Serpinc1 iRNA compositions and methods of use thereof
The invention relates to iRNA, e.g., double-stranded ribonucleic acid (dsRNA), compositions targeting the Serpinc1 gene, and methods of using such iRNA, e.g., dsRNA, compositions to inhibit expression of Serpinc1 and methods of treating subjects having a bleeding disorder, such as a hemophilia.
1 . A double-stranded ribonucleic acid (dsRNA) for inhibiting expression of Serpinc1, wherein said dsRNA double-stranded ribonucleic acid comprises a sense strand and an antisense strand, wherein the sense strand comprises the nucleotide sequence of 5′-GfsgsUfuAfaCfaCfCfAfuUfuAfcUfuCfaAf-3′ (SEQ ID NO:941) and the antisense strand comprises the nucleotide sequence of 5′-usUfsgAfaGfuAfaAfuggUfgUfuAfaCfcsasg-3′ (SEQ ID NO:960), wherein a, g, c, and u are 2′-O-methyl(2′-OMe) A, U G, C, or G and U, respectively; Af, Cf Gf, Gf or Cf, and Uf are 2′-fluoro A, G, C or, and U, respectively; and s is a phosphorothioate linkage.
2 . The dsRNA double-stranded ribonucleic acid of claim 1 , further comprising a ligand.
3 . The dsRNA double-stranded ribonucleic acid of claim 2 , wherein the ligand is conjugated to the 3′ end 3′-end of the sense strand of the dsRNA double-stranded ribonucleic acid.
4 . The dsRNA double-stranded ribonucleic acid of claim 2 , wherein the ligand is an N-acetylgalactosamine (GalNAc) derivative.
5 . The dsRNA double-stranded ribonucleic acid of claim 4 , wherein the ligand is
6 . The dsRNA double-stranded ribonucleic acid of claim 4 3 , wherein the dsRNA double-stranded ribonucleic acid is conjugated to the ligand as shown in the following schematic
and, wherein X is O or S.
7 . The dsRNA double-stranded ribonucleic acid of claim 6 , wherein the X is O.
8 . A cell containing the dsRNA double-stranded ribonucleic acid of claim 1 .
9 . A pharmaceutical composition for inhibiting expression of a Serpinc1 gene comprising the dsRNA double-stranded ribonucleic acid of claim 1 .
10 . A method of inhibiting Serpinc1 expression in a cell, the method comprising:
(a) contacting the cell with the dsRNA double-stranded ribonucleic acid of claim 1 ; and
(b) maintaining the cell produced in step (a) for a time sufficient to obtain degradation of the mRNA transcript of a Serpinc1 gene, thereby inhibiting expression of the Serpinc1 gene in the cell.
11 . The method of claim 10 , wherein said cell is within a subject.
12 . The method of claim 11 , wherein the subject is a human.
13 . The method of claim 12 , wherein the human subject suffers from a bleeding disorder.
14 . The method of claim 13 , wherein the bleeding disorder is a hemophilia.
15 . The method of any one of claims 10-14 , wherein the Serpinc1 expression is inhibited by at least about 30%.
16 . A method of treating a subject having a disorder that would benefit from reduction in Serpinc1 expression, comprising administering to the subject a therapeutically prophylactically effective amount of the dsRNA double-stranded ribonucleic acid of claim 1 , thereby treating said subject.
17 . A method of preventing at least one symptom in a subject having a disorder that would benefit from reduction in Serpinc1 expression, comprising administering to the subject a therapeutically prophylactically effective amount of the dsRNA double-stranded ribonucleic acid of claim 1 , thereby preventing at least one symptom in the subject having a the disorder that would benefit from reduction in Serpinc1 expression.
18 . The method of claim 16 , wherein the disorder is a bleeding disorder.
19 . The method of claim 18 , wherein the bleeding disorder is a hemophilia.
20 . The method of claim 16 , wherein the administration of the dsRNA double-stranded ribonucleic acid to the subject causes an increase in blood clotting and/or a decrease in Serpinc1 protein accumulation.
