Angiopoietin-like 3 (ANGPTL3) iRNA compositions and methods of use thereof
The present invention relates to RNAi agents, e.g., double stranded RNA (dsRNA) agents, targeting the Angiopoietin-like 3 (ANGPTL3) gene. The invention also relates to methods of using such RNAi agents to inhibit expression of an ANGPTL3 gene and to methods of preventing and treating an ANGPTL3-associated disorder, e.g., a disorder of lipid metabolism, such as hyperlipidemia or hypertriglyceridemia.
1 . A method of inhibiting expression of a Angiopoietin-like 3 (ANGPTL3) gene in a cell, the method comprising contacting the cell with a dsRNA agent, or salt thereof,
wherein the dsRNA agent, or salt thereof, comprises a sense strand and an antisense strand forming a double stranded region,
wherein the sense strand comprises at least 19 contiguous nucleotides of the nucleotide sequence 5′-AAGCUCCUUCUUUUUAUUGUU-3′ of SEQ ID NO:46 and the antisense strand comprises at least 21 contiguous nucleotides of the nucleotide sequence of 5′-AACAAUAAAAAGAAGGAGCUUGG-3′ of SEQ ID NO:661,
wherein all of the nucleotides of the sense strand and all of the nucleotides of the antisense strand comprise a modification, and
wherein at least one of the modifications is a 2′-deoxy-nucleotide modification, thereby inhibiting expression of the ANGPTL3 gene in the cell.
2 . The method of claim 1 , wherein contacting the cell with the dsRNA agent, or salt thereof, inhibits the expression of ANGPTL3 by at least 50%.
3 . The method of claim 1 , wherein the cell is within a subject.
4 . The method of claim 3 , wherein the subject is a human.
5 . The method of claim 3 , wherein inhibiting expression of ANGPTL3 decreases ANGPTL3 protein level in serum of the subject by at least 50%.
6 . The method of claim 5 , wherein the subject has an ANGPTL3-associated disorder.
7 . The method of claim 6 , wherein the ANGPTL3-associated disorder is a disorder of lipid metabolism.
8 . The method of claim 7 , wherein the disorder of lipid metabolism is hyperlipidemia or hypertriglyceridemia.
9 . The method of claim 1 , wherein the double stranded region is 19-30 nucleotide pairs in length.
10 . The method of claim 1 , wherein the double stranded region is 21-23 nucleotide pairs in length.
11 . The method of claim 1 , wherein each strand is independently 19-30 nucleotides in length.
12 . The method of claim 1 , wherein at least one strand comprises a 3′ overhang of at least 1 nucleotide.
13 . The method of claim 1 , wherein the dsRNA agent, or salt thereof, further comprises a ligand.
14 . The method of claim 13 , wherein the ligand is conjugated to the 3′ end of the sense strand of the dsRNA agent, or salt thereof.
15 . The method of claim 13 , wherein the ligand is an N-acetylgalactosamine (GalNAc) derivative.
16 . The method of claim 13 , wherein the ligand is one or more GalNAc derivatives attached through a monovalent, bivalent, or trivalent linker.
17 . The method of claim 16 , wherein the ligand is
18 . The method of claim 17 , wherein the dsRNA agent, or salt thereof, is conjugated to the ligand as shown in the following schematic
wherein X is O or S.
19 . The method of claim 18 , wherein X is O.
20 . The method of claim 1 , wherein the dsRNA agent, or salt thereof, further comprises at least one phosphorothioate or methylphosphonate internucleotide linkage.
21 . The method of claim 20 , wherein the phosphorothioate or methylphosphonate internucleotide linkage is at the 3′-terminus of one strand; the 5′-terminus of one strand; or at both the 5′- and 3′-terminus of one strand.
22 . The method of claim 1 , wherein the sense strand is 21 nucleotides in length and the antisense strand is 23 nucleotides in length.
23 . The method of claim 1 , wherein the sense strand comprises the nucleotide sequence 5′-AAGCUCCUUCUUUUUAUUGUU-3′ of SEQ ID NO: 46.
24 . The method of claim 1 , wherein the sense strand comprises the nucleotide sequence 5′-AAGCUCCUUCUUUUUAUUGUU-3′ of SEQ ID NO: 46 and the antisense strand comprises the nucleotide sequence 5′-AACAAUAAAAAGAAGGAGCUUGG-3′ of SEQ ID NO:661.
25 . The method of claim 1 , wherein the sense strand consists of the nucleotide sequence 5′-AAGCUCCUUCUUUUUAUUGUU-3′ of SEQ ID NO:46 and the antisense strand consists of the nucleotide sequence 5′-AACAAUAAAAAGAAGGAGCUUGG-3′ of SEQ ID NO:661.
