Cyclic antisense therapeutics

Abstract

The present invention provides oligonucleotides referred to as cyclic structured oligonucleotides ("CSOs") comprising a functional domain, a cyclizing domain, and a linker segment as described herein, compositions comprising same, and methods of using same. This design of cyclic oligonucleotides maintains a cyclic form until it is in the presence of and hybridizes with a targeted RNA.

Description

Abstract This invention provides oligonucleotides called cyclic oligonucleotides (“CSOs”) comprising the functional domains, cyclization domains, and linker fragments described herein, compositions comprising the same, and methods of using them. These cyclic oligonucleotides are designed to remain cyclic until they hybridize with a target RNA in its presence.

Claims

What is Claimed:

1. A cyclic structured oligonucleotide (CSO) comprising a functional domain and a cyclizing domain, wherein the functional domain and the cyclizing domain are linked at their 5’ ends; wherein the cyclizing domain comprises an oligonucleotide between 4 and 12 nucleotides in length and is complementary to a sequence of nucleotides within the functional domain and of polarity opposite to the sequence of nucleotides within the functional domain to which it is complementary; wherein the cyclizing domain hybridizes with the functional domain, thereby forming a cyclic structure; wherein the functional domain comprises an antisense oligonucleotide, wherein the antisense oligonucleotide is at least 80% identical to an antisense oligonucleotide selected from Table 1 and comprising at least 12 contiguous nucleobases complementary to an equal length portion of a target RNA sequence.

2. The CSO according to claim 1, wherein the antisense oligonucleotide comprises a 3’ domain and a 5’ domain, which is contiguous with the 3’ domain, wherein the 3’ domain begins at the terminal nucleotide at the 3’ end and is 10 to 12 nucleotides in length and wherein each nucleotide is unmodified and wherein each internucleotide linkage between adjacent nucleotides is a phosphodiester or phosphorothioate internucleotide linkage or combinations thereof; and wherein the 5’ domain begins at the first nucleotide following the 3’ domain and continues to the terminal nucleotide at the 5’ end, wherein at least 3 nucleotides of 5’ domain comprise a modified sugar and / or backbone; wherein the modified deoxyribonucleotides and / or modified ribonucleotides of the 5’ domain need not be consecutive; and wherein the modified deoxyribonucleotides and / or modified ribonucleotides of the 5’ domain prevent RNase H cleavage in the 5’ domain.

3. The CSO according to claim 2, wherein at least half of the nucleotides of the 5’ domain comprise a modified sugar and / or a modified backbone.

4. The CSO according to claim 2, wherein all of the nucleotides of the 5’ domain comprise a modified sugar and / or backbone.

5. The CSO according to any one of claims 1-4, wherein the cyclizing domain is 4 to 8 nucleotides in length.

6. The CSO according to any one of claims 1-5, wherein the nucleotides of the cyclizing domain comprises unmodified deoxyribonucleotides, unmodified ribonucleotides, modified deoxyribonucleotides, modified ribonucleotides, or combinations thereof.

7. The CSO according to claim 6, wherein the nucleotides of the cyclizing comprise unmodified ribonucleotides.

8. The CSO according to claim 6, wherein the nucleotides of the cyclizing comprise unmodified deoxyribonucleotides.

9. The CSO according to claim 6, wherein the nucleotides of the cyclizing comprise modified ribonucleotides.

10. The CSO according to claim 9, wherein the modified ribonucleotides comprise 2’- substituted ribonucleotides.

11. The CSO according to claim 10, wherein the 2 ’-substituted ribonucleotides are 2’- OMe ribonucleotides or 2’ -MOE ribonucleotides.

12. The CSO according to any one of claims 1-11, wherein the internucleotidic linkages of the oligonucleotides of the functional domain and / or the cyclizing domain comprise phosphorothioate intemucleotide linkages, phosphodiester internucleotide linkages, or combinations thereof.

13. The CSO according to claim 12, wherein the oligonucleotide of the functional domain comprises at least one phosphorothioate internucleotide linkage.

14. The CSO according to claim 13, wherein at least half of the intemucleotide linkages are phosphorothioate linkages.

15. The CSO according to claim 13, wherein all the intemucleotide linkages are phosphorothioate linkages.

16. The CSO according to claim 12, wherein all the intemucleotide linkages are phosphodiester linkages.

17. The CSO according to claim 12, wherein the oligonucleotide of the cyclizing domain comprises at least one phosphorothioate intemucleotide linkage.

18. The CSO according to claim 17, wherein at least half of the intemucleotide linkages are phosphorothioate linkages.

19. The CSO according to claim 17, wherein all the intemucleotide linkages are phosphorothioate linkages.

20. The CSO according to claim 12, wherein all the intemucleotide linkages are phosphodiester linkages.

21. The CSO according to any one of claims 1-19, wherein the functional domain comprises an antisense oligonucleotide comprising at least 80% sequence identity to an antisense oligonucleotide selected from Table 2.

22. The CSO according to any one of claims 1-21, wherein the cyclizing domain is selected from Table 3.

23. The CSO according to any one of claims 1-22, wherein the functional domain and the cyclizing domain are linked at their 5’ ends through a direct bond.

24. The CSO according to any one of claims 1-23, wherein the CSO comprises an antisense oligonucleotide comprising at least 80% identical to an antisense oligonucleotide selected from Table 4.

25. A pharmaceutical composition comprising the antisense oligonucleotide according to any one of claims 1-24 and a pharmaceutically acceptable carrier.

26. A method for inhibiting gene expression comprising administering a CSO according to any one of claims 1-24 or a composition according to claim 25.

27. The method according to claim 26, wherein in the method is useful for treating a subject having disease or disorder wherein inhibiting expression of a gene would be beneficial.

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