Antisense nucleic acid inducing skipping of exon 51

Abstract

The present specification provides a drug that causes highly-efficient skipping of exon 51 in the human dystrophin gene. The present specification provides an antisense oligomer having an activity to induce skipping of exon 51 in the human dystrophin gene.

1 . An antisense oligomer which is selected from the group consisting of (a1) to (d1) below:
 (a1) an antisense oligomer comprising a base sequence of any of SEQ ID NOs: 1 to 89 and 91 to 93;   (b1) an antisense oligomer which comprises a base sequence having deletion, substitution, insertion and/or addition of 1 to 5 base(s) in the base sequence of any of SEQ ID NOs: 1 to 89 and 91 to 93, and has an activity to induce skipping of exon  51  in the human dystrophin gene;   (c1) an antisense oligomer which comprises a base sequence having at least 80% sequence identity to a base sequence of any of SEQ ID NOs: 1 to 89 and 91 to 93, and has an activity to induce skipping of exon  51  in the human dystrophin gene; and   (d1) an antisense oligomer that hybridizes under stringent conditions to an oligonucleotide consisting of a base sequence complementary to the base sequence of any of SEQ ID NOs: 1 to 89 and 91 to 93, and has an activity to induce skipping of exon  51  in the human dystrophin gene,   (except for an antisense oligomer which consists of a base sequence of any of SEQ ID NOs: 90 and 97 to 126) or a pharmaceutically acceptable salt or hydrate thereof.   
     
     
         2 . The antisense oligomer according to  claim 1 , wherein the antisense oligomer is selected from the group consisting of (e) to (h) below:
 (e) an antisense oligomer which consists of a base sequence of any of SEQ ID NOs: 1 to 89 and 91 to 93;   (f) an antisense oligomer which consists of a base sequence having deletion and/or substitution of 1 to 5 base(s) in the base sequence of any of SEQ ID NOs: 1 to 89 and 91 to 93, and has an activity to induce skipping of exon  51  in the human dystrophin gene;   (g) an antisense oligomer which consists of a base sequence having at least 80% sequence identity to a base sequence of any of SEQ ID NOs: 1 to 89 and 91 to 93, and has an activity to induce skipping of exon  51  in the human dystrophin gene; and   (h) an antisense oligomer that hybridizes under high stringent conditions to an oligonucleotide consisting of a base sequence complementary to the base sequence of any of SEQ ID NOs: 1 to 89 and 91 to 93, and has an activity to induce skipping of exon  51  in the human dystrophin gene,   (except for an antisense oligomer which consists of a base sequence of any of SEQ ID NOs: 90 and 97 to 126) or a pharmaceutically acceptable salt or hydrate thereof.   
     
     
         3 . The antisense oligomer or the pharmaceutically acceptable salt or hydrate thereof according to  claim 1 , wherein
 the antisense oligomer is   an antisense oligomer that comprises a base sequence having at least 90% sequence identity to a base sequence of any of SEQ ID NOs: 1 to 89 and 91 to 93, and has an activity to induce skipping of exon  51  in the human dystrophin gene.   
     
     
         4 . The antisense oligomer or the pharmaceutically acceptable salt or hydrate thereof according to  claim 1 , wherein the antisense oligomer is an antisense oligomer selected from the group consisting of:
 (a2) an antisense oligomer comprising the base sequence of any of SEQ ID NOs: 7, 8, 10, 16, 21, 24, 31, 42, 67 and 76;   (b2) an antisense oligomer which comprises a base sequence having deletion, substitution, insertion and/or addition of 1 to 5 base(s) in the base sequence of any of SEQ ID NOs: 7, 8, 10, 16, 21, 24, 31, 42, 67 and 76, and has an activity to induce skipping of exon  51  in the human dystrophin gene;   (c2) an antisense oligomer which comprises a base sequence having at least 80% sequence identity to the base sequence of any of SEQ ID NOs: 7, 8, 10, 16, 21, 24, 31, 42, 67 and 76, and has an activity to induce skipping of exon  51  in the human dystrophin gene; and   (d2) an antisense oligomer that hybridizes under stringent conditions to an oligonucleotide consisting of a base sequence complementary to the base sequence of any of SEQ ID NOs: 7, 8, 10, 16, 21, 24, 31, 42, 67 and 76, and has an activity to induce skipping of exon  51  in the human dystrophin gene.   
     
     
         5 . The antisense oligomer or the pharmaceutically acceptable salt or hydrate thereof according to  claim 1 , wherein the antisense oligomer is an oligonucleotide. 
     
     
         6 . The antisense oligomer or the pharmaceutically acceptable salt or hydrate thereof according to  claim 5 , wherein the sugar moiety and/or the phosphate bond moiety of at least one nucleotide constituting the oligonucleotide are/is modified. 
     
     
         7 . The antisense oligomer or the pharmaceutically acceptable salt or hydrate thereof according to  claim 5 , 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, NH 2 , NHR, NR 2 , N 3 , CN, F, Cl, Br and I (wherein R is an alkyl or an aryl, and R′ is an alkylene). 
     
     
         8 . The antisense oligomer or the pharmaceutically acceptable salt or hydrate thereof according to  claim 5 , wherein the phosphate bond moiety 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. 
     
     
         9 . The antisense oligomer or the pharmaceutically acceptable salt or hydrate thereof according to  claim 1 , wherein the antisense oligomer is a morpholino oligomer. 
     
     
         10 . The antisense oligomer or the pharmaceutically acceptable salt or hydrate thereof according to  claim 9 , wherein the antisense oligomer is a phosphorodiamidate morpholino oligomer. 
     
     
         11 . The antisense oligomer or the pharmaceutically acceptable salt or hydrate thereof according to  claim 9 , wherein the 5′ end is any one of chemical formulae (1) to (3) below: 
       
         
           
           
               
               
           
         
       
     
     
         12 . A pharmaceutical composition for the treatment of muscular dystrophy, comprising the antisense oligomer or the pharmaceutically acceptable salt or hydrate thereof according to  claim 1 . 
     
     
         13 . The pharmaceutical composition according to  claim 12 , further comprising a pharmaceutically acceptable carrier. 
     
     
         14 - 18 . (canceled): 
     
     
         19 . A method for treatment of muscular dystrophy, which comprises administering to a patient with muscular dystrophy an effective amount of the antisense oligomer or the pharmaceutically acceptable salt or hydrate thereof according to  claim 1 , or a pharmaceutical composition comprising the antisense oligomer or the pharmaceutically acceptable salt or hydrate thereof according to  claim 1 . 
     
     
         20 . The method for treatment according to  claim 19 , wherein the patient is a human. 
     
     
         21 - 22 . (canceled): 
     
     
         23 . The method for treatment according to  claim 19 , wherein the patient carries a mutation in the dystrophin gene to be converted to in-frame by exon  51  skipping. 
     
     
         24 . The method for treatment according to  claim 23 , wherein the mutation is a frameshift mutation caused by deletion of an exon in the vicinity of exon  51 . 
     
     
         25 . The method for treatment according to  claim 24 , wherein the frameshift mutation is caused by a deletion of exons  13 - 50 , a deletion of exons  29 - 50 , a deletion of exons  40 - 50 , a deletion of exons  43 - 50 , a deletion of exons  45 - 50 , a deletion of exons  47 - 50 , a deletion of exons  48 - 50 , a deletion of exons  49 - 50 , a deletion of exon  50 , a deletion of exon  52 , or a deletion of exons  52 - 63  in the dystrophin gene.