Chimeric double-stranded nucleic acid
A method of reducing the level of a transcription product in a cell comprising contacting with the cell a composition comprising a double-stranded nucleic acid complex comprising a first nucleic acid strand annealed to a second nucleic acid strand, wherein: (i) the first nucleic acid strand hybridizes to the transcription product and comprises (a) a region consisting of at least 4 consecutive nucleotides that are recognized by RNase H when the strand is hybridized to the transcription product, (b) one or more nucleotide analogs located on 5′ terminal side of the region, (c) one or more nucleotide analogs located on 3′ terminal side of the region and (d) a total number of nucleotides and nucleotide analogs ranging from 8 to 35 nucleotides and (ii) the second nucleic acid strand comprises (a) nucleotides and optionally nucleotide analogs and (b) at least 4 consecutive RNA nucleotides.
1. A pharmaceutical composition for reducing the level of a transcription product in a cell comprising:
a double-stranded nucleic acid complex comprising a first nucleic acid strand, a second nucleic acid strand which has a sequence complementary to the first nucleic acid strand, wherein:
(i) the first nucleic acid strand hybridizes to the transcription product, and is (a) a nucleic acid strand which comprises at least 4 consecutive DNA nucleotides or modified DNA nucleotides that are recognized by RNase H when the strand is hybridized to the transcription product and the total number of nucleotides is 12 to 25; and
(ii) the second nucleic acid strand is a nucleic acid strand which comprises (a) at least 4 consecutive RNA nucleotides, and (b) one or more modified nucleotides, nucleotide analogs and/or modified nucleotide analogs located 5′ to the at least 4 consecutive RNA nucleotides, and/or (c) one or more modified nucleotides, nucleotide analogs and/or modified nucleotide analogs located 3′ to the at least 4 consecutive RNA nucleotides and the total number of nucleotides is 12 to 31.
2. The pharmaceutical composition according to claim 1 , wherein the first nucleic acid strand is further comprises (b) a 5′ wing region which comprises one or more nucleotide analogs or modified nucleotide analogs located 5′ to the at least 4 consecutive DNA nucleotides or modified DNA nucleotides that are recognized by RNase H; and/or (c) a 3′ wing region which comprises one or more nucleotide analogs or modified nucleotide analogs located 3′ to the at least 4 consecutive DNA nucleotides or modified DNA nucleotides that are recognized by RNase H.
3. The pharmaceutical composition according to claim 1 , wherein the double stranded nucleic acid complex further comprises a third nucleic acid strand annealed to the second nucleic acid strand.
4. The pharmaceutical composition according to claim 3 , wherein the third nucleic acid strand (i) comprises nucleotides and optionally nucleotide analogs and total number of the nucleotides and nucleotide analogs which are optionally comprised is 8 to 100, (ii) comprises at least 4 consecutive nucleotides that are recognized by RNase H when the strand is hybridized to a second transcription product, and (iii) hybridizes to the second transcription product.
5. The pharmaceutical composition according to claim 3 , wherein the third nucleic acid strand hybridizes to a transcription product that is identical to the transcription product to which the first nucleic acid strand hybridizes.
6. The pharmaceutical composition according to claim 3 , wherein the third nucleic acid strand hybridizes to a transcription product different from the transcription product to which the first nucleic acid strand hybridizes.
7. The pharmaceutical composition according to claim 1 , wherein the second nucleic acid strand further comprises a functional moiety having a function selected from a labeling function, a purification function, and a targeted delivery function.
8. The pharmaceutical composition according to claim 1 , wherein the nucleotide analogs in the first nucleic acid strand are bridged nucleotides.
9. The pharmaceutical composition according to claim 1 , wherein at least one of the nucleotides and the nucleotide analogs in the first nucleic acid strand is phosphorothioated.
10. The pharmaceutical composition according to claim 7 , wherein the functional moiety is a molecule selected from the group consisting of a lipid, a glycolipid, a glyceride, a sugar, a peptide, and a protein.
11. The pharmaceutical composition according to claim 7 , wherein the functional moiety is a lipid selected from the group consisting of a fatty acid, a lipid-soluble vitamin, a glycolipid, and a glyceride.
12. The pharmaceutical composition according to claim 7 , wherein the functional moiety is a molecule selected from the group consisting of cholesterol, a tocopherol, a tocotrienol, glucose, sucrose, acylcarintine, acyl-CoA, and an antibody.
13. The pharmaceutical composition according to claim 1 , wherein at least one of the nucleotide analogs is a molecule selected from the group consisting of a hexitol nucleic acid (HNA), a cyclohexane nucleic acid (CeNA), a peptide nucleic acid (PNA), a glycol nucleic acid (GNA), a threose nucleic acid (TNA), a morpholino nucleic acid, a tricyclo-DNA (tcDNA), a 2′-O-methylated nucleic acid, a 2′-MOE (2′-O-methoxyethyl) lated nucleic acid, a 2′-AP (2′-O-aminopropyl) lated nucleic acid, a 2′-fluorinated nucleic acid, a 2′-F-arabinonucleic acid (2′-FANA), and a bridged nucleic acid (BNA).
14. The pharmaceutical composition according to claim 8 , wherein the BNA is a molecule selected from the group consisting of α-L-methyleneoxy (4′-CH 2--O-2′ ) BNA, β-D-methyleneoxy (4′-CH 2--O-2′ ) BNA, ethyleneoxy (4′-CH 2-2-O-2′ ) BNA, β-D-thio (4′-CH 2--S-2′ ) BNA, aminooxy (4′-CH 2--O--N(R3) -2′) BNA, oxyamino (4′-CH 2′N(R3) --O-2′) BNA, 2′, 4′-BNA-COC, 3′-amino-2′, 4′-BNA, 5′-methyl BNA, 4′-CH(CH 3 )--O-2′-BNA, 4′-CH(CH 2OCH3 )--O-2′-BNA, amide (4′-C(O)--N(H)-2′) BNA, and amide (4′-C(O)--N(Me)-2′) BNA.