibiting double stranded ribonucleic acid (dsRNA) comprising:
i. a sense strand molecule which is 19-23 nucleotides in length comprising:
A. at least 19 contiguous nucleotides of SEQ ID NO:62;
B. three phosphorothioate linkages;
C. a ligand attached to the 5′ end wherein said ligand comprises at least one carbohydrate conjugate;
D. at least one inverted base;
E. all modified nucleotides;
F. at least 10 nucleotides comprising a 2′-O-methyl modification; and
ii. a separate antisense strand molecule which is 19-23 nucleotides in length comprising:
A. at least 19 contiguous nucleotides of SEQ ID NO: 106;
B. three phosphorothioate linkages;
C. all modified nucleotides;
D. at least 10 nucleotides comprising a 2′-O-methyl modification; and
wherein the sense strand and antisense strand are two separate molecules.
20. The method of claim 19 , wherein the disorder is Alpha 1 anti-trypsin deficiency liver disease.
21. The method of claim 19 , wherein the administration of the dsRNA to the patient causes a decrease in cirrhosis or fibrosis.
22. The method of claim 19 , wherein the likelihood of hepatocellular carcinoma occurring in the patient is reduced.
23. The method of claim 19 , wherein the dsRNA is administered at a concentration of 0.01 mg/kg 5 mg/kg body-weight of the patient.
24. The method of claim 1 , wherein said sense strand comprises a 2′-O-methyl modified nucleotide at positions 1, 3, 4, 5, 9, 11, 13, 14, 15, and 18 of the 19 contiguous nucleotides of SEQ ID NO 62.
25. The method of claim 1 , wherein said antisense strand comprises a 2′-O-methyl modified nucleotide at position 8 of the 19 contiguous nucleotides of SEQ ID NO:106.
26. The method of claim 1 , wherein all uridine nucleotides of the sense strand comprise a 2′-O-methyl modification.
27. The method of claim 1 , wherein all adenine nucleotides of the sense strand comprise a 2′-O-methyl modification.
28. The method of claim 1 , wherein said expression of the Serpina 1 gene is inhibited by at least 30%.
29. The method of claim 19 , wherein the administration of the dsRNA to the patient causes a decrease in Serpina 1 protein accumulation in the liver.