COMPOSITIONS AND METHODS FOR MODULATION OF SMN2 SPLICING

• US 20130109091A1
• Filed: 12/19/2012
• Published: 05/02/2013
• Est. Priority Date: 06/23/2005
• Status: Active Grant

• Current Assignee: Biogen MA Inc. (Biogen, Inc.), Cold Spring Harbor Laboratory
• Original Assignee: Cold Spring Harbor Laboratory, ISIS Pharmaceuticals Incorporated
• Inventors: Baker, Brenda F., Krainer, Adrian R., Hua, Yimin
• Granted Patent: US 8,946,183 B2
• Time in Patent Office: Days
• US Class Current: 435/375
• CPC Class Codes: A61K 48/00 Medicinal preparations containing genetic material which is inserted into cells of the living body to treat genetic diseases; Gene therapy
• CPC Class Codes: C07H 21/02 with ribosyl as saccharide radical
• 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 2310/321 2'-O-R Modification
• CPC Class Codes: C12N 2310/3341 5-Methylcytosine
• CPC Class Codes: C12N 2310/3525 MOE, methoxyethoxy
• CPC Class Codes: C12N 2320/33 Alteration of splicing

Abstract

Disclosed herein are compounds, compositions and methods for modulating splicing of SMN2 mRNA in a cell, tissue or animal. Also provided are uses of disclosed compounds and compositions in the manufacture of a medicament for treatment of diseases and disorders, including spinal muscular atrophy.

Claims

1. An antisense oligonucleotide targeted to intron 6, exon 7 or intron 7 of a nucleic acid molecule encoding SMN2, wherein said oligonucleotide is 12 to 20 nucleotides in length and comprises a 2′

-O-methoxyethyl sugar modification at each position.

2. The oligonucleotide of claim 1 which is 12 nucleotides in length.

3. The oligonucleotide of claim 1 which is 15 nucleotides in length.

4. The oligonucleotide of claim 1 which is 18 nucleotides in length.

5. The oligonucleotide of claim 1 which is targeted to an intronic splicing silencer element.

6. The oligonucleotide of claim 1 which is targeted to an exonic splicing silencer element.

7. The oligonucleotide of claim 1 which is targeted to intron 6.

8. The oligonucleotide of claim 7, wherein the target site of said oligonucleotide is nucleotide 5, 6, 7 or 8 of SEQ ID NO:

1.

9. The oligonucleotide of claim 7, wherein the nucleotide sequence of said oligonucleotide comprises at least an 8-nucleobase portion of SEQ ID NO:

6, 7, 8, 9, 10, 11, 12 or 13.

10. The oligonucleotide of claim 1 which is targeted to exon 7.

11. The oligonucleotide of claim 10 wherein the target site of said oligonucleotide is nucleotide 64, 65, 66, 67, 68, 94, 95, 96 or 97 of SEQ ID NO:

1.

12. The oligonucleotide of claim 10, wherein the nucleotide sequence of said oligonucleotide comprises at least an 8-nucleobase portion of SEQ ID NO:

37, 38, 41, 59, 62, 63, 64, 66, 67 or 68.

13. The oligonucleotide of claim 1 which is targeted to intron 7.

14. The oligonucleotide of claim 13 wherein the target site of said oligonucleotide is nucleotide 121, 122, 123, 124, 125, 126, 127, 128 or 129 of SEQ ID NO:

1.

15. The oligonucleotide of claim 13, wherein the nucleotide sequence of said oligonucleotide comprises at least an 8-nucleobase portion of SEQ ID NO:

79, 80, 81, 82, 83, 84, 85, 86, 87, 89, 91 or 93.

16. A method of promoting inclusion of exon 7 in SMN2 transcripts in a cell, tissue or organ, comprising contacting said cell, tissue or organ with the antisense oligonucleotide of claim 1.

17. The method of claim 16 wherein said oligonucleotide is targeted to an intronic splicing silencer element.

18. The method of claim 16 wherein said oligonucleotide is targeted to an exonic splicing silencer element.

19. An antisense oligonucleotide of claim 1 for use in therapy.

20. Use of an antisense oligonucleotide of claim 1 for the preparation of a medicament for modulating splicing of an SMN2 pre-mRNA.

21. The use of claim 20 wherein modulating splicing results in an increase in exon 7 inclusion.

22. Use of an antisense oligonucleotide of claim 1 for the preparation of a medicament for the treatment of spinal muscular atrophy.

23. A pharmaceutical composition comprising the antisense oligonucleotide of claim 1.