ANTISENSE NUCLEIC ACIDS

• US 20140343266A1
• Filed: 12/27/2012
• Published: 11/20/2014
• Est. Priority Date: 12/28/2011
• Status: Active Grant

• Current Assignee: National Center of Neurology and Psychiatry, Nippon Shinyaku Company Limited
• Original Assignee: National Center of Neurology and Psychiatry, Nippon Shinyaku Company Limited
• Inventors: Takeda, Shin'ichi, Nagata, Tetsuya, Watanabe, Naoki, Seo, Haruna
• Granted Patent: US 9,512,424 B2
• Time in Patent Office: Days
• US Class Current: 536/23.1
• CPC Class Codes: A61P 19/00 Drugs for skeletal disorders
• CPC Class Codes: A61P 21/00 Drugs for disorders of the muscular or neuromuscular system
• CPC Class Codes: A61P 21/04 for myasthenia gravis
• CPC Class Codes: A61P 43/00 Drugs for specific purposes, not provided for in groups A61P1/00-A61P41/00
• 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 2310/314 Phosphoramidates
• CPC Class Codes: C12N 2310/315 Phosphorothioates
• CPC Class Codes: C12N 2310/321 2'-O-R Modification
• CPC Class Codes: C12N 2310/322 2'-R Modification
• CPC Class Codes: C12N 2310/3233 Morpholino-type ring
• CPC Class Codes: C12N 2320/33 Alteration of splicing

Abstract

The present invention provides a pharmaceutical agent which causes skipping of the 55th, 45th, 50th or 44th exon in the human dystrophin gene with a high efficiency.

The present invention provides an oligomer which efficiently enables to cause skipping of the 55th, 45th, 50th or 44th exon in the human dystrophin gene.

Claims

1. An antisense oligomer which causes skipping of the 55th exon in the human dystrophin gene, consisting of a nucleotide sequence complementary to any one of the nucleotide sequences consisting of the 14th to the 34th, the −

2nd to the 19th, the −

2nd to the 20th, the −

2nd to the 21st, the −

2nd to the 22nd, the −

2nd to the 23rd, the −

1st to the 19th, the −

1st to the 20th, the −

1st to the 21st, the −

1st to the 22nd, the −

1st to the 23rd, the 1st to the 19th, the 1st to the 20th, the 1st to the 21st, the 1st to the 22nd, the 1st to the 23rd, the 2nd to the 19th, the 2nd to the 20th, the 2nd to the 21st, the 2nd to the 22nd, the 2nd to the 23rd, the 3rd to the 19th, the 3rd to the 20th, the 3rd to the 21st, the 3rd to the 22nd, the 3rd to the 23rd, the 9th to the 29th, the 9th to the 30th, the 9th to the 31st, the 9th to the 32nd, the 9th to the 33rd, the 10th to the 29th, the 10th to the 30th, the 10th to the 31st, the 10th to the 32nd, the 10th to the 33rd, the 11th to the 29th, the 11th to the 30th, the 11th to the 31st, the 11th to the 32nd, the 11th to the 33rd, the 12th to the 29th, the 12th to the 30th, the 12th to the 31st, the 12th to the 32nd, the 12th to the 33rd, the 13th to the 29th, the 13th to the 30th, the 13th to the 31st, the 13th to the 32nd, the 13th to the 33rd, the 12th to the 34th, the 12th to the 35th, the 12th to the 36th, the 13th to the 34th, the 13th to the 35th, the 13th to the 36th, the 14th to the 32nd, the 14th to the 33rd, 14th to the 35th, the 14th to the 36th, the 15th to the 32nd, the 15th to the 33rd, the 15th to the 34th, the 15th to the 35th, the 15th to the 36th, the 16th to the 32nd, the 16th to the 33rd, the 16th to the 34th, the 16th to the 35th, or the 16th to the 36th nucleotides, from the 5′

end of the 55th exon in the human dystrophin gene.

5. The antisense oligomer according to claim 1, which consists of a complementary sequence to the nucleotide sequences consisting of the 1st to the 21st, the 11th to the 31st, or the 14th to the 34th nucleotides, from the 5′

end of the 55th exon in the human dystrophin gene.

