Double stranded constructs comprising one or more short strands hybridized to a longer strand

• US 8,569,474 B2
• Filed: 03/04/2005
• Issued: 10/29/2013
• Est. Priority Date: 03/09/2004
• Status: Active Grant

• Current Assignee: Ionis Pharmaceuticals, Inc.
• Original Assignee: ISIS Pharmaceuticals Incorporated
• Inventors: Baker, Brenda F., Kraynack, Bryan A.
• Primary Examiner(s): SHIN, DANA H
• Application Number:
  US11/072,806
• Publication Number:
  US 20050245474A1
• Time in Patent Office: 3,161 Days
• Field of Search: 536/24.5, 536/23.1, 514/44, 435/6, 435/455, 435/375, 800/8
• US Class Current: 536/24.5
• CPC Class Codes: C12N 15/111 General methods applicable to biologically active non-coding nucleic acids
• CPC Class Codes: C12N 2310/14 interfering N.A.
• CPC Class Codes: C12N 2310/315 Phosphorothioates
• CPC Class Codes: C12N 2310/3183 Diol linkers, e.g. glycols or propanediols
• CPC Class Codes: C12N 2310/321 2'-O-R Modification
• CPC Class Codes: C12N 2310/322 2'-R Modification
• CPC Class Codes: C12N 2310/3231 having an additional ring, e.g. LNA, ENA
• CPC Class Codes: C12N 2310/3341 5-Methylcytosine
• CPC Class Codes: C12N 2310/341 Gapmers, i.e. of the type ===---===
• CPC Class Codes: C12N 2310/346 having a combination of backbone and sugar modifications
• CPC Class Codes: C12N 2310/3519 Fusion with another nucleic acid
• CPC Class Codes: C12N 2310/3525 MOE, methoxyethoxy
• CPC Class Codes: C12N 2310/3533 Halogen
• CPC Class Codes: C12N 2310/51 in polymeric form, e.g. multimers, concatemers
• CPC Class Codes: C12N 2330/30 chemically synthesised

Abstract

The present invention provides compositions comprising an antisense oligomeric compound hybridized to at least one shorter sense oligomeric compound. The sense oligomeric compounds can be covalently linked to the antisense oligomeric compounds. At least a portion of the antisense oligomeric compound is complementary to and hybridizes with a nucleic acid target. Methods for modulating gene expression are also provided using the compositions of the present invention.

Claims

1. A composition comprising an antisense oligomeric compound and two sense oligomeric compounds wherein:

each of the sense oligomeric compounds is complementary to a separate region of the antisense oligomeric compound and one of the sense oligomeric compounds forms a duplex region with the antisense oligomeric compound starting at the 5′

-end of the antisense oligomeric compound and the other of the sense oligomeric compounds forms a duplex region with the antisense oligomeric compound starting at the 3′

-end of the antisense oligomeric compound, and the duplex regions are separate and non-overlapping and include each nucleoside of the antisense oligomeric compound; and

the antisense oligomeric compound comprises from 20 to 24 linked nucleosides and each of the sense oligomeric compounds comprises from 10 to 12 linked nucleosides.

2. The composition of claim 1 wherein the conformational geometry of each of said linked nucleosides is 3′

-endo.

3. The composition of claim 2 wherein each of said nucleosides having 3′

-endo conformational geometry comprises a 2′

-substituent group.

4. The composition of claim 3 wherein each 2′

-substituent group is, independently, —

F, —

OH, —

O—

CH₂CH₂—

O—

CH₃, —

O—

CH₃, —

O—

CH₂—

CH═

CH₂ or —

O—

CH₂—

CH—

CH₂—

NH(R_j) where R_j is H or C₁-C₁₀ alkyl.

5. The composition of claim 4 wherein each of said 2′

-substituent groups is, independently, —

F, —

OH, —

O—

CH₂CH₂—

O—

CH₃ or —

O—

CH₃.

6. The composition of claim 2 wherein at least one of said nucleosides has a 2′

-OH group.

7. The composition of claim 2 wherein each of said nucleosides are modified nucleosides having other than 2′

-OH groups.

8. The composition of claim 2 wherein at least one of said nucleosides is a bicyclic sugar nucleoside having a 2′

-O—

(CH₂)_n-4′

bridge wherein n is 1 or 2.

9. The composition of claim 2 wherein at least one of said nucleosides is a 4′

-S sugar modified nucleoside.

10. The composition of claim 2 wherein each of said linked nucleosides is independently linked by a phosphodiester or a phosphorothioate internucleoside linking group.

11. The composition of claim 1 wherein at least one of said antisense oligomeric compound and said two sense oligomeric compounds further comprises at least one terminal phosphate group, a terminal stabilizing or capping group, a 3′

-overhang or a conjugate group.

12. The composition of claim 11 wherein the 5′

-terminus of said antisense oligomeric compound comprises a 5′

-phosphate group.

13. The composition of claim 1 wherein each of said sense oligomeric compounds is 10 nucleosides in length.

14. The composition of claim 1 wherein said antisense oligomeric compound is 20 nucleosides in length.

15. The composition of claim 14 wherein each of said linked nucleosides of said antisense oligomeric compound is linked by a phosphodiester internucleoside linking group.

16. The composition of claim 14 wherein each of said linked nucleosides of said antisense oligomeric compound independently comprises a 2′

-OH or a 2′

-F substituent group.

17. The composition of claim 1 wherein said antisense oligomeric compound is not covalently linked to either of said sense oligomeric compounds.

18. A method of inhibiting gene expression comprising contacting one or more cells, a tissue or an animal with the composition of claim 1.