Antisense nucleic acids

• US 10,647,741 B2
• Filed: 06/24/2019
• Issued: 05/12/2020
• Est. Priority Date: 09/01/2010
• Status: Active Grant

• Current Assignee: National Center of Neurology and Psychiatry, Nippon Shinyaku Company Limited
• Original Assignee: Nippon Shinyaku Company Limited
• Inventors: Watanabe, Naoki, Satou, Youhei, Takeda, Shin'ichi, Nagata, Tetsuya
• Primary Examiner(s): McGarry, Sean
• Application Number:
  US16/449,537
• Publication Number:
  US 20190315796A1
• Time in Patent Office: 323 Days
• Field of Search: None
• 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: C07H 21/00 Compounds containing two or more mononucleotide units having separate phosphate or polyphosphate groups linked by saccharide radicals of nucleoside groups, e.g. nucleic acids
• CPC Class Codes: C07H 21/04 with deoxyribosyl as saccharide radical
• CPC Class Codes: C07K 14/4708 Duchenne dystrophy
• 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 2310/3145 with the nitrogen in 3' or 5'-position
• CPC Class Codes: C12N 2310/315 Phosphorothioates
• CPC Class Codes: C12N 2310/321 2'-O-R Modification
• CPC Class Codes: C12N 2310/3525 MOE, methoxyethoxy
• CPC Class Codes: C12N 2320/33 Alteration of splicing

Abstract

The present invention provides an oligomer which efficiently enables to cause skipping of the 53rd exon in the human dystrophin gene. Also provided is a pharmaceutical composition which causes skipping of the 53rd exon in the human dystrophin gene with a high efficiency.

Claims

1. A method comprising administering to a patient with DMD an antisense phosphorodiamidate morpholino oligomer (PMO) consisting of a 25-mer oligomer that is 100% complementary to the 36^th to the 60th nucleotides from the 5′

end of the 53^rd exon in a human dystrophin pre-mRNA, wherein the 53^rd exon in said human dystrophin pre-mRNA consists of a nucleotide sequence corresponding to SEQ ID NO;

1, wherein said PMO hybridizes to said human dystrophin pre-mRNA with Watson-Crick base pairing, and wherein skipping of the 53^rd exon is induced in said patient.

2. The method according to claim 1, wherein each phosphorodiamidate morpholino monomer of said PMO has the formula:

3. The method according to claim 2, wherein the 5′

end of said PMO has a formula selected from the group consisting of;

4. A method of inducing exon 53 skipping in a patient with DMD comprising administering to said patient an antisense phosphorodiamidate morpholino oligomer (PMO) consisting of a 25-mer oligomer that is 100% complementary to the 36^th to the 60^th nucleotides from the 5′

end of the 53rd exon in a human dystrophin pre-mRNA, wherein the 53^rd exon in said human dystrophin pre-mRNA consists of a nucleotide sequence corresponding to SEQ ID NO;

1, and wherein said PMO hybridizes to said human dystrophin pre-mRNA with Watson-Crick base pairing.

5. The method according to claim 4, wherein each phosphorodiamidate morpholino monomer of said PMO has the formula:

6. The method according to claim 5, wherein the 5′

end of said PMO has a formula selected from the group consisting of;

7. A method comprising administering to a patient with DMD an antisense phosphorodiamidate morpholino oligomer (PMO) consisting of a 25-mer oligomer that is 100% complementary to the target sequence 5′

-GAACACCUUCAGAACCGGAGGCAAC-3′

(SEQ ID NO;

     124), wherein the 53^rd exon in a human dystrophin pre-mRNA consists of a nucleotide sequence corresponding to SEQ ID NO;

1, wherein said PMO hybridizes to said human dystrophin pre-mRNA with Watson-Crick base pairing, and wherein skipping of the 53^rd exon is induced in said patient.

8. The method according to claim 7, wherein each phosphorodiamidate morpholino monomer of said PMO has the formula:

9. The method according to claim 8, wherein the 5′

end of said PMO has a formula selected from the group consisting of;

10. A method of inducing exon 53 skipping in a patient with DMD comprising administering to said patient an antisense phosphorodiamidate morpholino oligomer (PMO) consisting of a 25-mer oligomer that is 100% complementary to the target sequence 5′

-GAACACCUUCAGAACCGGAGGCAAC-3′

(SEQ ID NO;

     124), wherein the 53rd exon in a human dystrophin pre-mRNA consists of a nucleotide sequence corresponding to SEQ ID NO;

1, and wherein said PMO hybridizes to said human dystrophin pre-mRNA with Watson-Crick base pairing.

11. The method according to claim 10, wherein each phosphorodiamidate morpholino monomer of said PMO has the formula:

12. The method according to claim 11, wherein the 5′

end of said PMO has a formula selected from the group consisting of;