26 . The method of claim 1 , wherein the nucleotide sequence of the sense strand differs by no more than 4 bases from the nucleotide sequence 5′-asasgcuccuUfCfUfuuuuauuguu-3′ of SEQ ID NO: 18 and the nucleotide sequence of the antisense strand differs by no more than 4 bases from the nucleotide sequence 5′-asdAscadAudAaaaadGaAfggagcuusgsg-3′ of SEQ ID NO:19,
wherein a, g, c and u are 2′-O-methyl (2′-OMe) A, G, C, and U, respectively; dA and dG, are 2′-deoxy A and G, respectively; Af, Cf and Uf are 2′-fluoro (2′-F) A, C and U, respectively; and s is a phosphorothioate linkage.
27 . The method of claim 1 , wherein the nucleotide sequence of the sense strand differs by no more than 3 bases from the nucleotide sequence 5′-asasgcuccuUfCfUfuuuuauuguu-3′ of SEQ ID NO: 18 and the nucleotide sequence of the antisense strand differs by no more than 3 bases from the nucleotide sequence 5′-asdAscadAudAaaaadGaAfggagcuusgsg-3′ of SEQ ID NO: 19,
wherein a, g, c and u are 2′-O-methyl (2′-OMe) A, G, C, and U, respectively; dA and dG, are 2′-deoxy A and G, respectively; Af, Cf and Uf are 2′-fluoro (2′-F) A, C and U, respectively; and s is a phosphorothioate linkage.
28 . The method of claim 1 , wherein the nucleotide sequence of the sense strand differs by no more than 2 bases from the nucleotide sequence 5′-asasgcuccuUfCfUfuuuuauuguu-3′ of SEQ ID NO: 18 and the nucleotide sequence of the antisense strand differs by no more than 2 bases from the nucleotide sequence 5′-asdAscadAudAaaaadGaAfggagcuusgsg-3′ of SEQ ID NO:19,
wherein a, g, c and u are 2′-O-methyl (2′-OMe) A, G, C, and U, respectively; dA and dG, are 2′-deoxy A and G, respectively; Af, Cf and Uf are 2′-fluoro (2′-F) A, C and U, respectively; and s is a phosphorothioate linkage.
29 . The method of claim 1 , wherein the nucleotide sequence of the sense strand differs by no more than 1 base from the nucleotide sequence 5′-asasgcuccuUfCfUfuuuuauuguu-3′ of SEQ ID NO: 18 and the nucleotide sequence of the antisense strand differs by no more than 1 base from the nucleotide sequence 5′-asdAscadAudAaaaadGaAfggagcuusgsg-3′ of SEQ ID NO:19,
wherein a, g, c and u are 2′-O-methyl (2′-OMe) A, G, C, and U, respectively; dA and dG, are 2′-deoxy A and G, respectively; Af, Cf and Uf are 2′-fluoro (2′-F) A, C and U, respectively; and s is a phosphorothioate linkage.
30 . The method of claim 1 , wherein the sense strand comprises the nucleotide sequence 5′-asasgcuccuUfCfUfuuuuauuguu-3′ of SEQ ID NO:18 and the antisense strand comprises the nucleotide sequence 5′-asdAscadAudAaaaadGaAfggagcuusgsg-3′ of SEQ ID NO:19,
wherein a, g, c and u are 2′-O-methyl (2′-OMe) A, G, C, and U, respectively; dA and dG, are 2′-deoxy A and G, respectively; Af, Cf and Uf are 2′-fluoro (2′-F) A, C and U, respectively; and s is a phosphorothioate linkage.
31 . The method of claim 1 , wherein the sense strand consists of the nucleotide sequence 5′-asasgcuccuUfCfUfuuuuauuguu-3′ of SEQ ID NO:18 and the antisense strand consists of the nucleotide sequence 5′-asdAscadAudAaaaadGaAfggagcuusgsg-3′ of SEQ ID NO:19,
wherein a, g, c and u are 2′-O-methyl (2′-OMe) A, G, C, and U, respectively; dA and dG, are 2′-deoxy A and G, respectively; Af, Cf and Uf are 2′-fluoro (2′-F) A, C and U, respectively; and s is a phosphorothioate linkage.
