2′-Fluoro substituted oligomeric compounds and compositions for use in gene modulations

• US 7,812,149 B2
• Filed: 11/04/2003
• Issued: 10/12/2010
• Est. Priority Date: 06/06/1996
• Status: Expired due to Fees

• Current Assignee: ISIS Pharmaceuticals Incorporated
• Original Assignee: ISIS Pharmaceuticals Incorporated
• Inventors: Bhat, Balkrishen, Swayze, Eric E., Baker, Brenda F., Prakash, Thazha P., Manoharan, Muthiah, Crooke, Stanley T., Eldrup, Anne B., Griffey, Richard
• Primary Examiner(s): Bowman; Amy
• Application Number:
  US10/700,689
• Publication Number:
  US 20040171029A1
• Time in Patent Office: 2,534 Days
• Field of Search: 514/44, 536/23.1, 536/24.3, 536/24.31, 536/24.33, 536/24.5
• 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 2310/321 2'-O-R Modification
• CPC Class Codes: C12N 2310/322 2'-R Modification
• 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 2310/3521 Methyl
• CPC Class Codes: C12N 2310/3533 Halogen
• CPC Class Codes: C12N 2310/531 Stem-loop; Hairpin
• CPC Class Codes: C12N 2320/51 modulating the chemical stability, e.g. nuclease-resistance

Abstract

Compositions comprising first and second oligomers are provided wherein at least a portion of the first oligomer is capable of hybridizing with at least a portion of the second oligomer, at least a portion of the first oligomer is complementary to and capable of hybridizing to a selected target nucleic acid, and at least one of the first or second oligomers includes a modified sugar and/or backbone modification. In some embodiments the modification is a 2′-F substituent group on a sugar moiety. Oligomer/protein compositions are also provided comprising an oligomer complementary to and capable of hybridizing to a selected target nucleic acid and at least one protein comprising at least a portion of an RNA-induced silencing complex (RISC), wherein at least one nucleoside of the oligomer has a modified sugar and/or backbone modification.

Claims

1. A composition comprising first and second chemically synthesized oligomeric compounds, wherein:

the first and second chemically synthesized oligomeric compounds are not covalently linked to each other;

each of the first and second oligomeric compounds consists of 20 to 24 linked nucleosides;

the first oligomeric compound is 100% complementary to the second oligomeric compound and to a selected target messenger RNA;

the first oligomeric compound has the formula;

(Z₁)_n—

X₁—

Y₁—

X₂—

Y₂—

X₃—

Y₃—

X₄—

Y₄—

(Z₂)_m

wherein;

each Z₁ is, independently, a natural or modified nucleoside;

X₁ is a region of two 2′

-F modified nucleosides;

Y₁ is a region of one 2′

-O-methyl modified nucleoside;

X₂ is a region of two 2′

-F modified nucleosides;

Y₂ is a region of two 2′

-O-methyl modified nucleosides;

X₃ is a region of five 2′

-F modified nucleosides;

Y₃ is a region of three 2′

-O-methyl modified nucleosides;

X₄ is a region of three 2′

-F modified nucleosides;

Y₄ is a region of two 2′

-O-methyl modified nucleosides;

each Z₂ is, independently, a natural or modified nucleoside; and

n and m are each, independently, 0, 1, or 2; and

the second oligomeric comprises a plurality of ribose nucleoside subunits.

2. The composition of claim 1 comprising a conjugate.

3. The composition of claim 2, wherein the conjugate is attached to either the 5′

or 3′

terminal nucleoside of either the first or the second oligomeric compound.

4. The composition of claim 1, wherein the target messenger RNA is a mammalian messenger RNA.

5. The composition of claim 4, wherein the target messenger RNA is a human messenger RNA.

6. The composition of claim 1 wherein Z₁ is deoxythymidine.

7. The composition of claim 1 wherein Z₂ is deoxythymidine.

8. The composition of claim 1 wherein Z₁ and Z₂ are each deoxythymidine.

9. The composition of claim 1 wherein n and m are each 0.

10. The composition of claim 1 wherein n and m are each 1.

11. The composition of claim 1 wherein n and m are each 2.

12. The composition of claim 1 wherein n is 0 and m is 1.

13. The composition of claim 1 wherein n is 0 and m is 2.

14. The composition of claim 1 wherein n is 1 and m is 0.

15. The composition of claim 1 wherein n is 1 and m is 2.

16. The composition of claim 1 wherein n is 2 and m is 0.

17. The composition of claim 1 wherein n is 2 and m is 1.

18. A composition comprising first and second chemically synthesized oligomeric compounds, wherein:

the first and second chemically synthesized oligomeric compounds are not covalently linked to each other;

each of the first and second oligomeric compounds consists of 20 linked nucleosides;

the first oligomeric compound is 100% complementary to the second oligomeric compound and to a selected target messenger RNA;

the first oligomeric compound has the formula;

X₁—

Y₁—

X₂—

Y₂—

X₃—

Y₃—

X₄—

Y₄

wherein;

X₁ is a region of two 2′

-F modified nucleosides;

Y₁ is a region of one 2′

-O-methyl modified nucleoside;

X₂ is a region of two 2′

-F modified nucleosides;

Y₂ is a region of two 2′

-O-methyl modified nucleosides;

X₃ is a region of five 2′

-F modified nucleosides;

Y₃ is a region of three 2′

-O-methyl modified nucleosides;

X₄ is a region of three 2′

-F modified nucleosides; and

Y₄ is a region of two 2′

-O-methyl modified nucleosides; and

the second oligomeric comprises a plurality of ribose nucleoside subunits.

19. The composition of claim 18 comprising a conjugate.

20. The composition of claim 19, wherein the conjugate is attached to either the 5′

or 3′

terminal nucleoside of either the first or the second oligomeric compound.

21. The composition of claim 18, wherein the target messenger RNA is a mammalian messenger RNA.

22. The composition of claim 21, wherein the target messenger RNA is a human messenger RNA.