Title - ISOLATED DOUBLE-STRANDED RNAi MOLECULE THAT INHIBITS AGT EXPRESSION IN A CELL

Abstract

Claim 1: A double-stranded ribonucleic acid (RNAi)-based agent for inhibiting angiotensinogen (AGT) expression in a cell, characterized in that it comprises a frame-directed strand and an antisense strand forming a double-stranded region, wherein the frame-directed strand comprises at least 15 contiguous nucleotides, of which no more than 3 differ from the nucleotide sequence depicted in SEQ ID No. 1, and the antisense strand comprises at least 15 contiguous nucleotides, of which no more than 3 differ from the nucleotide sequence depicted in SEQ ID No. 2, wherein substantially all nucleotides in the frame-directed strand and substantially all nucleotides in the antisense strand are modified nucleotides,and wherein the frame-directed strand is conjugated with a ligand attached at the end 3. Claim 32: The double-stranded RNAi-based agent according to any of claims 1-4 and 10, characterized in that the ligand is of formula (1). Claim 69: A pharmaceutical composition characterized in that it comprises a double-stranded RNAi-based agent according to any one of claims 1-4, 10, 57, 58 and 60-65, a composition comprising an agent that is a modified antisense polynucleotide according to claim 66 or a vector according to claim 67. Claim 75: A method for inhibiting the expression of angiotensinogen (AGT) in a cell, characterized in that it comprises (a) contacting the cell with a double-stranded RNAi-based agent according to any one of claims 1-4, 10, 57, 58 and 60-65,with a composition comprising an agent that is a modified antisense polynucleotide according to claim 63, with a vector according to claim 67, or with a pharmaceutical composition according to any of claims 69-74; and (b) maintaining the cell produced in step (a) for a period of time sufficient to induce degradation of the mRNA transcribed from a gene encoding AGT, so as to inhibit expression of the gene encoding AGT. Claim 81: A method for treating an angiotensinogen-associated disorder (AGT) in a subject, characterized in that it comprises administering to the subject subcutaneously a therapeutically effective amount of a double-stranded ribonucleic acid (RNAi)-based agent, wherein the double-stranded RNAi-based agent comprises a frame-sense strand and an antisense strand forming a double-stranded region,wherein the frame-directed strand comprises at least 15 contiguous nucleotides, of which no more than 3 differ from the nucleotide sequence depicted in SEQ ID No. 1, and the antisense strand comprises at least 15 contiguous nucleotides, of which no more than 3 differ from the nucleotide sequence depicted in SEQ ID No. 2, wherein substantially all nucleotides in the frame-directed strand have been modified with groups selected from 2-O-methyl and 2-fluoro, wherein the antisense strand comprises two phosphorothioate bonds between nucleotides at the 5' end and two phosphorothioate bonds between nucleotides at the 3' end, wherein substantially all nucleotides in the antisense strand have been modified with groups selected from 2-O-methyl and 2-fluoro,where the frame-directed chain comprises two phosphorothioate linkages between nucleotides at the 5' end, and where the frame-directed chain is conjugated with one or more GalNAc derivatives that are linked via a bivalent or trivalent branched connector at the 3' end, so as to effect the treatment in the subject.

Claims

1. An isolated double-stranded ribonucleic acid (RNAi) molecule, or a pharmaceutically acceptable salt thereof, that inhibits angiotensinogen (AGT) expression in a cell, characterized in that the double-stranded ribonucleic acid (RNAi) molecule comprises a frame-sense strand and an antisense strand forming a double-stranded region, wherein the sense strand comprises at least 18 contiguous nucleotides of the nucleotide sequence 5'-UCAUCCACAAUGAGAGUAA-3' of SEQ ID NO: 679 and the antisense strand comprises at least 18 contiguous nucleotides of the nucleotide sequence 5'-UUACUCUCAUUGUGGAUGA-3' of SEQ ID NO: 876, wherein each of the sense and antisense strands is independently 19 to 23 nucleotides in length,wherein all nucleotides in the sense strand and all nucleotides in the antisense strand comprise a nucleotide modification selected from the group consisting of a 2'-O-methyl nucleotide modification, a 2'-fluoro nucleotide modification, and a glycolic nucleic acid (GNA) nucleotide modification; and wherein the 3' end of the sense strand is conjugated with a ligand comprising one or more GalNAc derivatives linked via a bivalent or trivalent branched connector.