32 . The method of claim 1 , wherein the sense strand comprises the nucleotide sequence 5′-asasgcuccuUfCfUfuuuuauuguu-L96-3′ of SEQ ID NO:20 and the antisense strand comprises the nucleotide sequence 5′-asdAscadAudAaaaadGaAfggagcuusgsg-3′ of SEQ ID NO: 19,
wherein a, g, c and u are 2′-O-methyl (2′-OMe) A, G, C, and U, respectively; dA and dG, are 2′-deoxy A and G, respectively; Af, Cf and Uf are 2′-fluoro (2′-F) A, C and U, respectively; and s is a phosphorothioate linkage, and
wherein L96 is a ligand conjugated to the 3′-end of the sense strand as shown in the following schematic
wherein X is O.
33 . A method of treating a subject having a disorder that would benefit from reduction in Angiopoietin-like 3 (ANGPTL3) expression, comprising administering to the subject a therapeutically effective amount of a dsRNA agent, or salt thereof,
wherein the dsRNA agent, or salt thereof comprises a sense strand and an antisense strand forming a double stranded region,
wherein the sense strand comprises at least 19 contiguous nucleotides of the nucleotide sequence 5′-AAGCUCCUUCUUUUUAUUGUU-3′ of SEQ ID NO:46 and the antisense strand comprises at least 21 contiguous nucleotides of the nucleotide sequence of 5′-AACAAUAAAAAGAAGGAGCUUGG-3′ of SEQ ID NO:661,
wherein all of the nucleotides of the sense strand and all of the nucleotides of the antisense strand comprise a modification, and
wherein at least one of the modifications is a 2′-deoxy-nucleotide modification, thereby treating the subject having the disorder that would benefit from reduction in ANGPTL3 expression.
34 . The method of claim 33 , wherein the subject is a human.
35 . The method of claim 33 , wherein the disorder is an ANGPTL3-associated disorder.
36 . The method of claim 35 , wherein the ANGPTL3-associated disorder is a disorder of lipid metabolism.
37 . The method of claim 36 , wherein the disorder of lipid metabolism is hyperlipidemia or hypertriglyceridemia.
38 . The method of claim 33 , wherein the administration of the dsRNA agent, or salt thereof, to the subject causes a decrease in one or more serum lipid and/or a decrease in ANGPTL3 protein accumulation.
39 . The method of claim 33 , wherein the dsRNA agent, or salt thereof, is administered to the subject subcutaneously.
40 . The method of claim 33 , further comprising administering to the subject an additional therapeutic agent for treatment of an ANGPTL3-associated disorder.
41 . The method of claim 33 , wherein the double stranded region is 19-30 nucleotide pairs in length.
42 . The method of claim 33 , wherein the double stranded region is 21-23 nucleotide pairs in length.
43 . The method of claim 33 , wherein each strand is independently 19-30 nucleotides in length.
44 . The method of claim 33 , wherein at least one strand comprises a 3′ overhang of at least 1 nucleotide.
45 . The method of claim 33 , wherein the dsRNA agent, or salt thereof, further comprises a ligand.
46 . The method of claim 45 , wherein the ligand is conjugated to the 3′ end of the sense strand of the dsRNA agent, or salt thereof.
47 . The method of claim 45 , wherein the ligand is an N-acetylgalactosamine (GalNAc) derivative.
48 . The method of claim 45 , wherein the ligand is one or more GalNAc derivatives attached through a monovalent, bivalent, or trivalent linker.
49 . The method of claim 48 , wherein the ligand is
50 . The method of claim 49 , wherein the dsRNA agent, or salt thereof, is conjugated to the ligand as shown in the following schematic
wherein X is O or S.
51 . The method of claim 50 , wherein X is O.
52 . The method of claim 33 , wherein the dsRNA agent, or salt thereof, further comprises at least one phosphorothioate or methylphosphonate internucleotide linkage.
53 . The method of claim 52 , wherein the phosphorothioate or methylphosphonate internucleotide linkage is at the 3′-terminus of one strand; the 5′-terminus of one strand; or at both the 5′- and 3′-terminus of one strand.
54 . The method of claim 33 , wherein the sense strand is 21 nucleotides in length and the antisense strand is 23 nucleotides in length.
55 . The method of claim 33 , wherein the sense strand comprises the nucleotide sequence 5′-AAGCUCCUUCUUUUUAUUGUU-3′ of SEQ ID NO:46.
56 . The method of claim 33 , wherein the sense strand comprises the nucleotide sequence 5′-AAGCUCCUUCUUUUUAUUGUU-3′ of SEQ ID NO:46 and the antisense strand comprises the nucleotide sequence 5′-AACAAUAAAAAGAAGGAGCUUGG-3′ of SEQ ID NO:661.
57 . The method of claim 33 , wherein the sense strand consists of the nucleotide sequence 5′-AAGCUCCUUCUUUUUAUUGUU-3′ of SEQ ID NO:46 and the antisense strand consists of the nucleotide sequence 5′-AACAAUAAAAAGAAGGAGCUUGG-3′ of SEQ ID NO:661.