6. The antisense oligomer according to claim 1, consisting of the nucleotide sequence shown by any one selected from the group consisting of the 170th to the 190th, the 160th to the 180th, and the 157th to the 177th nucleotides of SEQ ID NO:

5.

13. The antisense oligomer according to claim 1, which is an oligonucleotide.

14. The antisense oligomer according to claim 13, wherein the sugar moiety and/or the phosphate-binding region of at least one nucleotide constituting the oligonucleotide is modified.

15. The antisense oligomer according to claim 14, wherein the sugar moiety of at least one nucleotide constituting the oligonucleotide is a ribose in which the 2′

—

OH group is replaced by any one selected from the group consisting of OR, R, R′

OR, SH, SR, NH₂, NHR, NR₂, N₃, CN, F, Cl, Br and I (wherein R is an alkyl or an aryl and R′

is an alkylene).

16. The antisense oligomer according to claim 14, wherein the phosphate-binding region of at least one nucleotide constituting the oligonucleotide is any one selected from the group consisting of a phosphorothioate bond, a phosphorodithioate bond, an alkylphosphonate bond, a phosphoramidate bond and a boranophosphate bond.

17. The antisense oligomer according to claim 1, which is a morpholino oligomer.

18. The antisense oligomer according to claim 17, which is a phosphorodiamidate morpholino oligomer.

19. The antisense oligomer according to claim 17, wherein the 5′

end is any one of the groups of chemical formulae (1) to (3) below;

20. A pharmaceutical composition for the treatment of muscular dystrophy, comprising as an active ingredient the antisense oligomer according to claim 1, or a pharmaceutically acceptable salt or hydrate thereof.

2. An antisense oligomer which causes skipping of the 45th exon in the human dystrophin gene, consisting of a nucleotide sequence complementary to any one of the nucleotide sequences consisting of the −

3rd to the 19th, the −

3rd to the 20th, the −

3rd to the 21st, the −

3rd to the 22nd, the −

3rd to the 23rd, the −

2nd to the 19th, the −

2nd to the 20th, the −

2nd to the 21st, the −

2nd to the 22nd, the −

2nd to the 23rd, the −

1st to the 19th, the −

1st to the 20th, the −

1st to the 21st, the −

1st to the 22nd, the −

1st to the 23rd, the 1st to the 19th, the 1st to the 20th, the 1st to the 21st, the 1st to the 22nd, the 1st to the 23rd, the 2nd to the 19th, the 2nd to the 20th, the 2nd to the 21st, the 2nd to the 22nd, the 2nd to the 23rd, the −

2nd to the 24th, the −

2nd to the 25th, the −

2nd to the 26th, the −

2nd to the 27th, the −

1st to the 24th, the −

1st to the 25th, the −

1st to the 26th, the −

1st to the 27th, the 1st to the 24th, the 1st to the 25th, the 1st to the 26th, the 1st to the 27th, the 2nd to the 24th, the 2nd to the 25th, the 2nd to the 26th, the 2nd to the 27th, the 3rd to the 23rd, the 3rd to the 24th, the 3rd to the 25th, the 3rd to the 26th, the 3rd to the 27th, the 4th to the 28th, the 4th to the 29th, the 4th to the 30th, the 4th to the 31st, the 4th to the 32nd, the 5th to the 28th, the 5th to the 29th, the 5th to the 30th, the 5th to the 31st, the 5th to the 32nd, the 6th to the 28th, the 6th to the 29th, the 6th to the 30th, the 6th to the 31st, the 6th to the 32nd, the 7th to the 28th, the 7th to the 29th, the 7th to the 30th, the 7th to the 31st, the 7th to the 32nd, the 8th to the 28th, the 8th to the 29th, the 8th to the 30th, the 8th to the 31st, or the 8th to the 32nd nucleotides, from the 5′

end of the 45th exon in the human dystrophin gene.

