Antisense nucleic acid for treating amyotrophy

Abstract

There has been a demand for a novel antisense nucleic acid or the like capable of inhibiting myostatin at the mRNA level. The present invention provides a specific antisense oligomer which allows exon 2 skipping in the myostatin gene or induces degradation of mRNA of the myostatin gene.

1 .- 22 . (canceled) 
     
     
         23 . Any one antisense oligomer selected from the group consisting of (A) to (H) shown below or a pharmaceutically acceptable salt or hydrate thereof:
 (A) an antisense oligomer consisting of a nucleotide sequence complementary to a nucleotide sequence of contiguous 14 to 30 bases in length selected from a nucleotide sequence located at positions −10 to 45 from the 5′-terminal end of exon 2 in the human myostatin gene;   (B) an antisense oligomer consisting of a nucleotide sequence complementary to a nucleotide sequence of contiguous 14 to 30 bases in length selected from a nucleotide sequence located at positions 91 to 145 from the 5′-terminal end of exon 2 in the human myostatin gene;   (C) an antisense oligomer consisting of a nucleotide sequence complementary to a nucleotide sequence of contiguous 14 to 30 bases in length selected from a nucleotide sequence located at positions 146 to 180 from the 5′-terminal end of exon 2 in the human myostatin gene;   (D) an antisense oligomer consisting of a nucleotide sequence complementary to a nucleotide sequence of contiguous 14 to 30 bases in length selected from a nucleotide sequence located at positions 331 to 374 from the 5′-terminal end of exon 2 in the human myostatin gene;   (E) an antisense oligomer consisting of a nucleotide sequence complementary to a nucleotide sequence of contiguous 14 to 30 bases in length selected from a nucleotide sequence located at positions −10 to 31 from the 5′-terminal end of exon 2 in the mouse myostatin gene;   (F) an antisense oligomer consisting of a nucleotide sequence complementary to a nucleotide sequence of contiguous 14 to 30 bases in length selected from a nucleotide sequence located at positions 111 to 162 from the 5′-terminal end of exon 2 in the mouse myostatin gene;   (G) an antisense oligomer consisting of a nucleotide sequence complementary to a nucleotide sequence of contiguous 14 to 30 bases in length selected from a nucleotide sequence located at positions 166 to 195 from the 5′-terminal end of exon 2 in the mouse myostatin gene; and   (H) an antisense oligomer consisting of a nucleotide sequence complementary to a nucleotide sequence of contiguous 14 to 30 bases in length selected from a nucleotide sequence located at positions 331 to 374 from the 5′-terminal end of exon 2 in the mouse myostatin gene.   
     
     
         24 . Any one antisense oligomer selected from the group consisting of (I) to (L) shown below or a pharmaceutically acceptable salt or hydrate thereof:
 (I) an antisense oligomer consisting of a nucleotide sequence complementary to a nucleotide sequence of contiguous 14 to 30 bases in length selected from a nucleotide sequence located at positions −10 to 31 from the 5′-terminal end of exon 2 in the human myostatin gene;   (J) an antisense oligomer consisting of a nucleotide sequence complementary to a nucleotide sequence of contiguous 14 to 30 bases in length selected from a nucleotide sequence located at positions 111 to 140 from the 5′-terminal end of exon 2 in the human myostatin gene, wherein the 3′-terminal base of the nucleotide sequence of 14 to 30 bases in length is a base located at position 140 from the 5′-terminal end of said exon 2;   (K) an antisense oligomer consisting of a nucleotide sequence complementary to a nucleotide sequence of contiguous 14 to 30 bases in length selected from a nucleotide sequence located at positions 146 to 180 from the 5′-terminal end of exon 2 in the human myostatin gene; and   (L) an antisense oligomer consisting of a nucleotide sequence complementary to a nucleotide sequence of contiguous 14 to 30 bases in length selected from a nucleotide sequence located at positions 331 to 374 from the 5′-terminal end of exon 2 in the human myostatin gene.   
     
     
         25 . The antisense oligomer or pharmaceutically acceptable salt or hydrate thereof according to  claim 23 , wherein the antisense oligomer (A) consists of any one nucleotide sequence selected from the group consisting of SEQ ID NO: 13 (NMS-17), SEQ ID NO: 76 (NMS-138), SEQ ID NO: 68 (NMS-120), SEQ ID NO: 75 (NMS-137), SEQ ID NO: 51 (NMS-76), SEQ ID NO: 52 (NMS-79), SEQ ID NO: 54 (NMS-81), SEQ ID NO: 55 (NMS-82), SEQ ID NO: 56 (NMS-83), SEQ ID NO: 53 (NMS-80), SEQ ID NO: 33 (NMS-49), SEQ ID NO: 63 (NMS-114), SEQ ID NO: 69 (NMS-124), SEQ ID NO: 70 (NMS-125), SEQ ID NO: 61 (NMS-110), SEQ ID NO: 31 (NMS-46), SEQ ID NO: 34 (NMS-50), SEQ ID NO: 50 (NMS-75), SEQ ID NO: 45 (NMS-67) and SEQ ID NO: 64 (NMS-115). 
     
