Angiotensinogen (AGT) iRNA compositions and methods of use thereof

Patent metadata

Field	Value
Publication number	US-10238749-B2
Application number	US-201515313145-A
Country	US
Kind code	B2
Filing date	May 22, 2015
Priority date	May 22, 2014
Publication date	Mar 26, 2019
Grant date	Mar 26, 2019

Abstract

The present invention relates to RNAi agents, e.g., double-stranded RNAi agents, targeting the angiotensinogen (AGT) gene, and methods of using such RNAi agents to inhibit expression of AGT and methods of treating subjects having an AGT-associated disorder, e.g., hypertension.

First claim

We claim:

1. A double-stranded ribonucleic acid (RNAi) agent for inhibiting expression of angiotensinogen (AGT) in a cell, wherein said double-stranded RNAi agent comprises a sense strand and an antisense strand forming a double-stranded region, wherein the sense strand comprises at least 15 contiguous nucleotides differing by no more than 3 nucleotides from nucleotides 2081-2104 of the nucleotide sequence of SEQ ID NO: 1, wherein the antisense strand is substantially complementary to the sense strand, wherein each of the sense strand and the antisense strand is independently 15 to 30 nucleotides in length, and wherein the RNAi agent comprises at least one modified nucleotide.

2. The double-stranded RNAi agent of claim 1 , wherein substantially all of the nucleotides of said sense strand are modified nucleotides, substantially all of the nucleotides of said antisense strand are modified nucleotides, or substantially all of the nucleotides of said sense strand and substantially all of the nucleotides of said antisense strand are modified nucleotides.

3. The double-stranded RNAi agent of claim 1 , wherein said sense strand is conjugated to a ligand attached at the 3′-terminus.

4. The double-stranded RNAi agent of claim 1 , wherein all of the nucleotides of said sense strand and all of the nucleotides of said antisense strand are modified nucleotides.

5. The double-stranded RNAi agent of claim 1 , wherein the antisense strand comprises the nucleotide sequence of 5′-AUUAGAAGAAAAGGUGGGAGACU-3′ (SEQ ID NO:537).

6. The double-stranded RNAi agent of claim 1 , wherein at least one of said modified nucleotides is selected from the group consisting of a 3′-terminal deoxy-thymine (dT) nucleotide, a 2′-O-methyl modified nucleotide, a 2′-fluoro modified nucleotide, a 2′-deoxy-modified nucleotide, a locked nucleotide, an unlocked nucleotide, a conformationally restricted nucleotide, a constrained ethyl nucleotide, an abasic nucleotide, a 2′-amino-modified nucleotide, a 2′-alkyl-modified nucleotide, a morpholino nucleotide, a phosphoramidate, a non-natural base comprising nucleotide, a nucleotide comprising a 5′-phosphorothioate group, and a terminal nucleotide linked to a cholesteryl derivative or a dodecanoic acid bisdecylamide group.

7. The double-stranded RNAi agent of claim 1 , wherein at least one strand comprises a 3′ overhang of at least 1 nucleotide; or at least one strand comprises a 3′ overhang of at least 2 nucleotides.

8. The double-stranded RNAi agent of claim 1 , wherein the double-stranded region is 15-30 nucleotide pairs in length; 17-23 nucleotide pairs in length; 17-25 nucleotide pairs in length; 23-27 nucleotide pairs in length; 19-21 nucleotide pairs in length; or 21-23 nucleotide pairs in length.

9. The double-stranded RNAi agent of claim 1 , wherein each of the sense strand and the antisense strand is independently 19 to 30 nucleotides in length or 21 to 23 nucleotides in length.

10. The double-stranded RNAi agent of claim 3 , wherein the ligand is one or more GalNAc derivatives attached through a bivalent or trivalent branched linker.

11. The double-stranded RNAi agent of claim 3 , wherein the ligand is

12. The double-stranded RNAi agent of claim 11 , wherein the RNAi agent is conjugated to the ligand as shown in the following schematic

wherein X is O or S.

13. The double-stranded RNAi agent of claim 1 , wherein said RNAi agent further comprises at least one phosphorothioate or methylphosphonate internucleotide linkage.

