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

• US 8,178,506 B2
• Filed: 12/30/2008
• Issued: 05/15/2012
• Est. Priority Date: 07/31/2003
• Status: Expired due to Fees

• Current Assignee: Regulus Therapeutics Inc.
• Original Assignee: Regulus Therapeutics Inc.
• Inventors: Lollo, Bridget, Bennett, C. Frank, Freier, Susan M., Griffey, Richard H.
• Primary Examiner(s): VIVLEMORE, TRACY ANN
• Application Number:
  US12/345,780
• Publication Number:
  US 20090298174A1
• Time in Patent Office: 1,232 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 modulating the level of a precursor miRNA comprising:

contacting a cell with an oligomeric compound consisting of 15 to 30 monomeric subunits;

wherein the oligomeric compound is complementary to the precursor miRNA and at least one monomeric subunit is a modified nucleoside or a nucleoside mimetic; and

thereby modulating the level of the precursor miRNA.

2. The method of claim 1 wherein the contacting comprises inhibiting processing of the precursor miRNA.

3. The method of claim 1 wherein the level of the precursor miRNA is decreased.

4. The method of claim 1 wherein the oligomeric compound is complementary to a region flanking a Drosha cleavage site within the precursor miRNA.

5. The method of claim 1 wherein the precursor miRNA is a a pri-miRNA or a pre-miRNA.

6. The method of claim 5 wherein the oligomeric compound is complementary to a region of a pri-miRNA comprising a 5′

Drosha cleavage site.

7. The method of claim 5 wherein the oligomeric compound is complementary to a region of a pri-miRNA comprising a 3′

Drosha cleavage site.

8. The method of claim 1 wherein the oligomeric compound is complementary to a region comprising the miRNA.

9. The method of claim 1 wherein the oligomeric compound is complementary to a region comprising a loop structure.

10. The method of claim 1 wherein the level of a pri-miRNA is increased.

11. The method of claim 1 wherein the precursor miRNA comprises two or more miRNAs.

12. The method of claim 11 wherein the contacting comprises inhibiting the processing of two or more miRNAs.

13. The method of claim 11 wherein the precursor miRNA is a polycistronic precursor, and the polycistronic precursor is contacted with a first oligomeric compound complementary to a first region of the polycistronic precursor and a second oligomeric compound complementary to a second region of the polycistronic precursor.

14. The method of claim 1 wherein at least one monomeric subunit comprises at least one modified sugar moiety.

15. The method of claim 14 wherein the modified sugar moiety is chosen from 2′

-F, 2′

-O-methyl, 2′

-O-methoxyethyl, and a bicyclic sugar moiety.

16. The method of claim 1 wherein the modified nucleoside or nucleoside mimetic comprises at least one modified internucleoside linkage.

17. The method of claim 16 wherein the modified internucleoside linkage is a phosphorothioate linkage.

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

19. The method of claim 18 wherein the modified sugar moiety is 2′

-O-methoxyethyl.

20. The method of claim 1 wherein the oligomeric compound consists of 17 to 25 monomeric subunits.

21. The method of claim 1 wherein the oligomeric compound is a single-stranded oligonucleotide.