     
         26 . The antisense oligomer or pharmaceutically acceptable salt or hydrate thereof according to  claim 23 , wherein the antisense oligomer (B) consists of any one nucleotide sequence selected from the group consisting of SEQ ID NO: 3 (NMS-6), SEQ ID NO:
 66 (NMS-118), SEQ ID NO: 67 (NMS-119), SEQ ID NO: 28 (NMS-33), SEQ ID NO: 72 (NMS-127), SEQ ID NO: 16 (NMS-20), SEQ ID NO: 82 (NMS-187) and SEQ ID NO: 25 (NMS-30).   
     
     
         27 . The antisense oligomer or pharmaceutically acceptable salt or hydrate thereof according to  claim 23 , wherein the antisense oligomer (C) consists of a nucleotide sequence shown in SEQ ID NO: 12 (NMS-16). 
     
     
         28 . The antisense oligomer or pharmaceutically acceptable salt or hydrate thereof according to  claim 23 , wherein the antisense oligomer (D) consists of a nucleotide sequence shown in SEQ ID NO: 4 (NMS-7). 
     
     
         29 . The antisense oligomer or pharmaceutically acceptable salt or hydrate thereof according to  claim 23 , wherein the antisense oligomer (E) consists of a nucleotide sequence shown in SEQ ID NO: 90 (NMS-51). 
     
     
         30 . The antisense oligomer or pharmaceutically acceptable salt or hydrate thereof according to  claim 23 , wherein the antisense oligomer (F) consists of any one nucleotide sequence selected from the group consisting of SEQ ID NO: 91 (NMS-52), SEQ ID NO: 28 (NMS-33) and SEQ ID NO: 25 (NMS-30), SEQ ID NO: 41 (NMS-61), SEQ ID NO: 24 (NMS-29), SEQ ID NO: 42 (NMS-62), SEQ ID NO: 43 (NMS-63), SEQ ID NO: 11 (NMS-15), SEQ ID NO: 67 (NMS-119), SEQ ID NO: 80 (NMS-161) and SEQ ID NO: 82 (NMS-187). 
     
     
         31 . The antisense oligomer or pharmaceutically acceptable salt or hydrate thereof according to  claim 23 , wherein the antisense oligomer (G) consists of a nucleotide sequence shown in SEQ ID NO: 7 (NMS-10). 
     
     
         32 . The antisense oligomer or pharmaceutically acceptable salt or hydrate thereof according to  claim 23 , wherein the antisense oligomer (H) consists of any one nucleotide sequence selected from the group consisting of SEQ ID NO: 4 (NMS-7), SEQ ID NO:
 9 (NMS-12), SEQ ID NO: 10 (NMS-14) and SEQ ID NO: 14 (NMS-18).   
     
     
         33 . The antisense oligomer or pharmaceutically acceptable salt or hydrate thereof  claim 23 , wherein the antisense oligomer is an oligonucleotide. 
     
     
         34 . The antisense oligomer or pharmaceutically acceptable salt or hydrate thereof according to  claim 33 , wherein at least one sugar moiety and/or at least one phosphate bond moiety in nucleotides constituting the oligonucleotide is modified. 
     
     
         35 . The antisense oligomer or pharmaceutically acceptable salt or hydrate thereof according to  claim 34 , wherein the at least one sugar moiety in nucleotides constituting the oligonucleotide is a ribose in which the —OH group at the 2′-position is substituted with any group 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 represents alkyl or aryl, and R′ represents alkylene). 
     
     
         36 . The antisense oligomer or pharmaceutically acceptable salt or hydrate thereof according to  claim 34 , wherein the at least one phosphate bond moiety in nucleotides constituting the oligonucleotide is any one selected from the group consisting of a phosphorothioate bond, a phosphorodithioate bond, an alkylphosphonate bond, a phosphoroamidate bond and a boranophosphate bond. 
     
     
         37 . The antisense oligomer or pharmaceutically acceptable salt or hydrate thereof according to  claim 23 , wherein the antisense oligomer is a morpholino oligomer. 
     
     
         38 . The antisense oligomer or pharmaceutically acceptable salt or hydrate thereof according to  claim 37 , wherein the morpholino oligomer is a phosphorodiamidate morpholino oligomer. 
     
     
         39 . The antisense oligomer or pharmaceutically acceptable salt or hydrate thereof according to  claim 37 , whose 5′-terminal end is any one of the groups represented by chemical formulae (1) to (3) shown below: 
       
         
           
           
               
               
           
         
       
     
     
         40 . A pharmaceutical composition comprising the antisense oligomer or pharmaceutically acceptable salt or hydrate thereof according to  claim 23 . 
     
     
         41 . The pharmaceutical composition according to  claim 40 , which further comprises a pharmaceutically acceptable carrier. 
     
     
         42 . The pharmaceutical composition according to  claim 40  or  41  for use in the treatment of an amyotrophic disease or a muscle wasting disease. 
     
     
         43 . The pharmaceutical composition according to  claim 42  for use in the treatment of muscular dystrophy. 
     
     
         44 . A method for prevention or treatment of an amyotrophic disease or a muscle wasting disease, which comprises administering a subject in need of prevention or treatment of an amyotrophic disease or a muscle wasting disease with a therapeutically effective amount of the antisense oligomer or pharmaceutically acceptable salt or hydrate thereof according to  claim 23 . 
     
     
         45 . The method according to  claim 44 , wherein the amyotrophic disease or muscle wasting disease is muscular dystrophy. 
     
     
         46 . The method according to  claim 44 , wherein the subject is a human subject. 
     
     
         47 - 48 . (canceled)