14. The double-stranded RNAi agent of claim 13 , 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.

15. The double-stranded RNAi agent of claim 14 , wherein said strand is the antisense strand.

16. The double-stranded RNAi agent of claim 14 , wherein said strand is the sense strand.

17. The double-stranded RNAi agent of claim 13 , wherein said RNAi agent comprises 6-8 phosphorothioate internucleotide linkages.

18. The double-stranded RNAi of claim 17 , wherein the antisense strand comprises two phosphorothioate internucleotide linkages at the 5′-terminus and two phosphorothioate internucleotide linkages at the 3′-terminus, and the sense strand comprises at least two phosphorothioate internucleotide linkages at either the 5′-terminus or the 3′-terminus.

19. The double-stranded RNAi agent of claim 1 , wherein the base pair at the 1 position of the 5′-end of the antisense strand is an AU base pair.

20. The double-stranded RNAi agent of claim 5 , wherein the sense strand of the RNAi agent comprises the nucleotide sequence of 5′-UCUCCCACCUUUUCUUCUAAU 3′ (SEQ ID NO:499) and the antisense strand comprises the nucleotide sequence of 5′-AUUAGAAGAAAAGGUGGGAGACU 3′ (SEQ ID NO:537).

21. A double-stranded ribonucleic acid (RNAi) agent for inhibiting expression of angiotensinogen (AGT),
wherein said double-stranded RNAi agent comprises a sense strand and an antisense strand forming a double-stranded region, wherein said sense strand comprises at least 15 contiguous nucleotides differing by no more than 3 nucleotides from nucleotides 2081-2104 of SEQ ID NO:1, and said antisense strand comprises at least 15 contiguous nucleotides differing by no more than 3 nucleotides from the nucleotide sequence of SEQ ID NO: 2,
wherein each of the sense strand and the antisense strand is independently 15 to 30 nucleotides in length,
wherein substantially all of the nucleotides of said sense strand comprise a modification selected from the group consisting of a 2′-O-methyl modification and a 2′-fluoro modification,
wherein said sense strand comprises two phosphorothioate internucleotide linkages at the 5′-terminus,
wherein substantially all of the nucleotides of said antisense strand comprise a modification selected from the group consisting of a 2′-O-methyl modification and a 2′-fluoro modification,
wherein said antisense strand comprises two phosphorothioate internucleotide linkages at the 5′-terminus and two phosphorothioate internucleotide linkages at the 3′-terminus, and
wherein said sense strand is conjugated to one or more GalNAc derivatives attached through a branched bivalent or trivalent linker at the 3′-terminus.

22. The double-stranded RNAi agent of claim 21 , wherein all of the nucleotides of said sense strand and all of the nucleotides of said antisense strand comprise a modification.

23. A double-stranded ribonucleic acid (RNAi) agent comprising a sense strand and an antisense strand, wherein said sense strand comprises the nucleotide sequence of 5′-UCUCCCACCUUUUCUUCUAAU-3′ (SEQ ID NO:499), and said antisense strand comprises the nucleotide sequence of 5′-AUUAGAAGAAAAGGUGGGAGACU-3′ (SEQ ID NO:537).

24. A composition comprising a modified antisense polynucleotide agent, wherein the agent comprises about 14 to about 30 contiguous nucleotides, wherein said agent is capable of inhibiting the expression of AGT in a cell, and comprises a nucleotide sequence substantially complementary to the nucleotide sequence of 5′-UCUCCCACCUUUUCUUCUAAU-3′ (SEQ ID NO:499).

25. An isolated cell containing the double-stranded RNAi agent of claim 1 .

26. A pharmaceutical com

Assignees

• Alnylam Pharmaceuticals Inc

Inventors

• Foster Donald
• Bettencourt Brian
• Charisse Klaus
• Hinkle Gregory
• Kuchimanchi Satyanarayana
• Maier Martin
• Milstein Stuart

Classifications

• A61P43/00

  Drugs for specific purposes, not provided for in groups A61P1/00-A61P41/00 · CPC title

• A61P9/12

  Antihypertensives · CPC title

• A61P9/10

  for treating ischaemic or atherosclerotic diseases, e.g. antianginal drugs, coronary vasodilators, drugs for myocardial infarction, retinopathy, cerebrovascula insufficiency, renal arteriosclerosis · CPC title

• A61P9/04

  Inotropic agents, i.e. stimulants of cardiac contraction; Drugs for heart failure · CPC title

• A61P9/00

  Drugs for disorders of the cardiovascular system · CPC title