ANTISENSE NUCLEIC ACID FOR TREATING AMYOTROPHY

• US 20180251760A1
• Filed: 09/16/2016
• Published: 09/06/2018
• Est. Priority Date: 09/16/2015
• Status: Active Grant

• Current Assignee: Kawasaki Gakuen Educational Foundation, Nippon Shinyaku Company Limited
• Original Assignee: Kawasaki Gakuen Educational Foundation, Nippon Shinyaku Company Limited
• Inventors: NAKAGAWA, Shinichiro, NAGATA, Seigo, SUNADA, Yoshihide, OHSAWA, Yutaka, NISHIMATSU, Shin-ichiro
• Granted Patent: US 10,563,199 B2
• Time in Patent Office: Days
• CPC Class Codes: A61K 31/7088 Compounds having three or more nucleosides or nucleotides
• CPC Class Codes: A61P 21/00 Drugs for disorders of the muscular or neuromuscular system
• CPC Class Codes: A61P 21/02 Muscle relaxants, e.g. for tetanus or cramps
• CPC Class Codes: A61P 21/04 for myasthenia gravis
• 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 15/1136 against growth factors, growth regulators, cytokines, lymphokines or hormones
• CPC Class Codes: C12N 2310/11 Antisense
• CPC Class Codes: C12N 2310/31 of the backbone
• CPC Class Codes: C12N 2310/32 of the sugar
• CPC Class Codes: C12N 2310/3233 Morpholino-type ring
• CPC Class Codes: C12N 2310/3519 Fusion with another nucleic acid
• CPC Class Codes: C12N 2320/33 Alteration of splicing

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.

Claims

1. An antisense oligomer of 14 to 30 bases in length comprising the following unit oligomers connected together:

(a) a first unit oligomer comprising a nucleotide sequence complementary to a first nucleotide sequence consisting of contiguous 7 to 15 bases in exon 2 of the human or mouse myostatin gene; and

(b) a second unit oligomer comprising a nucleotide sequence complementary to a second nucleotide sequence consisting of contiguous 7 to 15 bases in said exon 2,

wherein the first nucleotide sequence and the second nucleotide sequence are not contiguous to each other or do not overlap with each other,

or a pharmaceutically acceptable salt or hydrate thereof.

2. The antisense oligomer or pharmaceutically acceptable salt or hydrate thereof according to claim 1, wherein the antisense oligomer is (c) or (d) shown below:

(c) an antisense oligomer of 14 to 30 bases in length comprising connected two unit oligomers selected from the group consisting of unit oligomers (c-1) to (c-6) shown below;

(c-1) a unit oligomer consisting of a nucleotide sequence complementary to a nucleotide sequence consisting of contiguous 7 to 15 bases selected from a nucleotide sequence located at positions −

10 to 45 from the 5′

-terminal end of exon 2 in the human myostatin gene;

(c-2) a unit oligomer consisting of a nucleotide sequence complementary to a nucleotide sequence consisting of contiguous 7 to 15 bases selected from a nucleotide sequence located at positions 91 to 135 from the 5′

-terminal end of exon 2 in the human myostatin gene;

(c-3) a unit oligomer consisting of a nucleotide sequence complementary to a nucleotide sequence consisting of contiguous 7 to 15 bases selected from a nucleotide sequence located at positions 121 to 155 from the 5′

-terminal end of exon 2 in the human myostatin gene;

(c-4) a unit oligomer consisting of a nucleotide sequence complementary to a nucleotide sequence consisting of contiguous 7 to 15 bases selected from a nucleotide sequence located at positions 166 to 210 from the 5′

-terminal end of exon 2 in the human myostatin gene;

(c-5) a unit oligomer consisting of a nucleotide sequence complementary to a nucleotide sequence consisting of contiguous 7 to 15 bases selected from a nucleotide sequence located at positions 204 to 233 from the 5′

-terminal end of exon 2 in the human myostatin gene; and

(c-6) a unit oligomer consisting of a nucleotide sequence complementary to a nucleotide sequence consisting of contiguous 7 to 15 bases selected from a nucleotide sequence located at positions 326 to 374 from the 5′

-terminal end of exon 2 in the human myostatin gene;

or

(d) an antisense oligomer of 14 to 30 bases in length comprising connected two unit oligomers selected from the group consisting of unit oligomers (d-1) to (d-7) shown below;

(d-1) a unit oligomer consisting of a nucleotide sequence complementary to a nucleotide sequence consisting of contiguous 7 to 15 bases selected from a nucleotide sequence located at positions −

10 to 65 from the 5′

-terminal end of exon 2 in the mouse myostatin gene;

