Double-stranded agents for delivering therapeutic oligonucleotides

A double-stranded nucleic acid agent comprising a first nucleic acid strand annealed to a second nucleic acid strand, wherein:

• the first nucleic acid strand comprises
• (i) an RNA region with 8-11 consecutive natural RNA nucleotides that can be recognized by RNase H when the first nucleic acid strand is hybridized to the second nucleic acid strand,
• (ii) a therapeutic oligonucleotide region wherein at least one internucleotide linkage at the 3′ end and at the 5′ end of the therapeutic oligonucleotide region is more nuclease-resistant than a natural internucleotide linkage, wherein the at least one nuclease-resistant internucleotide linkage is a phosphorothioate group, and wherein the therapeutic oligonucleotide region is
• (a) a single-stranded region that hybridizes to a precursor mRNA (pre-mRNA) or a micro-RNA, but does not hybridize to the second nucleic acid strand,
• (b) a splice-switching oligonucleotide, an intron targeting gapmer, or an antagomir,
• (c) linked to the 3′ end or 5′ end of the RNA region of the first nucleic acid strand, and
• (d) released from the double-stranded nucleic acid agent in vivo via RNase H-mediated cleavage of the RNA region of (c), and
• (e) comprised of one or more of modified nucleotides, nucleotide analogues, or DNA nucleotides comprising a phosphorothioate group, and
• (iii) at least one internucleotide linkage at the 3′ end and at the 5′ end of the RNA region of the first nucleic acid strand is more nuclease-resistant than a natural internucleotide linkage, wherein the at least one nuclease-resistant internucleotide linkage is a phosphorothioate group, and
• the second nucleic acid strand comprises
• (i) a 13 consecutive DNA/LNA gapmer or DNA/LNA mixmer region that is hybridized to the RNA region of the first nucleic acid strand and can promote the recognition of the 8-11 consecutive natural RNA nucleotides in the first nucleic acid strand by RNase H, and
• (ii) at least one internucleotide linkage at the 3′ end and at the 5′ end of the second nucleic acid strand is more nuclease-resistant than a natural internucleotide linkage, wherein the at least one nuclease-resistant internucleotide linkage is a phosphorothioate group,
• and wherein
• the double-stranded nucleic acid agent further comprises a targeting moiety which is (a) selected from a lipid, a sugar, a peptide, and a protein and (b) joined to the 3′-terminal nucleotide or the 5′-terminal nucleotide of the second nucleic acid strand, or to the 3′-terminal nucleotide or the 5′-terminal nucleotide of the first nucleic acid strand.