7. The antisense oligomer according to claim 2, which consists of a complementary sequence to the nucleotide sequences consisting of the −

2nd to the 19th, the 1st to the 21st, the 1st to the 25th, or the 6th to the 30th nucleotides, from the 5′

end of the 45th exon in the human dystrophin gene.

8. The antisense oligomer according to claim 2, consisting of the nucleotide sequence shown by any one selected from the group consisting of the 158th to the 178th, the 156th to the 176th, the 152nd to the 176th, and the 147th to the 171st nucleotides of SEQ ID NO:

6.

3. An antisense oligomer which causes skipping of the 50th exon in the human dystrophin gene, consisting of a nucleotide sequence complementary to any one of the nucleotide sequences consisting of the 105th to the 125th, the 105th to the 126th, the 105th to the 127th, the 105th to the 128th, the 105th to the 129th, the 106th to the 125th, the 106th to the 126th, the 106th to the 127th, the 106th to the 128th, the 106th to the 129th, the 107th to the 125th, the 107th to the 126th, the 107th to the 127th, the 107th to the 128th, the 107th to the 129th, the 108th to the 125th, the 108th to the 126th, the 108th to the 127th, the 108th to the 128th, the 108th to the 129th, the 109th to the 125th, the 109th to the 126th, the 109th to the 127th, the 109th to the 128th, or the 109th to the 129th nucleotides, from the 5′

end of the 50th exon in the human dystrophin gene.

9. The antisense oligomer according to claim 3, which consists of a complementary sequence to the nucleotide sequences consisting of the 106th to the 126th or the 107th to the 127th nucleotides, from the 5′

end of the 50th exon in the human dystrophin gene.

10. The antisense oligomer according to claim 3, consisting of the nucleotide sequence shown by any one selected from the group consisting of the 4th to the 24th and the 3rd to the 23rd nucleotides of SEQ ID NO:

7.

4. An antisense oligomer which causes skipping of the 44th exon in the human dystrophin gene, consisting of a nucleotide sequence complementary to any one of the nucleotide sequences consisting of the 9th to the 30th, 9th to the 31st, the 9th to the 32nd, the 9th to the 33rd, the 9th to the 34th, the 10th to the 30th, the 10th to the 31st, the 10th to the 32nd, the 10th to the 33rd, the 10th to the 34th, the 11th to the 30th, the 11th to the 31st, the 11th to the 32nd, the 11th to the 33rd, the 11th to the 34th, the 12th to the 30th, the 12th to the 31st, the 12th to the 32nd, the 12th to the 33rd, the 12th to the 34th, the 13th to the 30th, the 13th to the 31st, the 13th to the 32nd, the 13th to the 33rd, the 13th to the 34th, the 24th to the 45th, the 24th to the 46th, the 24th to the 47th, the 24th to the 48th, the 24th to the 49th, the 25th to the 45th, the 25th to the 46th, the 25th to the 47th, the 25th to the 48th, the 25th to the 49th, the 26th to the 45th, the 26th to the 46th, the 26th to the 47th, the 26th to the 48th, the 26th to the 49th, the 27th to the 45th, the 27th to the 46th, the 27th to the 47th, the 27th to the 48th, the 27th to the 49th, the 28th to the 45th, the 28th to the 46th, the 28th to the 47th, the 28th to the 48th, the 28th to the 49th, the 29th to the 45th, the 29th to the 46th, the 29th to the 47th, the 29th to the 48th, or the 29th to the 49th nucleotides, from the 5′

end of the 44th exon in the human dystrophin gene.

11. The antisense oligomer according to claim 4, which consists of a complementary sequence to the nucleotide sequences consisting of the 11th to the 32nd, the 25th to the 45th, the 26th to the 46th, the 26th to the 47th or the 27th to the 47th nucleotides, from the 5′

end of the 44th exon in the human dystrophin gene.

12. The antisense oligomer according to claim 4, consisting of the nucleotide sequence shown by any one selected from the group consisting of the 117th to the 138th, the 104th to the 124th, the 103rd to the 123rd, the 102nd to the 123rd and the 102nd to the 122nd nucleotides of SEQ ID NO:

8.