(d-2) a unit oligomer consisting of a nucleotide sequence complementary to a nucleotide sequence consisting of contiguous 7 to 15 bases selected from a nucleotide sequence located at positions 91 to 135 from the 5′

-terminal end of exon 2 in the mouse myostatin gene;

(d-3) a unit oligomer consisting of a nucleotide sequence complementary to a nucleotide sequence consisting of contiguous 7 to 15 bases selected from a nucleotide sequence located at positions 121 to 155 from the 5′

-terminal end of exon 2 in the mouse myostatin gene;

(d-4) a unit oligomer consisting of a nucleotide sequence complementary to a nucleotide sequence consisting of contiguous 7 to 15 bases selected from a nucleotide sequence located at positions 151 to 210 from the 5′

-terminal end of exon 2 in the mouse myostatin gene;

(d-5) a unit oligomer consisting of a nucleotide sequence complementary to a nucleotide sequence consisting of contiguous 7 to 15 bases selected from a nucleotide sequence located at positions 196 to 233 from the 5′

-terminal end of exon 2 in the mouse myostatin gene;

(d-6) a unit oligomer consisting of a nucleotide sequence complementary to a nucleotide sequence consisting of contiguous 7 to 15 bases selected from a nucleotide sequence located at positions 266 to 295 from the 5′

-terminal end of exon 2 in the mouse myostatin gene; and

(d-7) a unit oligomer consisting of a nucleotide sequence complementary to a nucleotide sequence consisting of contiguous 7 to 15 bases selected from a nucleotide sequence located at positions 326 to 374 from the 5′

-terminal end of exon 2 in the mouse myostatin gene.

3. The antisense oligomer or pharmaceutically acceptable salt or hydrate thereof according to claim 1, wherein the antisense oligomer is (e) or (f) shown below:

(e) an antisense oligomer of 14 to 30 bases in length comprising connected two unit oligomers selected from the group consisting of unit oligomers (e-1) to (e-6) shown below;

(e-1) a unit oligomer consisting of a nucleotide sequence complementary to a nucleotide sequence consisting of contiguous 7 to 15 bases selected from a nucleotide sequence located at positions −

4 to 25 from the 5′

-terminal end of exon 2 in the human myostatin gene;

(e-2) a unit oligomer consisting of a nucleotide sequence complementary to a nucleotide sequence consisting of contiguous 7 to 15 bases selected from a nucleotide sequence located at positions 112 to 131 from the 5′

-terminal end of exon 2 in the human myostatin gene;

(e-3) a unit oligomer consisting of a nucleotide sequence complementary to a nucleotide sequence consisting of contiguous 7 to 15 bases selected from a nucleotide sequence located at positions 128 to 142 from the 5′

-terminal end of exon 2 in the human myostatin gene;

(e-4) a unit oligomer consisting of a nucleotide sequence complementary to a nucleotide sequence consisting of contiguous 7 to 15 bases selected from a nucleotide sequence located at positions 169 to 206 from the 5′

-terminal end of exon 2 in the human myostatin gene;

(e-5) a unit oligomer consisting of a nucleotide sequence complementary to a nucleotide sequence consisting of contiguous 7 to 15 bases selected from a nucleotide sequence located at positions 209 to 228 from the 5′

-terminal end of exon 2 in the human myostatin gene; and

(e-6) a unit oligomer consisting of a nucleotide sequence complementary to a nucleotide sequence consisting of contiguous 7 to 15 bases selected from a nucleotide sequence located at positions 331 to 365 from the 5′

-terminal end of exon 2 in the human myostatin gene;

or

(f) an antisense oligomer of 14 to 30 bases in length comprising connected two unit oligomers selected from the group consisting of unit oligomers (f-1) to (f-7) shown below;

(f-1) a unit oligomer consisting of a nucleotide sequence complementary to a nucleotide sequence consisting of contiguous 7 to 15 bases selected from a nucleotide sequence located at positions 1 to 18 from the 5′

-terminal end of exon 2 in the mouse myostatin gene;

(f-2) a unit oligomer consisting of a nucleotide sequence complementary to a nucleotide sequence consisting of contiguous 7 to 15 bases selected from a nucleotide sequence located at positions 116 to 131 from the 5′

-terminal end of exon 2 in the mouse myostatin gene;

(f-3) a unit oligomer consisting of a nucleotide sequence complementary to a nucleotide sequence consisting of contiguous 7 to 15 bases selected from a nucleotide sequence located at positions 130 to 140 from the 5′

-terminal end of exon 2 in the mouse myostatin gene;