21 . The method of claim 16 , wherein the dsRNA double-stranded ribonucleic acid is conjugated to a ligand.
22 . The method of claim 21 , wherein the ligand is conjugated to the 3′-end of the sense strand of the dsRNA double-stranded ribonucleic acid.
23 . The method of claim 22 , wherein the ligand is an N-acetylgalactosamine (GalNAc) derivative.
24 . The method of claim 17 , wherein the dsRNA is administered at a dose of about 0.01 mg/kg to about 50 mg/kg.
25 . The method of claim 24 , wherein the dsRNA is administered at a dose of about 10 mg/kg to about 30 mg/kg.
26 . The method of claim 24 , wherein the dsRNA is administered at a dose selected from the group consisting of 0.3 mg/kg 0.5 mg/kg 1 mg/kg, 1.5 mg/kg, 3 mg/kg, 10 mg/kg, and 30 mg/kg.
27 . The method of claim 25 or 26 17 , wherein the dsRNA double-stranded ribonucleic acid is administered to the subject once a week.
28 . The method of claim 25 or 26 17 , wherein the dsRNA double-stranded ribonucleic acid is administered to the subject twice a month.
29 . The method of claim 16 , further comprising measuring thrombin levels in said subject.
30 . The method of claim 17 , wherein the dsRNA is administered to the subject subcutaneously at a cumulative weekly dose of about 0.5 mg/kg to about 5 mg/kg.
31 . A method of inhibiting the expression of Serpinc1 in a subject, the method comprising
administering to said subject a therapeutically prophylactically effective amount of the dsRNA double-stranded ribonucleic acid of claim 1 , thereby inhibiting the expression of Serpinc1 in said subject.
32 . The method of claim 31 , wherein the dsRNA double-stranded ribonucleic acid is conjugated to a ligand.
33 . The method of claim 32 , wherein the ligand is conjugated to the 3′-end of the sense strand of the dsRNA double-stranded ribonucleic acid.
34 . The method of claim 33 , wherein the ligand is an N-acetylgalactosamine (GalNAc) derivative.
35 . The method of claim 31 , wherein the dsRNA is administered at a dose of about 0.01 mg/kg to about 50 mg/kg.
36 . The method of claim 35 , wherein the dsRNA is administered at a dose of about 10 mg/kg to about 30 mg/kg.
37 . The method of claim 35 , wherein the dsRNA is administered at a dose selected from the group consisting of 1 mg/kg, 3 mg/kg, 10 mg/kg, and 30 mg/kg.
38 . The method of claim 37 31 , wherein the dsRNA double-stranded ribonucleic acid is administered to the subject once a week.
39 . The method of claim 37 31 , wherein the dsRNA double-stranded ribonucleic acid is administered to the subject twice a month.
40 . The method of claim 31 , wherein the dsRNA is administered to the subject subcutaneously at a cumulative weekly dose of about 0.5 mg/kg to about 5 mg/kg.
41 . The method of claim 31 , further comprising measuring thrombin levels in said subject.
42 . The method of claim 21 , wherein the ligand is
43 . The method of claim 21 22 , wherein the dsRNA double-stranded ribonucleic acid is conjugated to the ligand as shown in the following schematic
and, wherein X is O or S.
44 . The dsRNA method of claim 43 , wherein the X is O.
45 . The method of claim 16 , wherein the dsRNA is administered at a dose of about 0.01 mg/kg to about 50 mg/kg.
46 . The method of claim 45 , wherein the dsRNA is administered at a dose of about 10 mg/kg to about 30 mg/kg.
47 . The method of claim 45 , wherein the dsRNA is administered at a dose selected from the group consisting of 0.3 mg/kg 0.5 mg/kg 1 mg/kg, 1.5 mg/kg, 3 mg/kg, 10 mg/kg, and 30 mg/kg.
