Oligomeric compounds and compositions for use in modulation of small non-coding RNAS

• US 8,110,558 B2
• Filed: 12/31/2008
• Issued: 02/07/2012
• Est. Priority Date: 07/31/2003
• Status: Active Grant

• Current Assignee: Regulus Therapeutics Inc.
• Original Assignee: Regulus Therapeutics Inc.
• Inventors: Bennett, C. Frank, Freier, Susan M., Griffey, Richard H.
• Primary Examiner(s): VIVLEMORE, TRACY ANN
• Application Number:
  US12/346,919
• Publication Number:
  US 20110224277A1
• Time in Patent Office: 1,133 Days
• Field of Search: None
• US Class Current: 514/44
• CPC Class Codes: A61P 3/00 Drugs for disorders of the metabolism of the blood or the extracellular fluid A61P7/00
• CPC Class Codes: A61P 37/00 Drugs for immunological or allergic disorders
• CPC Class Codes: C12N 15/111 General methods applicable to biologically active non-coding nucleic acids
• CPC Class Codes: C12N 15/113 Non-coding nucleic acids modulating the expression of genes, e.g. antisense oligonucleotides; Antisense DNA or RNA; Triplex- forming oligonucleotides; Catalytic nucleic acids, e.g. ribozymes; Nucleic acids used in co-suppression or gene silencing when used in plants C12N15/8218
• CPC Class Codes: C12N 2310/11 Antisense
• CPC Class Codes: C12N 2310/113 targeting other non-coding nucleic acids, e.g. antagomirs
• CPC Class Codes: C12N 2310/14 interfering N.A.
• CPC Class Codes: C12N 2310/141 MicroRNAs, miRNAs
• CPC Class Codes: C12N 2310/31 of the backbone
• CPC Class Codes: C12N 2310/315 Phosphorothioates
• CPC Class Codes: C12N 2310/317 with an inverted bond, e.g. a cap structure
• CPC Class Codes: C12N 2310/3181 Peptide nucleic acid, PNA
• CPC Class Codes: C12N 2310/32 of the sugar
• CPC Class Codes: C12N 2310/321 2'-O-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 2310/351 Conjugate
• CPC Class Codes: C12N 2310/3515 Lipophilic moiety, e.g. cholesterol
• CPC Class Codes: C12N 2310/3525 : MOE, methoxyethoxy
• CPC Class Codes: C12N 2320/11 : for the determination of target sites, i.e. of active nucleic acids
• CPC Class Codes: C12N 2320/30 : Special therapeutic applications
• CPC Class Codes: C12N 2320/50 : Methods for regulating/modulating their activity

Abstract

Compounds, compositions and methods are provided for modulating the expression and function of small non-coding RNAs. The compositions comprise oligomeric compounds, targeted to small non-coding RNAs. Methods of using these compounds for modulation of small non-coding RNAs as well as downstream targets of these RNAs and for diagnosis and treatment of disease associated with small non-coding RNAs are also provided.

Claims

1. A method of inhibiting the activity of a microRNA comprising contacting a cell comprising the microRNA with a compound comprising an oligomeric compound, wherein:

the oligomeric compound is at least 90% complementary to a miRNA or a precursor thereof;

the oligomeric compound consists of 15 to 30 linked monomeric subunits;

at least one monomeric subunit is a modified nucleoside; and

the miRNA is selected from miR-21 (SEQ ID NO;

     64) and miR-1 (SEQ ID NO;

     29).

2. The method of claim 1, wherein the miRNA is miR-21 (SEQ ID NO:

64).

3. The method of claim 1, wherein the miRNA is miR-1 (SEQ ID NO:

29).

4. The method of claim 1 wherein the oligomeric compound is at least 95% complementary to the miRNA.

5. The method of claim 1 wherein the oligomeric compound is 100% complementary to the miRNA.

6. The method of claim 1 wherein the oligomeric compound is 100% complementary to at least an 8-nucleobase portion of the miRNA.

7. The method of claim 1 wherein the oligomeric compound consists of 19 to 23 linked monomeric subunits.

8. The method of claim 1 wherein the oligomeric compound consists of an oligonucleotide.

9. The method of claim 8 wherein the oligonucleotide consists of 19 to 23 linked nucleosides.

10. The method of claim 1 wherein the oligomeric compound comprises at least one modified sugar moiety.

11. The method of claim 10 wherein the modified sugar moiety is selected from 2′

-F, 2′

-O-methyl, 2′

-O-methoxyethyl, and a bicyclic sugar moiety.

12. The method of claim 11 wherein the bicyclic sugar moiety comprises a 4′

-CH₂—

O-2′

bridge.

13. The method of claim 1 wherein the oligomeric compound comprises at least one modified internucleoside linkage.

14. The method of claim 13 wherein the modified internucleoside linkage is a phosphorothioate linkage.

15. The method of claim 1 wherein each internucleoside linkage of the oligomeric compound is a phosphorothioate linkage.

16. The method of claim 1 wherein each monomeric subunit comprises a modified sugar moiety.

17. The method of claim 1 wherein the oligonucleotide comprises two or more chemically distinct regions.

18. The method of claim 17 wherein each nucleoside of the two or more chemically distinct regions is independently selected from a 2′

-fluoro nucleoside, a 2′

-O-methyl nucleoside, a 2′

-O-methoxyethyl nucleoside, a 2′

-deoxynucleoside, and a bicyclic sugar nucleoside.

19. The method of claim 17 wherein each nucleoside of the two or more chemically distinct regions is independently selected from a 2′

-deoxynucleoside and a bicyclic sugar nucleoside.

20. The method of claim 19 wherein the bicyclic nucleoside comprises a 4′

-CH₂—

O-2′

bridge.

21. The method of claim 1 wherein the oligomeric compound comprises at least one modified nucleobase.

22. The method of claim 21 wherein the modified nucleobase is 5-methylcytosine.

23. The method of claim 1 wherein the oligomeric compound is attached to a conjugate group.

24. The method of claim 23 wherein the conjugate group is cholesterol.

25. A composition comprising a compound comprising an oligomeric compound, wherein:

the oligomeric compound is at least 90% complementary to a miRNA or a precursor thereof;

the oligomeric compound consists of 15 to 30 linked monomeric subunits;

at least one monomeric subunit is a modified nucleoside; and

the miRNA is selected from miR-21 (SEQ ID NO;

     64) and miR-1 (SEQ ID NO;

     29);

and a pharmaceutically acceptable diluent or carrier.