(f-4) a unit oligomer consisting of a nucleotide sequence complementary to a nucleotide sequence consisting of contiguous 7 to 15 bases selected from a nucleotide sequence located at positions 160 to 206 from the 5′

-terminal end of exon 2 in the mouse myostatin gene;

(f-5) a unit oligomer consisting of a nucleotide sequence complementary to a nucleotide sequence consisting of contiguous 7 to 15 bases selected from a nucleotide sequence located at positions 211 to 225 from the 5′

-terminal end of exon 2 in the mouse myostatin gene;

(f-6) a unit oligomer consisting of a nucleotide sequence complementary to a nucleotide sequence consisting of contiguous 7 to 15 bases selected from a nucleotide sequence located at positions 271 to 282 from the 5′

-terminal end of exon 2 in the mouse myostatin gene; and

(f-7) a unit oligomer consisting of a nucleotide sequence complementary to a nucleotide sequence consisting of contiguous 7 to 15 bases selected from a nucleotide sequence located at positions 341 to 365 from the 5′

-terminal end of exon 2 in the mouse myostatin gene.

4. The antisense oligomer or pharmaceutically acceptable salt or hydrate thereof according to claim 2, wherein the antisense oligomer (c) consists of any one nucleotide sequence selected from the group consisting of SEQ ID NO:

103 (NMS-48), SEQ ID NO;

116 (NMS-89), SEQ ID NO;

117 (NMS-90), SEQ ID NO;

120 (NMS-93), SEQ ID NO;

128 (NMS-101), SEQ ID NO;

131 (NMS-104), SEQ ID NO;

136 (NMS-113), SEQ ID NO;

137 (NMS-117), SEQ ID NO;

140 (NMS-123), SEQ ID NO;

145 (NMS-136), SEQ ID NO;

146 (NMS-139), SEQ ID NO;

147 (NMS-140), SEQ ID NO;

148 (NMS-141), SEQ ID NO;

149 (NMS-142), SEQ ID NO;

152 (NMS-145), SEQ ID NO;

155 (NMS-148), SEQ ID NO;

156 (NMS-149), SEQ ID NO;

157 (NMS-150), SEQ ID NO;

159 (NMS-152), SEQ ID NO;

162 (NMS-156), SEQ ID NO;

163 (NMS-157), SEQ ID NO;

165 (NMS-162), SEQ ID NO;

166 (NMS-163), SEQ ID NO;

167 (NMS-164), SEQ ID NO;

168 (NMS-166), SEQ ID NO;

169 (NMS-167), SEQ ID NO;

170 (NMS-168), SEQ ID NO;

171 (NMS-169), SEQ ID NO;

176 (NMS-174), SEQ ID NO;

177 (NMS-175), SEQ ID NO;

178 (NMS-176), SEQ ID NO;

179 (NMS-177), SEQ ID NO;

180 (NMS-178), SEQ ID NO;

183 (NMS-181), SEQ ID NO;

187 (NMS-185), SEQ ID NO;

189 (NMS-188), SEQ ID NO;

190 (NMS-189), SEQ ID NO;

191 (NMS-190), SEQ ID NO;

192 (NMS-191), SEQ ID NO;

193 (NMS-192), SEQ ID NO;

196 (NMS-195), SEQ ID NO;

199 (NMS-198), SEQ ID NO;

200 (NMS-199), SEQ ID NO;

201 (NMS-200), SEQ ID NO;

203 (NMS-202), SEQ ID NO;

204 (NMS-203), SEQ ID NO;

206 (NMS-206), SEQ ID NO;

208 (NMS-208), SEQ ID NO;

212 (NMS-212), SEQ ID NO;

213 (NMS-213), SEQ ID NO;

214 (NMS-214), SEQ ID NO;

215 (NMS-215), SEQ ID NO;

217 (NMS-217), SEQ ID NO;

225 (NMS-225), SEQ ID NO;

226 (NMS-228), SEQ ID NO;

228 (NMS-230), SEQ ID NO;

229 (NMS-231), SEQ ID NO;

231 (NMS-233), SEQ ID NO;

232 (NMS-234), SEQ ID NO;

233 (NMS-235), SEQ ID NO;

236 (NMS-240), SEQ ID NO;

237 (NMS-241), SEQ ID NO;

240 (NMS-244), SEQ ID NO;

243 (NMS-247), SEQ ID NO;

244 (NMS-248), SEQ ID NO;

245 (NMS-249), SEQ ID NO;

246 (NMS-250), SEQ ID NO;

247 (NMS-251), SEQ ID NO;

248 (NMS-252), SEQ ID NO;

252 (NMS-256), SEQ ID NO;

261 (NMS-272), SEQ ID NO;

273 (NMS-284), SEQ ID NO;

274 (NMS-285), SEQ ID NO;

275 (NMS-286) and SEQ ID NO;

277 (NMS-297).