48 . The method of claim 46 or 47 16 , wherein the dsRNA double-stranded ribonucleic acid is administered to the subject once a week.
49 . The method of claim 46 or 47 16 , wherein the dsRNA double-stranded ribonucleic acid is administered to the subject twice a month.
50 . The method of claim 17 , wherein the disorder is a bleeding disorder.
51 . The method of claim 50 , wherein the bleeding disorder is a hemophilia.
52 . The method of claim 17 , wherein the administration of the dsRNA double-stranded ribonucleic acid to the subject causes an increase in blood clotting and/or a decrease in Serpinc1 protein accumulation.
53 . The method of claim 17 , wherein the dsRNA double-stranded ribonucleic acid is conjugated to a ligand.
54 . The method of claim 53 , wherein the ligand is conjugated to the 3′-end of the sense strand of the dsRNA double-stranded ribonucleic acid.
55 . The method of claim 54 , wherein the ligand is an N-acetylgalactosamine (GalNAc) derivative.
56 . The method of claim 55 , wherein the ligand is
57 . The method of claim 55 , wherein the dsRNA double-stranded ribonucleic acid is conjugated to the ligand as shown in the following schematic
and, wherein X is O or S.
58 . The method of claim 57 , wherein the X is O.
59 . The method of claim 17 , further comprising measuring thrombin levels in said subject.
60 . The method of claim 34 , wherein the ligand is
61 . The method of claim 34 , wherein the dsRNA double-stranded ribonucleic acid is conjugated to the ligand as shown in the following schematic
and, wherein X is O or S.
62 . The method of claim 61 , wherein the X is O.
63. A double-stranded ribonucleic acid comprising a sense strand, an antisense strand, and a ligand, wherein the nucleotide sequence of the sense strand consists of 5′-GfsgsUfuAfaCfaCfCfAfuUfuAfcUfuCfaAf-3′ (SEQ ID NO:941) and the nucleotide sequence of the antisense strand consists of 5′-usUfsgAfaGfuAfaAfuggUfgUfuAfaCfcsasg-3′ (SEQ ID NO:960), wherein a, g, c, and u are 2′-O-methyl A, G, C, and U, respectively; Af, Gf, Cf, and Uf are 2′-fluoro A, G, C, and U, respectively; and s is a phosphorothioate linkage, wherein the ligand is conjugated to the 3′ end of the sense strand as shown in the schematic
and wherein X is O.
64. A pharmaceutical composition comprising the double-stranded ribonucleic acid of claim 63 and phosphate buffered saline.
65. The pharmaceutical composition of claim 9 , wherein the pharmaceutical composition comprises phosphate buffered saline.
66. A method of treating hemophilia A or hemophilia B prophylactically in a subject in need thereof, comprising administering the double-stranded ribonucleic acid of claim 63 in a prophylactically effective amount to the subject in need thereof.
67. The method of claim 66 , wherein the subject is a hemophilia A or hemophilia B patient with inhibitors.
68. A method of treating hemophilia A or hemophilia B prophylactically in a subject in need thereof, comprising administering the pharmaceutical composition of claim 64 in a prophylactically effective amount to the subject in need thereof.
69. The method of claim 68 , wherein the subject is a hemophilia A or hemophilia B patient with inhibitors.
70. A method of treating hemophilia A or hemophilia B prophylactically in a subject in need thereof, comprising administering the pharmaceutical composition of claim 65 in a prophylactically effective amount to the subject in need thereof.
71. The method of any one of claims 44, 58, 62, 66, 68, or 70 , wherein the subject is a hemophilia A patient with inhibitors.
72. The method of any one of claims 44, 58, 62, 66, 68, and 70 , wherein the subject is a hemophilia B patient with inhibitors.
73. The method of any one of claims 44, 58, 62, 66, 68, and 70 , wherein the subject is a hemophilia A patient without inhibitors.
74. The method of any one of claims 44, 58, 62, 66, 68, and 70 , wherein the subject is a hemophilia B patient without inhibitors.