58 . The method of claim 33 , wherein the nucleotide sequence of the sense strand differs by no more than 4 bases from the nucleotide sequence 5′-asasgcuccuUfCfUfuuuuauuguu-3′ of SEQ ID NO: 18 and the nucleotide sequence of the antisense strand differs by no more than 4 bases from the nucleotide sequence 5′-asdAscadAudAaaaadGaAfggagcuusgsg-3′ of SEQ ID NO:19,
wherein a, g, c and u are 2′-O-methyl (2′-OMe) A, G, C, and U, respectively; dA and dG, are 2′-deoxy A and G, respectively; Af, Cf and Uf are 2′-fluoro (2′-F) A, C and U, respectively; and s is a phosphorothioate linkage.
59 . The method of claim 33 , wherein the nucleotide sequence of the sense strand differs by no more than 3 bases from the nucleotide sequence 5′-asasgcuccuUfCfUfuuuuauuguu-3′ of SEQ ID NO: 18 and the nucleotide sequence of the antisense strand differs by no more than 3 bases from the nucleotide sequence 5′-asdAscadAudAaaaadGaAfggagcuusgsg-3′ of SEQ ID NO: 19,
wherein a, g, c and u are 2′-O-methyl (2′-OMe) A, G, C, and U, respectively; dA and dG, are 2′-deoxy A and G, respectively; Af, Cf and Uf are 2′-fluoro (2′-F) A, C and U, respectively; and s is a phosphorothioate linkage.
60 . The method of claim 33 , wherein the nucleotide sequence of the sense strand differs by no more than 2 bases from the nucleotide sequence 5′-asasgcuccuUfCfUfuuuuauuguu-3′ of SEQ ID NO: 18 and the nucleotide sequence of the antisense strand differs by no more than 2 bases from the nucleotide sequence 5′-asdAscadAudAaaaadGaAfggagcuusgsg-3′ of SEQ ID NO:19,
wherein a, g, c and u are 2′-O-methyl (2′-OMe) A, G, C, and U, respectively; dA and dG, are 2′-deoxy A and G, respectively; Af, Cf and Uf are 2′-fluoro (2′-F) A, C and U, respectively; and s is a phosphorothioate linkage.
61 . The method of claim 33 , wherein the nucleotide sequence of the sense strand differs by no more than 1 base from the nucleotide sequence 5′-asasgcuccuUfCfUfuuuuauuguu-3′ of SEQ ID NO: 18 and the nucleotide sequence of the antisense strand differs by no more than 1 base from the nucleotide sequence 5′-asdAscadAudAaaaadGaAfggagcuusgsg-3′ of SEQ ID NO:19,
wherein a, g, c and u are 2′-O-methyl (2′-OMe) A, G, C, and U, respectively; dA and dG, are 2′-deoxy A and G, respectively; Af, Cf and Uf are 2′-fluoro (2′-F) A, C and U, respectively; and s is a phosphorothioate linkage.
62 . The method of claim 33 , wherein the sense strand comprises the nucleotide sequence 5′-asasgcuccuUfCfUfuuuuauuguu-3′ of SEQ ID NO:18 and the antisense strand comprises the nucleotide sequence 5′-asdAscadAudAaaaadGaAfggagcuusgsg-3′ of SEQ ID NO:19,
wherein a, g, c and u are 2′-O-methyl (2′-OMe) A, G, C, and U, respectively; dA and dG, are 2′-deoxy A and G, respectively; Af, Cf and Uf are 2′-fluoro (2′-F) A, C and U, respectively; and s is a phosphorothioate linkage.
63 . The method of claim 33 , wherein the sense strand consists of the nucleotide sequence 5′-asasgcuccuUfCfUfuuuuauuguu-3′ of SEQ ID NO: 18 and the antisense strand consists of the nucleotide sequence 5′-asdAscadAudAaaaadGaAfggagcuusgsg-3′ of SEQ ID NO:19,
wherein a, g, c and u are 2′-O-methyl (2′-OMe) A, G, C, and U, respectively; dA and dG, are 2′-deoxy A and G, respectively; Af, Cf and Uf are 2′-fluoro (2′-F) A, C and U, respectively; and s is a phosphorothioate linkage.