5. The antisense oligomer or pharmaceutically acceptable salt or hydrate thereof according to claim 2, wherein the antisense oligomer (d) consists of any one nucleotide sequence selected from the group consisting of SEQ ID NO:

95 (NMS-38), SEQ ID NO;

96 (NMS-39), SEQ ID NO;

107 (NMS-66), SEQ ID NO;

223 (NMS-223), SEQ ID NO;

234 (NMS-238), SEQ ID NO;

235 (NMS-239), SEQ ID NO;

242 (NMS-246), SEQ ID NO;

249 (NMS-253), SEQ ID NO;

250 (NMS-254), SEQ ID NO;

251 (NMS-255), SEQ ID NO;

257 (NMS-268), SEQ ID NO;

(NMS-280), SEQ ID NO;

(NMS-281), SEQ ID NO;

(NMS-282), SEQ ID NO;

(NMS-288), SEQ ID NO;

(NMS-289), SEQ ID NO;

(NMS-290), SEQ ID NO;

(NMS-292), SEQ ID NO;

(NMS-293), SEQ ID NO;

(NMS-294), SEQ ID NO;

(NMS-295), SEQ ID NO;

(NMS-298), SEQ ID NO;

(NMS-299), SEQ ID NO;

(NMS-300), SEQ ID NO;

(NMS-302) and SEQ ID NO;

(NMS-303).

6. The antisense oligomer or pharmaceutically acceptable salt or hydrate thereof according to claim 1, wherein the antisense oligomer is an oligonucleotide.

7. The antisense oligomer or pharmaceutically acceptable salt or hydrate thereof according to claim 6, wherein at least one sugar moiety and/or at least one phosphate bond moiety in nucleotides constituting the oligonucleotide is modified.

8. The antisense oligomer or pharmaceutically acceptable salt or hydrate thereof according to claim 7, 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, NH₂, NHR, NR₂, N₃, CN, F, Cl, Br and I (wherein R represents alkyl or aryl, and R′

represents alkylene).

9. The antisense oligomer or pharmaceutically acceptable salt or hydrate thereof according to claim 7, 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.

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

11. The antisense oligomer or pharmaceutically acceptable salt or hydrate thereof according to claim 10, wherein the morpholino oligomer is a phosphorodiamidate morpholino oligomer.

12. The antisense oligomer or pharmaceutically acceptable salt or hydrate thereof according to claim 10, whose 5′

-terminal end is any one of the groups represented by chemical formulae (1) to (3) shown below.

13. The antisense oligomer or pharmaceutically acceptable salt or hydrate thereof according to claim 4, wherein the antisense oligomer consists of any one nucleotide sequence selected from the group consisting of SEQ ID NO:

171 (NMS-169), SEQ ID NO;

192 (NMS-191), SEQ ID NO;

245 (NMS-249) and SEQ ID NO;

231 (NMS-233).

14. A pharmaceutical composition comprising the antisense oligomer or pharmaceutically acceptable salt or hydrate thereof according to claim 3.

15. The pharmaceutical composition according to claim 14, which further comprises a pharmaceutically acceptable carrier.

16. The pharmaceutical composition according to claim 14 for use in the treatment of an amyotrophic disease or a muscle wasting disease.

17. The pharmaceutical composition according to claim 16 for use in the treatment of muscular dystrophy.

18. 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 1.

19. The method according to claim 18, wherein the amyotrophic disease or muscle wasting disease is muscular dystrophy.

20. The method according to claim 18, wherein the subject is a human subject.

21. Use of the antisense oligomer or pharmaceutically acceptable salt or hydrate thereof according to claim 1 in the manufacture of a pharmaceutical composition for treatment of an amyotrophic disease or a muscle wasting disease.

22. The antisense oligomer or pharmaceutically acceptable salt or hydrate thereof according to claim 1 for use in the treatment of an amyotrophic disease or a muscle wasting disease.

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.

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 according to 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, NH₂, NHR, NR₂, N₃, 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 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. Use of the antisense oligomer or pharmaceutically acceptable salt or hydrate thereof according to claim 23 in the manufacture of a pharmaceutical composition for treatment of an amyotrophic disease or a muscle wasting disease.

48. The antisense oligomer or pharmaceutically acceptable salt or hydrate thereof according to claim 23 for use in the treatment of an amyotrophic disease or a muscle wasting disease.

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.