Antisense nucleic acids

• US 9,988,629 B2
• Filed: 03/11/2015
• Issued: 06/05/2018
• Est. Priority Date: 03/12/2014
• 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: Wakayama, Tatsushi, Seo, Haruna, Satou, Youhei, Takeda, Shin'ichi, Nagata, Tetsuya
• Primary Examiner(s): Schnizer, Richard A
• Application Number:
  US15/122,435
• Publication Number:
  US 20170067048A1
• Time in Patent Office: 1,182 Days
• Field of Search: None
• CPC Class Codes: A61K 31/713 Double-stranded nucleic acids or oligonucleotides
• 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: 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 2310/3521 Methyl
• CPC Class Codes: C12N 2310/3525 MOE, methoxyethoxy
• CPC Class Codes: C12N 2310/3533 Halogen
• CPC Class Codes: C12N 2310/3535 Nitrogen
• CPC Class Codes: C12N 2320/33 Alteration of splicing

Abstract

Provided is a drug that allows highly-efficient skipping of exon 51 in the human dystrophin gene. The present invention provides an antisense oligomer which enables exon 51 in the human dystrophin gene to be skipped.

Claims

1. An antisense oligomer of 25-35 nucleobases, or a pharmaceutically acceptable salt or hydrate thereof, wherein the antisense oligomer is a morpholino oligomer, a peptide nucleic acid (PNA), or an oligonucleotide comprising at least one nucleotide having:

(i) a modified sugar moiety, wherein the 2′

—

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

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

is an alkylene), or

(ii) a modified phosphate-binding region selected from the group consisting of a phosphorothioate bond, a phosphorodithioate bond, an alkylphosphonate bond, a phosphoramidate bond, and a boranophosphate bond,

and wherein the antisense oligomer is;

(a) an antisense oligomer comprising the nucleobase sequence of SEQ ID NO;

1or 2;

or

(b) an antisense oligomer which i) consists of a nucleobase sequence that differs from the nucleobase sequence of SEQ ID NO;

1 or 2 by no more than 5nucleobases, wherein each difference independently may be a deletion, a substitution, an insertion, or an addition of a nucleobase, and ii) has an activity to cause skipping of the 51st exon of a human dystrophin gene.

2. The antisense oligomer or a pharmaceutically acceptable salt or hydrate thereof according to claim 1, wherein the antisense oligomer is:

(c) an antisense oligomer which consists of the nucleobase sequence of SEQ ID NO;

1 or 2;

or

(d) an antisense oligomer which consists of a nucleobase sequence that differs from the nucleobase sequence of SEQ ID NO;

1 or 2 by no more than 3 nucleobases.

3. The antisense oligomer according to claim 1, which is a morpholino oligomer, or a pharmaceutically acceptable salt or hydrate thereof.

4. The antisense oligomer according to claim 3, which is a phosphorodiamidate morpholino oligomer, or a pharmaceutically acceptable salt or hydrate thereof.

5. The antisense oligomer according to claim 3, or a pharmaceutically acceptable salt or hydrate thereof, wherein the 5′

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

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

7. The pharmaceutical composition according to claim 6, comprising a pharmaceutically acceptable carrier.

8. A method for treatment of muscular dystrophy, which comprises administering to a patient with muscular dystrophy an antisense oligomer of 25-35nucleobases, or a pharmaceutically acceptable salt or hydrate thereof, wherein the antisense oligomer is:

(a) an antisense oligomer comprising the nucleobase sequence of SEQ ID NO;

1or 2;

or

(b) an antisense oligomer which i) consists of a nucleobase sequence that differs from the nucleobase sequence of SEQ ID NO;

1 or 2 by no more than 5nucleobases, wherein each difference independently may be a deletion, a substitution, an insertion, or an addition of a nucleobase, and ii) has an activity to cause skipping of the 51st exon of a human dystrophin gene;

wherein the antisense oligomer is a morpholino oligomer, a peptide nucleic acid (PNA), or an oligonucleotide that the sugar moiety and/or the phosphate-binding region of at least one nucleotide constituting the oligonucleotide is modified, and wherein the patient with muscular dystrophy has a mutation converting to in-frame by exon 51 skipping.

9. The method for treatment according to claim 8, wherein the patient with muscular dystrophy is a patient with deletions of exons 29-50, 50, 45-50, 48-50, 49-50, 52, 52-63, 13-50, 19-50, 43-50 or 47-50.

10. The method for treatment according to claim 8, wherein the patient is a human.