64 . The method of claim 33 , wherein the sense strand comprises the nucleotide sequence 5′-asasgcuccuUfCfUfuuuuauuguu-L96-3′ of SEQ ID NO:20 and the antisense strand comprises the nucleotide sequence 5′-asdAscadAudAaaaadGaAfggagcuusgsg-3′ of SEQ ID NO:19,
wherein a, g, c and u are 2′-O-methyl (2′-OMe) A, G, C, and U, respectively; dA and dG, are 2′-deoxy A and G, respectively; Af, Cf and Uf are 2′-fluoro (2′-F) A, C and U, respectively; and s is a phosphorothioate linkage, and
wherein L96 is a ligand conjugated to the 3′-end of the sense strand as shown in the following schematic
wherein X is O.
65 . A method of treating a subject having a disorder that would benefit from reduction in Angiopoietin-like 3 (ANGPTL3) expression, comprising administering to the subject a therapeutically effective amount of a dsRNA agent, or salt thereof,
wherein the dsRNA agent, or salt thereof, comprises a sense strand and an antisense strand forming a double stranded region,
wherein the sense strand comprises the nucleotide sequence 5′-asasgcuccuUfCfUfuuuuauuguu-L96-3′ of SEQ ID NO:20 and the antisense strand comprises the nucleotide sequence 5′-asdAscadAudAaaaadGaAfggagcuusgsg-3′ of SEQ ID NO:19,
wherein a, g, c and u are 2′-O-methyl (2′-OMe) A, G, C, and U, respectively; dA and dG, are 2′-deoxy A and G, respectively; Af, Cf and Uf are 2′-fluoro (2′-F) A, C and U, respectively; and s is a phosphorothioate linkage, and
wherein L96 is a ligand conjugated to the 3′-end of the sense strand as shown in the following schematic
wherein X is O, thereby treating the subject.
66 . The method of claim 65 , wherein the subject is human.
67 . The method of claim 66 , wherein the disorder is an ANGPTL3-associated disorder.
68 . The method of claim 67 , wherein the ANGPTL3-associated disorder is a disorder of lipid metabolism.
69 . The method of claim 68 , wherein the disorder of lipid metabolism is hyperlipidemia or hypertriglyceridemia.
70 . The method of claim 65 , wherein the administration of the agent to the subject causes a decrease in one or more serum lipid and/or a decrease in ANGPTL3 protein accumulation.
71 . The method of claim 65 , wherein the dsRNA agent, or salt thereof, is administered to the subject subcutaneously.
72 . The method of claim 65 , further comprising administering to the subject an additional therapeutic agent for treatment of an ANGPTL3-associated disorder.
73 . The method of claim 65 , wherein the dsRNA agent is in a salt form.
74 . The method of claim 65 , wherein the dsRNA agent, or salt thereof, is present in a pharmaceutical composition.
75 . A method of treating a subject having a disorder that would benefit from reduction in Angiopoietin-like 3 (ANGPTL3) expression, comprising administering to the subject a therapeutically effective amount of a dsRNA agent, or salt thereof,
wherein the dsRNA agent, or salt thereof, comprises a sense strand and an antisense strand forming a double stranded region,
wherein the sense strand consists of the nucleotide sequence 5′-asasgcuccuUfCfUfuuuuauuguu-L96-3′ of SEQ ID NO:20 and the antisense strand consists of the nucleotide sequence 5′-asdAscadAudAaaaadGaAfggagcuusgsg-3′ of SEQ ID NO:19,
wherein a, g, c and u are 2′-O-methyl (2′-OMe) A, G, C, and U, respectively; dA and dG, are 2′-deoxy A and G, respectively; Af, Cf and Uf are 2′-fluoro (2′-F) A, C and U, respectively; and s is a phosphorothioate linkage, and
wherein L96 is a ligand conjugated to the 3′-end of the sense strand as shown in the following schematic
wherein X is O, thereby treating the subject.
76 . The method of claim 75 , wherein the subject is human.
77 . The method of claim 76 , wherein the disorder is an ANGPTL3-associated disorder.
78 . The method of claim 77 , wherein the ANGPTL3-associated disorder is a disorder of lipid metabolism.
79 . The method of claim 78 , wherein the disorder of lipid metabolism is hyperlipidemia or hypertriglyceridemia.
80 . The method of claim 75 , wherein the administration of the agent to the subject causes a decrease in one or more serum lipid and/or a decrease in ANGPTL3 protein accumulation.
81 . The method of claim 75 , wherein the dsRNA agent, or salt thereof, is administered to the subject subcutaneously.
82 . The method of claim 75 , further comprising administering to the subject an additional therapeutic agent for treatment of an ANGPTL3-associated disorder.
83 . The method of claim 75 , wherein the dsRNA agent is in a salt form.
84 . The method of claim 75 , wherein the dsRNA agent, or salt thereof, is present in a pharmaceutical composition.