Oligonucleotide duplexes comprising DNA-like and RNA-like nucleotides and uses thereof
View Patent ↗Novel oligonucleotide pairs which can form a duplex comprising one or more DNA-like nucleotides (e.g., 2′-substituted arabinonucleotides (ANA)); in combination with one or more RNA-like nucleotides (e.g., 2′-substituted ribonucleotides (RNA) and/or locked nucleic acid nucleotides (LNA)), are disclosed. The use of such oligonucleotide duplexes, such as for silencing the expression of a nucleic acid or gene of interest using small interfering RNA (siRNA) technologies, is also disclosed.
1. A chemically-modified siRNA comprising an oligonucleotide duplex comprising a sense strand and an antisense strand, each respectively comprising:
(i) Sense: [(2′F-ANA) 3 (2′F-RNA) 3 ] 2 [(2′F-ANA)(2′F-RNA)] 3 (2′F-ANA)
Antisense: (RNA) 19 ,
(ii) Sense: [(2′F-ANA) 3 (2′F-RNA) 3 ] 2 [(2′F-ANA)(2′F-RNA)] 3 (2′F-ANA)
Antisense: (2′F-RNA) 19 ,
(iii) Sense: [(2′F-ANA)(2′F-RNA)] 9 (2′F-ANA)
Antisense: (RNA) 19 ,
(iv) Sense: [(2′F-ANA)(2′F-RNA)] 9 (2′F-ANA)
Antisense: (2′F-RNA) 19 ,
(v) Sense: [(2′F-ANA) 3 (2′F-RNA) 3 ] 3 (2′F-ANA)
Antisense: (RNA) 19 ,
(vi) Sense: [(2′F-ANA) 3 (2′F-RNA) 3 ] 3 (2′F-ANA)
Antisense: (2′F-RNA) 19
(vii) Sense: (2′F-ANA) 19 ,
Antisense: (2′F-RNA) 19 , or
(viii) Sense: (2′F-ANA) 14 (RNA) 5
Antisense: (2′F-RNA) 19 .
2. The chemically modified siRNA of claim 1 , which comprises:
Sense: [(2′F-ANA) 3 (2′F-RNA) 3 ] 2 [(2′F-ANA)(2′F-RNA)] 3 (2′F-ANA)
Antisense: (RNA) 19 .
3. The chemically modified siRNA of claim 1 , which comprises:
Sense: [(2′F-ANA) 3 (2′F-RNA) 3 ] 2 [(2′F-ANA)(2′F-RNA)] 3 (2′F-ANA)
Antisense: (2′F-RNA) 19 .
4. The chemically modified siRNA of claim 1 , which comprises:
Sense: [(2′F-ANA)(2′F-RNA)] 9 (2′F-ANA)
Antisense: (RNA) 19 .
5. The chemically modified siRNA of claim 1 , which comprises:
Sense: [(2′F-ANA)(2′F-RNA)] 9 (2′F-ANA)
Antisense: (2′F-RNA) 19 .
6. The chemically modified siRNA of claim 1 , which comprises:
Sense: [(2′F-ANA) 3 (2′F-RNA) 3 ] 3 (2′F-ANA)
Antisense: (RNA) 19 .
7. The chemically modified siRNA of claim 1 , which comprises:
Sense: [(2′F-ANA) 3 (2′F-RNA) 3 ] 3 (2′F-ANA)
Antisense: (2′F-RNA) 19 .
8. The chemically modified siRNA of claim 1 , which comprises:
Sense: (2′F-ANA) 19 ,
Antisense: (2′F-RNA) 19 .
9. The chemically modified siRNA of claim 1 , which comprises:
Sense: (2′F-ANA) 14 (RNA) 5
Antisense: (2′F-RNA) 19 .
10. The chemically-modified siRNA of claim 1 , wherein the oligonucleotide duplex is fully modified with one or more 2′F-RNA and 2′F-ANA residues.
11. The chemically-modified siRNA of claim 1 , wherein the oligonucleotide duplex comprises an overhang.
12. The chemically-modified siRNA of claim 11 , wherein the overhang is a 1 to 5 residue overhang.
13. The chemically-modified siRNA of claim 12 , wherein the overhang is a 2 residue overhang.
14. The chemically-modified siRNA of claim 11 , wherein the overhang comprises DNA, 2′F-ANA, and/or 2′F-RNA residues.
15. The chemically-modified siRNA of claim 14 , wherein the overhang comprises two 2′F-ANA residues.
16. The chemically-modified siRNA of claim 15 , wherein the two 2′F-ANA residues are on the sense strand.
17. The chemically-modified siRNA of claim 15 , wherein the two 2′F-ANA residues are on the antisense strand.
18. The chemically-modified siRNA of claim 1 , wherein the sense strand and the antisense strand have a length of 19 to 23 nucleotide residues.
19. A pharmaceutical composition comprising the chemically-modified siRNA according to claim 1 , together with a pharmaceutically acceptable excipient or carrier.
20. A method of degrading or decreasing the level of a target nucleic acid, or for decreasing production of a polypeptide encoded by said target nucleic acid, in a cell, the method comprising contacting the cell with an oligonucleotide duplex of claim 1 , wherein the sense strand of the oligonucleotide duplex comprises a nucleobase sequence substantially identical to a nucleobase sequence of the target nucleic acid.